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Sample records for isolated human penile

  1. Isolated penile torsion in newborns

    PubMed Central

    Eroglu, Egemen; Gundogdu, Gokhan

    2015-01-01

    Introduction: We reported on the incidence of isolated penile torsion among our healthy children and our approach to this anomaly. Methods: Between 2011 and 2014, newborn babies with penile torsion were classified according to the angle of torsion. Surgical correction (penile degloving and reattachment for moderate cases and dorsal dartos flap technique in case of resistance) after 6 months was advised to the babies with rotations more than 45°. Results: Among 1000 newborn babies, 200 isolated penile torsions were found, and among these, 43 had torsions more than 45°, and 4 of these had angles greater than 90°. The mean angle of the rotations was found 30.45° (median: 20°). In total, 8 children with 60° torsions were previously circumcised. Surgery was performed on 19 patients, with a mean patient age of 12 ± 2 months. Of these 19, 13 babies were corrected with degloving and reattachment. This technique was not enough on the remaining 6 patients; therefore, derotational dorsal dartos flap was added to correct the torsion. After a mean of 15.6 ± 9.8 months, residual penile rotation, less than 15°, was found only in 2 children. Conclusion: The incidence of isolated penile torsion is 20% in newborns. However, rotation more than 45° angles are seen in 4.3% of male babies. Correction is not necessary in mild degrees, and penile degloving with reattachment is enough in most cases. If the initial correction is insufficient, dorsal dartos flap rotation is easy and effective. Prior circumcision neither disturbs the operative procedure nor affects the outcomes. PMID:26600889

  2. Penile degloving and dorsal dartos flap rotation approach for the management of isolated penile torsion

    PubMed Central

    Aykaç, Aykut; Baran, Özer; Yapıcı, Onur; Aygün, Bülent Alper; Aydın, Cemil; Çakan, Murat

    2016-01-01

    Objective Penile torsion is a counterclockwise rotational anomaly of the penile shaft or glans. We aimed to evaluate the outcomes of penile degloving and dorsal dartos flap rotation used for the repair of isolated penil torsion. Material and methods During evaluation of the patients admitted to our polyclinic for circumcision between January 2013-December 2014, 5 cases of isolated penile torsion were determined. Following the circumcision procedure performed with bipolar cautery, patients undergoing penile degloving were checked whether penile torsion was relieved or not. In case of insufficient improvement, torsion was corrected with application of dorsal dartos flap. Penile torsion was corrected with dartos flap in 2, and penile degloving in 3 cases. Results The mean age of the patients was 5.6 years (4–7), and the mean operative time 12 minutes (7–20), respectively. The mean operation time was 17.5 (15–20) minutes and 8.3 (7–10) minutes in the dorsal dartos flap and penile degloving groups, respectively. The preoperative mean degree of penile torsion was 50° (30°–70°). The mean degree of torsion was 65° and 40° in the dorsal dartos and penile degloving groups, respectively. During the postoperative follow up, 1 case of residual torsion (<10°) was observed in the dorsal dartos flap group. Residual torsion was not observed in other patients. Conclusion Exploration for isolated cases of penile torsion during the circumcision procedure should not be overlooked. Successful results can be obtained with penile degloving and dartos flap rotation in cases with low and moderate torsion. PMID:27011878

  3. A penile spine/vibrissa enhancer sequence is missing in modern and extinct humans but is retained in multiple primates with penile spines and sensory vibrissae.

    PubMed

    Reno, Philip L; McLean, Cory Y; Hines, Jasmine E; Capellini, Terence D; Bejerano, Gill; Kingsley, David M

    2013-01-01

    Previous studies show that humans have a large genomic deletion downstream of the Androgen Receptor gene that eliminates an ancestral mammalian regulatory enhancer that drives expression in developing penile spines and sensory vibrissae. Here we use a combination of large-scale sequence analysis and PCR amplification to demonstrate that the penile spine/vibrissa enhancer is missing in all humans surveyed and in the Neandertal and Denisovan genomes, but is present in DNA samples of chimpanzees and bonobos, as well as in multiple other great apes and primates that maintain some form of penile integumentary appendage and facial vibrissae. These results further strengthen the association between the presence of the penile spine/vibrissa enhancer and the presence of penile spines and macro- or micro- vibrissae in non-human primates as well as show that loss of the enhancer is both a distinctive and characteristic feature of the human lineage.

  4. Isolated penile degloving from milling machine injury in a child.

    PubMed

    Onumaegbu, O O; Okechukwu, O C

    2015-01-01

    Degloving injuries of the male perineum are an uncommon urological emergency often requiring reconstruction. They are usually related to industrial and/or agricultural machinery and tend to involve both the penile shaft and the scrotal skin. Adolescents and young adults are the usual victims. The treatment options commonly employed include the use of the degloved skin either as a flap or more often as a free skin graft, yielding variable results. Other modalities include the use of free split skin grafts or free full thickness skin grafts (FTSG) (Wolfe grafts). We report a case of degloving injury in a 10-year-old child, with injury exclusively affecting the skin of the penile shaft, who presented in the sub-acute phase. Optimization of the wound and subsequent cover of the defect with an FTSG yielded a good outcome and patient satisfaction.

  5. Human penile erection and organic impotence: normal histology and histopathology.

    PubMed

    Conti, G; Virag, R

    1989-01-01

    A very large amount of human material (7 embryos, 12 stillborns, 12 penes of males aged between 2 and 86 years, as well as bioptical material from 80 subjects affected by impotence problems) has been examined so as to study the penis arterial and venous walls, the blood flow regulation mechanisms and the intracavernal trabecular morphology. The amount of muscle tissue and of collagenous connective tissue has been numerically quantified by computer-assisted methods. This study enables the authors to underline three fundamental facts: (a) it confirms the normal penile erection mechanism, and the consequent theory, (b) it confirms that vascular sclerosis is a systemic phenomenon correlated to age, and that the penis is not exempt, and (c) in the case of impotence problems, the same sclerosis phenomenon may appear at an earlier age, and therefore induce pathological impotence. PMID:2800066

  6. Brain potentials related to the human penile erection.

    PubMed

    Ponseti, J; Kropp, P; Bosinski, H A

    2009-01-01

    The aim of this study was to elucidate the brain processes preceding penile responses. Electroencephalographic (EEG) potentials and penile circumference were recorded simultaneously while male subjects were exposed to visual sexual stimuli (VSS). The trials were sorted by the penile response of the subjects (erection, maintenance or detumescence). The corresponding EEG recordings were then subjected to independent component analysis. We found that 200 ms after VSS onset brain potentials differ according to the genital response to follow. Whereas early posterior negativity (EPN) was predominantly related to erection and maintenance, P3-like activity was found to precede detumescence. EPN indicates a more 'emotional' processing state of the brain, whereas P3-like activity related to detumescence indicates a more 'cognitive' processing state. The latter is assumed to reflect activity of the locus coeruleus-norepinephrine system. Further research should evaluate the contribution of P3-related brain activity to psychogenic erectile dysfunction.

  7. Human papillomavirus type 16 sequence variation in concurrent vulvar and penile carcinoma in a married couple.

    PubMed

    Mitamura, Takashi; Watari, Hidemichi; Yamashiro, Katsushige; Kato, Tatsuya; Hosaka, Masayoshi; Shimada, Chisa; Fukumoto, Shun; Noshiro, Kiwamu; Sasaki, Takayuki; Sakuragi, Noriaki

    2015-06-01

    We encountered an elderly married couple with concurrent vulvar and penile carcinoma with an Asian variant of human papillomavirus type 16. Asian variants might have an elevated risk of concurrent external genital carcinomas of a male and a female, and analysis of human papillomavirus variants might be important to understand the mechanism of carcinogenesis. PMID:25970313

  8. Torsion/detorsion of the testis does not modify responses to nitric oxide in rat isolated penile bulb.

    PubMed

    Ozkan, Melike H; Vural, I Mert; Moralioglu, Serdar; Uma, Serdar; Sarioglu, Yusuf

    2007-08-01

    Ischaemia-reperfusion damage induced by torsion/detorsion of the testicles may be a causative factor leading to erectile dysfunction through oxidative stress-dependent changes in the responses of the penile bulb, an erectile tissue of the penis. We aimed at investigating the effects of unilateral testicular torsion/detorsion (2 or 24 hr) treatment on relaxations induced by electrical field stimulation and sodium nitroprusside in rat isolated penile bulb. Male Sprague-Dawley rats used in the study were divided into two groups. The treatment group was subjected to unilateral torsion followed by detorsion for 2 or 24 hr, while the control group underwent only sham operation. For in vitro organ bath experiments, penile bulbs were isolated and responses to relaxant agents and electrical field stimulation (70 V, 1 msec., 0.5-8 Hz, 5 sec.) were recorded on a computer-based data acquisition system via a force displacement transducer. In tissues precontracted with phenylephrine (3 x 10(-6 )M), relaxations induced by electrical field stimulation were not significantly different before and after 2 or 24 hr of detorsion. Similarly sodium nitroprusside- (10(-8)-3 x 10(-6 )M) and papaverine-induced (10(-7)-10(-4 )M) relaxations were also found unchanged in the detorsion group compared to control. In conclusion, spermatic cord torsion did not lead to impairment in nitric oxide-mediated relaxant responses of the rat isolated penile bulb.

  9. Male circumcision is associated with a lower prevalence of human papillomavirus-associated penile lesions among Kenyan men.

    PubMed

    Backes, Danielle M; Bleeker, Maaike C G; Meijer, Chris J L M; Hudgens, Michael G; Agot, Kawango; Bailey, Robert C; Ndinya-Achola, J O; Hayombe, Juma; Hogewoning, Cornelis J A; Moses, Stephen; Snijders, Peter J F; Smith, Jennifer S

    2012-04-15

    Human papillomavirus (HPV)-associated penile lesions in men may increase the risk of HPV transmission to their female partners. Risk factor data on HPV-associated penile lesions are needed from regions with a high burden of cervical cancer. Visual inspection of the penis was conducted using a colposcope at the 24-month visit among participants in a randomized controlled trial of male circumcision in Kenya, from May 2006 to October 2007. All photos were read independently by two observers for quality control. Penile exfoliated cells sampled from the glans/coronal sulcus and the shaft were tested for HPV DNA using GP5+/6+ PCR and for HPV16, 18 and 31 viral loads using a real time PCR assay. Of 275 men, 151 were circumcised and 124 uncircumcised. The median age was 22 years. Circumcised men had a lower prevalence of flat penile lesions (0.7%) versus uncircumcised (26.0%); adjusted odds ratio (OR) = 0.02; 95% confidence interval (CI) = 0.003-0.1. Compared to men who were HPV negative, men who were HPV DNA positive (OR = 6.5; 95% CI = 2.4-17.5) or who had high HPV16/18/31 viral load (OR = 5.2; 95% CI = 1.1-24.4) had higher odds of flat penile lesions. Among men with flat penile lesions, HPV56 (29.0%) and 16 (25.8%) were the most common types within single or multiple infections. Flat penile lesions are much more frequent in uncircumcised men and associated with higher prevalence of HPV and higher viral loads. This study suggests that circumcision reduces the prevalence of HPV-associated flat lesions and may ultimately reduce male-to-female HPV transmission. PMID:21618520

  10. Three-dimensional mapping and comparative analysis of the distal human corpus cavernosum and the inflatable penile prosthesis.

    PubMed

    Hakky, Tariq S; Ferguson, Daniel; Spiess, Philippe E; Bradley, Paul; Lue, Tom F; Carrion, Rafael E

    2013-07-01

    The intricate anatomy of the corpus cavernosum in both the flaccid and tumescent state has not been fully elucidated. We report our experience using a three-dimensional (3D) scanner to reconstruct cadaveric casts and compare them with 3D images of two prototypes of penile prosthesis. Two different models of the Titan Coloplast inflatable penile prosthesis were analyzed using a 3D scanner. The first was the standard model and the second was a newer model with a rounder silicone tip. Two cadaveric phalluses were harvested using Smooth-Cast 300Q polyurethane molding. The molds were excised and scanned along side the penile prosthesis. 3D scans were completed and analyzed using Leios Mesh software, and GOM Inspect software. The 3D scans demonstrated the mean human corporal radii 2 mm from the distal tip to be 36.51 mm (36.01-37.0 mm), which is an obtuse angle. The standard Titan penile prosthesis spherical radius at the same level was 202.52 mm, while the new silicone tip prosthesis had a radius of 139.33 mm. 3D mapping further demonstrated the trajectory of the cavernosa appeared curvilinear and the distal ends appeared blunt. The use of cadaveric cavernosal molds in combination with the 3D scanner allowed us to accurately image the corpus cavernosum for the first time. Our findings suggest that anatomically accurate corporal tips appear to be relatively blunt and that the new Titan silicone tip penile prosthesis more closely resembles the human corporal tip.

  11. Prevalence and risk factors of human papillomavirus infection by penile site in uncircumcised Kenyan men.

    PubMed

    Smith, Jennifer S; Backes, Danielle M; Hudgens, Michael G; Bailey, Robert C; Veronesi, Giovanni; Bogaarts, Martijn; Agot, Kawango; Ndinya-Achola, J O; Maclean, Ian; Agingu, Walter; Meijer, Chris J L M; Moses, Stephen; Snijders, Peter J F

    2010-01-15

    Human papillomavirus (HPV) prevalence was estimated from 2,705 sexually active, uncircumcised, human immunodeficiency virus seronegative men aged 17-28 years in Kisumu, Kenya. HPV prevalence was 51.1% (95% confidence interval: 49.2-53.0%) in penile cells from the glans/coronal sulcus and/or shaft. HPV prevalence varied by anatomical site, with 46.5% positivity in the glans/coronal sulcus compared with 19.1% in the shaft (p < 0.0001). High-risk HPV was detected in 31.2% of glans and 12.3% of shaft samples (p < 0.0001). HPV16 was the most common type and 29.2% of men were infected with more than one HPV type. Risk factors for HPV infection included presence of C. trachomatis, N. gonorrhea, self-reported sexually transmitted infections, and less frequent bathing. Lifetime number of sexual partners and herpes simplex virus type-2 seropositivity were also marginally associated with HPV infection. PMID:19626601

  12. DNA Copy Number Aberrations, and Human Papillomavirus Status in Penile Carcinoma. Clinico-Pathological Correlations and Potential Driver Genes

    PubMed Central

    Lambros, Maryou; Stankiewicz, Elzbieta; Ng, Charlotte K. Y.; Weigelt, Britta; Rajab, Ramzi; Tinwell, Brendan; Corbishley, Cathy; Watkin, Nick; Berney, Dan; Reis-Filho, Jorge S.

    2016-01-01

    Penile squamous cell carcinoma is a rare disease, in which somatic genetic aberrations have yet to be characterized. We hypothesized that gene copy aberrations might correlate with human papillomavirus status and clinico-pathological features. We sought to determine the spectrum of gene copy number aberrations in a large series of PSCCs and to define their correlations with human papillomavirus, histopathological subtype, and tumor grade, stage and lymph node status. Seventy formalin-fixed, paraffin embedded penile squamous cell carcinomas were centrally reviewed by expert uropathologists. DNA was extracted from micro-dissected samples, subjected to PCR-based human papillomavirus assessment and genotyping (INNO-LiPA human papillomavirus Genotyping Extra Assay) and microarray-based comparative genomic hybridization using a 32K Bacterial Artificial Chromosome array platform. Sixty-four samples yielded interpretable results. Recurrent gains were observed in chromosomes 1p13.3-q44 (88%), 3p12.3-q29 (86%), 5p15.33-p11 (67%) and 8p12-q24.3 (84%). Amplifications of 5p15.33-p11 and 11p14.1-p12 were found in seven (11%) and four (6%) cases, respectively. Losses were observed in chromosomes 2q33-q37.3 (86%), 3p26.3-q11.1 (83%) and 11q12.2-q25 (81%). Although many losses and gains were similar throughout the cohort, there were small significant differences observed at specific loci, between human papillomavirus positive and negative tumors, between tumor types, and tumor grade and nodal status. These results demonstrate that despite the diversity of genetic aberrations in penile squamous cell carcinomas, there are significant correlations between the clinico-pathological data and the genetic changes that may play a role in disease natural history and progression and highlight potential driver genes, which may feature in molecular pathways for existing therapeutic agents. PMID:26901676

  13. Penile size and penile enlargement surgery: a review.

    PubMed

    Dillon, B E; Chama, N B; Honig, S C

    2008-01-01

    Penile size is a considerable concern for men of all ages. Herein, we review the data on penile size and conditions that will result in penile shortening. Penile augmentation procedures are discussed, including indications, procedures and complications of penile lengthening procedures, penile girth enhancement procedures and penile skin reconstruction. PMID:18496548

  14. Involvement of large-conductance Ca(2+) -activated K(+) channels in both nitric oxide and endothelium-derived hyperpolarization-type relaxation in human penile small arteries.

    PubMed

    Király, István; Pataricza, János; Bajory, Zoltán; Simonsen, Ulf; Varro, András; Papp, Julius Gy; Pajor, Lászlo; Kun, Attila

    2013-07-01

    Large-conductance Ca(2+) -activated K(+) channels (BKC a ), located on the vascular smooth muscle, play an important role in regulation of vascular tone. In penile corpus cavernosum tissue, opening of BKC a channels leads to relaxation of corporal smooth muscle, which is essential during erection; however, there is little information on the role of BKC a channels located in penile vascular smooth muscle. This study was designed to investigate the involvement of BKC a channels in endothelium-dependent and endothelium-independent relaxation of human intracavernous penile arteries. In human intracavernous arteries obtained in connection with transsexual operations, change in isometric force was recorded in microvascular myographs, and endothelium-dependent [nitric oxide (NO) and endothelium-derived hyperpolarization (EDH)-type] and endothelium-independent (NO-donor) relaxations were measured in contracted arteries. In penile small arteries contracted with phenylephrine, acetylcholine evoked NO- and EDH-type relaxations, which were sensitive to iberiotoxin (IbTX), a selective blocker of BKC a channels. Iberiotoxin also inhibited relaxations induced by a NO-donor, sodium nitroprusside. NS11021, a selective opener of BKC a channels, evoked pronounced relaxations that were inhibited in the presence of IbTX. NS13558, a BKC a -inactive analogue of NS11021, failed to relax human penile small arteries. Our results show that BKC a channels are involved in both NO- and EDH-type relaxation of intracavernous penile arteries obtained from healthy men. The effect of a selective opener of BKC a channels also suggests that direct activation of the channel may be an advantageous approach for treatment of impaired endothelium-dependent relaxation often associated with erectile dysfunction.

  15. Penile Cancer

    PubMed Central

    Clark, Peter E.; Spiess, Philippe E.; Agarwal, Neeraj; Biagioli, Matthew C.; Eisenberger, Mario A.; Greenberg, Richard E.; Herr, Harry W.; Inman, Brant A.; Kuban, Deborah A.; Kuzel, Timothy M.; Lele, Subodh M.; Michalski, Jeff; Pagliaro, Lance; Pal, Sumanta K.; Patterson, Anthony; Plimack, Elizabeth R.; Pohar, Kamal S.; Porter, Michael P.; Richie, Jerome P.; Sexton, Wade J.; Shipley, William U.; Small, Eric J.; Trump, Donald L.; Wile, Geoffrey; Wilson, Timothy G.; Dwyer, Mary; Ho, Maria

    2014-01-01

    Squamous cell carcinoma of the penis represents approximately 0.5% of all cancers among men in the United States and other developed countries. Although rare, it is associated with significant disfigurement, and only half of the patients survive beyond 5 years. Proper evaluation of both the primary lesion and lymph nodes is critical, because nodal involvement is the most important factor of survival. The NCCN Clinical Practice Guidelines in Oncology for Penile Cancer provide recommendations on the diagnosis and management of this devastating disease based on evidence and expert consensus. PMID:23667209

  16. 21 CFR 876.5020 - External penile rigidity devices.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false External penile rigidity devices. 876.5020 Section 876.5020 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... maintain sufficient penile rigidity for sexual intercourse. External penile rigidity devices include...

  17. 21 CFR 876.5020 - External penile rigidity devices.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false External penile rigidity devices. 876.5020 Section 876.5020 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... maintain sufficient penile rigidity for sexual intercourse. External penile rigidity devices include...

  18. 21 CFR 876.5020 - External penile rigidity devices.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false External penile rigidity devices. 876.5020 Section 876.5020 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... maintain sufficient penile rigidity for sexual intercourse. External penile rigidity devices include...

  19. Antibody responses following incident anal and penile infection with human papillomavirus in teenage men who have sex with men.

    PubMed

    Zou, Huachun; Tabrizi, Sepehr N; Grulich, Andrew E; Hocking, Jane S; Garland, Suzanne M; Bradshaw, Catriona S; Cornall, Alyssa M; Fairley, Christopher K; Chen, Marcus Y

    2016-08-01

    Men who have sex with men (MSM) are at risk for human papillomavirus (HPV)-related anal cancer. Few data exist on antibody responses following incident anogenital infection with HPV in teenage MSM. A cohort of 200 MSM aged 16-20 years from Melbourne, Australia were assessed at baseline, 3, 6 and 12 months. At each visit anal and penile swabs were collected for HPV DNA and serum for HPV antibodies for genotypes 6, 11, 16 and 18 (Merck's Multiplex Assays using Luminex). The main outcome, seroconversion, was defined as the detection of HPV antibodies following a negative antibody result for the same HPV type at baseline. The seroincidence rates for HPV types 6, 11, 16 and 18 were: 19 (95% CI 12-26), 7 (3-12), 4 (1-8) and 6 (3-11) per 100 person-years, respectively. Men who experienced incident anal HPV infections from types 6/11 were significantly more likely to develop serum antibodies to the same HPV type(s) than those who experienced incident anal infections from types 16/18 [73 vs. 18%, odds ratio (OR) = 15, 95% CI: 2-118]. The median time between incident anal HPV infection and seroconversion for HPV 6, 11, 16 and 18 was: 91, 38, 161 and 182 days, respectively. Antibody responses against HPV types 6/11 were significantly more likely to occur following incident anal compared with incident penile infection with HPV types 6/11 (OR = 6, 95% CI: 2-21). The likelihood of antibody responses following anogenital HPV infections depends on the HPV type and site of infection. PMID:26991809

  20. Ethical issues in penile transplantation.

    PubMed

    Zhang, Li-Chao; Zhao, Yong-Bing; Hu, Wei-Lie

    2010-11-01

    This article provides an overview of the ethical issues associated with penile transplantation, a form of composite tissue allografting. There is only one reported case of human penile transplantation, and, as such, this technique is considered to be experimental. The ethical issues at stake involve both the graft donor and the graft recipient. With regard to the recipient, there are significant concerns relating to surgical risks and benefits, informed consent, body image (including surgical expectations and outcomes) and compliance. Donor issues may include family consent and privacy, as well as graft harvesting (leaving the donor cadaver without a penis). Many of these ethical issues can be explored during the recipient's assessment and consent process. Because no medium-term or long-term outcome data for this procedure exist-only one such operation has ever been performed-the burdens and ethical issues concerning penile transplantation remain unknown.

  1. Evolution of penile prosthetic devices.

    PubMed

    Le, Brian; Burnett, Arthur L

    2015-03-01

    Penile implant usage dates to the 16th century yet penile implants to treat erectile dysfunction did not occur until nearly four centuries later. The modern era of penile implants has progressed rapidly over the past 50 years as physicians' knowledge of effective materials for penile prostheses and surgical techniques has improved. Herein, we describe the history of penile prosthetics and the constant quest to improve the technology. Elements of the design from the first inflatable penile prosthesis by Scott and colleagues and the Small-Carrion malleable penile prosthesis are still found in present iterations of these devices. While there have been significant improvements in penile prosthesis design, the promise of an ideal prosthetic device remains elusive. As other erectile dysfunction therapies emerge, penile prostheses will have to continue to demonstrate a competitive advantage. A particular strength of penile prostheses is their efficacy regardless of etiology, thus allowing treatment of even the most refractory cases. PMID:25763121

  2. 21 CFR 876.3350 - Penile inflatable implant.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Penile inflatable implant. 876.3350 Section 876.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3350 Penile inflatable...

  3. 21 CFR 876.3630 - Penile rigidity implant.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Penile rigidity implant. 876.3630 Section 876.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3630 Penile rigidity implant....

  4. 21 CFR 876.3350 - Penile inflatable implant.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Penile inflatable implant. 876.3350 Section 876.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3350 Penile inflatable...

  5. 21 CFR 876.3350 - Penile inflatable implant.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Penile inflatable implant. 876.3350 Section 876.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3350 Penile inflatable...

  6. 21 CFR 876.3630 - Penile rigidity implant.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Penile rigidity implant. 876.3630 Section 876.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3630 Penile rigidity implant....

  7. 21 CFR 876.3630 - Penile rigidity implant.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Penile rigidity implant. 876.3630 Section 876.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3630 Penile rigidity implant....

  8. 21 CFR 876.3350 - Penile inflatable implant.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Penile inflatable implant. 876.3350 Section 876.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3350 Penile inflatable...

  9. 21 CFR 876.3630 - Penile rigidity implant.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Penile rigidity implant. 876.3630 Section 876.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3630 Penile rigidity implant....

  10. 21 CFR 876.3630 - Penile rigidity implant.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Penile rigidity implant. 876.3630 Section 876.3630 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3630 Penile rigidity implant....

  11. 21 CFR 876.3350 - Penile inflatable implant.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Penile inflatable implant. 876.3350 Section 876.3350 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Prosthetic Devices § 876.3350 Penile inflatable...

  12. General Information about Penile Cancer

    MedlinePlus

    ... Treatment Penile Cancer Treatment (PDQ®)–Patient Version General Information About Penile Cancer Go to Health Professional Version ... the PDQ Adult Treatment Editorial Board . Clinical Trial Information A clinical trial is a study to answer ...

  13. The penile tourniquet

    PubMed Central

    Parsons, Brian A; Kalejaiye, Odunayo; Mohammed, Mustafa; Persad, Raj A

    2013-01-01

    Tourniquets are commonly used in penile surgery to achieve a bloodless operating field or produce artificial erections intraoperatively. Several techniques have been described, but there is a paucity of data and a lack of guidelines to direct their safe use. In penile surgery, it is the local rather than systemic effects of tourniquet use that are the main concern. Tourniquet time should be kept to a minimum, as the limited data available suggests that reperfusion injury can occur even after short periods of ischaemia. High risk groups such as diabetics and arteriopaths are at particular risk. Further studies are needed to determine safe tourniquet times and pressures. PMID:23542140

  14. Overexpression of ANXA1 in Penile Carcinomas Positive for High-Risk HPVs

    PubMed Central

    Calmon, Marilia Freitas; Mota, Mânlio Tasso de Oliveira; Babeto, Érica; Candido, Natália Maria; Girol, Ana Paula; Mendiburu, Carlos Fabian; Bonilha, Jane Lopes; Silvestre, Rodrigo Vellasco Duarte; Rosa, Bruno Miziara; Thomé, Jorge Alberto; Medeiros, Gustavo Hernandez Américo; Soares, Fernando Augusto; Guimarães, Gustavo Cardoso; de Arruda, José Germano Ferraz; Oliani, Sonia Maria; Villa, Luisa Lina; Vassallo, José; Rahal, Paula

    2013-01-01

    The incidence of penile cancer varies between populations but is rare in developed nations. Penile cancer is associated with a number of established risk factors and associated diseases including phimosis with chronic inflammation, human papillomavirus (HPV) infection, poor hygiene and smoking. The objective of this study was to identify genes related to this type of cancer. The detection of HPV was analyzed in 47 penile squamous cell carcinoma samples. HPV DNA was detected in 48.9% of penile squamous cell carcinoma cases. High-risk HPV were present in 42.5% of cases and low-risk HPV were detected in 10.6% of penile squamous cell carcinomas. The RaSH approach identified differential expression of Annexin A1 (ANXA1), p16, RPL6, PBEF1 and KIAA1033 in high-risk HPV positive penile carcinoma; ANXA1 and p16 were overexpressed in penile squamous cells positive for high-risk HPVs compared to normal penile samples by qPCR. ANXA1 and p16 proteins were significantly more expressed in the cells from high-risk HPV-positive penile carcinoma as compared to HPV-negative tumors (p<0.0001) independently of the subtype of the carcinoma. Overexpression of ANXA1 might be mediated by HPV E6 in penile squamous cell carcinoma of patients with high-risk HPVs, suggesting that this gene plays an important role in penile cancer. PMID:23341933

  15. Overexpression of ANXA1 in penile carcinomas positive for high-risk HPVs.

    PubMed

    Calmon, Marilia Freitas; Mota, Mânlio Tasso de Oliveira; Babeto, Érica; Candido, Natália Maria; Girol, Ana Paula; Mendiburu, Carlos Fabian; Bonilha, Jane Lopes; Silvestre, Rodrigo Vellasco Duarte; Rosa, Bruno Miziara; Thomé, Jorge Alberto; Medeiros, Gustavo Hernandez Américo; Soares, Fernando Augusto; Guimarães, Gustavo Cardoso; de Arruda, José Germano Ferraz; Oliani, Sonia Maria; Villa, Luisa Lina; Vassallo, José; Rahal, Paula

    2013-01-01

    The incidence of penile cancer varies between populations but is rare in developed nations. Penile cancer is associated with a number of established risk factors and associated diseases including phimosis with chronic inflammation, human papillomavirus (HPV) infection, poor hygiene and smoking. The objective of this study was to identify genes related to this type of cancer. The detection of HPV was analyzed in 47 penile squamous cell carcinoma samples. HPV DNA was detected in 48.9% of penile squamous cell carcinoma cases. High-risk HPV were present in 42.5% of cases and low-risk HPV were detected in 10.6% of penile squamous cell carcinomas. The RaSH approach identified differential expression of Annexin A1 (ANXA1), p16, RPL6, PBEF1 and KIAA1033 in high-risk HPV positive penile carcinoma; ANXA1 and p16 were overexpressed in penile squamous cells positive for high-risk HPVs compared to normal penile samples by qPCR. ANXA1 and p16 proteins were significantly more expressed in the cells from high-risk HPV-positive penile carcinoma as compared to HPV-negative tumors (p<0.0001) independently of the subtype of the carcinoma. Overexpression of ANXA1 might be mediated by HPV E6 in penile squamous cell carcinoma of patients with high-risk HPVs, suggesting that this gene plays an important role in penile cancer. PMID:23341933

  16. Penile Epithelioid Angiomatoid Nodule.

    PubMed

    Pirpiris, Athina; Gilbourd, Daniel; Ranasinghe, Anudini; Dill, Tony; Lynnhtun, Kyaw; Rindani, Rahul

    2015-10-01

    Cutaneous epithelioid angiomatoid nodule is a rare clinical entity that is common on the trunk and limbs. This is the first report of penile cutaneous epithelioid angiomatoid nodule. Although it is a benign entity, it must be differentiated from vascular neoplasms, as it can bear similar clinical and pathologic features. PMID:26171823

  17. Penile incerceration—A tight affair

    PubMed Central

    Alkizim, Faraj O.; Kanyata, Daniel; Githaiga, Joseph; Oliech, Joseph

    2015-01-01

    A patient presents with penile ring incarceration after using it for penile enlargement and prolonging tumescence. We present a case study of removal of the penile ring under local anaesthesia in a setting where cutting tools were inadequate. PMID:26519807

  18. [A case report of progressive penile necrosis].

    PubMed

    Haba, Tomomi; Koike, Hiroshi

    2014-05-01

    The penis is provided with blood by multiple arteries. Penile necrosis is uncommon. Penile necrosis sporadically occurs in patients with progressive diabetes mellitus and/or end stage renal failure. Penile necrosis is often considered a poor prognostic feature. We present a case of penile necrosis in a patient with mild diabetes mellitus.

  19. Epidemiology and natural history of penile cancer.

    PubMed

    Pow-Sang, Mariela R; Ferreira, Ubirajara; Pow-Sang, Julio M; Nardi, Aguinaldo C; Destefano, Victor

    2010-08-01

    An extensive literature search was performed using the key words squamous cell carcinoma of the penis, phimosis, circumcision, chronic balanitis, cigarette smoking, genital warts and human papillomavirus (HPV) infection. All selected studies were classified according to the level of evidence (LE). The final grades of recommendation were assigned after discussion by the full panel of the International Consultation on Penile Cancer in November 2008. The factors positively associated with invasive penile cancers include the presence of phimosis (LE 3a), tobacco smoking (LE 3a-4), chewing tobacco (LE 3a), injury to the penis (LE 3a), balanitis (LE 3a), genital warts (LE 3a), and high-risk HPV infection (LE 3a-4). PMID:20691882

  20. Human Adaptation To Isolated And Confined Environments

    NASA Technical Reports Server (NTRS)

    Evans, Gary W.; Stokols, Daniel; Carrere, Sna Sybil

    1992-01-01

    Data from Antarctic research station analyzed. Report describes study of physiology and psychology of humans in isolated and confined environment. Suggests ways in which such environments made more acceptable to human inhabitants.

  1. Use of Human Hybridoma Technology To Isolate Human Monoclonal Antibodies.

    PubMed

    Smith, Scott A; Crowe, James E

    2015-02-01

    The human hybridoma technique offers an important approach for isolation of human monoclonal antibodies. A diversity of approaches can be used with varying success. Recent technical advances in expanding the starting number of human antigen-specific B cells, improving fusion efficiency, and isolating new myeloma partners and new cell cloning methods have enabled the development of protocols that make the isolation of human monoclonal antibodies from blood samples feasible. Undoubtedly, additional innovations that could improve efficiency are possible.

  2. A review of penile metastasis

    PubMed Central

    Mearini, Luigi; Colella, Renato; Zucchi, Alessandro; Nunzi, Elisabetta; Porrozzi, Carlo; Porena, Massimo

    2012-01-01

    Penile cancer as primary disease is relatively rare in developed countries. The penis is a rare site of metastases in spite of its rich vascularization. Approximately 500 cases have been reported in the literature; almost 70% of primary lesions are of pelvic origin (from genitourinary or recto-sigmoid primary tumors). We describe a case of penile metastasis from lung cancer. The rarity of the event prompted us to also explore related reviews and discuss the incidence, physiopathology, diagnosis and therapy of penile secondary cancer. PMID:25992200

  3. Penile enhancement procedures with simultaneous penile prosthesis placement.

    PubMed

    Hakky, Tariq S; Suber, Jessica; Henry, Gerard; Smith, David; Bradley, Paul; Martinez, Daniel; Carrion, Rafael E

    2012-01-01

    Here we present an overview of various techniques performed concomitantly during penile prosthesis surgery to enhance penile length and girth. We report on the technique of ventral phalloplasty and its outcomes along with augmentation corporoplasty, suprapubic lipectomy, suspensory ligament release, and girth enhancement procedures. For the serious implanter, outcomes can be improved by combining the use of techniques for each scar incision. These adjuvant procedures are a key addition in the armamentarium for the serious implant surgeon. PMID:22811703

  4. Sonographic Assessment of the Fetal Penile Development.

    PubMed

    Akpinar, Funda; Yilmaz, Saynur; Akdag Cirik, Derya; Kayikcioglu, Fulya; Dilbaz, Berna; Yucel, Husniye; Gelisen, Orhan

    2016-01-01

    The intrauterine diagnosis of micropenis is an important clue in the discernment of some syndromes and hormonal deficiencies. In this study, we tried to establish reference ranges for the fetal penile length and penile width. This prospective cross-sectional study included 179 healthy singleton male fetus pregnancies that were between 17 and 37 weeks of gestation. Of these pregnancies, the fetal penile length and width were measured using trans-abdominal ultrasound. The correlation coefficients of gestational age with penile measurements were calculated. We observed that as the gestational age increased both the penile length and width increased (p < .0001, correlation coefficients R(2) = 0.854 and R(2) = 0.883; respectively). Reference values of the penile length in the Turkish Population were similar to previously evaluated populations including English, American and Israeli populations. The penile width measurement is a convenient way to diagnose micropenis, but penile width measurement alone might miss some penile abnormalities including chordee and hypospadias.

  5. Drugs Approved for Penile Cancer

    Cancer.gov

    This page lists cancer drugs approved by the Food and Drug Administration (FDA) for penile cancer. The list includes generic names and brand names. The drug names link to NCI’s Cancer Drug Information summaries.

  6. How Is Penile Cancer Diagnosed?

    MedlinePlus

    ... These might include a biopsy and imaging tests. Biopsy A biopsy is needed to diagnose penile cancer. ... depends on the nature of the abnormality. Incisional biopsy For an incisional biopsy only a part of ...

  7. Immunohistochemical profile of the penile urethra and differential expression of GATA3 in urothelial versus squamous cell carcinomas of the penile urethra.

    PubMed

    Chaux, Alcides; Han, Jeong S; Lee, Stephen; Gonzalez-Roibon, Nilda; Sharma, Rajni; Burnett, Arthur L; Cubilla, Antonio L; Netto, George J

    2013-12-01

    The penile urethra has a distinctive morphology not yet fully characterized by immunohistochemistry. In addition, both urothelial and squamous cell carcinomas have been reported in the penile urethra, and the distinction between these 2 tumors might be difficult. The purposes of this study are to assess the histology and immunohistochemical profile (CK20, CK7, p63, and GATA3) of the penile urethra and to assess the usefulness of Trans-acting T-cell-specific transcription factor (GATA3) and human papillomavirus detection in distinguishing urothelial versus squamous cell carcinomas. Normal penile urethra was evaluated in 11 total penectomies. The penile urethra was lined by 2 cell layers: a superficial single layer of CK7+, CK20-, and p63- columnar cells and a deep stratified layer of CK7-, CK20-, and p63+ cubical cells. Both layers were GATA3+, supporting urothelial differentiation. In addition, 2 tissue microarrays and 6 surgical specimens of primary tumors of the penile urethra (3 urothelial and 3 squamous cell carcinomas) were evaluated for GATA3 expression. In the tissue microarrays, 22 of 25 upper tract urothelial carcinomas and 0 of 38 penile squamous cell carcinomas were GATA3+. In the surgical specimens, GATA3 was positive in all urothelial carcinomas and negative in all squamous cell carcinomas. Human papillomavirus was detected in 2 of 3 squamous cell carcinomas and in 0 of 3 of the urothelial carcinomas. In conclusion, the penile urethra is covered by epithelial cells that are unique in morphology and immunohistochemical profile. In addition, our study suggests that GATA3 and human papillomavirus detection are useful markers for distinguishing urothelial carcinomas from squamous cell carcinomas of the penile urethra.

  8. Penile cancer in patient with a ‘Bouglou’ penile adornment

    PubMed Central

    Teixeira, Thiago; Souza, Gustavo; Campos, Raiza; Pena, Luis; Hallak, Jorge; Tongu, Maira

    2014-01-01

    Implantation of penile nodules under the foreskin aims at improving male and female sexual pleasures during intercourse. This case discusses the characteristics of the South American penile adornment ‘Bouglou’ as well as the risk factors for penile cancer in this region of Amazonia. This is a case report of a 48-year-old man whose penis presented a destructive gland/penile shaft lesion and three adornments in the penile shaft, reported as ‘Bouglou’. The diagnosis of penile cancer was confirmed after total penectomy. This study is the first that suggests a possible association between squamous cell carcinoma and these penile adornments. PMID:25452262

  9. Magnetic resonance imaging of penile cancer.

    PubMed

    Gupta, Sumit; Rajesh, Arumugam

    2014-05-01

    Penile cancer is a rare neoplasm that, although rare in the developed world, has devastating physical and psychological consequences for the patient. Novel MR imaging techniques such as lymphotropic nanoparticle-enhanced MR imaging may help identify metastatic lymph node disease. This article reviews the normal penile anatomy and MR imaging techniques and features of primary and metastatic penile cancer. Recent advances in penile cancer imaging are discussed.

  10. Penile Granuloma Annulare.

    PubMed

    Mercieca, Liam; Carabot, Philip; Boffa, Michael J

    2016-01-01

    A 50-year-old man presented to the genitourinary medicine clinic with a 3-year history of skin-colored circular papules over the shaft and glans of the penis. There were multiple lesions that were initially small, around 0.3 cm in diameter, and gradually enlarged. Physical examination revealed five nodules over the glans and shaft of the penis, with the largest lesion measuring 3×1 cm over the lateral aspect of the shaft (Figure 1). There were no similar lesions elsewhere. There was no lymphadenopathy and the rest of the examination was unremarkable. The patient complained of discomfort during sexual intercourse but the lesions were otherwise asymptomatic and nontender. There was no history of trauma to the area and no dermatological history. He had had the same sexual partner for the past 22 years, no significant medical history, and was not taking any medication. He was a smoker with a 32-pack-year history. His family history did not include any dermatological diseases. His father was diagnosed with type II diabetes at 65 years of age and his mother had hypertension since age 60 years. He had consulted his general practitioner regarding the penile eruption a year earlier and was treated for a presumed fungal infection with clotrimazole cream for 1 month with no effect. Results from genitourinary investigations for sexually transmitted diseases including syphilis were all negative. PMID:27502267

  11. 21 CFR 876.5020 - External penile rigidity devices.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false External penile rigidity devices. 876.5020 Section 876.5020 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5020 External...

  12. 21 CFR 876.5020 - External penile rigidity devices.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false External penile rigidity devices. 876.5020 Section 876.5020 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5020 External...

  13. Melanocortin Receptors, Melanotropic Peptides and Penile Erection

    PubMed Central

    King, Stephen H.; Mayorov, Alexander V.; Balse-Srinivasan, Preeti; Hruby, Victor J.; Vanderah, Todd W.; Wessells, Hunter

    2009-01-01

    Penile erection is a complex physiologic event resulting from the interactions of the nervous system on a highly specialized vascular organ. Activation of central nervous system melanocortinergic (MC) receptors with either endogenous or synthetic melanotropic ligands may initiate and/or facilitate spontaneous penile erection. While the CNS contains principally the MC3 and MC4 receptor subtypes, there is conflicting data as to which receptor mediates erection. Although the MC4R is emerging as the principle effector of MC induced erection, the role of the MC3R is poorly understood. Manipulation of each receptor subtype with newly synthesized receptor specific agonists and antagonists, as well as knockout mice, has elucidated their individual contributions. Novel data from our laboratories suggests that antagonism of forebrain MC3R may enhance melanocortin-induced erections. Furthermore, melanocortin agents may interact with better-studied systems such as oxytocinergic pathways at the hypothalamic, brainstem or spinal level. Current therapies for erectile dysfunction target end organ vascular tissue. Manipulation of MC receptors may provide an alternative, centrally mediated therapeutic approach for erectile and other sexual dysfunctions. The non-specific “superpotent” MC agonist, PT-141, which is the carboxylate derivative of MT-II, has reached phase II human trials. Through their centrally mediated activity, melanocortin agonists have potential to treat erectile dysfunction as well as possible applications to the unmet medical needs of decreased sexual motivation and loss of libido. PMID:17584130

  14. Melanocortin receptors, melanotropic peptides and penile erection.

    PubMed

    King, Stephen H; Mayorov, Alexander V; Balse-Srinivasan, Preeti; Hruby, Victor J; Vanderah, Todd W; Wessells, Hunter

    2007-01-01

    Penile erection is a complex physiologic event resulting from the interactions of the nervous system on a highly specialized vascular organ. Activation of central nervous system melanocortinergic (MC) receptors with either endogenous or synthetic melanotropic ligands may initiate and/or facilitate spontaneous penile erection. While the CNS contains principally the MC3 and MC4 receptor subtypes, there is conflicting data as to which receptor mediates erection. Although the MC4R is emerging as the principle effector of MC induced erection, the role of the MC3R is poorly understood. Manipulation of each receptor subtype with newly synthesized receptor specific agonists and antagonists, as well as knockout mice, has elucidated their individual contributions. Novel data from our laboratories suggests that antagonism of forebrain MC3R may enhance melanocortin-induced erections. Furthermore, melanocortin agents may interact with better-studied systems such as oxytocinergic pathways at the hypothalamic, brainstem or spinal level. Current therapies for erectile dysfunction target end organ vascular tissue. Manipulation of MC receptors may provide an alternative, centrally mediated therapeutic approach for erectile and other sexual dysfunctions. The non-specific "superpotent" MC agonist, PT-141, which is the carboxylate derivative of MT-II, has reached phase II human trials. Through their centrally mediated activity, melanocortin agonists have potential to treat erectile dysfunction as well as possible applications to the unmet medical needs of decreased sexual motivation and loss of libido.

  15. Isolation of human serum spreading factor.

    PubMed

    Barnes, D W; Silnutzer, J

    1983-10-25

    Serum spreading factor (SF) was isolated from human serum by a four-step procedure employing affinity chromatography on glass beads, concanavalin A-Sepharose, DEAE-agarose, and heparin-agarose. The final product was purified approximately 260-fold from the starting material and was maximally active in assays of cell spreading-promoting activity at 300 ng/ml. The isolated human SF preparation consisted of two proteins of apparent molecular weights approximately 65,000 (SF65) and 75,000 (SF75). Both SF65 and SF75 have been shown previously to exhibit cell spreading-promoting activity and to bind monoclonal antibody to human serum SF. PMID:6630199

  16. Dysfunctional penile cholinergic nerves in diabetic impotent men

    SciTech Connect

    Blanco, R.; Saenz de Tejada, I.; Goldstein, I.; Krane, R.J.; Wotiz, H.H.; Cohen, R.A. )

    1990-08-01

    Impotence in the diabetic man may be secondary to a neuropathic condition of the autonomic penile nerves. The relationship between autonomic neuropathy and impotence in diabetes was studied in human corporeal tissue obtained during implantation of a penile prosthesis in 19 impotent diabetic and 15 nondiabetic patients. The functional status of penile cholinergic nerves was assessed by determining their ability to accumulate tritiated choline (34), and synthesize (34) and release (19) tritiated-acetylcholine after incubation of corporeal tissue with tritiated-choline (34). Tritiated-choline accumulation, and tritiated-acetylcholine synthesis and release were significantly reduced in the corporeal tissue from diabetic patients compared to that from nondiabetic patients (p less than 0.05). The impairment in acetylcholine synthesis worsened with the duration of diabetes (p less than 0.025). No differences in the parameters measured were found between insulin-dependent (11) and noninsulin-dependent (8) diabetic patients. The ability of the cholinergic nerves to synthesize acetylcholine could not be predicted clinically with sensory vibration perception threshold testing. It is concluded that there is a functional penile neuropathic condition of the cholinergic nerves in the corpus cavernosum of diabetic impotent patients that may be responsible for the erectile dysfunction.

  17. Testicular shielding in penile brachytherapy

    PubMed Central

    Bindal, Arpita; Tambe, Chandrashekhar M.; Ghadi, Yogesh; Murthy, Vedang; Shrivastava, Shyam Kishore

    2015-01-01

    Purpose Penile cancer, although rare, is one of the common genitourinary cancers in India affecting mostly aged uncircumcised males. For patients presenting with small superficial lesions < 3 cm restricted to glans, surgery, radical external radiation or brachytherapy may be offered, the latter being preferred as it allows organ and function preservation. In patients receiving brachytherapy, testicular morbidity is not commonly addressed. With an aim to minimize and document the doses to testis after adequate shielding during radical interstitial brachytherapy for penile cancers, we undertook this study in 2 patients undergoing brachytherapy and forms the basis of this report. Material and methods Two patients with early stage penile cancer limited to the glans were treated with radical high-dose-rate (HDR) brachytherapy using interstitial implant. A total of 7-8 tubes were implanted in two planes, parallel to the penile shaft. A total dose of 44-48 Gy (55-60 Gy EQD2 doses with α/β = 10) was delivered in 11-12 fractions of 4 Gy each delivered twice daily. Lead sheets adding to 11 mm (4-5 half value layer) were interposed between the penile shaft and scrotum. The testicular dose was measured using thermoluminescent dosimeters. For each patient, dosimetry was done for 3 fractions and mean calculated. Results The cumulative testicular dose to left and right testis was 31.68 cGy and 42.79 cGy for patient A, and 21.96 cGy and 23.28 cGy for patient B. For the same patients, the mean cumulative dose measured at the posterior aspect of penile shaft was 722.15 cGy and 807.72 cGy, amounting to 16.4% and 16.8% of the prescribed dose. Hence, the application of lead shield 11 mm thick reduced testicular dose from 722-808 cGy to 21.96-42.57 cGy, an “absolute reduction” of 95.99 ± 1.5%. Conclusions With the use of a simple lead shield as described, we were able to effectively reduce testicular dose from “spermicidal” range to “oligospermic” range with possible

  18. Signs, symptoms and treatment of penile fracture.

    PubMed

    Bhoil, Rohit; Sood, Dinesh

    2015-10-01

    Penile fracture is an uncommon injury and requires urgent treatment, therefore emergency nurses should be aware of the signs and symptoms and understand the importance of immediate surgical referral. This article describes the anatomy and physiology of penile erection and the ways in which penile fracture can occur. It also outlines the management of patients and includes a case study of a fracture caused by vigorous masturbation. PMID:26451939

  19. Penile perception of Koro patients.

    PubMed

    Chowdhury, A N

    1989-08-01

    Koro, a state where the perception of penis length decreases due to shrinkage from intra-abdominal traction, constitutes the central pathology. The present study was an attempt to explore the penis perception of the Koro patients by devising a graphomotor projective test, the Draw-a-penis Test (DAPT). DAPT elicited the penis image of the drawer, both of a normal penis and own penis perception. It also elicited the nature of perceptual process concerning the penile state changes, i.e. from flaccid to extended state. This controlled DAPT investigation showed a perceptual abnormality of penis image of Koro patients. They perceived less penis length, both for the penis of a normal person and their own penis and failed to perceive effectively the morphological change of penile states in terms of length increase. This perceptual abnormality in organ image is discussed in relation to the Koro vulnerability and psychopathology.

  20. [Hydrogen sulfide and penile erection].

    PubMed

    Huang, Yi-Ming; Cheng, Yong; Jiang, Rui

    2012-09-01

    Hydrogen sulfide (H2S) is the third type of active endogenous gaseous signal molecule following nitric oxide (NO) and carbon monoxide (CO). In mammalians, H2S is mainly synthesized by two proteases, cystathionine-beta-synthase (CBS) and cystathionine-gamma-lyase (CSE). H2S plays an essential function of physiological regulation in vivo, and promotes penile erection by acting on the ATP-sensitive potassium channels to relax the vascular smooth muscle as well as by the synergistic effect with testosterone and NO to relax the corpus cavernosum smooth muscle (CCSM). At present, the selective phosphodiesterase type 5 (PDE5) inhibitor is mainly used for the treatment of erectile dysfunction (ED), but some ED patients fail to respond. Therefore, further studies on the mechanism of H2S regulating penile erection may provide a new way for the management of erectile dysfunction.

  1. Genetic insights into human isolated gonadotropin deficiency.

    PubMed

    Trarbach, Ericka Barbosa; Silveira, Leticia Gontijo; Latronico, Ana Claudia

    2007-01-01

    The identification of naturally occurring genetic mutations has provided unique insight into the current knowledge of the human hypothalamic-pituitary-gonadal axis. In the past decade, several monogenic causes have been reported in patients with isolated gonadotropin deficiency. Kallmann Syndrome is a clinically and genetically heterogeneous disorder, characterized by isolated hypogonadotropic hypogonadism and anosmia or hyposmia. To date, loss-of-function mutations in the genes encoding anosmin-1 (KAL1) and fibroblast growth factor receptor 1 (FGFR1) have been described in the X-linked and autosomal dominant forms of this syndrome, respectively. More recently, several heterozygous, homozygous or compound heterozygous mutations in the G protein-coupled prokineticin receptor-2 (PROKR2) and one of its ligands, prokineticin-2 (PROK2) were described in Kallmann syndrome. In addition, complex genetic transmission (digenic inheritance) was recently demonstrated in this condition. Regarding isolated hypogonadotropic hypogonadism without olfactory abnormalities, loss-of-function mutations in the Gonadotropin-releasing hormone (GnRH) receptor (GnRH-R) or the G-protein coupled receptor 54 (GPR54) genes, both encoding transmembrane receptors, have been described, as well as FGFR1 mutations. Finally, mutations of the beta sub-units of LH and FSH have been described in patients with selective gonadotropin deficiency. We review the role of these distinct genetic factors in human isolated hypogonadotropic hypogonadism.

  2. Buried penis: An unrecognized risk factor in the development of invasive penile cancer

    PubMed Central

    Abdulla, Alym; Daya, Dean; Pinthus, Jehonathan; Davies, Timothy

    2012-01-01

    One of the documented benefits of neonatal circumcision is protection against invasive penile cancer. To date there have been a handful of published cases of invasive penile cancer in men circumcised as neonates. We report a case of a 73-year-old man, with a history of neonatal circumcision with no evidence of previous human papillomavirus exposure, who developed a buried penis secondary to obesity. He was diagnosed with Grade 2, pT3N0 squamous cell carcinoma of the penis. This report suggests that buried penis may pose a risk factor for the development of penile cancer despite the protective effects of neonatal circumcision. Thus periodic examination of a buried penis is warranted even in patients with no risk factors for penile cancer. A review of the literature is provided. PMID:23093645

  3. Neurophysiological basis of penile erection.

    PubMed

    Priviero, Fernanda B M; Leite, Romulo; Webb, R Clinton; Teixeira, Cleber E

    2007-06-01

    Penile erection involves a complex interaction between the central nervous system and local factors. It is a neurovascular event modulated by psychological and hormonal factors. The discovery of nitric oxide (NO) as an intercellular messenger or neurotransmitter paved the way for identifying important mechanisms underlying physiological and pathophysiological events in the penis, in addition to providing the knowledge for the development of new therapeutics based on a novel concept of molecule and cell interaction. Despite the fact that sinusoidal endothelial cells also produce and release NO in response to chemical and possibly physical stimuli, roles of neurogenic NO in penile erection appear to be more attractive and convincing, since the pharmacological neuromodulation represents an essential step to attaining penile erection. Erectile dysfunction (ED) is caused by a variety of pathogenic factors, particularly impaired formation and action of NO. Hence, a thorough knowledge of the physiology of erection is essential for future pharmacological innovations in the field of male ED, particularly targeting NO or intracellular cyclic GMP, which represent the most promising therapeutic approach to treat patients with ED.

  4. Isolation of Human Skin Dendritic Cell Subsets.

    PubMed

    Gunawan, Merry; Jardine, Laura; Haniffa, Muzlifah

    2016-01-01

    Dendritic cells (DCs) are specialized leukocytes with antigen-processing and antigen-presenting functions. DCs can be divided into distinct subsets by anatomical location, phenotype and function. In human, the two most accessible tissues to study leukocytes are peripheral blood and skin. DCs are rare in human peripheral blood (<1 % of mononuclear cells) and have a less mature phenotype than their tissue counterparts (MacDonald et al., Blood. 100:4512-4520, 2002; Haniffa et al., Immunity 37:60-73, 2012). In contrast, the skin covering an average total surface area of 1.8 m(2) has approximately tenfold more DCs than the average 5 L of total blood volume (Wang et al., J Invest Dermatol 134:965-974, 2014). DCs migrate spontaneously from skin explants cultured ex vivo, which provide an easy method of cell isolation (Larsen et al., J Exp Med 172:1483-1493, 1990; Lenz et al., J Clin Invest 92:2587-2596, 1993; Nestle et al., J Immunol 151:6535-6545, 1993). These factors led to the extensive use of skin DCs as the "prototype" migratory DCs in human studies. In this chapter, we detail the protocols to isolate DCs and resident macrophages from human skin. We also provide a multiparameter flow cytometry gating strategy to identify human skin DCs and to distinguish them from macrophages. PMID:27142012

  5. HPV type concordance in sexual couples determines the effect of condoms on regression of flat penile lesions.

    PubMed

    Bleeker, M C G; Berkhof, J; Hogewoning, C J A; Voorhorst, F J; van den Brule, A J C; Starink, T M; Snijders, P J F; Meijer, C J L M

    2005-04-25

    We earlier demonstrated, in a randomised clinical trial, that the regression time of flat penile lesions in male sexual partners of women with cervical intraepithelial neoplasia (CIN) was shorter in men who used condoms compared to those who did not. To further evaluate this finding, we examined whether the effect of condom use on the regression of flat penile lesions depends on the presence of human papillomavirus (HPV) type concordance in sexual couples, as determined in cervical and penile scrapes by GP5+/6+ PCR testing. A Cox model with time-dependent covariates showed a beneficial effect of condoms on regression of flat penile lesions in concordant couples (hazard ratio 2.63, 95% CI 1.07-6.48) but not in those who were nonconcordant. When both partners harboured different HPV types, no effect of condoms was found (hazard ratio 0.90, 95% CI 0.27-2.96). Delayed regression of flat penile lesions was associated with either stable lesions or with new penile lesions developing at sites surrounding pre-existing lesions suggesting reinfection of the penile epithelium. We conclude that condom use blocks sexual HPV transmission by preventing reinfection and development of new penile lesions in men who are susceptible to the same type as present in the female partner.

  6. Human blood glycosaminoglycans: isolation and analysis.

    PubMed

    Anower-E-Khuda, Md Ferdous; Kimata, Koji

    2015-01-01

    Glycosaminoglycans (GAGs) are linear polysaccharides having disaccharide building blocks consisting of an amino sugar (N-acetylglucosamine, or N-acetylgalactosamine) and a uronic acid (glucuronic acid or iduronic acid) or galactose. Glycosaminoglycans have sulfated residues at various positions except for hyaluronan, and those sulfated residues regulate the biological functions of a wide variety of proteins, primarily through high-affinity interactions mediated by specific patterns/densities of sulfation and sugar sequences. Alteration of GAG structure is associated with a number of disease conditions and therefore the analyses of GAG structures and their sulfation patterns are important for the development of disease biomarkers and for treatment options. Extensive structural and quantitative analyses of GAGs from human blood are largely unexplored which may be due to the exhaustive isolation process because of the presence of too much interfering proteins and lipids such as serum albumin. Therefore we established a new GAG isolation method using the least amount (~200 μl) of human blood, consisting of a combination of proteolytic digestion and selective ethanol precipitation of GAGs, digestion of GAGs recovered on the filter cup by direct addition of GAG lyase reaction solution, and subsequent high-pressure liquid chromatography of unsaturated disaccharide products that enable to analyze GAG structures and contents. This isolation method offers an 80 % recovery of GAGs and can be applied to analyze a minute GAG content (≥1 nmol) from the least amount of biological fluids. Hence the method could be useful for the development of disease biomarkers.

  7. [A cylinder aneurysm of a penile prosthesis].

    PubMed

    Pannek, J; Bartel, P; Göcking, K

    2011-07-01

    Herniation of a penile prosthesis (cylinder aneurysm) is an extremely rare complication of penile prosthesis surgery. We report the first case of such an aneurysm in a patient with spinal cord injury. The treatment of choice is surgical revision with replacement of the faulty device. Filling of the implanted system with contrast media facilitates preoperative diagnostic workup. PMID:21567276

  8. Penetration of aflatoxins through isolated human epidermis

    SciTech Connect

    Riley, R.T.; Kemppainen, B.W.; Norred, W.P.

    1985-01-01

    The purpose of this study was to determine if aflatoxin B1 (AFB1) could penetrate through isolated human epidermis (stratum corneum plus viable epidermis). ( UC)AFB1 (7.5-9.3 micrograms) was applied to the stratum corneum of epidermal disks mounted in Teflon diffusion cells. ( UC)AFB1 penetrated chemically unaltered through the isolated epidermis. Chloroform-extractable radioactivity accounted for 82.5 +/- 3.7% of the total penetrating radioactivity in the receptor fluid of the diffusion cells. The rate of penetration was very slow when experiments were conducted under nonoccluded conditions, but was approximately 40 times greater under conditions of occlusion. Penetration after 46 h was less than 0.05% and 3.41% of the applied dose under nonoccluded and occluded conditions, respectively. Total recovery expressed as a percentage of the applied radioactivity was 98.6 +/- 6.4%.

  9. Isolation of osteocytes from human trabecular bone.

    PubMed

    Prideaux, Matthew; Schutz, Christine; Wijenayaka, Asiri R; Findlay, David M; Campbell, David G; Solomon, Lucian B; Atkins, Gerald J

    2016-07-01

    Osteocytes are essential regulators of bone homeostasis. However, they are difficult to study due to their location within the bone mineralised matrix. Although several techniques have been published for the isolation of osteocytes from mouse bone, no such technique has been described for human osteocytes. We have therefore developed a protocol for the isolation of osteocytes from human trabecular bone samples acquired during surgery. The cells were digested from the bone matrix by sequential collagenase and ethylenediaminetetraacetic acid (EDTA) digestions and the cells from later digests displayed characteristic dendritic osteocyte morphology when cultured ex vivo. Furthermore, the cells expressed characteristic osteocyte marker genes, such as E11, dentin matrix protein 1 (DMP1), SOST, matrix extracellular phosphoglycoprotein (MEPE) and phosphate regulating endopeptidase homologue, X-linked (PHEX). In addition, genes associated with osteocyte perilacunar remodelling, including matrix metallopeptidase-13 (MMP13), cathepsin K (CTSK) and carbonic anhydrase 2 (CAR2) were expressed. The cells also responded to parathyroid hormone (PTH) by downregulating SOST mRNA expression and to 1α,25-dihydroxyvitamin D3 (1,25D) by upregulating fibroblast growth factor 23 (FGF23) mRNA expression. Therefore, the cells behave in a similar manner to osteocytes in vivo. These cells represent an important tool in enhancing current knowledge in human osteocyte biology. PMID:27109824

  10. Genetics of human isolated hereditary nail disorders.

    PubMed

    Khan, S; Basit, S; Habib, R; Kamal, A; Muhammad, N; Ahmad, W

    2015-10-01

    Human hereditary nail disorders constitute a rare and heterogeneous group of ectodermal dysplasias. They occur as isolated and/or syndromic ectodermal conditions where other ectodermal appendages are also involved, and can occur associated with skeletal dysplasia. 'Nail disorder, nonsyndromic congenital' (OMIM; Online Mendelian Inheritance in Man) is subclassified into 10 different types. The underlying genes identified thus far are expressed in the nail bed and play important roles in nail development and morphogenesis. Here, we review the current literature on nail disorders and present a coherent review on the genetics of nail disorders. This review will pave the way to identifying putative genes and pathways involved in nail development and morphogenesis.

  11. Patient satisfaction and penile morphology changes with postoperative penile rehabilitation 2 years after Coloplast Titan prosthesis

    PubMed Central

    Pryor, Michael B; Carrion, Rafael; Wang, Run; Henry, Gerard

    2016-01-01

    A common complaint after inflatable penile prosthesis surgery is reduced penile length. We previously reported how using the Coloplast Titan inflatable penile prosthesis with aggressive new length measurement technique (NLMT) coupled with postoperative IPP rehabilitation of the implant for 1-year helped to improve patient satisfaction and erectile penile measurements. This is a 2 years follow-up of a prospective, three-center, study of 40 patients who underwent Titan prosthesis placement, with new length measurement technique for erectile dysfunction. Patient instructions were to inflate daily for 6 months and then inflate maximally for 1–2 h daily for 6–24 months. Fifteen penile measurements were taken before and immediately after surgery and at follow-up visits. Measurement changes were improved at 24 months as compared to immediately postoperative and at 12 months. 67.8% of subjects were satisfied with their length at 2 years, and 77% had perceived penile length that was longer (30.8%) or the same (46.2%) as prior to the surgery. 64.3% and 17.9% of subjects had increased and unchanged satisfaction, respectively, with penile length as compared to prior to penile implant surgery. All but one subject (96.5%) was satisfied with the overall function of his implant. This study suggests using the Coloplast Titan with aggressive cylinder sizing, and a postoperative penile rehabilitation inflation protocol can optimize patient satisfaction and erectile penile measurements at 2 years postimplant. PMID:26459782

  12. Patient satisfaction and penile morphology changes with postoperative penile rehabilitation 2 years after Coloplast Titan prosthesis.

    PubMed

    Pryor, Michael B; Carrion, Rafael; Wang, Run; Henry, Gerard

    2016-01-01

    A common complaint after inflatable penile prosthesis surgery is reduced penile length. We previously reported how using the Coloplast Titan inflatable penile prosthesis with aggressive new length measurement technique (NLMT) coupled with postoperative IPP rehabilitation of the implant for 1-year helped to improve patient satisfaction and erectile penile measurements. This is a 2 years follow-up of a prospective, three-center, study of 40 patients who underwent Titan prosthesis placement, with new length measurement technique for erectile dysfunction. Patient instructions were to inflate daily for 6 months and then inflate maximally for 1-2 h daily for 6-24 months. Fifteen penile measurements were taken before and immediately after surgery and at follow-up visits. Measurement changes were improved at 24 months as compared to immediately postoperative and at 12 months. 67.8% of subjects were satisfied with their length at 2 years, and 77% had perceived penile length that was longer (30.8%) or the same (46.2%) as prior to the surgery. 64.3% and 17.9% of subjects had increased and unchanged satisfaction, respectively, with penile length as compared to prior to penile implant surgery. All but one subject (96.5%) was satisfied with the overall function of his implant. This study suggests using the Coloplast Titan with aggressive cylinder sizing, and a postoperative penile rehabilitation inflation protocol can optimize patient satisfaction and erectile penile measurements at 2 years postimplant.

  13. A Contemporary Review of HPV and Penile Cancer.

    PubMed

    Stratton, Kelly L; Culkin, Daniel J

    2016-03-01

    Human papillomavirus (HPV) is a widespread sexually transmitted infection. In both men and women, HPV infection can result in a spectrum of genitourinary manifestations ranging from genital warts to cancer. Cervical cancer is nearly always associated with high-risk HPV infection. For men, penile cancer can develop following or independently of HPV infection. Basaloid and warty subtypes of penile squamous cell carcinoma are most frequently associated with HPV infection. Further research into the molecular alterations caused by HPV infection may provide prognostic markers and future treatment targets. Until an effective treatment for HPV infection is developed, prevention will remain the focus of disease control. For women, vaccination is increasingly utilized to prevent HPV infection and subsequent cervical cancer development. New recommendations for routine male vaccination may further reduce cancers for both men and women.

  14. Lymphatic Dissemination of Simian Immunodeficiency Virus after Penile Inoculation

    PubMed Central

    Ma, Zhong-Min; Dutra, Joseph; Fritts, Linda

    2016-01-01

    ABSTRACT The human immunodeficiency virus (HIV) is primarily transmitted by heterosexual contact, and approximately equal numbers of men and women worldwide are infected with the virus. Understanding the biology of HIV acquisition and dissemination in men exposed to the virus by insertive penile intercourse is likely to help with the rational design of vaccines that can limit or prevent HIV transmission. To characterize the target cells and dissemination pathways involved in establishing systemic simian immunodeficiency virus (SIV) infection, we necropsied male rhesus macaques at 1, 3, 7, and 14 days after penile SIV inoculation and quantified the levels of unspliced SIV RNA and spliced SIV RNA in tissue lysates and the number of SIV RNA-positive cells in tissue sections. We found that penile (glans, foreskin, coronal sulcus) T cells and, to a lesser extent, macrophages and dendritic cells are primary targets of infection and that SIV rapidly reaches the regional lymph nodes. At 7 days after inoculation, SIV had disseminated to the blood, systemic lymph nodes, and mucosal lymphoid tissues. Further, at 7 days postinoculation (p.i.), spliced SIV RNA levels were the highest in the genital lymph nodes, indicating that this is the site where the infection is initially amplified. By 14 days p.i., spliced SIV RNA levels were high in all tissues, but they were the highest in the gastrointestinal tract, indicating that the primary site of virus replication had shifted from the genital lymph nodes to the gut. The stepwise pattern of virus replication and dissemination described here suggests that vaccine-elicited immune responses in the genital lymph nodes could help prevent infection after penile SIV challenge. IMPORTANCE To be the most effective, vaccines should produce antiviral immune responses in the anatomic sites of virus replication. Thus, understanding the path taken by HIV from the mucosal surfaces, which are the site of virus exposure, to the deeper tissues where

  15. Penile anthropometry in North Indian children

    PubMed Central

    Bhat, Amilal; Upadhyay, Ravi; Bhat, Mahakshit; Sabharwal, Karamveer; Singla, Manish; Kumar, Vinay

    2015-01-01

    Introduction: Physicians frequently encounter questions by parents regarding the normal size of a child's penis. We evaluated normal variations of penile dimensions, correlation of penile length with height, weight, and body mass index (BMI) of boys and analyzed the differences in penile dimensions from those reported from other countries. Materials and Methods: A cross-sectional study was conducted at our institution during October 2012-December 2012. A total of 250 subjects (birth to 10 years) were evaluated and divided into 10 groups with 1-year interval taking 25 children in each. Penile dimensions measured twice by a single observer with Vernier calipers included the length of flaccid penis fully stretched and diameters at mid-shaft and corona. Diameters were multiplied by pi (π = 3.14) to calculate circumferences. Mean, standard deviation, and range were calculated. Height, weight, and BMI were noted and statistically correlated with the penile length using the Pearson correlation coefficient. Data were compared with similar studies reported on other populations in the past and individually evaluated with every study using Student's t-test. Results: The mean values for the penile length, mid-shaft circumference, and coronal circumference were 3.34, 3.05, 3.29 cm during infancy, 4.28, 3.86, 4.11 cm during 4-5 years, and 5.25, 4.78, 5.05 cm during 9-10 years, respectively. The penile length increased with advancing age in successive age groups, but it did not have a direct correlation with either height, weight, or BMI. Penile dimensions in North Indian children were found to be statistically smaller in comparison with most studies from other countries. Conclusion: We provide the normal range and variations of penile dimensions in North Indian children. PMID:25878409

  16. Acute penile trauma and associated morbidity: 9-year experience at a tertiary care center.

    PubMed

    Phillips, Elizabeth A; Esposito, Anthony J; Munarriz, Ricardo

    2015-05-01

    Penile fracture is an uncommon urologic emergency, defined as traumatic rupture of the tunica albuginea of the corpus cavernosum. It occurs mainly in young adults during sexual activity. In the United States, urethral injury is associated with 10-38% of all penile fractures. Diagnosis can be made clinically with the classic triad of an audible crack, detumescence, and appearance of hematoma. We sought to identify characteristics associated with true penile fracture vs. other diagnoses, and determine associated morbidity and risk factors for complications. Retrospective operative chart review identified 39 patients (mean age 39.4 years) with clinical features of penile fracture presenting to Boston Medical Center from June 2004 to May 2013. Average time from injury to presentation was 76 h (range 0.5 h-9 days) and the mechanism of injury was coital in 32 (82%) patients. Thirty-two patients (82%) had confirmed penile fracture, 7 (18%) had isolated vascular injury. Of confirmed fractures, 4 (13%) had bilateral corporal injury and associated urethral injury. Imaging was utilized in a total of 21 cases, penoscrotal ultrasound (US) in 17 cases, retrograde urethrogram (RUG) in 3 cases, and magnetic resonance imaging (MRI) in 1 case. Penile exploration was carried out via degloving (n = 5, 13%) or penoscrotal (n = 34, 87%) incisions. At follow-up, six patients (15%) had complications: 2 wound infections, 2 new-onset erectile dysfunction (ED), 1 urethral stricture, 1 fistula and 1 wound dehiscence. Urethral injury increased the risk of post-operative complications (p = 0.015). Penile fracture is primarily a clinical diagnosis, however imaging may be helpful if diagnosis is uncertain. Urethral injury should be suspected in cases of bilateral corporal injury and may be associated with increased morbidity. Surgical approach does not affect morbidity, but may facilitate surgical repair.

  17. Current Treatment Options for Penile Fractures

    PubMed Central

    Jack, Gregory S; Garraway, Isla; Reznichek, Richard; Rajfer, Jacob

    2004-01-01

    The diagnosis of “penile fracture” describes the traumatic rupture of the tunica albuginea of an erect penis. Penile fractures typically occur when the engorged penile corpora are forced to buckle and literally “pop” under the pressure of a blunt sexual trauma. Patients typically describe immediate detumescence, severe pain, and swelling as a result of the injury. Prompt surgical exploration and corporal repair is the most efficacious therapy. Although a majority of cases can be diagnosed from the history and physical examination alone, radiographic studies, including retrograde urethrography and corporal cavernosography can aid in the diagnosis of unusual cases. PMID:16985591

  18. Penile abscess secondary to neglected penile fracture after intracavernosal vasoactive drug injection.

    PubMed

    Song, Wan; Ko, Kwang Jin; Shin, Seung Jea; Ryu, Dong Soo

    2012-12-01

    Penile abscesses are rare, but can develop after trauma, injection therapy, or surgery of the penis, or as an unusual presentation of sexually transmitted diseases. We report a case of penile abscess in a 51-year-old diabetic man, presented 9 days after neglected penile fracture following intracavernosal injection therapy and sexual intercourse. Penile ultrasonography and surgical exploration confirmed the physical examination findings of involvement of the corpus cavernosum. The pus culture from the abscess revealed Enterococcous faecalis. The patient was successfully treated by surgical drainage of the abscess and primary closure of the ruptured tunica albuginea. PMID:23596611

  19. Atypical Chest Pain: An Unusual Presentation of Spinal Metastasis due to Penile Carcinoma

    PubMed Central

    Pywell, Sarah; Dott, Cameron; Khan, Mohammad Taimur; Sivanadarajah, Naveethan

    2016-01-01

    Spinal metastases may present in a myriad of ways, most commonly back pain with or without neurology. We report an unusual presentation of isolated atypical chest pain preceding metastatic cord compression, secondary to penile carcinoma. Spinal metastasis from penile carcinoma is rare with few cases reported. This unusual presentation highlights the need for a heightened level of clinical suspicion for spinal metastases as a possible cause for chest pain in any patients with a history of carcinoma. The case is discussed with reference to the literature. PMID:27429829

  20. PENILE AGENESIS WITH CROSSED FUSED RENAL ECTOPIA.

    PubMed

    Derbew, Milliard; Ketema, Elbet

    2015-04-01

    Penile agenesis is one of the rarest urogenital anomalies with only less than 100 cases reported worldwide so far. Only 3 cases have been reported from Africa and to our knowledge none has been reported from our country Ethiopia. Viability depends on associated anomalies. Urogenital anomalies are the most common associated ones accounting for 54% of cases. This case report is unusual presentation, which is the first reported case of penile agenesis associated with left to right, crossed fused renal ectopia.

  1. Genetic analysis of human clinical isolates of Lactococcus garvieae: Relatedness with isolates from foods.

    PubMed

    Reguera-Brito, Mercedes; Galán-Sánchez, Fátima; Blanco, M Mar; Rodríguez-Iglesias, Manuel; Domínguez, Lucas; Fernández-Garayzábal, José F; Gibello, Alicia

    2016-01-01

    Lactococcus garvieae is a Gram-positive bacterium well-known as an important pathogen in aquaculture, and it is also a human pathogen of increasing clinical significance. Forty-three human L. garvieae isolates from clinical specimens were characterized by Multilocus Sequence Typing (MLST). Twenty-six different sequence types (STs) were identified among the human isolates, of which 20 were novel STs. Most human isolates clustered into four clonal complexes, with a predominance of CC3. Within CC3, ST10 was the most common genotype, indicating the existence of a circulating genetic lineage among the human isolates analyzed. The four CCs also grouped L. garvieae strains isolated from meat, dairy and fish, indicating a genetic overlap between isolates from human and these foods. Genetic relatedness among human and food L. garvieae isolates was confirmed by phylogenetic analysis based on the concatenated sequences of the seven MLST genes. These results represent the first evidence of genetic relatedness between isolates of L. garvieae of human and those isolated meat, milk and dairy products and suggest that, in addition to fish and seafood, these foods might represent important sources of human L. garvieae infections.

  2. The pericyte as a cellular regulator of penile erection and a novel therapeutic target for erectile dysfunction.

    PubMed

    Yin, Guo Nan; Das, Nando Dulal; Choi, Min Ji; Song, Kang-Moon; Kwon, Mi-Hye; Ock, Jiyeon; Limanjaya, Anita; Ghatak, Kalyan; Kim, Woo Jean; Hyun, Jae Seog; Koh, Gou Young; Ryu, Ji-Kan; Suh, Jun-Kyu

    2015-06-05

    Pericytes are known to play critical roles in vascular development and homeostasis. However, the distribution of cavernous pericytes and their roles in penile erection is unclear. Herein we report that the pericytes are abundantly distributed in microvessels of the subtunical area and dorsal nerve bundle of mice, followed by dorsal vein and cavernous sinusoids. We further confirmed the presence of pericytes in human corpus cavernosum tissue and successfully isolated pericytes from mouse penis. Cavernous pericyte contents from diabetic mice and tube formation of cultured pericytes in high glucose condition were greatly reduced compared with those in normal conditions. Suppression of pericyte function with anti-PDGFR-β blocking antibody deteriorated erectile function and tube formation in vivo and in vitro diabetic condition. In contrast, enhanced pericyte function with HGF protein restored cavernous pericyte content in diabetic mice, and significantly decreased cavernous permeability in diabetic mice and in pericytes-endothelial cell co-culture system, which induced significant recovery of erectile function. Overall, these findings showed the presence and distribution of pericytes in the penis of normal or pathologic condition and documented their role in the regulation of cavernous permeability and penile erection, which ultimately explore novel therapeutics of erectile dysfunction targeting pericyte function.

  3. Microvascular temporalis fascia transfer for penile girth enhancement.

    PubMed

    Küçükçelebi, A; Ertaş, N M; Aydin, A; Eroğlu, A; Ozmen, E; Velidedeoğlu, H

    2001-07-01

    The authors report a 44-year-old man with inadequate penile girth that caused psychological problems. Using microvascular temporalis fascia transfer, they achieved satisfactory penile girth enhancement based on reliable vascularity in a single stage. PMID:11756810

  4. Physiological regulation of penile arteries and veins.

    PubMed

    Prieto, D

    2008-01-01

    Recent experimental evidence suggests that arterial insufficiency precedes the structural and functional changes in corpora cavernosa (CC) leading to organic erectile dysfunction (ED). The present review gives an overview of the physiological factors involved in the regulation of penile vasculature. Sympathetic nerves maintain flaccidity and tonically released noradrenaline induces vasoconstriction of both arteries and veins through alpha(1)- and alpha(2)-postsynaptic receptors and downregulates its own release and that of nitric oxide (NO) through alpha(2)-presynaptic receptors. The sympathetic cotransmitter neuropeptide Y (NPY) modulates noradrenergic vasoconstriction in penile small arteries by both enhancing and depressing noradrenaline contractions through Y(1)- and Y(2)-postsynaptic and a NO-independent atypical endothelial receptor, respectively. Activation of alpha(1)-adrenoceptors involves both Ca(2+) influx through L-type and receptor-operated Ca(2+) channels (ROC) and Ca(2+) sensitization mechanisms mediated by protein kinase C (PKC), tyrosine kinases (TKs) and Rho kinase (RhoK). In addition, RhoK can regulate Ca(2+) entry in penile arteries upon receptor stimulation. Vasodilatation of penile arteries and large veins during erection is mediated by neurally released NO. The subsequent increased arterial inflow to the cavernosal sinoids and shear stress on the endothelium lining penile arteries activates endothelial NO production through Akt phosphorylation of endothelial NO synthase (eNOS). NO stimulates guanylate cyclase and increased cyclic guanin 3'-monophosphate (cGMP) levels in turn activate protein kinase G (PKG), which enhances K(+) efflux through Ca(2+)-activated (K(Ca)) and voltage-dependent Ca(2+) (K(v)) channels in penile arteries and veins, respectively. PKG-mediated decrease in Ca(2+) sensitivity and its regulation by RhoK remains to be clarified in penile vasculature. Phosphodiesterase type 5 (PDE5) inhibitors are potent vasodilators of

  5. Penile rehabilitation after pelvic cancer surgery.

    PubMed

    Aoun, Fouad; Peltier, Alexandre; van Velthoven, Roland

    2015-01-01

    Erectile dysfunction is the most common complication after pelvic radical surgery. Rehabilitation programs are increasingly being used in clinical practice but there is no high level of evidence supporting its efficacy. The principle of early penile rehabilitation stems from animal studies showing early histological and molecular changes associated with penile corporal hypoxia after cavernous nerve injury. The concept of early penile rehabilitation was developed in late nineties with a subsequent number of clinical studies supporting early pharmacologic penile rehabilitation. These studies included all available phosphodiesterase type 5 inhibitors, intracavernosal injection and intraurethral use of prostaglandin E1 and to lesser extent vacuum erectile devices. However, these studies are of small number, difficult to interpret, and often with no control group. Furthermore, no studies have proven an in vivo derangement of endothelial or smooth muscle cell metabolism secondary to a prolonged flaccid state. The purpose of the present report is a synthetic overview of the literature in order to analyze the concept and the rationale of rehabilitation program of erectile dysfunction following radical pelvic surgery and the evidence of such programs in clinical practice. Emphasis will be placed on penile rehabilitation programs after radical cystoprostatectomy, radical prostatectomy, and rectal cancer treatment. Future perspectives are also analyzed. PMID:25785286

  6. Penile Rehabilitation after Pelvic Cancer Surgery

    PubMed Central

    Aoun, Fouad; Peltier, Alexandre; van Velthoven, Roland

    2015-01-01

    Erectile dysfunction is the most common complication after pelvic radical surgery. Rehabilitation programs are increasingly being used in clinical practice but there is no high level of evidence supporting its efficacy. The principle of early penile rehabilitation stems from animal studies showing early histological and molecular changes associated with penile corporal hypoxia after cavernous nerve injury. The concept of early penile rehabilitation was developed in late nineties with a subsequent number of clinical studies supporting early pharmacologic penile rehabilitation. These studies included all available phosphodiesterase type 5 inhibitors, intracavernosal injection and intraurethral use of prostaglandin E1 and to lesser extent vacuum erectile devices. However, these studies are of small number, difficult to interpret, and often with no control group. Furthermore, no studies have proven an in vivo derangement of endothelial or smooth muscle cell metabolism secondary to a prolonged flaccid state. The purpose of the present report is a synthetic overview of the literature in order to analyze the concept and the rationale of rehabilitation program of erectile dysfunction following radical pelvic surgery and the evidence of such programs in clinical practice. Emphasis will be placed on penile rehabilitation programs after radical cystoprostatectomy, radical prostatectomy, and rectal cancer treatment. Future perspectives are also analyzed. PMID:25785286

  7. Penile rehabilitation after pelvic cancer surgery.

    PubMed

    Aoun, Fouad; Peltier, Alexandre; van Velthoven, Roland

    2015-01-01

    Erectile dysfunction is the most common complication after pelvic radical surgery. Rehabilitation programs are increasingly being used in clinical practice but there is no high level of evidence supporting its efficacy. The principle of early penile rehabilitation stems from animal studies showing early histological and molecular changes associated with penile corporal hypoxia after cavernous nerve injury. The concept of early penile rehabilitation was developed in late nineties with a subsequent number of clinical studies supporting early pharmacologic penile rehabilitation. These studies included all available phosphodiesterase type 5 inhibitors, intracavernosal injection and intraurethral use of prostaglandin E1 and to lesser extent vacuum erectile devices. However, these studies are of small number, difficult to interpret, and often with no control group. Furthermore, no studies have proven an in vivo derangement of endothelial or smooth muscle cell metabolism secondary to a prolonged flaccid state. The purpose of the present report is a synthetic overview of the literature in order to analyze the concept and the rationale of rehabilitation program of erectile dysfunction following radical pelvic surgery and the evidence of such programs in clinical practice. Emphasis will be placed on penile rehabilitation programs after radical cystoprostatectomy, radical prostatectomy, and rectal cancer treatment. Future perspectives are also analyzed.

  8. Penile strangulation: report of a fatal case.

    PubMed

    Morentin, Benito; Biritxinaga, Begoña; Crespo, Lourdes

    2011-12-01

    Penile strangulation or entrapment is an unusual entity that requires urgent treatment due to its potential complications. Several cases have been reported in the medical literature, some of them describing serious injuries such as necrosis, gangrene, and amputation of the penis. However, as far as we know, no fatal cases have been described before. We present the death of an adult male secondary to the complications due to penile strangulation with a plastic bottle neck. The time of incarceration was unknown, but according to a witness it could be about 10 to 14 days. The findings of autopsy were penile strangulation, necrosis of the penis, acute pyelonephritis, and bronchopneumonia. The subject's refusal to ask for medical help was the cause of this atypical evolution. PMID:22101437

  9. Patient satisfaction with Mentor inflatable penile prosthesis.

    PubMed

    Whalen, R K; Merrill, D C

    1991-06-01

    Patient satisfaction with the Mentor inflatable penile prosthesis was assessed by sending a thirty-six-item questionnaire to 251 patients who had undergone implantation of the device by the senior author (D.C.M.). A total of 152 (61%) of the patients responded. Recovery time, satisfaction, reasons for dissatisfaction, perceptions of erection quality, and psychosexual parameters were evaluated. Eight-eight percent of the patients were engaging in regular sexual activity. Depending on the definition of satisfaction, 81-89 percent of the respondents reported that they were satisfied with the prosthesis. Sixty-eight percent of the survey group were satisfied with the length, width, and firmness of their prosthetic-induced erection. The majority of patients reported improvement in psychosexual functioning after implantation. Reasons for dissatisfaction included inadequate penile length, insufficient firmness, and difficulty with inflation and deflation of the penile cylinders. PMID:2038786

  10. Identification and ribotypes of Staphylococcus caprae isolates isolated as human pathogens and from goat milk.

    PubMed

    Vandenesch, F; Eykyn, S J; Bes, M; Meugnier, H; Fleurette, J; Etienne, J

    1995-04-01

    We report five cases of human infection with Staphylococcus caprae. Two were community acquired (one case each of endocarditis and urinary tract infection); the other three were acquired in a hospital (two cases of bacteremia associated with intravenous access and one case of urinary tract infection). Analysis of human isolates and goat isolates from eight herds showed that they could be misidentified by some commercial identification systems but were clearly identified as S. caprae by ribotyping, according to their species-specific ribotype. Phylogenetic methods applied to the ribotypes did not reveal two distinct lineages corresponding to the goat and human origins of the isolates, although human ribotypes were clearly distinguishable by the presence of a core of four specific bands. The latter observation may reflect some degree of evolutionary change within the species between human and goat isolates.

  11. Stretch due to Penile Prosthesis Reservoir Migration

    PubMed Central

    Baten, E.; Vandewalle, T.; van Renterghem, K.

    2016-01-01

    A 43-year old patient presented to the emergency department with stretch, due to impossible deflation of the penile prosthesis, 4 years after successful implant. A CT-scan showed migration of the reservoir to the left rectus abdominis muscle. Refilling of the reservoir was inhibited by muscular compression, causing stretch. Removal and replacement of the reservoir was performed, after which the prosthesis was well-functioning again. Migration of the penile prosthesis reservoir is extremely rare but can cause several complications, such as stretch. PMID:26793592

  12. Severe Penile Curvature following Otis Urethrotomy.

    PubMed

    Karaguzel, Ersagun; Gur, Metin; Tok, Dogan S; Kazaz, Ilke O; Eren, Huseyin; Kutlu, Omer; Ozgur, Guner K

    2013-01-01

    Urethral stricture is a common urological pathology with a high recurrence rate after treatment. Urethral manipulations are among its main causes. In this paper, urethral stricture developed secondary to urethral catheterization and was treated with cold-knife internal urethrotomy and the Otis urethrotomy procedure. During the follow-up period, severe ventral penile curvature preventing sexual intercourse developed due to fibrosis of the corpus spongiosum and tunica albuginea of the penis. This ventral penile curvature was corrected with a separate operation using a tunica vaginalis flap harvested from the left scrotum.

  13. Stretch due to Penile Prosthesis Reservoir Migration.

    PubMed

    Baten, E; Vandewalle, T; van Renterghem, K

    2016-03-01

    A 43-year old patient presented to the emergency department with stretch, due to impossible deflation of the penile prosthesis, 4 years after successful implant. A CT-scan showed migration of the reservoir to the left rectus abdominis muscle. Refilling of the reservoir was inhibited by muscular compression, causing stretch. Removal and replacement of the reservoir was performed, after which the prosthesis was well-functioning again. Migration of the penile prosthesis reservoir is extremely rare but can cause several complications, such as stretch.

  14. Penile Epidermal Cyst: A Case Report

    PubMed Central

    Kumaraguru, Veerapandian; Prabhu, Ravi

    2016-01-01

    Epidermal cysts also known as epidermoid cysts, is one of the common benign tumours presenting anywhere in the body. However, epidermal cyst in the penis is very rare. This condition in children is usually congenital due to abnormal embryologic closure of the median raphe; hence, it is termed as median raphe cysts (MRCs). Penile epidermal cysts may occur in adults following trauma or surgery due to epidermal elements being trapped within closed space. During wound healing, trapped squamous epithelium, undergoing keratinisation leads to cyst formation. Here, we report a rare case of patient with a penile epidermoid cyst whose main complaints was discomfort during coitus. PMID:27437298

  15. Genetics of human isolated acromesomelic dysplasia.

    PubMed

    Khan, Saadullah; Basit, Sulman; Khan, Muzammil Ahmad; Muhammad, Noor; Ahmad, Wasim

    2016-04-01

    Acromesomelic dysplasia is a type of skeletal malformation affecting distal and middle segments of the extremities. It occurs in both isolated (non-syndromic) and syndromic forms. In later case, it shows association with cardiac, respiratory, neurological and genital abnormalities. Acromesomelic dysplasia segregates in autosomal recessive mode. Mutations in three genes (GDF5, NPR2, BMPR1B) have been reported to cause different forms of acromesomelic dysplasia. In the present review, we have discussed clinical spectrum, genetics and signalopathies of isolated acromesomelic dysplasias. PMID:26926249

  16. Genetics of human isolated acromesomelic dysplasia.

    PubMed

    Khan, Saadullah; Basit, Sulman; Khan, Muzammil Ahmad; Muhammad, Noor; Ahmad, Wasim

    2016-04-01

    Acromesomelic dysplasia is a type of skeletal malformation affecting distal and middle segments of the extremities. It occurs in both isolated (non-syndromic) and syndromic forms. In later case, it shows association with cardiac, respiratory, neurological and genital abnormalities. Acromesomelic dysplasia segregates in autosomal recessive mode. Mutations in three genes (GDF5, NPR2, BMPR1B) have been reported to cause different forms of acromesomelic dysplasia. In the present review, we have discussed clinical spectrum, genetics and signalopathies of isolated acromesomelic dysplasias.

  17. Heterogeneity of the neuropeptide Y (NPY) contractile and relaxing receptors in horse penile small arteries.

    PubMed

    Prieto, Dolores; Arcos, Luis Rivera de Los; Martínez, Pilar; Benedito, Sara; García-Sacristán, Albino; Hernández, Medardo

    2004-12-01

    The distribution of neuropeptide Y (NPY)-immunorective nerves and the receptors involved in the effects of NPY upon electrical field stimulation (EFS)- and noradrenaline (NA)-elicited contractions were investigated in horse penile small arteries. NPY-immunoreactive nerves were widely distributed in the erectile tissues with a particularly high density around penile intracavernous small arteries. In small arteries isolated from the proximal part of the corpora cavernosa, NPY (30 nM) produced a variable modest enhancement of the contractions elicited by both EFS and NA. At the same concentration, the NPY Y(1) receptor agonist, [Leu(31), Pro(34)]NPY, markedly potentiated responses to EFS and NA, whereas the NPY Y(2) receptor agonist, NPY(13-36), enhanced exogenous NA-induced contractions. In arteries precontracted with NA, NPY, peptide YY (PYY), [Leu(31), Pro(34)]NPY and the NPY Y(2) receptor agonists, N-acetyl[Leu(28,31)]NPY (24-36) and NPY(13-36), elicited concentration-dependent contractile responses. Human pancreatic polypeptide (hPP) evoked a biphasic response consisting of a relaxation followed by contraction. NPY(3-36), the compound 1229U91 (Ile-Glu-Pro-Dapa-Tyr-Arg-Leu-Arg-Tyr-NH2, cyclic(2,4')diamide) and eventually NPY(13-36) relaxed penile small arteries. The selective NPY Y(1) receptor antagonist BIBP3226 ((R)-N(2)-(diphenacetyl)-N-[(4-hydroxyphenyl)methyl]D-arginineamide) (0.3 microM) shifted to the right the concentration-response curves to both NPY and [Leu(31), Pro(34)]NPY and inhibited the contractions induced by the highest concentrations of hPP but not the relaxations observed at lower doses. In the presence of the selective NPY Y(2) receptor antagonist BIIE0246 ((S)-N2-[[1-[2-[4-[(R,S)-5,11-dihydro-6(6h)-oxodibenz[b,e]azepin-11-y1]-1-piperazinyl]-2-oxoethyl]cyclo-pentyl-N-[2-[1,2-dihydro-3,5 (4H)-dioxo-1,2-diphenyl-3H-1,2, 4-triazol-4-yl]ethyl]-argininamide) (0.3 microM), the Y(2) receptor agonists NPY(13-36) and N-acetyl[Leu(28,31)]NPY (24

  18. Heterogeneity of the neuropeptide Y (NPY) contractile and relaxing receptors in horse penile small arteries

    PubMed Central

    Prieto, Dolores; Arcos, Luis Rivera de los; Martínez, Pilar; Benedito, Sara; García-Sacristán, Albino; Hernández, Medardo

    2004-01-01

    The distribution of neuropeptide Y (NPY)-immunorective nerves and the receptors involved in the effects of NPY upon electrical field stimulation (EFS)- and noradrenaline (NA)-elicited contractions were investigated in horse penile small arteries. NPY-immunoreactive nerves were widely distributed in the erectile tissues with a particularly high density around penile intracavernous small arteries. In small arteries isolated from the proximal part of the corpora cavernosa, NPY (30 nM) produced a variable modest enhancement of the contractions elicited by both EFS and NA. At the same concentration, the NPY Y1 receptor agonist, [Leu31, Pro34]NPY, markedly potentiated responses to EFS and NA, whereas the NPY Y2 receptor agonist, NPY(13–36), enhanced exogenous NA-induced contractions. In arteries precontracted with NA, NPY, peptide YY (PYY), [Leu31, Pro34]NPY and the NPY Y2 receptor agonists, N-acetyl[Leu28,31]NPY (24–36) and NPY(13–36), elicited concentration-dependent contractile responses. Human pancreatic polypeptide (hPP) evoked a biphasic response consisting of a relaxation followed by contraction. NPY(3–36), the compound 1229U91 (Ile-Glu-Pro-Dapa-Tyr-Arg-Leu-Arg-Tyr-NH2, cyclic(2,4′)diamide) and eventually NPY(13–36) relaxed penile small arteries. The selective NPY Y1 receptor antagonist BIBP3226 ((R)-N2-(diphenacetyl)-N-[(4-hydroxyphenyl)methyl]D-arginineamide) (0.3 μM) shifted to the right the concentration–response curves to both NPY and [Leu31, Pro34]NPY and inhibited the contractions induced by the highest concentrations of hPP but not the relaxations observed at lower doses. In the presence of the selective NPY Y2 receptor antagonist BIIE0246 ((S)-N2-[[1-[2-[4-[(R,S)-5,11-dihydro-6(6h)-oxodibenz[b,e]azepin-11-y1]-1-piperazinyl]-2-oxoethyl]cyclo-pentyl-N-[2-[1,2-dihydro–3,5 (4H)-dioxo-1,2-diphenyl-3H-1,2, 4-triazol-4-yl]ethyl]-argininamide) (0.3 μM), the Y2 receptor agonists NPY(13–36) and N-acetyl[Leu28,31]NPY (24–36) evoked potent slow

  19. Penises not required: a systematic review of the potential for human papillomavirus horizontal transmission that is non-sexual or does not include penile penetration.

    PubMed

    Liu, Zhiyue; Rashid, Tasnuva; Nyitray, Alan G

    2016-02-01

    The primary mode of human papillomavirus (HPV) transmission is through penetrative sex; however, there is evidence of other modes of transmission. No systematic review was found that focussed on HPV horizontal transmission that is not penocentric. A systematic review of the literature by searching Medline (Ovid), PubMed (NLM) and Embase (Ovid) was conducted to retrieve articles published from 1946 to March 2014. Studies that suggested evidence of non-sexual or non-penetrative sexual transmission of α-HPV genotypes were included. After review of 2061 titles and abstracts, 51 studies were abstracted. Fifteen studies examined HPV fomites from medical settings or public environments, and 36 examined HPV in humans. Human papillomavirus DNA was detected in the genital tract of female virgins, with prevalence estimates ranging from 0% to 51.1%. HPV transmission from hands to genitals or genitals to hands was reported for both sexes and heterosexual couples. Other studies commonly found HPV on surfaces in medical settings and public environments. Further studies on non-sexual and non-penetrative sexual transmission are needed to understand the complexity of HPV transmission. Health-care policies may need to be reassessed/established to ensure the safety of medical instruments and to reduce the risk of HPV nosocomial infection.

  20. Penile intracavernosal pillars: lessons from anatomy and potential implications for penile prosthesis placement.

    PubMed

    Pagano, M J; Weinberg, A C; Deibert, C M; Hernandez, K; Alukal, J; Zhao, L; Wilson, S K; Egydio, P H; Valenzuela, R J

    2016-05-01

    The objective of this study was to anatomically describe the relationship of penile intracavernosal pillars to penile surgery, specifically corporal dilation during penile prosthesis placement. Corpora cavernosa from four embalmed male cadavers were dissected and subjected to probe dilation. Corpora were cross-sectioned and examined for the gross presence and location of pillars and dilated spaces. Infrapubic penile prosthesis insertion was performed on one fresh-frozen cadaveric male pelvis, followed by cross-sectioning. A single patient had intracavernosal pillars examined intraoperatively during Peyronie's plaque excision and penile prosthesis insertion. Intracavernosal pillars were identified in all cadavers and one surgical patient, passing obliquely from the dorsolateral tunica albuginea across the sinusoidal space to the ventral intercorporal septum. This delineated each corpus into two potential compartments for dilation: dorsomedial and ventrolateral. Dorsal dilation seated instruments and prosthetics satisfactorily in the dorsal mid glans and provided additional tissue coverage over weak ventral areas of the tunica albuginea, while ventrolateral dilation appeared to result in ventral seating and susceptibility to perforation. Intracavernosal pillars are an important anatomic consideration during penile prosthesis placement. Dorsal dilation appears to result in improved distal seating of cylinder tips, which may be protective against tip malposition, perforation or subsequent erosion. PMID:27053154

  1. Comparison of Staphylococcus aureus isolates associated with food intoxication with isolates from human nasal carriers and human infections.

    PubMed

    Wattinger, L; Stephan, R; Layer, F; Johler, S

    2012-04-01

    Staphylococcus aureus represents an organism of striking versatility. While asymptomatic nasal colonization is widespread, it can also cause serious infections, toxinoses and life-threatening illnesses in humans and animals. Staphylococcal food poisoning (SFP), one of the most prevalent causes of foodborne intoxication worldwide, results from oral intake of staphylococcal enterotoxins leading to violent vomiting, diarrhea and cramps shortly upon ingestion. The aim of the present study was to compare isolates associated with SFP to isolates collected from cases of human nasal colonization and clinical infections in order to investigate the role of S. aureus colonizing and infecting humans as a possible source of SFP. Spa typing and DNA microarray profiling were used to characterize a total of 120 isolates, comprising 50 isolates collected from the anterior nares of healthy donors, 50 isolates obtained from cases of clinical infections in humans and 20 isolates related to outbreaks of staphylococcal food poisoning. Several common spa types were found among isolates of all three sources (t015, t018, t056, t084). DNA microarray results showed highly similar virulence gene profiles for isolates from all tested sources. These results suggest contamination of foodstuff with S. aureus colonizing and infecting food handlers to represent a source of SFP.

  2. Isolation and characterization of Thy 1 homologue from human thymus.

    PubMed

    Bonewald, L F; Goust, J M; Sade, R M; Wang, A C

    1985-01-01

    A 40 000 M.W. glycoprotein was isolated from human thymus. This molecule binds lentil lectin, reacts with an antiserum made against the p25 antigen (the human Thy 1 homologue) and possesses almost identical amino acid composition as the p25 antigen and its 40 000 M.W. dimer. PMID:2864756

  3. Limited Diversity among Human Isolates of Bartonella henselae

    PubMed Central

    Dillon, B.; Valenzuela, J.; Don, R.; Blanckenberg, D.; Wigney, D. I.; Malik, R.; Morris, A. J.; Robson, J. M.; Iredell, J.

    2002-01-01

    A study of 59 isolates of Bartonella henselae reveals relatively limited diversity among those of human origin (n = 28). Either of two distinct alleles of both gltA and 16S ribosomal DNA (rDNA) was found in all isolates, with a high level of congruity between 16S and gltA inheritance among proven human pathogens. Human isolates from all over Eastern Australia were most commonly 16S rDNA (Bergmans) type I, with the same gltA allele as the type strain (Houston-1). Comparable feline isolates were more commonly 16S type II, with less congruity of inheritance between 16S and gltA alleles. Previously described arbitrarily primed PCR and EagI-HhaI infrequent restriction site PCR fingerprinting techniques separated Bartonella species effectively but lacked discriminating power within B. henselae. Examination of the 16-23S intergenic spacer region revealed for several strains several point mutations as well as a repeat sequence of unknown significance which is readily detected by HaeIII restriction fragment length polymorphism analysis. The bacteriophage-associated papA gene was present in all isolates. Enterobacterial repetitive intergenic consensus PCR proved to be a useful and robust typing tool and clearly separated human isolates (including imported strains) from the majority of feline isolates. Our data are consistent with published evidence and with previous suggestions of intragenomic rearrangements in the type strain and suggest that human isolates come from a limited subset of B. henselae strains. They strengthen arguments for careful exploration of genotype-phenotype relationships and for the development of a multilocus enzyme electrophoresis and multilocus sequence typing-based approach to the phylogeny of B. henselae. PMID:12454174

  4. Molecular analysis of environmental and human isolates of Salmonella typhi.

    PubMed Central

    Thong, K L; Cordano, A M; Yassin, R M; Pang, T

    1996-01-01

    Molecular characterization of a total of 54 isolates of Salmonella typhi from Santiago, Chile, was performed by pulsed-field gel electrophoresis (PFGE) after digestion of chromosomal DNA with three restriction endonucleases: XbaI (5'-TCTAGA-3'), AvrII (5'-CCTAGG-3'), and SpeI (5'-ACTAGT-3'). Thirteen of the 54 isolates were obtained from environmental sources (sewage and river water), and the rest were isolates from clinical cases of typhoid fever. Considerable genetic diversity was detected among the human isolates obtained in 1994, as evidenced by the presence of 14 to 19 different PFGE patterns among 20 human isolates, with F (coefficient of similarity) values ranging from 0.69 to 1.0 (XbaI), 0.61 to 1.0 (AvrII), and 0.70 to 1.0 (SpeI). A total of eight phage types were detected among these 20 isolates, with 50% possessing the E1 or 46 phage type. There was no correlation between PFGE pattern and phage types. Similar diversity was seen among 21 isolates obtained in 1983, with 17 to 19 PFGE patterns detected and F values of 0.56 to 1.0 (XbaI), 0.55 to 1.0 (AvrII), and 0.67 to 1.0 (SpeI). Comparison of these two groups of human isolates obtained 11 years apart indicated that certain molecular types of S. typhi are shared and are able to persist for considerable periods. A similar degree of genetic diversity was also detected among the environmental isolates of S. typhi, for which 10 to 12 different PFGE patterns were detected among the 13 isolates analyzed, with F values ranging from 0.56 to 1.0 (XbaI), 0.52 to 1.0 (AvrII), and 0.69 to 1.0 (SpeI). Certain molecular types present among the environmental isolates of S. typhi were also found among the human isolates from the same time period, providing evidence for the epidemiological link between environmental reservoirs and human infection. PMID:8572705

  5. Mesenchymal stem cell isolation and characterization from human spinal ligaments.

    PubMed

    Asari, Toru; Furukawa, Ken-Ichi; Tanaka, Sunao; Kudo, Hitoshi; Mizukami, Hiroki; Ono, Atsushi; Numasawa, Takuya; Kumagai, Gentaro; Motomura, Shigeru; Yagihashi, Soroku; Toh, Satoshi

    2012-01-27

    Mesenchymal stem cells (MSCs) have a fibroblast-like morphology, multilineage potential, long-term viability and capacity for self-renewal. While several articles describe isolating MSCs from various human tissues, there are no reports of isolating MSCs from human spinal ligaments, and their localization in situ. If MSCs are found in human spinal ligaments, they could be used to investigate hypertrophy or ossification of spinal ligaments. To isolate and characterize MSCs from human spinal ligaments, spinal ligaments were harvested aseptically from eight patients during surgery for lumbar spinal canal stenosis and ossification of the posterior longitudinal ligament. After collagenase digestion, nucleated cells were seeded at an appropriate density to avoid colony-to-colony contact. Cells were cultured in osteogenic, adipogenic or chondrogenic media to evaluate their multilineage differentiation potential. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Spinal ligaments were processed for immunostaining using MSC-related antibodies. Cells from human spinal ligaments could be extensively expanded with limited senescence. They were able to differentiate into osteogenic, adipogenic or chondrogenic cells. Flow cytometry revealed that their phenotypic characteristics met the minimum criteria of MSCs. Immunohistochemistry revealed the localization of CD90-positive cells in the collagenous matrix of the ligament, and in adjacent small blood vessels. We isolated and expanded MSCs from human spinal ligaments and demonstrated localization of MSCs in spinal ligaments. These cells may play an indispensable role in elucidating the pathogenesis of numerous spinal diseases.

  6. Molecular characterization of Cryptosporidium isolates from humans in Equatorial Guinea.

    PubMed

    Blanco, María Alejandra; Iborra, Asunción; Vargas, Antonio; Nsie, Eugenia; Mbá, Luciano; Fuentes, Isabel

    2009-12-01

    The aim of the study was to perform a molecular characterization of clinical isolates of Cryptosporidium species from Equatorial Guinea. Standard laboratory methods were used to identify 35 cryptosporidiosis cases among 185 patients. PCR-RFLP successfully identified 34 Cryptosporidium species from these 35 cases, comprising C. parvum (52.9%), C. hominis (44.1%) and C. meleagridis (2.9%); over 90% of the species were isolated from HIV-positive patients. This is the first report of the molecular characterization of Cryptosporidium species isolated from humans in Equatorial Guinea and shows that zoonotic and anthroponotic transmission is present in this country.

  7. Molecular Characterization of First Human Bartonella Strain Isolated in Italy

    PubMed Central

    Ciervo, Alessandra; Petrucca, Andrea; Ciarrocchi, Simonetta; Pinto, Antonella; Bonazzi, Lucio; Fabio, Anna; Farnetti, Enrico; Chomel, Bruno B.; Ciceroni, Lorenzo

    2001-01-01

    The aim of this study was to characterize a Bartonella strain (BA-1) isolated from a blood culture of an Italian, human immunodeficiency virus-positive patient with bacillary angiomatosis. We analyzed the isolate using molecular biology methods such as whole-cell fatty acid analysis, PCR-restriction fragment length polymorphism analysis, type-specific 16S rRNA PCRs, sequence analysis of the 16S rRNA, pulsed-field gel electrophoresis, and arbitrarily primed PCR. The BA-1 isolate turned out to be a Bartonella quintana strain, similar but not identical to B. quintana Oklahoma, which was used as a control strain. PMID:11724882

  8. Human Adaptation to Isolated and Confined Environments

    NASA Technical Reports Server (NTRS)

    Evans, Gary W.; Stokols, Daniel; Carrere, Sybil

    1987-01-01

    A study was conducted over seven months in a winter Antarctic isolated and confined environment (ICE). Physiological and psychological data was collected several times a week. Information was collected on a monthly basis on behavior and the use of physical facilities. Adaptation and information indicated that there was a significant decrease in epinephrine and norepinephrine during the middle trimester of the winter. No vital changes were found for blood pressure. Self reports of hostility and anxiety show a linear increase. There were no significant changes in depression during ICE. The physiological and psychological data do not move in a synchronous fashion over time. The data also suggest that both ambient qualities of an ICE and discrete social environmental events, such as the arrival of the summer crew, have an impact on the outcome measures used. It may be most appropiate to develop a model for ICE's that incorporates not only global chronic stressors common to all ICE's but also the role of discrete environmental effects which can minimize or enhance the influence of more chronic stressors. Behavioral adjustment information highlight the importance of developing schedules which balance work and recreational activities.

  9. The effect of vacuum devices on penile hemodynamics

    SciTech Connect

    Katz, P.G.; Haden, H.T.; Mulligan, T.; Zasler, N.D. )

    1990-01-01

    External vacuum devices are being used increasingly for the management of erectile dysfunction. There is limited information regarding the effect of vacuum devices on penile blood flow and potential for ischemic penile injury. The penile xenon washout rate was measured before and after application of 2 vacuum systems in 15 subjects. Compared to flaccid state measurements the xenon washout rate did not change significantly with the Synergist Erection System but it was significantly reduced with the Osbon ErecAid System. However, the degree and duration of decrease in penile blood flow that may result in ischemic changes are unknown.

  10. Isolation of human lactate dehydrogenase isoenzyme X by affinity chromatography.

    PubMed Central

    Kolk, A H; van Kuyk, L; Boettcher, B

    1978-01-01

    Human isoenzyme LDH-X (lactate dehydrogenase isoenzyme X) was isolated from seminal fluid of frozen semen samples by affinity chromatography by using oxamate-Sepharose and AMP-Sepharose. In the presence of 1.6 mM-NAD+, isoenzyme LDH-X does not bind to AMP-Sepharose, whereas the other lactate dehydrogenase isoenzymes do. This is the crucial point in the isolation of isoenzyme LDH-X from the other isoenzymes. The purified human isoenzyme LDH-X had a specific activity of 146 units/mg of protein. Images Fig. 2. Fig. 3. PMID:213050

  11. Current penile-rehabilitation strategies: Clinical evidence.

    PubMed

    Segal, Robert L; Bivalacqua, Trinity J; Burnett, Arthur L

    2013-09-01

    We review the current strategies used for penile rehabilitation (PR) after a radical prostatectomy, where PR is defined as the attempt to restore spontaneous erectile function so that the patient can generate erections with no need for erectile aids. We searched PubMed for relevant reports, using the keywords 'radical prostatectomy', 'penile rehabilitation', 'phosphodiesterase inhibitors', 'vacuum erection device', 'injection therapy', 'urethral suppository', and 'erectile dysfunction'. In all, 155 articles were identified and reviewed, and had a level of evidence ranging from 1b-4. The use of PR strategies should be based on the patient's goals after a thorough explanation of realistic expectations, and the risks and consequences of the various treatment options. While a multitude of studies suggest a benefit with PR strategies, there are no established, proven regimens. Further research is needed to establish the optimal approaches to PR. PMID:26558087

  12. Large Penile Mass With Unusual Benign Histopathology.

    PubMed

    Johnson, Nate; Voznesensky, Maria; VerLee, Graham

    2015-09-01

    Pseudoepitheliomatous hyperplasia is an extremely rare condition presenting as a lesion on the glans penis in older men. Physical exam without biopsy cannot differentiate malignant from nonmalignant growth. We report a case of large penile mass in an elderly male with a history of lichen sclerosis, highly suspicious for malignancy. Subsequent surgical removal and biopsy demonstrated pseudoepitheliomatous hyperplasia, an unusual benign histopathologic diagnosis with unclear prognosis. We review the literature and discuss options for treatment and surveillance. PMID:26793536

  13. Isolation & characterization of Brucella melitensis isolated from patients suspected for human brucellosis in India

    PubMed Central

    Barua, Anita; Kumar, Ashu; Thavaselvam, Duraipandian; Mangalgi, Smita; Prakash, Archana; Tiwari, Sapana; Arora, Sonia; Sathyaseelan, Kannusamy

    2016-01-01

    Background & objectives: Brucellosis is endemic in the southern part of India. A combination of biochemical, serological and molecular methods is required for identification and biotyping of Brucella. The present study describes the isolation and biochemical, molecular characterization of Brucella melitensis from patients suspected for human brucellosis. Methods: The blood samples were collected from febrile patients suspected to have brucellosis. A total of 18 isolates were obtained from 102 blood samples subjected to culture. The characterization of these 18 isolates was done by growth on Brucella specific medium, biochemical reactions, CO2 requirement, H2S production, agglutination with A and M mono-specific antiserum, dye sensitivity to basic fuchsin and thionin. Further, molecular characterization of the isolates was done by amplification of B. melitensis species specific IS711 repetitive DNA fragment and 16S (rRNA) sequence analysis. PCR-restriction fragment length polymorphism (RFLP) analysis of omp2 locus and IS711 gene was also done for molecular characterization. Results: All 102 suspected samples were subjected to bacteria isolation and of these, 18 isolates could be recovered on blood culture. The biochemical, PCR and PCR-RFLP and 16s rRNA sequencing revealed that all isolates were of B. melitensis and matched exactly with reference strain B. melitensis 16M. Interpretation & conclusions: The present study showed an overall isolation rate of 17.64 per cent for B. melitensis. There is a need to establish facilities for isolation and characterization of Brucella species for effective clinical management of the disease among patients as well as surveillance and control of infection in domestic animals. Further studies are needed from different geographical areas of the country with different level of endemicity to plan and execute control strategies against human brucellosis. PMID:27488010

  14. Penile Injuries in Wild and Domestic Pigs.

    PubMed

    Weiler, Ulrike; Isernhagen, Marie; Stefanski, Volker; Ritzmann, Mathias; Kress, Kevin; Hein, Charlotte; Zöls, Susanne

    2016-03-25

    In boars, sexually motivated mounting can not only cause problems such as lameness, but penile injuries are also reported. The relevance of penis biting in boars is discussed controversially, but reliable data is missing. In the present study, boars ( n = 435) and barrows ( n = 85) from experimental farms were therefore evaluated for scars, fresh wounds and severe injuries of the penis. Similarly, 321 boars from 11 farms specializing in pork production with boars, and 15 sexually mature wild boars from the hunting season of 2015/16 were included in the study. In domestic boars, a high incidence of penile injuries was obvious (76.6%-87.0% of animals with scars and/or wounds at experimental farms, 64.0%-94.9% at commercial farms). The number of boars with severe injuries was in a similar range in both groups (7.3% vs. 9.3%). At commercial farms, the number of scars but not that of fresh wounds increased per animal with age by 0.3 per week. Moreover, raising boars in mixed groups led to about a 1.5 times higher number of scars than in single-sex groups. In wild boars, a considerable proportion of animals (40%) revealed penile injuries, which were even severe in three animals. We therefore conclude that penis biting is a highly relevant and severe welfare problem in the male pig population, but this phenomenon is not limited to intensive production systems.

  15. [Less mutilating treatment of penile carcinoma].

    PubMed

    Kroon, B K; Horenblas, S; Nieweg, O E

    2005-10-29

    Four patients with penile carcinoma are described. A 60-year-old man with a T1-tumour underwent penis-conserving laser treatment. Two men, aged 52 and 65 years old, with T2-tumors and clinically node-negative groins underwent penile amputation. Sentinel-node biopsy (SNB) revealed no metastases in the 52-year-old patient. High-resolution ultrasound-guided fine-needle aspiration cytology revealed bilateral metastases in the other patient, who underwent bilateral inguinal lymphadenectomy. In the fourth patient, a 73-year-old man with a T3-tumor, a pathological lymph node was palpated in one groin. Inguinal lymphadenectomy revealed 3 positive nodes and an additional pelvic lymphadenectomy was performed. SNB on the other side was positive and inguinal lymphadenectomy followed. No additional positive nodes were found in the dissection specimen. All patients were alive without evidence of disease 4, 3, 3 and 4 years later, respectively. New developments in the management of penile cancer such as laser treatment, high-resolution ultrasonography and SNB result in a more tailored approach with less morbidity without reducing survival rates.

  16. Organ-preserving surgery for penile carcinoma.

    PubMed

    Martins, Francisco E; Rodrigues, Raul N; Lopes, Tomé M

    2008-01-01

    Introduction. Penile carcinoma has traditionally been treated by either surgical amputation or radical radiotherapy, both associated with devastating anatomical, functional, and psychological impact on the patient's life. Innovative surgical techniques have focused on penile preservation in well-selected patients to minimize physical disfigurement and consequently maximize quality of life. The objective of this article is to define the current status of these organ-preserving surgical options for penile carcinoma. Materials and Methods. An extensive review of the Pubmed literature was performed to find articles discussing only reconstructive surgery which have contributed significantly to change traditional, frequently mutilating treatments, to develop less disfiguring surgery, and to improve patients' quality of life over the last two decades. Results. Several articles were included in this analysis in which a major contribution to the change in therapy was thought to have occurred and was documented as beneficial. Some articles reported novel techniques of less-mutilating surgery involving different forms of glans reconstruction with the use of flaps or grafts. The issue of safe surgical margins was also addressed. Conclusion. The development of less-disfiguring techniques allowing phallus preservation has reduced the negative impact on functional and cosmetic outcomes of amputation without sacrificing oncological objectives in appropriately selected patients based on stage, grade, and location of the tumour. Until more prospective studies are available and solid evidence is documented, organ preservation should be offered with caution.

  17. Penile Injuries in Wild and Domestic Pigs.

    PubMed

    Weiler, Ulrike; Isernhagen, Marie; Stefanski, Volker; Ritzmann, Mathias; Kress, Kevin; Hein, Charlotte; Zöls, Susanne

    2016-01-01

    In boars, sexually motivated mounting can not only cause problems such as lameness, but penile injuries are also reported. The relevance of penis biting in boars is discussed controversially, but reliable data is missing. In the present study, boars ( n = 435) and barrows ( n = 85) from experimental farms were therefore evaluated for scars, fresh wounds and severe injuries of the penis. Similarly, 321 boars from 11 farms specializing in pork production with boars, and 15 sexually mature wild boars from the hunting season of 2015/16 were included in the study. In domestic boars, a high incidence of penile injuries was obvious (76.6%-87.0% of animals with scars and/or wounds at experimental farms, 64.0%-94.9% at commercial farms). The number of boars with severe injuries was in a similar range in both groups (7.3% vs. 9.3%). At commercial farms, the number of scars but not that of fresh wounds increased per animal with age by 0.3 per week. Moreover, raising boars in mixed groups led to about a 1.5 times higher number of scars than in single-sex groups. In wild boars, a considerable proportion of animals (40%) revealed penile injuries, which were even severe in three animals. We therefore conclude that penis biting is a highly relevant and severe welfare problem in the male pig population, but this phenomenon is not limited to intensive production systems. PMID:27023619

  18. DIFFERENTIATING HUMAN FROM ANIMAL ISOLATES OF CRYPTOSPORIDIUM PARVUM

    EPA Science Inventory

    We analyzed 9s Cryptosporidium parvum isolates from humans and animals by a polymerase chain reaction/restriction fragment length polymorphism method based on the thrombospondin-related anonymous protein 2 gene sequence. Used as a molecular marker, this method can differentiate ...

  19. Human Streptococcus agalactiae isolate in Nile tilapia (Oreochromis niloticus)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Streptococcus agalactiae, the Lancefield group B Streptococcus (GBS), long recognized as a mammalian pathogen, is an emerging pathogen to fish. We show that a GBS serotype Ia, multilocus sequence type ST-7 isolate from a human neonatal meningitis clinical case causes disease signs and mortality in N...

  20. Isolation of Cancer Stem Cells From Human Prostate Cancer Samples

    PubMed Central

    Vidal, Samuel J.; Quinn, S. Aidan; de la Iglesia-Vicente, Janis; Bonal, Dennis M.; Rodriguez-Bravo, Veronica; Firpo-Betancourt, Adolfo; Cordon-Cardo, Carlos; Domingo-Domenech, Josep

    2014-01-01

    The cancer stem cell (CSC) model has been considerably revisited over the last two decades. During this time CSCs have been identified and directly isolated from human tissues and serially propagated in immunodeficient mice, typically through antibody labeling of subpopulations of cells and fractionation by flow cytometry. However, the unique clinical features of prostate cancer have considerably limited the study of prostate CSCs from fresh human tumor samples. We recently reported the isolation of prostate CSCs directly from human tissues by virtue of their HLA class I (HLAI)-negative phenotype. Prostate cancer cells are harvested from surgical specimens and mechanically dissociated. A cell suspension is generated and labeled with fluorescently conjugated HLAI and stromal antibodies. Subpopulations of HLAI-negative cells are finally isolated using a flow cytometer. The principal limitation of this protocol is the frequently microscopic and multifocal nature of primary cancer in prostatectomy specimens. Nonetheless, isolated live prostate CSCs are suitable for molecular characterization and functional validation by transplantation in immunodeficient mice. PMID:24686446

  1. Penile inversion through a penoscrotal incision for the treatment of penile urethral strictures

    PubMed Central

    Tracey, James M; Zhumkhawala, Ali A; Chan, Kevin G.; Lau, Clayton S.

    2016-01-01

    Purpose This article describes a novel technique for the repair of penile urethral strictures and establishes the safety, feasibility, and efficacy of this innovative surgical approach. Materials and Methods Patients with urethral strictures underwent a one-sided anterior dorsal oral mucosal graft urethroplasty through a penoscrotal inversion technique. The clinical outcome was considered a failure when any instrumentation was needed postoperatively, including dilatation. Results Five patients underwent the novel procedure. The patients' mean age was 58 years. The cause of stricture was instrumentation in 2 cases (40%), lichen sclerosis in 1 case (20%), and failed hypospadias repair in 2 cases (40%). The mean stricture length was 3 cm. The overall mean (range) follow-up was 6 months (range, 3–9 months). Of the 5 patients, 4 (80%) had a successful outcome and 1 (20%) had a failed outcome. The failure was successfully treated by use of a meatotomy. Conclusions The penile inversion technique through a penoscrotal incision is a viable option for the management of penile urethral strictures with several advantages to other techniques: namely, no penile skin incision, a single-stage operation, and supine positioning. PMID:26981596

  2. Therapeutic strategies for patients with micropenis or penile dysmorphic disorder.

    PubMed

    Kayes, Oliver; Shabbir, Majid; Ralph, David; Minhas, Suks

    2012-09-01

    Micropenis in adults is defined as a stretched length of <7.5 cm. Many aetiologies exist, including congenital and endocrinological causes as well as pathological conditions, such as penile lichen sclerosus, trauma and genital cancer. The resulting reduction in functional penile length can lead to considerable psychosexual morbidity. Furthermore, the subset of patients with micropenis who also suffer from penile dysmorphic disorder require careful and intensive psychological counselling. Corrective surgery for micropenis can be performed in patients with realistic expectations. Total phalloplasty using radial-artery-based forearm skin flaps can offer restoration of normal penile length in selected patients. More-conservative surgical techniques to improve length or girth are limited by minimal enhancement but associated with a significantly lower rate of complications and comorbidity compared to total phalloplasty. Emerging tissue engineering techniques might represent a suitable alternative to penile replacement surgery in the future. PMID:22890302

  3. Three-piece Inflatable Penile Prosthesis: Surgical Techniques and Pitfalls

    PubMed Central

    Al-Enezi, Ahmad; Al-Khadhari, Sulaiman; Al-Shaiji, Tariq F.

    2011-01-01

    Penile prosthesis surgery plays a vital role in the treatment of erectile dysfunction (ED). As far as outcome is concerned, it is one of the most rewarding procedures for both patients and surgeons. We describe our surgical technique for implantation of the three-piece inflatable penile prosthesis and point out the major surgical pitfalls accompanying this procedure and their specific management. The psychological outcome of penile prosthesis surgery is also discussed. Different surgical approaches are available when performing the procedure. A number of procedure-related problems can be encountered and a thorough knowledge of these is of paramount importance. Penile prosthesis surgery has a favorable psychological outcome. Surgery for implantation of an inflatable penile prosthesis is a rewarding procedure, with a high yield of patient satisfaction. Urologists should have thorough understanding of the surgical pitfalls peculiar to this procedure and their management. PMID:22413049

  4. Therapeutic strategies for patients with micropenis or penile dysmorphic disorder.

    PubMed

    Kayes, Oliver; Shabbir, Majid; Ralph, David; Minhas, Suks

    2012-09-01

    Micropenis in adults is defined as a stretched length of <7.5 cm. Many aetiologies exist, including congenital and endocrinological causes as well as pathological conditions, such as penile lichen sclerosus, trauma and genital cancer. The resulting reduction in functional penile length can lead to considerable psychosexual morbidity. Furthermore, the subset of patients with micropenis who also suffer from penile dysmorphic disorder require careful and intensive psychological counselling. Corrective surgery for micropenis can be performed in patients with realistic expectations. Total phalloplasty using radial-artery-based forearm skin flaps can offer restoration of normal penile length in selected patients. More-conservative surgical techniques to improve length or girth are limited by minimal enhancement but associated with a significantly lower rate of complications and comorbidity compared to total phalloplasty. Emerging tissue engineering techniques might represent a suitable alternative to penile replacement surgery in the future.

  5. MicroRNA Expression Profile in Penile Cancer Revealed by Next-Generation Small RNA Sequencing

    PubMed Central

    Zhang, Yuanwei; Xu, Bo; Zhou, Jun; Fan, Song; Hao, Zongyao; Shi, Haoqiang; Zhang, Xiansheng; Kong, Rui; Xu, Lingfan; Gao, Jingjing; Zou, Duohong; Liang, Chaozhao

    2015-01-01

    Penile cancer (PeCa) is a relatively rare tumor entity but possesses higher morbidity and mortality rates especially in developing countries. To date, the concrete pathogenic signaling pathways and core machineries involved in tumorigenesis and progression of PeCa remain to be elucidated. Several studies suggested miRNAs, which modulate gene expression at posttranscriptional level, were frequently mis-regulated and aberrantly expressed in human cancers. However, the miRNA profile in human PeCa has not been reported before. In this present study, the miRNA profile was obtained from 10 fresh penile cancerous tissues and matched adjacent non-cancerous tissues via next-generation sequencing. As a result, a total of 751 and 806 annotated miRNAs were identified in normal and cancerous penile tissues, respectively. Among which, 56 miRNAs with significantly different expression levels between paired tissues were identified. Subsequently, several annotated miRNAs were selected randomly and validated using quantitative real-time PCR. Compared with the previous publications regarding to the altered miRNAs expression in various cancers and especially genitourinary (prostate, bladder, kidney, testis) cancers, the most majority of deregulated miRNAs showed the similar expression pattern in penile cancer. Moreover, the bioinformatics analyses suggested that the putative target genes of differentially expressed miRNAs between cancerous and matched normal penile tissues were tightly associated with cell junction, proliferation, growth as well as genomic instability and so on, by modulating Wnt, MAPK, p53, PI3K-Akt, Notch and TGF-β signaling pathways, which were all well-established to participate in cancer initiation and progression. Our work presents a global view of the differentially expressed miRNAs and potentially regulatory networks of their target genes for clarifying the pathogenic transformation of normal penis to PeCa, which research resource also provides new insights

  6. [Isolation of lymphocytic choriomeningitis virus from human individuals].

    PubMed

    Saavedra, M C; Ambrosio, A M; Riera, L; Levis, S; Sottosanti, J; Sabattini, M

    2001-01-01

    The activity of lymphocytic choriomeningitis virus (LCMv) in Argentina has been previously reported on the basis of serological evidence in rodents and humans and the isolation of only one strain of LCMv from a Mus domesticus captured in the province of Córdoba. The aim of this paper was to register patients with serological diagnosis of LCM, to isolate and to identify human strains of LCMv in Argentina. During the last 19 years, 15 cases were diagnosed as LCM by immunoflourescent indirect assay (IFI) and enzyme-linked immunosorbent assay (ELISA) but when neutralizing assay (NT) was incorporated, eight cases were classified as confirmed, three as probable and four as negative. The geographic distribution of the cases included three provinces: Córdoba, Buenos Aires and Santa Fe. Viral isolation was attempted in five patients classified as confirmed and only two resulted positive (P5226 and P8573). They were identified as LCMv by IFI and NT. The coexistence of LCMv with other arenaviruses, such as Junin and Oliveros viruses, in the same area, raises the probability of interactions between them, which could modify the virulence and/or pathogenicity for humans associated to genomic changes. Future studies of antigenic, genomic and virulence variability of different Argentine strains of LCMv, as well as the systematic search for human infection, will contribute to define the importance of this viral agent in our country and to implement control measures.

  7. Selective medium for isolation of Clostridium butyricum from human feces.

    PubMed

    Popoff, M R

    1984-09-01

    A selective medium, Clostridium butyricum isolation medium (BIM), is described for the isolation of C. butyricum from human feces. The BIM is a synthetic minimal medium and contains trimethoprim (16 micrograms/ml), D-cycloserine (10 micrograms/ml), and polymyxin B sulfate (20 micrograms/ml) as selective inhibitory agents. Qualitative tests indicated that C. butyricum and other butyric acid-producing clostridia grew on BIM, Clostridium sphenoides and Bacillus cereus produced small colonies, and other clostridia and other obligate anaerobic or facultatively anerobic bacteria were inhibited. Quantitative recovery of C. butyricum from cultures or seeded fecal samples was comparable with BIM and with complex medium, but the quantitative recovery of the other butyric acid-producing clostridia tested (C. beijerinckii, C. acetobutylicum) was lower with BIM than with complex medium. The BIM should aid the rapid isolation of C. butyricum from fecal samples and should be useful for bacteriological investigation of neonatal necrotizing enterocolitis.

  8. Selective medium for isolation of Clostridium butyricum from human feces.

    PubMed Central

    Popoff, M R

    1984-01-01

    A selective medium, Clostridium butyricum isolation medium (BIM), is described for the isolation of C. butyricum from human feces. The BIM is a synthetic minimal medium and contains trimethoprim (16 micrograms/ml), D-cycloserine (10 micrograms/ml), and polymyxin B sulfate (20 micrograms/ml) as selective inhibitory agents. Qualitative tests indicated that C. butyricum and other butyric acid-producing clostridia grew on BIM, Clostridium sphenoides and Bacillus cereus produced small colonies, and other clostridia and other obligate anaerobic or facultatively anerobic bacteria were inhibited. Quantitative recovery of C. butyricum from cultures or seeded fecal samples was comparable with BIM and with complex medium, but the quantitative recovery of the other butyric acid-producing clostridia tested (C. beijerinckii, C. acetobutylicum) was lower with BIM than with complex medium. The BIM should aid the rapid isolation of C. butyricum from fecal samples and should be useful for bacteriological investigation of neonatal necrotizing enterocolitis. PMID:6490827

  9. First human isolate of Hantavirus (Andes virus) in the Americas.

    PubMed

    Galeno, Hector; Mora, Judith; Villagra, Eliecer; Fernandez, Jorge; Hernandez, Jury; Mertz, Gregory J; Ramirez, Eugenio

    2002-07-01

    We isolated Andes virus (formal name: Andes virus [ANDV], a species in the genus Hantavirus), from serum of an asymptomatic 10-year-old Chilean boy who died 6 days later of hantavirus pulmonary syndrome (HPS). The serum was obtained 12 days after his grandmother died from HPS and 2 days before he became febrile. No hantavirus immunoglobulin (Ig) G or IgM antibodies were detected in the serum sample. After three blind passages, ANDV antigens were detected in Vero E6 cells by immunofluorescence assay and enzyme-linked immunosorbent assay, and ANDV RNA was detected by reverse transcription-polymerase chain reaction. A fragment of the virus genome showed 96.2% nucleotide identity with that of prototype ANDV. To our knowledge, this is the first isolation of any agent of hemorrhagic fever with HPS from a human and the first such isolation of hantavirus before symptoms of that syndrome or HPS began.

  10. Penile nodules in the penal system.

    PubMed

    Griffith, Jack; Horowitz, David

    2012-05-01

    The insertion of inert spherical objects under the skin of the penile shaft with the intent of enhancing the sexual experience of one's partner has been reported mostly among Southeast Asian men. This practice is gaining popularity among the Hispanic jail population and prison inmates in southern California. We present a case series of 4 inmate patients with genital modifications (artificial penis nodules [APNs]), including one patient with vitiligo that was induced by his APN. Additionally, we review the literature pertaining to this practice and the relevant clinical implications.

  11. Does tunica anatomy matter in penile implant?

    PubMed Central

    Chen, Heng-Shuen; Huang, Sheng-Jean

    2015-01-01

    Background Overall prosthesis survival is important in penile implant, which remains the final viable solution to many patients with refractory erectile dysfunction (ED). This paper is to retrospectively study the role of the anatomy of tunica albuginea (TA). Methods From March 1987 to March 1991 while the TA was regarded as a circumferential single layer, 21 organically ED men, aged from 27 to 77, received penile prosthesis implantation and were allocated to conventional group. From August 1992 to March 2013 while the tip of Hegar’s dilator was categorically directed medial-dorsally during corporal dilatation derived from newfound TA as a bi-layered structure with a 360° complete inner circular layer and a 300° incomplete outer longitudinal coat, 196 ED males, aged from 35 to 83, underwent penile implant and were categorized to advanced group. The model of prosthesis was recorded. Prosthesis loss rate and survival time were analyzed and the follow up period ranged from 22.4-26.4 (average 24.3) years and 0.4-20.6 (average 15.8) years to the conventional and advanced group respectively. Results To the conventional and advanced group, the number of inflatable and rigid type prosthesis used were 2, 19 and 15, 181 respectively, whereas the prosthesis loss was encountered in 50.0% (1/2), 15.8% (3/19) and 0.0% (0/15), 0.6% (1/181) respectively. And the prosthesis survival time were 5.1-6.3 (5.7) years, 1.3-26.4 (15.2) years and 6.1-16.2 (11.2) years, 0.4-20.6 (15.3) years to the conventional and advanced group respectively. Statistical significance was noted on prosthesis loss in groups (P=0.01) while the Mentor Acuform stood out in prosthesis survival. Conclusions Anatomy-based managing maneuver appears to deliver better surgery success in penile implant. Tunica anatomy is significant in performing implant surgery. PMID:26816839

  12. Penile brachytherapy: Results for 49 patients

    SciTech Connect

    Crook, Juanita M. . E-mail: juanita.crook@rmp.uhn.on.ca; Jezioranski, John; Grimard, Laval; Esche, Bernd; Pond, G.

    2005-06-01

    Purpose: To report results for 49 men with squamous cell carcinoma (SCC) of the penis treated with primary penile interstitial brachytherapy at one of two institutions: the Ottawa Regional Cancer Center, Ottawa, and the Princess Margaret Hospital, Toronto, Ontario, Canada. Methods and Materials: From September 1989 to September 2003, 49 men (mean age, 58 years; range, 22-93 years) had brachytherapy for penile SCC. Fifty-one percent of tumors were T1, 33% T2, and 8% T3; 4% were in situ and 4% Tx. Grade was well differentiated in 31%, moderate in 45%, and poor in 2%; grade was unspecified for 20%. One tumor was verrucous. All tumors in Toronto had pulsed dose rate (PDR) brachytherapy (n = 23), whereas those in Ottawa had either Iridium wire (n 22) or seeds (n = 4). Four patients had a single plane implant with a plastic tube technique, and all others had a volume implant with predrilled acrylic templates and two or three parallel planes of needles (median, six needles). Mean needle spacing was 13.5 mm (range, 10-18 mm), mean dose rate was 65 cGy/h (range, 33-160 cGy/h), and mean duration was 98.8 h (range, 36-188 h). Dose rates for PDR brachytherapy were 50-61.2 cGy/h, with no correction in total dose, which was 60 Gy in all cases. Results: Median follow-up was 33.4 months (range, 4-140 months). At 5 years, actuarial overall survival was 78.3% and cause-specific survival 90.0%. Four men died of penile cancer, and 6 died of other causes with no evidence of recurrence. The cumulative incidence rate for never having experienced any type of failure at 5 years was 64.4% and for local failure was 85.3%. All 5 patients with local failure were successfully salvaged by surgery; 2 other men required penectomy for necrosis. The soft tissue necrosis rate was 16% and the urethral stenosis rate 12%. Of 8 men with regional failure, 5 were salvaged by lymph node dissection with or without external radiation. All 4 men with distant failure died of disease. Of 49 men, 42 had an intact

  13. [Methods for evaluation of penile erection hardness].

    PubMed

    Yuan, Yi-Ming; Zhou, Su; Zhang, Kai

    2010-07-01

    Penile erection hardness is one of the key factors for successful sexual intercourse, as well as an important index in the diagnosis and treatment of erectile dysfunction (ED). This article gives an overview on the component and impact factors of erection hardness, summarizes some commonly used evaluation methods, including those for objective indexes, such as Rigiscan, axial buckling test and color Doppler ultrasonography, and those for subjective indexes of ED patients, such as IIEF, the Erectile Function Domain of IIEF (IIEF-EF), and Erection Hardness Score (EHS), and discusses the characteristics of these methods.

  14. Isolation, characterization, and differentiation of human multipotent dermal stem cells.

    PubMed

    Li, Ling; Fukunaga-Kalabis, Mizuho; Herlyn, Meenhard

    2013-01-01

    Skin, as the body's largest organ, has been extensively used to study adult stem cells. Most previous skin-related studies have focused on stem cells isolated from hair follicles and from keratinocytes. Here we present a protocol to isolate multipotent neural crest stem-like dermis-derived stem cells (termed dermal stem cells or DSCs) from human neonatal foreskins. DSCs grow like neural spheres in human embryonic stem cell medium and gain the ability to self-renew and differentiate into several cell lineages including melanocytes, neuronal cells, Schwann cells, smooth muscle cells, adipocytes, and chondrocytes. These cells express neural crest stem cell markers (NGFRp75 and nestin) as well as an embryonic stem cell marker (OCT4).

  15. Isolation and characterization of endothelial progenitor cells from human blood.

    PubMed

    Mead, Laura E; Prater, Daniel; Yoder, Mervin C; Ingram, David A

    2008-07-01

    Circulating endothelial progenitor cells (EPCs) in adult human peripheral blood were originally identified in 1997 by Asahara et al., which challenged the paradigm that vasculogenesis is a process restricted to embryonic development. Since their original identification, EPCs have been extensively studied as biomarkers to assess the risk of cardiovascular disease in human subjects and as a potential cell therapeutic for vascular regeneration. Endothelial colony-forming cells (ECFCs), which are a subtype of EPCs, were recently identified from circulating adult and human umbilical cord blood. In contrast to other types of EPCs, which display various monocyte/macrophage phenotypes and functions, ECFCs are characterized by robust proliferative potential, secondary and tertiary colony formation upon replating, and de novo blood vessel formation in vivo when transplanted into immunodeficient mice. In this unit, we describe detailed methodologies for isolation and characterization of ECFCs from both human peripheral and umbilical cord blood.

  16. AB020. Penile rehabilitation with PDE5 inhibitors in men following radical prostatectomy

    PubMed Central

    Jiann, Bang-Ping

    2015-01-01

    Post-radical prostatectomy (RP) erectile dysfunction (ED) remains a challenge for the urologist. Despite the improvements in surgical technique, ED occurs between 20% and 90% in patients treated with bilateral nerve-sparing RP. Patient factors, cancer selection, type of surgery, surgical techniques, and surgeon factors represent the key significant contributors to erectile function recovery. The aim of a penile rehabilitation program is to preserve the functional smooth-muscle content of the corpus cavernosum during the neuropraxia period. Phosphodiesterase type 5 (PDE5) inhibitors are commonly used in rehabilitation programs. In animal models, such an approach could promote erectile function recovery, improve smooth muscle-to-collagen penile ration, reduce penile apoptotic index, preserve penile endothelial function and promote neuroprotection during nerve damage. Despite the strong basic science support from animal studies, discordant results have been reached in humans. The previous randomized trials comparing chronic versus on-demand PDE-5 inhibitors use after RP may be affected by improper patient selection in that only men at low risk of postoperative ED were included. These patients would recover erectile function regardless of the type of PDE5 inhibitor administration because of their excellent baseline profile. Prospective, randomized trials have shown a significant benefit of daily PDE5-I administration as compared with placebo in terms of postoperative EF recovery. Patients with intermediate risk of ED after surgery are the best candidates for daily treatment with PDE5 inhibitor after bilateral nerve-sparing RP. The maximal effect of penile rehabilitation may be found in those men with a certain (but not high) degree of systemic and erectile impairment preoperatively. In conclusion, penile rehabilitation could achieve faster and better natural erectile function after RP and should be started as early as possible. Chronic use of PDE5-I may confer the

  17. Central control of penile erection: role of the paraventricular nucleus of the hypothalamus.

    PubMed

    Argiolas, Antonio; Melis, Maria Rosaria

    2005-05-01

    sexual behaviour and penile physiology, which has largely increased in the last years our knowledge of peripheral and central mechanisms controlling erectile function (drugs that induce penile erection in male rats usually do so also in man), the above results may have great significance in terms of a human perspective for the treatment of erectile dysfunction.

  18. Isolation and Quantification of Glycosaminoglycans from Human Hair Shaft

    PubMed Central

    Bonovas, Stefanos; Sitaras, Nikolaos

    2016-01-01

    Background There is evidence that glycosaminoglycans (GAGs) are present in the hair shaft within the follicle but there are no studies regarding GAGs isolation and measurement in the human hair shaft over the scalp surface, it means, in the free hair shaft. Objective The purpose of our research was to isolate and measure the total GAGs from human free hair shaft. Methods Seventy-five healthy individuals participated in the study, 58 adults, men and women over the age of 50 and 17 children (aged 4~9). GAGs in hair samples, received from the parietal and the occipital areas, were isolated with 4 M guanidine HCl and measured by the uronic acid-carbazole reaction assay. Results GAGs concentration was significantly higher in the occipital area than in the parietal area, in all study groups. GAG levels from both areas were significantly higher in children than in adults. GAG levels were not associated with gender, hair color or type. Conclusion We report the presence of GAGs in the human free hair shaft and the correlation of hair GAG levels with the scalp area and participants' age. PMID:27746630

  19. Isolation, identification and differentiation of human embryonic cartilage stem cells.

    PubMed

    Fu, Changhao; Yan, Zi; Xu, Hao; Zhang, Chen; Zhang, Qi; Wei, Anhui; Yang, Xi; Wang, Yi

    2015-07-01

    We isolated human embryonic cartilage stem cells (hECSCs), a novel stem cell population, from the articular cartilage of eight-week-old human embryos. These stem cells demonstrated a marker expression pattern and differentiation potential intermediate to those of human embryonic stem cells (hESCs) and human adult stem cells (hASCs). hECSCs expressed markers associated with both hESCs (OCT4, NANOG, SOX2, SSEA-3 and SSEA-4) and human adult stem cells (hASCs) (CD29, CD44, CD90, CD73 and CD10). These cells also differentiated into adipocytes, osteoblasts, chondrocytes, neurons and islet-like cells under specific inducing conditions. We identified N(6), 2'-O-dibutyryl cyclic adenosine 3':5'-monophosphate (Bt2cAMP) as an inducer of chondrogenic differentiation in hECSCs. Similar results using N(6), 2'-O-dibutyryl cyclic adenosine 3':5'-monophosphate (Bt2cAMP) were obtained for two other types of human embryonic tissue-derived stem cells, human embryonic hepatic stem cells (hEHSCs) and human embryonic amniotic fluid stem cells (hEASCs), both of which exhibited a marker expression pattern similar to that of hECSCs. The isolation of hECSCs and the discovery that N(6), 2'-O-dibutyryl cyclic adenosine 3':5'-monophosphate (Bt2cAMP) induces chondrogenic differentiation in different stem cell populations might aid the development of strategies in tissue engineering and cartilage repair.

  20. A new surgical method for penile girth enhancement

    PubMed Central

    Li, Xiaoge; Tao, Ling; Cao, Chuan; Shi, Haishan; Li, Le; Chen, Liang; Li, Shirong

    2015-01-01

    Objective: We developed a new surgical model of penile girth enhancement in dog, with minimal damage, fewer complications, and high success rate, to enable the experimental investigation of penile implants. Methods: We obtained materials for penile girth enhancement by processing the pericardium and blood vessel wall collected from pigs. Incisions were made at the penile bulb for the implantation of the materials, and facilitate observation and data collection, based on the anatomical features of dog’s penis. We measured the girth of the flaccid penis before and after the operation, and erectile function at 1-month postoperation. In addition to evaluation of recovery from the incision and local pathological changes, ultrasonic examination was performed to monitor the long-term changes associated with implantation. Results: The mean girth of the flaccid penis significantly increased from 7.37±0.40 cm before the operation, to 8.70±0.56 cm postoperation. Dogs resumed normal mating at 1 month after the operation, without any significant change in the mating time. Ultrasonic examination clearly illustrated the implants, and helped in the measurement of the distance between the materials and the baculum. Conclusion: Chinese Rural dog is a promising animal model for penile girth enhancement surgery. The findings demonstrated that surgical implantation into penile bulb was associated with less damage, faster postoperative recovery, and higher success. For the first time, ultrasonic examination provided objective data on the surgical outcomes of penile girth enhancement. PMID:26379868

  1. Cutaneous myeloid sarcoma of the penile foreskin.

    PubMed

    Afrose, Ruquiya; Nebhnani, Deepa; Wadhwa, Neelam

    2015-01-01

    Myeloid sarcoma, considered to herald the onset of a blast crisis in the setting of chronic myeloproliferative neoplasm/dysplasia, typically presents during the course of the disorder. Cutaneous involvement is uncommon and lesions on genital skin are seldom seen. We present a case of a well-differentiated myeloid sarcoma in the penile foreskin in an apparently healthy 29-year-old male presenting with phimosis. The unusual composition of the inflammatory cell infiltrate, and characteristic sparing of dermal blood vessels, nerves and smooth muscle fibres led to the correct diagnosis. Absence of commonly observed changes in the circumcision skin like those of balanitis xerotica was also helpful. Detailed hematological work up revealed a previously undiagnosed chronic myeloid leukemia in chronic phase. The patient also had simultaneous priapism, another rare presentation of chronic myeloid leukemia. One year hence, the patient is in hematological remission with no evidence of extramedullary disease. Although priapism has been described as a rare presenting symptom in chronic myeloid leukemia, the present case is unique as this is the first time a cutaneous myeloid sarcoma has been documented in the penile foreskin. PMID:24913300

  2. Penile Implants among Prisoners—A Cause for Concern?

    PubMed Central

    Yap, Lorraine; Butler, Tony; Richters, Juliet; Malacova, Eva; Wand, Handan; Smith, Anthony M. A.; Grant, Luke; Richards, Alun; Donovan, Basil

    2013-01-01

    Background We report the prevalence of penile implants among prisoners and determine the independent predictors for having penile implants. Questions on penile implants were included in the Sexual Health and Attitudes of Australian Prisoners (SHAAP) survey following concerns raised by prison health staff that increasing numbers of prisoners reported having penile implants while in prison. Methods Computer-Assisted Telephone Interviewing (CATI) of a random sample of prisoners was carried out in 41 prisons in New South Wales and Queensland (Australia). Men were asked, “Have you ever inserted or implanted an object under the skin of your penis?” If they responded Yes: “Have you ever done so while you were in prison?” Univariate logistic regression and logistic regression were used to determine the factors associated with penile implants. Results A total of 2,018 male prisoners were surveyed, aged between 18 and 65 years, and 118 (5.8%) reported that they had inserted or implanted an object under the skin of their penis. Of these men, 87 (73%) had this done while they were in prison. In the multivariate analysis, a younger age, birth in an Asian country, and prior incarceration were all significantly associated with penile implants (p<0.001). Men with penile implants were also more likely to report being paid for sex (p<0.001), to have had body piercings (p<0.001) or tattoos in prison (p<0.001), and to have taken non-prescription drugs while in prison (p<0.05). Conclusions Penile implants appear to be fairly common among prisoners and are associated with risky sexual and drug use practices. As most of these penile implants are inserted in prison, these men are at risk of blood borne viruses and wound infection. Harm reduction and infection control strategies need to be developed to address this potential risk. PMID:23326383

  3. Erectile dysfunction: the role of penile Doppler ultrasound in diagnosis.

    PubMed

    Halls, James; Bydawell, Gareth; Patel, Uday

    2009-11-01

    Erectile dysfunction (ED) is a common and debilitating condition with physical, psychological, and pharmacological aetiologies. The physical causes can be divided into problems with arterial inflow, structural penile abnormalities, or problems with the venous occlusion mechanism. Penile Doppler sonography is a specialized technique requiring a thorough knowledge of the topic in order to aid diagnosis and direct subsequent treatment. This technique is indicated in those patients with erectile dysfunction who do not respond to oral pharmacological agents (e.g., PDE-5 inhibitors). This pictorial essay will visit the anatomy and physiology of penile erection, the technique for performing the procedure, and review the imaging features for specific causes of ED.

  4. Azoreductase activity of anaerobic bacteria isolated from human intestinal microflora.

    PubMed Central

    Rafii, F; Franklin, W; Cerniglia, C E

    1990-01-01

    A plate assay was developed for the detection of anaerobic bacteria that produce azoreductases. With this plate assay, 10 strains of anaerobic bacteria capable of reducing azo dyes were isolated from human feces and identified as Eubacterium hadrum (2 strains), Eubacterium spp. (2 species), Clostridium clostridiiforme, a Butyrivibrio sp., a Bacteroides sp., Clostridium paraputrificum, Clostridium nexile, and a Clostridium sp. The average rate of reduction of Direct Blue 15 dye (a dimethoxybenzidine-based dye) in these strains ranged from 16 to 135 nmol of dye per min per mg of protein. The enzymes were inactivated by oxygen. In seven isolates, a flavin compound (riboflavin, flavin adenine dinucleotide, or flavin mononucleotide) was required for azoreductase activity. In the other three isolates and in Clostridium perfringens, no added flavin was required for activity. Nondenaturing polyacrylamide gel electrophoresis showed that each bacterium expressed only one azoreductase isozyme. At least three types of azoreductase enzyme were produced by the different isolates. All of the azoreductases were produced constitutively and released extracellularly. Images PMID:2202258

  5. Isolation of human umbilical arterial smooth muscle cells (HUASMC).

    PubMed

    Ribeiro, Maximiano P; Relvas, Ricardo; Chiquita, Samuel; Correia, Ilídio J

    2010-07-03

    The human umbilical cord (UC) is a biological sample that can be easily obtained just after birth. This biological sample is, most of the time, discarded and their collection does not imply any added risk to the newborn or mother s health. Moreover no ethical concerns are raised. The UC is composed by one vein and two arteries from which both endothelial cells (ECs) and smooth muscle cells (SMCs), two of the main cellular components of blood vessels, can be isolated. In this project the SMCs were obtained after enzymatic treatment of the UC arteries accordingly the experimental procedure previously described by Jaffe et al. After cell isolation they were kept in t-flash with DMEM-F12 supplemented with 5% of fetal bovine serum and were cultured for several passages. Cells maintained their morphological and other phenotypic characteristics in the different generations. The aim of this study was to isolate smooth muscle cells in order to use them as models for future assays with constrictor drugs, isolate and structurally characterize L-type calcium channels, to study cellular and molecular aspects of the vascular function and to use them in tissue engineering.

  6. Isolation and characterization of the human MRE11 homologue

    SciTech Connect

    Petrini, J.H.J.; Walsh, M.E.; DiMare, C.

    1995-09-01

    Mutation of the Saccharomyces cerevisiae RAD52 epistasis group gene, MRE11, blocks meiotic recombination, confers profound sensitivity to double-strand break damage, and has a hyperrecombinational phenotype in mitotic cells. We isolated a highly conserved human MRE11 homologue using a two-hybrid screen for DNA ligase I-interacting proteins. Human MRE11 shares approximately 50% identity with its yeast counterpart over the N-terminal half of the protein. MRE11 is expressed at the highest levels in proliferating tissues, but is also observed in other tissues. The MRE11 locus maps to human chromosome 11q21 in a region frequently associated with cancer-related chromosomal abnormalities. A MRE11-related locus was found on chromosome 7q11.2-q11.3. 60 refs., 4 figs.

  7. Isolation of Mouse and Human Tumor-Associated Macrophages.

    PubMed

    Cassetta, Luca; Noy, Roy; Swierczak, Agnieszka; Sugano, Gaël; Smith, Harriet; Wiechmann, Lisa; Pollard, Jeffrey W

    2016-01-01

    The tumor microenvironment is a complex network of cells that support tumor progression and malignancy. It has been demonstrated that tumor cells can educate the immune system to promote a tumor-friendly environment. Among all these immune cells, tumor-associated macrophages (TAMs) are well represented and their presence in mouse models has been shown to promote tumor progression and metastasis. These effects are through the stimulation of angiogenesis, enhancement of tumor cell invasion and intravasation, immunosuppression, and at the metastatic site tumor cell extravasation and growth. However, the precise mechanisms are not fully understood. Furthermore there is limited information on TAMs derived from human cancers. For this reason it is important to be able to extract TAMs from tumors in order to compare their phenotypes, functions, and transcriptomes with normal resident tissue macrophages. Isolation of these cells is challenging due to the lack of markers and standardized protocols. Here we show an optimized protocol for the efficient isolation and extraction of resident macrophages and TAMs from human and mouse tissues by using multicolor flow cytometry. These protocols allow for the extraction of thousands of macrophages in less than 5 h from tissues as small as half a gram. The isolated macrophages can then be used for both "omics" and in vitro studies. PMID:27325269

  8. Isolation of Mouse and Human Tumor-Associated Macrophages

    PubMed Central

    Cassetta, Luca; Noy, Roy; Swierczak, Agnieszka; Sugano, Gaël; Smith, Harriet; Wiechmann, Lisa; Pollard, Jeffrey W.

    2016-01-01

    The tumor microenvironment is a complex network of cells that support tumor progression and malignancy. It has been demonstrated that tumor cells can educate the immune system to promote a tumor-friendly environment. Among all these immune cells, tumor-associated macrophages (TAMs) are well represented and their presence in mouse models has been shown to promote tumor progression and metastasis. These effects are through the stimulation of angiogenesis, enhancement of tumor cell invasion and intravasation, immunosuppression, and at the metastatic site tumor cell extravasation and growth. However, the precise mechanisms are not fully understood. Furthermore there is limited information on TAMs derived from human cancers. For this reason it is important to be able to extract TAMs from tumors in order to compare their phenotypes, functions, and transcriptomes with normal resident tissue macrophages. Isolation of these cells is challenging due to the lack of markers and standardized protocols. Here we show an optimized protocol for the efficient isolation and extraction of resident macrophages and TAMs from human and mouse tissues by using multicolor flow cytometry. These protocols allow for the extraction of thousands of macrophages in less than 5 h from tissues as small as half a gram. The isolated macrophages can then be used for both “omics” and in vitro studies. PMID:27325269

  9. Multipotent progenitor cells isolated from adult human pancreatic tissue.

    PubMed

    Todorov, I; Nair, I; Ferreri, K; Rawson, J; Kuroda, A; Pascual, M; Omori, K; Valiente, L; Orr, C; Al-Abdullah, I; Riggs, A; Kandeel, F; Mullen, Y

    2005-10-01

    The supply of islet cells is a limiting factor for the widespread application of islet transplantation of type-1 diabetes. Islets constitute 1% to 2% of pancreatic tissue, leaving approximately 98% as discard after islet isolation and purification. In this report we present our data on the isolation of multipotent progenitor cells from discarded adult human pancreatic tissue. The collected cells from discarded nonislet fractions, after enzymatic digestion and gradient purification of islets, were dissociated for suspension culture in a serum-free medium. The cell clusters grown to a size of 100 to 150 mum contained cells staining for stage-specific embryonic antigens, but not insulin or C-peptide. To direct cell differentiation toward islets, clusters were recultured in a pancreatic differentiation medium. Insulin and C-peptide-positive cells by immunocytochemistry appeared within a week, reaching over 10% of the cell population. Glucagon and somatostatin-positive cells were also detected. The cell clusters were found to secrete insulin in response to glucose stimulation. Cells from the same clusters also had the capacity for differentiation into neural cells, as documented by staining for neural and glial cell markers when cultured as monolayers in media containing neurotrophic factors. These data suggest that multipotent pancreatic progenitor cells exist within the human pancreatic tissue that is typically discarded during islet isolation procedures. These adult progenitor cells can be successfully differentiated into insulin-producing cells, and thus they have the potential for treatment of type-1 diabetes mellitus. PMID:16298614

  10. Acrolein generation stimulates hypercontraction in isolated human blood vessels

    SciTech Connect

    Conklin, D.J. . E-mail: dj.conklin@louisville.edu; Bhatnagar, A.; Cowley, H.R.; Johnson, G.H.; Trent, M.B.; Boor, P.J.

    2006-12-15

    Increased risk of vasospasm, a spontaneous hyperconstriction, is associated with atherosclerosis, cigarette smoking, and hypertension-all conditions involving oxidative stress, lipid peroxidation, and inflammation. To test the role of the lipid peroxidation- and inflammation-derived aldehyde, acrolein, in human vasospasm, we developed an ex vivo model using human coronary artery bypass graft (CABG) blood vessels and a demonstrated acrolein precursor, allylamine. Allylamine induces hypercontraction in isolated rat coronary artery in a semicarbazide-sensitive amine oxidase activity (SSAO) dependent manner. Isolated human CABG blood vessels (internal mammary artery, radial artery, saphenous vein) were used to determine: (1) vessel responses and sensitivity to acrolein, allylamine, and H{sub 2}O{sub 2} exposure (1 {mu}M-1 mM), (2) SSAO dependence of allylamine-induced effects using SSAO inhibitors (semicarbazide, 1 mM; MDL 72274-E, active isomer; MDL 72274-Z, inactive isomer; 100 {mu}M), (3) the vasoactive effects of two other SSAO amine substrates, benzylamine and methylamine, and (4) the contribution of extracellular Ca{sup 2+} to hypercontraction. Acrolein or allylamine but not H{sub 2}O{sub 2}, benzylamine, or methylamine stimulated spontaneous and pharmacologically intractable hypercontraction in CABG blood vessels that was similar to clinical vasospasm. Allylamine-induced hypercontraction and blood vessel SSAO activity were abolished by pretreatment with semicarbazide or MDL 72274-E but not by MDL 72274-Z. Allylamine-induced hypercontraction also was significantly attenuated in Ca{sup 2+}-free buffer. In isolated aorta of spontaneously hypertensive rat, allylamine-induced an SSAO-dependent contraction and enhanced norepinephrine sensitivity but not in Sprague-Dawley rat aorta. We conclude that acrolein generation in the blood vessel wall increases human susceptibility to vasospasm, an event that is enhanced in hypertension.

  11. Postnatal penile growth concurrent with mini-puberty predicts later sex-typed play behavior: Evidence for neurobehavioral effects of the postnatal androgen surge in typically developing boys.

    PubMed

    Pasterski, Vickie; Acerini, Carlo L; Dunger, David B; Ong, Ken K; Hughes, Ieuan A; Thankamony, Ajay; Hines, Melissa

    2015-03-01

    The masculinizing effects of prenatal androgens on human neurobehavioral development are well established. Also, the early postnatal surge of androgens in male infants, or mini-puberty, has been well documented and is known to influence physiological development, including penile growth. However, neurobehavioral effects of androgen exposure during mini-puberty are largely unknown. The main aim of the current study was to evaluate possible neurobehavioral consequences of mini-puberty by relating penile growth in the early postnatal period to subsequent behavior. Using multiple linear regression, we demonstrated that penile growth between birth and three months postnatal, concurrent with mini-puberty, significantly predicted increased masculine/decreased feminine behavior assessed using the Pre-school Activities Inventory (PSAI) in 81 healthy boys at 3 to 4years of age. When we controlled for other potential influences on masculine/feminine behavior and/or penile growth, including variance in androgen exposure prenatally and body growth postnally, the predictive value of penile growth in the early postnatal period persisted. More specifically, prenatal androgen exposure, reflected in the measurement of anogenital distance (AGD), and early postnatal androgen exposure, reflected in penile growth from birth to 3months, were significant predictors of increased masculine/decreased feminine behavior, with each accounting for unique variance. Our findings suggest that independent associations of PSAI with AGD at birth and with penile growth during mini-puberty reflect prenatal and early postnatal androgen exposures respectively. Thus, we provide a novel and readily available approach for assessing effects of early androgen exposures, as well as novel evidence that early postnatal aes human neurobehavioral development.

  12. Postnatal penile growth concurrent with mini-puberty predicts later sex-typed play behavior: Evidence for neurobehavioral effects of the postnatal androgen surge in typically developing boys.

    PubMed

    Pasterski, Vickie; Acerini, Carlo L; Dunger, David B; Ong, Ken K; Hughes, Ieuan A; Thankamony, Ajay; Hines, Melissa

    2015-03-01

    The masculinizing effects of prenatal androgens on human neurobehavioral development are well established. Also, the early postnatal surge of androgens in male infants, or mini-puberty, has been well documented and is known to influence physiological development, including penile growth. However, neurobehavioral effects of androgen exposure during mini-puberty are largely unknown. The main aim of the current study was to evaluate possible neurobehavioral consequences of mini-puberty by relating penile growth in the early postnatal period to subsequent behavior. Using multiple linear regression, we demonstrated that penile growth between birth and three months postnatal, concurrent with mini-puberty, significantly predicted increased masculine/decreased feminine behavior assessed using the Pre-school Activities Inventory (PSAI) in 81 healthy boys at 3 to 4years of age. When we controlled for other potential influences on masculine/feminine behavior and/or penile growth, including variance in androgen exposure prenatally and body growth postnally, the predictive value of penile growth in the early postnatal period persisted. More specifically, prenatal androgen exposure, reflected in the measurement of anogenital distance (AGD), and early postnatal androgen exposure, reflected in penile growth from birth to 3months, were significant predictors of increased masculine/decreased feminine behavior, with each accounting for unique variance. Our findings suggest that independent associations of PSAI with AGD at birth and with penile growth during mini-puberty reflect prenatal and early postnatal androgen exposures respectively. Thus, we provide a novel and readily available approach for assessing effects of early androgen exposures, as well as novel evidence that early postnatal aes human neurobehavioral development. PMID:25597916

  13. Reconstructive surgery for idealising penile shape and restoring erectile function in patients with penile dysmorphology and erectile dysfunction

    PubMed Central

    Hsu, Geng-Long; Molodysky, Eugen; Liu, Shih-Ping; Chang, Hong-Chiang; Hsieh, Cheng-Hsing; Hsu, Chih-Yuan

    2013-01-01

    Objective To report an innovative combination of two surgical procedures to treat patients with erectile dysfunction and penile deviation, arising from advances in penile anatomy. Patients and methods From October 1998 to October 2011, 132 men (aged 23–39 years) underwent penile venous stripping and corporoplasty. Of these, 37 were allocated to a transverse and 95 to a longitudinal group, with an infrapubic transverse or pubic median longitudinal approach, respectively. The abridged five-item version of the International Index of Erectile Function (IIEF-5) and cavernosography were used for assessment, as necessary. Under acupuncture-aided local anaesthesia, and after a circumferential incision, the deep dorsal vein and cavernous veins were completely stripped, with 6-0 Nylon sutures for ligation, followed by tunical surgery for correcting the penile shape. Results In the transverse and longitudinal groups the mean (SD) duration of surgery was 4.6 (0.2) and 4.8 (0.3) h, respectively. Before surgery the mean (SD) IIEF-5 score was 9.4 (2.3) and 9.6 (2.1), which increased to 20.6 (2.4) and 20.8 (2.7), respectively, after surgery. The penile shape (<15°) was deemed satisfactory in 92% (34/37) and 96% (91/95) of patients in the transverse and longitudinal groups, respectively. The cavernosograms consistently showed a good penile shape. There were significant differences in the mean (SD) duration of penile oedema, at 3.2 (1.6) vs. 11.9 (2.1) days, the overall satisfaction rate and the prevalence of hypertrophied scarring (all P < 0.001). Conclusion This combination of unique penile venous stripping with a pubic median longitudinal approach and an anatomy-based corporoplasty is ideally suited to the simultaneous restoration of penile erectile function and morphological reconstruction. PMID:26558108

  14. AVE 0991, a non-peptide Mas-receptor agonist, facilitates penile erection.

    PubMed

    da Costa Gonçalves, Andrey C; Fraga-Silva, Rodrigo A; Leite, Romulo; Santos, Robson A S

    2013-03-01

    The renin-angiotensin system plays a crucial role in erectile function. It has been shown that elevated levels of angiotensin II contribute to the development of erectile dysfunction both in humans and in aminals. On the contrary, the heptapeptide angiotensin-(1-7) appears to mediate penile erection by activation of the Mas receptor. Recently, we have shown that the erectile function of Mas gene-deleted mice was substantially reduced, which was associated with a marked increase in fibrous tissue in the corpus cavernosum. We have hypothesized that the synthetic non-peptide Mas agonist, AVE 0991, would potentiate penile erectile function. We showed that intracavernosal injection of AVE 0991 potentiated the erectile response of anaesthetized Wistar rats, measured as the ratio between corpus cavernosum pressure and mean arterial pressure, upon electrical stimulation of the major pelvic ganglion. The facilitatory effect of AVE 0991 on erectile function was dose dependent and completely blunted by the nitric oxide synthesis inhibitor, l-NAME. Importantly, concomitant intracavernosal infusion of the specific Mas receptor blocker, A-779, abolished the effect of AVE 0991. We demonstrated that AVE 0991 potentiates the penile erectile response through Mas in an NO-dependent manner. Importantly, these results suggest that Mas agonists, such as AVE 0991, might have significant therapeutic benefits for the treatment of erectile dysfunction.

  15. Phosphorylated endothelial nitric oxide synthase mediates vascular endothelial growth factor-induced penile erection.

    PubMed

    Musicki, Biljana; Palese, Michael A; Crone, Julie K; Burnett, Arthur L

    2004-02-01

    The objective of the present study was to evaluate whether vascular endothelial growth factor (VEGF)-induced penile erection is mediated by activation of endothelial nitric oxide synthase (eNOS) through its phosphorylation. We assessed the role of constitutively activated eNOS in VEGF-induced penile erection using wild-type (WT) and eNOS-knockout (eNOS(-/-)) mice with and without vasculogenic erectile dysfunction. Adult WT and eNOS(-/-) mice were subjected to sham operation or bilateral castration to induce vasculogenic erectile dysfunction. At the time of surgery, animals were injected intracavernosally with a replication-deficient adenovirus expressing human VEGF145 (10(9) particle units) or with empty virus (Ad.Null). After 7 days, erectile function was assessed in response to cavernous nerve electrical stimulation. Total and phosphorylated protein kinase B (Akt) as well as total and phosphorylated eNOS were quantitatively assessed in mice penes using Western immunoblot and immunohistochemistry. In intact WT mice, VEGF145 significantly increased erectile responses, and in WT mice after castration, it completely recovered penile erection. However, VEGF145 failed to increase erectile responses in intact eNOS(-/-) mice and only partially recovered erectile function in castrated eNOS(-/-) mice. In addition, VEGF145 significantly increased phosphorylation of eNOS at Serine 1177 by approximately 2-fold in penes of both intact and castrated WT mice. The data provide a molecular explanation for VEGF stimulatory effect on penile erection, which involves phosphorylated eNOS (Serine 1177) mediation. PMID:14522830

  16. Timing of androgen receptor disruption and estrogen exposure underlies a spectrum of congenital penile anomalies.

    PubMed

    Zheng, Zhengui; Armfield, Brooke A; Cohn, Martin J

    2015-12-29

    Congenital penile anomalies (CPAs) are among the most common human birth defects. Reports of CPAs, which include hypospadias, chordee, micropenis, and ambiguous genitalia, have risen sharply in recent decades, but the causes of these malformations are rarely identified. Both genetic anomalies and environmental factors, such as antiandrogenic and estrogenic endocrine disrupting chemicals (EDCs), are suspected to cause CPAs; however, little is known about the temporal window(s) of sensitivity to EDCs, or the tissue-specific roles and downstream targets of the androgen receptor (AR) in external genitalia. Here, we show that the full spectrum of CPAs can be produced by disrupting AR at different developmental stages and in specific cell types in the mouse genital tubercle. Inactivation of AR during a narrow window of prenatal development results in hypospadias and chordee, whereas earlier disruptions cause ambiguous genitalia and later disruptions cause micropenis. The neonatal phase of penile development is controlled by the balance of AR to estrogen receptor α (ERα) activity; either inhibition of androgen or augmentation of estrogen signaling can induce micropenis. AR and ERα have opposite effects on cell division, apoptosis, and regulation of Hedgehog, fibroblast growth factor, bone morphogenetic protein, and Wnt signaling in the genital tubercle. We identify Indian hedgehog (Ihh) as a novel downstream target of AR in external genitalia and show that conditional deletion of Ihh inhibits penile masculinization. These studies reveal previously unidentified cellular and molecular mechanisms by which antiandrogenic and estrogenic signals induce penile malformations and demonstrate that the timing of endocrine disruption can determine the type of CPA.

  17. Timing of androgen receptor disruption and estrogen exposure underlies a spectrum of congenital penile anomalies

    PubMed Central

    Armfield, Brooke A.; Cohn, Martin J.

    2015-01-01

    Congenital penile anomalies (CPAs) are among the most common human birth defects. Reports of CPAs, which include hypospadias, chordee, micropenis, and ambiguous genitalia, have risen sharply in recent decades, but the causes of these malformations are rarely identified. Both genetic anomalies and environmental factors, such as antiandrogenic and estrogenic endocrine disrupting chemicals (EDCs), are suspected to cause CPAs; however, little is known about the temporal window(s) of sensitivity to EDCs, or the tissue-specific roles and downstream targets of the androgen receptor (AR) in external genitalia. Here, we show that the full spectrum of CPAs can be produced by disrupting AR at different developmental stages and in specific cell types in the mouse genital tubercle. Inactivation of AR during a narrow window of prenatal development results in hypospadias and chordee, whereas earlier disruptions cause ambiguous genitalia and later disruptions cause micropenis. The neonatal phase of penile development is controlled by the balance of AR to estrogen receptor α (ERα) activity; either inhibition of androgen or augmentation of estrogen signaling can induce micropenis. AR and ERα have opposite effects on cell division, apoptosis, and regulation of Hedgehog, fibroblast growth factor, bone morphogenetic protein, and Wnt signaling in the genital tubercle. We identify Indian hedgehog (Ihh) as a novel downstream target of AR in external genitalia and show that conditional deletion of Ihh inhibits penile masculinization. These studies reveal previously unidentified cellular and molecular mechanisms by which antiandrogenic and estrogenic signals induce penile malformations and demonstrate that the timing of endocrine disruption can determine the type of CPA. PMID:26598695

  18. Phosphorylated endothelial nitric oxide synthase mediates vascular endothelial growth factor-induced penile erection.

    PubMed

    Musicki, Biljana; Palese, Michael A; Crone, Julie K; Burnett, Arthur L

    2004-02-01

    The objective of the present study was to evaluate whether vascular endothelial growth factor (VEGF)-induced penile erection is mediated by activation of endothelial nitric oxide synthase (eNOS) through its phosphorylation. We assessed the role of constitutively activated eNOS in VEGF-induced penile erection using wild-type (WT) and eNOS-knockout (eNOS(-/-)) mice with and without vasculogenic erectile dysfunction. Adult WT and eNOS(-/-) mice were subjected to sham operation or bilateral castration to induce vasculogenic erectile dysfunction. At the time of surgery, animals were injected intracavernosally with a replication-deficient adenovirus expressing human VEGF145 (10(9) particle units) or with empty virus (Ad.Null). After 7 days, erectile function was assessed in response to cavernous nerve electrical stimulation. Total and phosphorylated protein kinase B (Akt) as well as total and phosphorylated eNOS were quantitatively assessed in mice penes using Western immunoblot and immunohistochemistry. In intact WT mice, VEGF145 significantly increased erectile responses, and in WT mice after castration, it completely recovered penile erection. However, VEGF145 failed to increase erectile responses in intact eNOS(-/-) mice and only partially recovered erectile function in castrated eNOS(-/-) mice. In addition, VEGF145 significantly increased phosphorylation of eNOS at Serine 1177 by approximately 2-fold in penes of both intact and castrated WT mice. The data provide a molecular explanation for VEGF stimulatory effect on penile erection, which involves phosphorylated eNOS (Serine 1177) mediation.

  19. Islet isolation from human pancreas with extended cold ischemia time.

    PubMed

    Kühtreiber, W M; Ho, L T; Kamireddy, A; Yacoub, J A W; Scharp, D W

    2010-01-01

    The general consensus among transplant centers is that a cold ischemia time (CIT) beyond 8 hours results in reduced yields and quality of human islets. We sought to optimize the isolation process and enzymes for pancreata with extended CIT. We processed 16 extended CIT pancreata (13.2 +/- 0.7 hours). Donors averaged 50.8 +/- 2.6 (standard error of the mean) years old with a body mass index of 28.6 +/- 1.5. Glands were shipped in cold organ preservation solution without oxygenated perfluorocarbon. Isolations were performed under a protocol optimized for digestion with the new cGMP collagenase from Roche. Purification used continuous Euroficoll/University of Wisconsin gradients. Islets were cultured in two types of Prodo cGMP islet culture media and/or in Miami 1A media. Glucose-stimulated insulin secretion assays were performed after 8 to 16 days of culture. Prepurification yield averaged 415 +/- 41 KIEQ postpurification, 359 +/- 29 KIEQ (purification loss 13.5%); and postculture 317 +/- 27 KIEQ (culture loss 11.7%). Our process liberated an average of 4278 IEQ/g of pancreas (97 +/- 5 g). Most islets were recovered in the purest fraction (purity 79.7% +/- 1.9%). Culture loss in our enhanced culture media was 11.7%. After 2 to 3 days in culture, viability was 92% +/- 1%. Islets exhibited compactness and dithizone staining. Glucose-stimulated insulin secretion assays performed after 3 to 23 days in our PIM(R) media resulted in a stimulation index of 6.8 +/- 1.7 (G50 to G350). We concluded that our human islet isolation process permitted the recovery of large numbers of high-quality human islets from extended CIT pancreata and that our cGMP islet culture media was superior to the current standard CMRL-based media. PMID:20692399

  20. Salmonella enterica serovar Senftenberg human clinical isolates lacking SPI-1.

    PubMed

    Hu, Qinghua; Coburn, Bryan; Deng, Wanyin; Li, Yuling; Shi, Xiaolu; Lan, Quanxue; Wang, Bing; Coombes, Brian K; Finlay, B Brett

    2008-04-01

    Nontyphoidal Salmonella species cause gastrointestinal disease worldwide. The prevailing theory of Salmonella enteropathogenesis is that bacterial invasion of the intestinal epithelium is essential for virulence and that this requires the virulence-associated genomic region Salmonella pathogenicity island 1 (SPI-1). Recent studies of Salmonella enterica infection models have demonstrated that enterocolitis and diarrhea in mice and cows can occur independently of SPI-1. In this study, we sought to confirm whether two S. enterica serovar Senftenberg clinical isolates lacked genes essential for SPI-1 function. Two clinical strains were isolated and identified as being S. enterica serovar Senftenberg from four stool samples from a food-borne disease outbreak affecting seven individuals in Shenzhen, Guangdong Province, China, using conventional methods, pulsed-field gel electrophoresis and multilocus sequence typing. The possibility of coinfection with other potential bacteria or usual viruses was excluded. Two isolates were analyzed for the presence of invA, sipA, ssaR, sifA, and sopE2 by PCR and Southern blotting and were then assayed for the presence of SPI-1 by PCR and long-range PCR for fhlA-hilA, hilA-spaP, and spaP-invH and Southern blot analysis. A long-range PCR fragment from fhlA to mutS covering the 5' and 3' flanks of SPI-1 was also amplified from the two clinical isolates and sequenced. In addition, the two clinical isolates were assayed for enteroinvasiveness in vitro. Murine infection models were also examined. Biochemical tests and serotyping confirmed that the two clinical isolates are S. enterica serovar Senftenberg. However, they lacked genes critical for SPI-1 function but contained SPI-2 genes and were attenuated for the invasion of cultured intestinal epithelial cells. In conclusion, clinical S. enterica serovar Senftenberg strains isolated from a food-borne disease outbreak lack the invasion-associated locus SPI-1, indicating that SPI-1 is not

  1. Clinical neuroanatomy and neurotransmitter-mediated regulation of penile erection.

    PubMed

    Jung, Junyang; Jo, Hyun Woo; Kwon, Hyunseob; Jeong, Na Young

    2014-06-01

    Erectile dysfunction (ED) has an adverse impact on men's quality of life. Penile erection, which is regulated by nerves that are innervated into the erectile tissue, can be affected by functional or anatomical trauma of the perineal region, including specific structures of the penis, causing ED. Penile erection is neurologically controlled by the autonomic nervous system. Therefore, it is of utmost importance to understand the neurogenic structure of the erectile tissue and the types of neurotransmitters involved in the penile erection process. Here, we highlight the basic clinical anatomy and erectile function of the penis. Understanding the clinical connotation of the relationship between penile erectile structure and function may provide fresh insights for identifying the main mechanisms involved in ED and help develop surgical techniques for the treatment of ED.

  2. Human senile cataractous lens protease. Isolation and some chemical characteristics

    PubMed Central

    Swanson, Arnold A.; Nichols, John T.

    1971-01-01

    A proteolytic enzyme was isolated from human senile cataractous lens by anion-exchange and gel-filtration chromatography. Sedimentation and zone-electrophoretic experiments indicated a high degree of homogeneity for the enzyme. A molecular weight of 27000 was calculated from measurements of sedimentation velocity and diffusion coefficient. Chelating agents decreased activity which could be restored by addition of certain bivalent metal ions. Di-isopropyl phosphorofluoridate and phenylmethanesulphonyl fluoride inhibit the proteolytic activities. Optimum rates of hydrolysis were observed at pH5.2. ImagesFig. 6. PMID:5144759

  3. Penile lichen sclerosus: An urologist's nightmare! - A single center experience

    PubMed Central

    Singh, Jitendra Pratap; Priyadarshi, Vinod; Goel, Hemant Kumar; Vijay, Mukesh Kumar; Pal, Dilip Kumar; Chakraborty, Sudip; Kundu, Anup Kumar

    2015-01-01

    Purpose: Penile lichen sclerosus (LS) is a nagging condition and its progression result in devastating urinary and sexual problems and reduction in the quality-of-life. This study has been carried out to present our experience about this disease with simultaneous review of the available literature. Materials and Methods: This retrospective study has been done at a tertiary care center of eastern India. The data of 306 patients affected with LS were analyzed for clinical presentation, physical examination, investigations, and treatment offered. Results: Presenting symptoms were non-specific. The prepuce was most commonly involved location followed by glans and meatus. Urethral involvement was not isolated as the primary site. Circumcision was done in 237 patients, while 63 patients underwent meatotomy. Thirty-six of 39 cases of LS induced stricture were treated with buccal mucosal graft (BMG) either in one stage or in two stages. Conclusion: LS varies from being a highly aggressive disease of the penis and anterior urethra to a burnt out condition affecting just the meatus and surrounding glans. Early diagnosis and treatment are required to prevent its complication and associated morbidity. Management depends on the anatomical location of lesion, extent of involvement, rapidity of progression and its severity. Use of BMG in LS induced urethral stricture has shown encouraging results. PMID:26229314

  4. Multimodality imaging of penile cancer: what radiologists need to know.

    PubMed

    Suh, Chong Hyun; Baheti, Akshay D; Tirumani, Sree Harsha; Rosenthal, Michael H; Kim, Kyung Won; Ramaiya, Nikhil H; Shinagare, Atul B

    2015-02-01

    The purpose of this article is to provide a comprehensive update on the role of imaging in the diagnosis and management of penile cancer. Imaging plays a major role in the initial assessment, treatment planning, and follow-up of patients with penile carcinoma. MRI helps in assessing the T staging of the primary and in detecting local recurrence. PET/CT and CT are useful for detecting regional nodal and distant metastases.

  5. Penile Sarcoma: Report of a Rare Malignancy

    PubMed Central

    Kumar, Vijay; Chaturvedi, Arun; Vishnoi, Jeevan Ram; Dontula, Prashant

    2016-01-01

    Penile cancer is an uncommon malignancy. Squamous cell carcinoma constitutes approximately 95% of all histology. Non-squamous malignancies are rare in penis. Sarcomas of penis are rarer among them. Spindle cell sarcoma is one of the extremely rare sarcoma of penis. To best of our knowledge, only two cases have been reported so far, one in English literature and other in Japanese. We are presenting this uncommon case of spindle cell sarcoma of penis, which was diagnosed with microscopy with its characteristic immunohistochemistry. The disease had an aggressive course with multiple recurrences in a short duration despite margin negative resection. Disease responded poorly with the chemotherapy and patient succumbed to the disease. PMID:27630937

  6. Penile Sarcoma: Report of a Rare Malignancy.

    PubMed

    Rajan, Shiv; Kumar, Vijay; Chaturvedi, Arun; Vishnoi, Jeevan Ram; Dontula, Prashant

    2016-07-01

    Penile cancer is an uncommon malignancy. Squamous cell carcinoma constitutes approximately 95% of all histology. Non-squamous malignancies are rare in penis. Sarcomas of penis are rarer among them. Spindle cell sarcoma is one of the extremely rare sarcoma of penis. To best of our knowledge, only two cases have been reported so far, one in English literature and other in Japanese. We are presenting this uncommon case of spindle cell sarcoma of penis, which was diagnosed with microscopy with its characteristic immunohistochemistry. The disease had an aggressive course with multiple recurrences in a short duration despite margin negative resection. Disease responded poorly with the chemotherapy and patient succumbed to the disease. PMID:27630937

  7. Isolation of Leukocytes from the Human Maternal-fetal Interface.

    PubMed

    Xu, Yi; Plazyo, Olesya; Romero, Roberto; Hassan, Sonia S; Gomez-Lopez, Nardhy

    2015-01-01

    Pregnancy is characterized by the infiltration of leukocytes in the reproductive tissues and at the maternal-fetal interface (decidua basalis and decidua parietalis). This interface is the anatomical site of contact between maternal and fetal tissues; therefore, it is an immunological site of action during pregnancy. Infiltrating leukocytes at the maternal-fetal interface play a central role in implantation, pregnancy maintenance, and timing of delivery. Therefore, phenotypic and functional characterizations of these leukocytes will provide insight into the mechanisms that lead to pregnancy disorders. Several protocols have been described in order to isolate infiltrating leukocytes from the decidua basalis and decidua parietalis; however, the lack of consistency in the reagents, enzymes, and times of incubation makes it difficult to compare these results. Described herein is a novel approach that combines the use of gentle mechanical and enzymatic dissociation techniques to preserve the viability and integrity of extracellular and intracellular markers in leukocytes isolated from the human tissues at the maternal-fetal interface. Aside from immunophenotyping, cell culture, and cell sorting, the future applications of this protocol are numerous and varied. Following this protocol, the isolated leukocytes can be used to determine DNA methylation, expression of target genes, in vitro leukocyte functionality (i.e., phagocytosis, cytotoxicity, T-cell proliferation, and plasticity, etc.), and the production of reactive oxygen species at the maternal-fetal interface. Additionally, using the described protocol, this laboratory has been able to describe new and rare leukocytes at the maternal-fetal interface.

  8. Genetics of human isolated hereditary hair loss disorders.

    PubMed

    Basit, S; Khan, S; Ahmad, W

    2015-09-01

    Hereditary hair loss in human is a group of clinically and genetically heterogeneous disorders. It is characterized by sparse to complete absence of hair on the scalp and other parts of the body. In few cases tightly curled twisted wooly hair (WH) on the scalp has been reported as well. The hair loss disorders, including both syndromic and non-syndromic (isolated) forms, segregate either in autosomal dominant or autosomal recessive pattern. To date, seven autosomal dominant and equal numbers of autosomal recessive isolated forms of hair loss disorders have been characterized. Genes responsible for causing most of these disorders have been identified. In this review, we have provided an update on clinical and genetic aspects of isolated hereditary hair loss disorders manifesting with hypotrichosis and/or WHs. Because most of the recessive genes have been mapped using consanguineous families of Pakistani origin, therefore emphasis is given to mutations identified in these families. OMIM nomenclature has been followed to indicate different forms of hair loss disorders.

  9. Penile rehabilitation following prostate cancer treatment: review of current literature

    PubMed Central

    Clavell-Hernandez, Jonathan; Wang, Run

    2015-01-01

    Radical prostatectomy (RP) and radiotherapy (RT) are highly effective in improving prostate cancer survival. However, both have a detrimental effect on erectile function (EF). Penile rehabilitation consists of understanding the mechanisms that cause erectile dysfunction (ED) and utilizing pharmacologic agents, devices or interventions to promote male sexual function. For the past decade, many researchers have pursued to define effective treatment modalities to improve ED after prostate cancer treatment. Despite the understanding of the mechanisms and well-established rationale for postprostate treatment penile rehabilitation, there is still no consensus regarding effective rehabilitation programs. This article reviews a contemporary series of trials that assess penile rehabilitation and explore treatment modalities that might play a role in the future. Published data and trials related to penile rehabilitation after RP and RT were reviewed and presented. Although recent trials have shown that most therapies are well-tolerated and aid in some degree on EF recovery, we currently do not have tangible evidence to recommend an irrefutable penile rehabilitation algorithm. However, advancements in research and technology will ultimately create and refine management options for penile rehabilitation. PMID:25851656

  10. Development of the penile urethra in the tammar wallaby.

    PubMed

    Leihy, M W; Shaw, G; Wilson, J D; Renfree, M B

    2011-01-01

    Hypospadias is increasingly common, and requires surgery to repair, but its aetiology is poorly understood. The marsupial tammar wallaby provides a unique opportunity to study hypospadias because penile differentiation occurs postnatally. Androgens are responsible for penile development in the tammar, but the majority of differentiation, in particular formation and closure of the urethral groove forming the penile urethra in males, occurs when there is no measurable sex difference in the concentrations of testosterone or dihydrotestosterone in either the gonads or the circulation [corrected]. Phalluses were examined morphologically from the sexually indifferent period (when androgens are high) to well after the time that the phallus becomes sexually dimorphic. We show that penile development and critical changes in the positioning of the urethra occur in the male phallus begin during an early window of time when androgens are high. Remodelling of the urethra in the male occurs between days 20-60. The critical period of time for the establishment urethral closure occurs during the earliest phases of penile development. This study suggests that there is an early window of time before day 60 when androgen imprinting must occur for normal penile development and closure of the urethral groove.

  11. Development of the penile urethra in the tammar wallaby.

    PubMed

    Leihy, M W; Shaw, G; Wilson, J D; Renfree, M B

    2011-01-01

    Hypospadias is increasingly common, and requires surgery to repair, but its aetiology is poorly understood. The marsupial tammar wallaby provides a unique opportunity to study hypospadias because penile differentiation occurs postnatally. Androgens are responsible for penile development in the tammar, but the majority of differentiation, in particular formation and closure of the urethral groove forming the penile urethra in males, occurs when there is no measurable sex difference in the concentrations of testosterone or dihydrotestosterone in either the gonads or the circulation [corrected]. Phalluses were examined morphologically from the sexually indifferent period (when androgens are high) to well after the time that the phallus becomes sexually dimorphic. We show that penile development and critical changes in the positioning of the urethra occur in the male phallus begin during an early window of time when androgens are high. Remodelling of the urethra in the male occurs between days 20-60. The critical period of time for the establishment urethral closure occurs during the earliest phases of penile development. This study suggests that there is an early window of time before day 60 when androgen imprinting must occur for normal penile development and closure of the urethral groove. PMID:22116535

  12. Isolating Mesangiogenic Progenitor Cells (MPCs) from Human Bone Marrow.

    PubMed

    Montali, Marina; Barachini, Serena; Pacini, Simone; Panvini, Francesca M; Petrini, Mario

    2016-01-01

    In a research study aimed to isolate human bone marrow (hBM)-derived Mesenchymal Stromal Cells (MSCs) for clinical applications, we identified a novel cell population specifically selected for growth in human serum supplemented medium. These cells are characterized by morphological, phenotypic, and molecular features distinct from MSCs and we named them Mesodermal Progenitor Cells (MPCs). MPCs are round, with a thick highly refringent core region; they show strong, trypsin resistant adherence to plastic. Failure to expand MPCs directly revealed that they are slow in cycling. This is as also suggested by Ki-67 negativity. On the other hand, culturing MPCs in standard medium designed for MSC expansion, gave rise to a population of exponentially growing MSC-like cells. Besides showing mesenchymal differentiation capacity MPCs retained angiogenic potential, confirming their multiple lineage progenitor nature. Here we describe an optimized highly reproducible protocol to isolate and characterize hBM-MPCs by flow cytometry (CD73, CD90, CD31, and CD45), nestin expression, and F-actin organization. Protocols for mesengenic and angiogenic differentiation of MPCs are also provided. Here we also suggest a more appropriate nomenclature for these cells, which has been re-named as "Mesangiogenic Progenitor Cells". PMID:27500428

  13. Isolation of Human Genomic DNA Sequences with Expanded Nucleobase Selectivity.

    PubMed

    Rathi, Preeti; Maurer, Sara; Kubik, Grzegorz; Summerer, Daniel

    2016-08-10

    We report the direct isolation of user-defined DNA sequences from the human genome with programmable selectivity for both canonical and epigenetic nucleobases. This is enabled by the use of engineered transcription-activator-like effectors (TALEs) as DNA major groove-binding probes in affinity enrichment. The approach provides the direct quantification of 5-methylcytosine (5mC) levels at single genomic nucleotide positions in a strand-specific manner. We demonstrate the simple, multiplexed typing of a variety of epigenetic cancer biomarker 5mC with custom TALE mixes. Compared to antibodies as the most widely used affinity probes for 5mC analysis, i.e., employed in the methylated DNA immunoprecipitation (MeDIP) protocol, TALEs provide superior sensitivity, resolution and technical ease. We engineer a range of size-reduced TALE repeats and establish full selectivity profiles for their binding to all five human cytosine nucleobases. These provide insights into their nucleobase recognition mechanisms and reveal the ability of TALEs to isolate genomic target sequences with selectivity for single 5-hydroxymethylcytosine and, in combination with sodium borohydride reduction, single 5-formylcytosine nucleobases. PMID:27429302

  14. Antigenic Relationships among Human Pathogenic Orientia tsutsugamushi Isolates from Thailand

    PubMed Central

    Nawtaisong, Pruksa; Tanganuchitcharnchai, Ampai; Smith, Derek J.; Day, Nicholas P. J.; Paris, Daniel H.

    2016-01-01

    Background Scrub typhus is a common cause of undiagnosed febrile illness in certain tropical regions, but can be easily treated with antibiotics. The causative agent, Orientia tsutsugamushi, is antigenically variable which complicates diagnosis and efforts towards vaccine development. Methodology/Principal Findings This study aimed to dissect the antigenic and genetic relatedness of O. tsutsugamushi strains and investigate sero-diagnostic reactivities by titrating individual patient sera against their O. tsutsugamushi isolates (whole-cell antigen preparation), in homologous and heterologous serum-isolate pairs from the same endemic region in NE Thailand. The indirect immunofluorescence assay was used to titrate Orientia tsutsugamushi isolates and human sera, and a mathematical technique, antigenic cartography, was applied to these data to visualise the antigenic differences and cross-reactivity between strains and sera. No functional or antigen-specific analyses were performed. The antigenic variation found in clinical isolates was much less pronounced than the genetic differences found in the 56kDa type-specific antigen genes. The Karp-like sera were more broadly reactive than the Gilliam-like sera. Conclusions/Significance Antigenic cartography worked well with scrub typhus indirect immunofluorescence titres. The data from humoral responses suggest that a Karp-like strain would provide broader antibody cross-reactivity than a Gilliam-like strain. Although previous exposure to O. tsutsugamushi could not be ruled out, scrub typhus patient serum antibody responses were characterised by strong homologous, but weak heterologous antibody titres, with little evidence for cross-reactivity by Gilliam-like sera, but a broader response from some Karp-like sera. This work highlights the importance of antigenic variation in O. tsutsugamushi diagnosis and determination of new serotypes. PMID:27248711

  15. Staphylococcus massiliensis sp. nov., isolated from a human brain abscess.

    PubMed

    Al Masalma, Mouhamad; Raoult, Didier; Roux, Véronique

    2010-05-01

    Gram-positive, catalase-positive, coagulase-negative, non-motile, non-fermentative and novobiocin-susceptible cocci were isolated from a human brain abscess sample (strain 5402776(T)). This novel strain was analysed by a polyphasic taxonomic approach. The respiratory quinones detected were MK-7 (93 %) and MK-6 (7 %) and the major fatty acids were C(15 : 0) iso (60.5 %), C(17 : 0) iso (8.96 %) C(15 : 0) anteiso (7.93 %) and C(19 : 0) iso (6.78 %). The peptidoglycan type was A3alpha l-Lys-Gly(2-3)-l-Ser-Gly. Based on cellular morphology and biochemical criteria, the new isolate was assigned to the genus Staphylococcus, although it did not correspond to any recognized species. The G+C content of the DNA was 36.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that the new isolate was most closely related to Staphylococcus piscifermentans, Staphylococcus condimenti, Staphylococcus carnosus subsp. carnosus, S. carnosus subsp. utilis and Staphylococcus simulans (97.7 %, 97.6 %, 97.6 %, 97.6 % and 96.5 % sequence similarity, respectively). Comparison of tuf, hsp60, rpoB, dnaJ and sodA gene sequences was also performed. In phylogenetic analysis inferred from tuf, dnaJ and rpoB gene sequence comparisons, strain 5402776(T) clustered with Staphylococcus pettenkoferi (93.7 %, 82.5 % and 89 % sequence similarity, respectively) and on phylogenetic analysis inferred from sodA gene sequence comparisons, it clustered with Staphylococcus chromogenes (82.8 %). On the basis of phenotypic and genotypic data, this isolate represents a novel species for which the name Staphylococcus massiliensis sp. nov. is proposed (type strain 5402776(T)=CCUG 55927(T)=CSUR P23(T)). PMID:19666814

  16. [Isolation and characterization of a human enterovirus 74 isolated in Shandong Province, China].

    PubMed

    Liu, Xiao-Lin; Tao, Ze-Xin; Wang, Hai-Yan; Liu, Yao; Song, Li-Zhi; Ji, Feng; Wang, Tong-Zhan; Lin, Xiao-Juan; Wang, Su-Ting; Xu, Ai-Qiang

    2013-06-01

    Human Enterovirus HEV 74 is a new member of species Human enterovirus B (HEV-B). To understand its evolution and restructuring characteristics, we report the complete genome sequence of a HEV74 strain 05293/SD/CHN/2005(abbreviated as 05293) isolated from an acute flaccid paralysis (AFP) case in Shangdong Province, China, 2005. Analysis of the complete genomic sequence of 05293 showed that its genome was collinear with that of previously described 2 HEV74 strains, except for insertions and deletions at the 5'NTR and the 3 NTR regions. The complete genome sequence of strain 05293 displayed 80. 8% nucleotide and 96% amino acid identity to the prototype strain USA/CA75-10213, and 80. 6% and 95. 9% to another isolated strain Rikaze-136. The P1, P2 and P3 coding regions of strain 05293 displayed 81. 5%, 80. 0%, 79. 7% nucleotide and 95. 9%, 96. 0%, 96.2% amino acid identity to the prototype strain USA/CA75-10213, and 81. 9%, 78. 8%, 79. 5% and 95. 9%, 96. 1%, 95. 7% to strain Rikaze-136, respectively. The phylogenetic tree and Simplot analysis on 05293 and HEV-B genome sequences were performed, and the result indicated frequent recombination within HEV-B.

  17. Human Chlamydia pneumoniae isolates demonstrate ability to recover infectivity following penicillin treatment whereas animal isolates do not.

    PubMed

    Chacko, Anu; Beagley, Kenneth W; Timms, Peter; Huston, Wilhelmina M

    2015-03-01

    Chlamydia pneumoniae strains have recently been demonstrated to have substantially different capacities to enter and recover from IFN-γ-induced persistence, depending on whether they are from human or animal host sources. Here, we examined the ability of two human and two animal strains to enter and be rescued from penicillin-induced persistence. The ability to form inclusions after the addition of penicillin was much reduced in the two animal isolates (koala LPCoLN, bandicoot B21) compared to the two human isolates (respiratory AR39 and heart A03). The penicillin treatment resulted in a dose-dependent loss of infectious progeny for all isolates, with the human strains failing to produce infectious progeny at lower doses of penicillin than the animal strains. The most remarkable finding however was the contrasting ability of the isolates to recover infectious progeny production after rescue by removal of the penicillin (at 72 h) and continued culture. The animal isolates both showed virtually no recovery from the penicillin treatment conditions. In contrast, the human isolates showed a significant ability to recovery infectivity, with the heart isolate (A03) showing the most marked recovery. Combined, these data further support the hypothesis that the ability to establish and recover from persistence appears to be enhanced in human C. pneumoniae strains compared to animal strains.

  18. Human Chlamydia pneumoniae isolates demonstrate ability to recover infectivity following penicillin treatment whereas animal isolates do not.

    PubMed

    Chacko, Anu; Beagley, Kenneth W; Timms, Peter; Huston, Wilhelmina M

    2015-03-01

    Chlamydia pneumoniae strains have recently been demonstrated to have substantially different capacities to enter and recover from IFN-γ-induced persistence, depending on whether they are from human or animal host sources. Here, we examined the ability of two human and two animal strains to enter and be rescued from penicillin-induced persistence. The ability to form inclusions after the addition of penicillin was much reduced in the two animal isolates (koala LPCoLN, bandicoot B21) compared to the two human isolates (respiratory AR39 and heart A03). The penicillin treatment resulted in a dose-dependent loss of infectious progeny for all isolates, with the human strains failing to produce infectious progeny at lower doses of penicillin than the animal strains. The most remarkable finding however was the contrasting ability of the isolates to recover infectious progeny production after rescue by removal of the penicillin (at 72 h) and continued culture. The animal isolates both showed virtually no recovery from the penicillin treatment conditions. In contrast, the human isolates showed a significant ability to recovery infectivity, with the heart isolate (A03) showing the most marked recovery. Combined, these data further support the hypothesis that the ability to establish and recover from persistence appears to be enhanced in human C. pneumoniae strains compared to animal strains. PMID:25663156

  19. Characterization of Listeria monocytogenes isolated from Ganges water, human clinical and milk samples at Varanasi, India.

    PubMed

    Soni, Dharmendra K; Singh, Rakesh K; Singh, Durg V; Dubey, Suresh K

    2013-03-01

    Listeria monocytogenes isolated from Ganges water, human clinical and milk samples were characterized by antibiotic susceptibility, serotype identification, detection of virulence genes and ERIC- and REP-PCR fingerprint analyses. All isolates were uniformly resistant to ampicillin, except two isolates, and showed variable resistance to gentamicin, cotrimoxazole, ofloxacin, rifampicin and tetracycline. Of the 20 isolates found positive for pathogens, seven (four human and three water isolates) belong to serogroups 4b, 4d and 4e; six (one human and five water isolates) belong to serogroups 1/2c and 3c; four milk isolates belong to serogroups 1/2b and 3b; and three milk isolates belong to serogroups 1/2a and 3a. Two water isolates, all human isolates, except one (Pb1) lacking inlJ gene, and three milk isolates possess inlA, inlC, plcA, prfA, actA, hlyA and iap genes. The remaining water and milk isolates showed variable presence of inlJ, plcA, prfA, and iap genes. ERIC- and REP-PCR based analyses collectively indicated that isolates of human clinical samples belong to identical or similar clone and isolates of water and milk samples belong to different clones. Overall study demonstrates the prevalence of pathogenic L. monocytogenes species in the environmental and clinical samples. Most of the isolates were resistant to commonly used antibiotics. PMID:23201044

  20. Shape of the isolated ex-vivo human crystalline lens

    PubMed Central

    Urs, Raksha; Manns, Fabrice; Ho, Arthur; Borja, David; Amelinckx, Adriana; Smith, Jared; Jain, Rakhi; Augusteyn, Robert; Parel, Jean-Marie

    2009-01-01

    Purpose To develop an age-dependent mathematical model of the isolated ex-vivo human crystalline lens shape to serve as basis for use in computational modeling. Methods Profiles of whole isolated human lenses (n=27) aged 6 to 82, were measured from shadow-photogrammetric images. Two methods were used to analyze the lenses. In the Two-Curves Method (TCM) the anterior and posterior surfaces of the lens were fit to 10th-order even polynomials and in the One-Curve Method (OCM) the contour of one half-meridional section of the lens was fit to 10th-order polynomials. The age-dependence of the polynomial coefficients was assessed. The analysis was used to produce an age-dependent polynomial model of the whole lens shape. Results The root mean squared errors for the fits ranged from 11 to 70 μm for the OCM, 9 to 27 μm for the posterior surface of the TCM and 8 to 134 μm for the anterior surface of the TCM. The coefficients of the OCM did not display a significant trend with age. The 2nd, 6th and 10th-order coefficients of the anterior surface of the TCM decreased with age while the 8th-order coefficient increased. For the posterior surface of the TCM, the 8th-order coefficient significantly decreased with age and the 10th-order coefficient increased. The age-dependent equations of both the models provide a reliable model from age 20 to 60. The OCM model can be used for lenses older than 60 as well. Conclusion The shape of the whole human crystalline lens can be accurately modeled with 10th-order polynomial functions. These models can serve to improve computational modeling, such as finite element (FE) modeling of crystalline lenses. PMID:18950656

  1. Characterization of Staphylococcus caprae Clinical Isolates Involved in Human Bone and Joint Infections, Compared with Goat Mastitis Isolates.

    PubMed

    d'Ersu, J; Aubin, G G; Mercier, P; Nicollet, P; Bémer, P; Corvec, S

    2016-01-01

    Staphylococcus caprae is an emerging microorganism in human bone and joint infections (BJI). The aim of this study is to describe the features of S. caprae isolates involved in BJI (H for human) compared with those of isolates recovered in goat mastitis (A for animal). Fourteen isolates of each origin were included. Identifications were performed using a Vitek 2 GP ID card, tuf gene sequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) Vitek MS. Molecular typing was carried out using pulsed-field gel electrophoresis (PFGE) and DiversiLab technology. The crystal violet method was used to determine biofilm-forming ability. Virulence factors were searched by PCR. Vitek MS technology provides an accurate identification for the two types of isolates compared to that of gold-standard sequencing (sensitivity, 96.4%), whereas the Vitek 2 GP ID card was more effective for H isolates. Molecular typing methods revealed two distinct lineages corresponding to the origin despite few overlaps: H and A. In our experimental conditions, no significant difference was observed in biofilm production ability between H and A isolates. Nine isolates (5 H isolates and 4 A isolates) behaved as weak producers while one A isolate was a strong producer. Concerning virulence factors, the autolysin atlC and the serine aspartate adhesin (sdrZ) genes were detected in 24 isolates (86%), whereas the lipase gene was always detected, except in one H isolate (96%). The ica operon was present in 23 isolates (82%). Fibrinogen-binding (fbe) or collagen-binding (cna) genes were not detected by using primers designed for Staphylococcus aureus or Staphylococcus epidermidis, even in low stringency conditions. Although S. caprae probably remains underestimated in human infections, further studies are needed to better understand the evolution and the adaptation of this species to its host.

  2. Characterization of Staphylococcus caprae Clinical Isolates Involved in Human Bone and Joint Infections, Compared with Goat Mastitis Isolates

    PubMed Central

    d'Ersu, J.; Aubin, G. G.; Mercier, P.; Nicollet, P.; Bémer, P.

    2015-01-01

    Staphylococcus caprae is an emerging microorganism in human bone and joint infections (BJI). The aim of this study is to describe the features of S. caprae isolates involved in BJI (H for human) compared with those of isolates recovered in goat mastitis (A for animal). Fourteen isolates of each origin were included. Identifications were performed using a Vitek 2 GP ID card, tuf gene sequencing, and matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) Vitek MS. Molecular typing was carried out using pulsed-field gel electrophoresis (PFGE) and DiversiLab technology. The crystal violet method was used to determine biofilm-forming ability. Virulence factors were searched by PCR. Vitek MS technology provides an accurate identification for the two types of isolates compared to that of gold-standard sequencing (sensitivity, 96.4%), whereas the Vitek 2 GP ID card was more effective for H isolates. Molecular typing methods revealed two distinct lineages corresponding to the origin despite few overlaps: H and A. In our experimental conditions, no significant difference was observed in biofilm production ability between H and A isolates. Nine isolates (5 H isolates and 4 A isolates) behaved as weak producers while one A isolate was a strong producer. Concerning virulence factors, the autolysin atlC and the serine aspartate adhesin (sdrZ) genes were detected in 24 isolates (86%), whereas the lipase gene was always detected, except in one H isolate (96%). The ica operon was present in 23 isolates (82%). Fibrinogen-binding (fbe) or collagen-binding (cna) genes were not detected by using primers designed for Staphylococcus aureus or Staphylococcus epidermidis, even in low stringency conditions. Although S. caprae probably remains underestimated in human infections, further studies are needed to better understand the evolution and the adaptation of this species to its host. PMID:26511738

  3. Characterization of Staphylococcus caprae Clinical Isolates Involved in Human Bone and Joint Infections, Compared with Goat Mastitis Isolates.

    PubMed

    d'Ersu, J; Aubin, G G; Mercier, P; Nicollet, P; Bémer, P; Corvec, S

    2016-01-01

    Staphylococcus caprae is an emerging microorganism in human bone and joint infections (BJI). The aim of this study is to describe the features of S. caprae isolates involved in BJI (H for human) compared with those of isolates recovered in goat mastitis (A for animal). Fourteen isolates of each origin were included. Identifications were performed using a Vitek 2 GP ID card, tuf gene sequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) Vitek MS. Molecular typing was carried out using pulsed-field gel electrophoresis (PFGE) and DiversiLab technology. The crystal violet method was used to determine biofilm-forming ability. Virulence factors were searched by PCR. Vitek MS technology provides an accurate identification for the two types of isolates compared to that of gold-standard sequencing (sensitivity, 96.4%), whereas the Vitek 2 GP ID card was more effective for H isolates. Molecular typing methods revealed two distinct lineages corresponding to the origin despite few overlaps: H and A. In our experimental conditions, no significant difference was observed in biofilm production ability between H and A isolates. Nine isolates (5 H isolates and 4 A isolates) behaved as weak producers while one A isolate was a strong producer. Concerning virulence factors, the autolysin atlC and the serine aspartate adhesin (sdrZ) genes were detected in 24 isolates (86%), whereas the lipase gene was always detected, except in one H isolate (96%). The ica operon was present in 23 isolates (82%). Fibrinogen-binding (fbe) or collagen-binding (cna) genes were not detected by using primers designed for Staphylococcus aureus or Staphylococcus epidermidis, even in low stringency conditions. Although S. caprae probably remains underestimated in human infections, further studies are needed to better understand the evolution and the adaptation of this species to its host. PMID:26511738

  4. Infection of human keratinocytes by Streptococcus dysgalactiae subspecies dysgalactiae isolated from milk of the bovine udder.

    PubMed

    Roma-Rodrigues, Catarina; Alves-Barroco, Cynthia; Raposo, Luís R; Costa, Mafalda N; Fortunato, Elvira; Baptista, Pedro Viana; Fernandes, Alexandra R; Santos-Sanches, Ilda

    2016-04-01

    Streptococcus dysgalactiae subsp. dysgalactiae (SDSD) are considered exclusive animal pathogens; however, a putative zoonotic upper limb cellulitis, a prosthetic joint infection and an infective endocarditis were described in humans. To unravel if bovine SDSD isolates are able to infect human cells, the adherence and internalization to human primary keratinocytes of two bovine SDSD strains isolated from milk collected from udder were analyzed. Bacterial adhesion assays and confocal microscopy indicate a high adherence and internalization of SDSD isolates to human cells, suggesting for the first time the ability of bovine isolates to infect human cells. PMID:26655883

  5. Penile cancer: Perspective from a Canadian tertiary care centre

    PubMed Central

    Beech, Benjamin; Izawa, Jonathan; Pautler, Stephen; Chin, Joseph; Power, Nicholas

    2015-01-01

    Introduction: Penile squamous cell carcinoma (SCC) is rare in North America; however, the morbidity can be devastating. This analysis represents the first reported penile cancer experience at a tertiary care centre in Canada. Methods: We carried out a retrospective review of all patients who received care at our centre for penile SCC from 2005 until the present time. Epidemiological and clinical data were collected for all patients. Survival analysis was performed using Kaplan-Meier methods with log-rank test and Cox regression for univariate and multivariate analysis, respectively. Results: We identified 42 patients who were treated at our centre for penile SCC. Of these, 29% underwent excisional biopsy, 38% had partial penectomy, and 33% had total penectomy. Five patients with high-risk tumours underwent modified inguinal lymph node dissection (ILND), while 7 patients had radical ILND for clinically palpable disease. Overall, the median cancer specific survival (CSS) was undefined, with a 60% survival at 102 months. However CSS was significantly correlated to pT stage, pN stage, and tumour grade. The median follow-up was 25 months (interquartile range: 11–48). Conclusion: These findings confirm the poor CSS of patients with positive lymph nodes in penile SCC. Patients with pN0 after ILND had a durable CSS. Risk factors for penile SCC were confirmed as elevated body mass index, positive smoking history, and lack of circumcision. This first epidemiologic report on penile SCC from a Canadian tertiary care centre should be expanded to other national centres. PMID:26644802

  6. Streptococcus rubneri sp. nov., isolated from the human throat.

    PubMed

    Huch, Melanie; De Bruyne, Katrien; Cleenwerck, Ilse; Bub, Achim; Cho, Gyu-Sung; Watzl, Bernhard; Snauwaert, Isabel; Franz, Charles M A P; Vandamme, Peter

    2013-11-01

    The novel, Gram-stain-positive, ovoid, lactic acid bacterial isolates LMG 27205, LMG 27206, LMG 27207(T) and MRI-F 18 were obtained from throat samples of healthy humans. 16S rRNA gene sequence analyses indicated that these isolates belong to the genus Streptococcus, specifically the Streptococcus mitis group, with Streptococcus australis and Streptococcus mitis as the nearest neighbours (99.45 and 98.56 % 16S rRNA gene sequence similarity to the respective type strains). Genotypic fingerprinting by fluorescent amplified fragment length polymorphism (FAFLP) and pulsed-field gel electrophoresis (PFGE), DNA-DNA hybridizations, comparative sequence analysis of pheS, rpoA and atpA and physiological and biochemical tests revealed that these bacteria formed a taxon well separated from its nearest neighbours and other species of the genus Streptococcus with validly published names and, therefore, represent a novel species, for which the name Streptococcus rubneri sp. nov. is proposed, with LMG 27207(T) ( = DSM 26920(T)) as the type strain.

  7. Isolation and Characterization of Human Lung Lymphatic Endothelial Cells.

    PubMed

    Lorusso, Bruno; Falco, Angela; Madeddu, Denise; Frati, Caterina; Cavalli, Stefano; Graiani, Gallia; Gervasi, Andrea; Rinaldi, Laura; Lagrasta, Costanza; Maselli, Davide; Gnetti, Letizia; Silini, Enrico M; Quaini, Eugenio; Ampollini, Luca; Carbognani, Paolo; Quaini, Federico

    2015-01-01

    Characterization of lymphatic endothelial cells from the respiratory system may be crucial to investigate the role of the lymphatic system in the normal and diseased lung. We describe a simple and inexpensive method to harvest, isolate, and expand lymphatic endothelial cells from the human lung (HL-LECs). Fifty-five samples of healthy lung selected from patients undergoing lobectomy were studied. A two-step purification tool, based on paramagnetic sorting with monoclonal antibodies to CD31 and Podoplanin, was employed to select a pure population of HL-LECs. The purity of HL-LECs was assessed by morphologic criteria, immunocytochemistry, flow cytometry, and functional assays. Interestingly, these cells retain in vitro several receptor tyrosine kinases (RTKs) implicated in cell survival and proliferation. HL-LECs represent a clinically relevant cellular substrate to study lymphatic biology, lymphoangiogenesis, interaction with microbial agents, wound healing, and anticancer therapy. PMID:26137493

  8. Isolation and Characterization of Human Lung Lymphatic Endothelial Cells

    PubMed Central

    Lorusso, Bruno; Falco, Angela; Madeddu, Denise; Frati, Caterina; Cavalli, Stefano; Graiani, Gallia; Gervasi, Andrea; Rinaldi, Laura; Lagrasta, Costanza; Maselli, Davide; Gnetti, Letizia; Silini, Enrico M.; Quaini, Eugenio; Ampollini, Luca; Carbognani, Paolo; Quaini, Federico

    2015-01-01

    Characterization of lymphatic endothelial cells from the respiratory system may be crucial to investigate the role of the lymphatic system in the normal and diseased lung. We describe a simple and inexpensive method to harvest, isolate, and expand lymphatic endothelial cells from the human lung (HL-LECs). Fifty-five samples of healthy lung selected from patients undergoing lobectomy were studied. A two-step purification tool, based on paramagnetic sorting with monoclonal antibodies to CD31 and Podoplanin, was employed to select a pure population of HL-LECs. The purity of HL-LECs was assessed by morphologic criteria, immunocytochemistry, flow cytometry, and functional assays. Interestingly, these cells retain in vitro several receptor tyrosine kinases (RTKs) implicated in cell survival and proliferation. HL-LECs represent a clinically relevant cellular substrate to study lymphatic biology, lymphoangiogenesis, interaction with microbial agents, wound healing, and anticancer therapy. PMID:26137493

  9. Expression of penile neuronal nitric oxide synthase variants in the rat and mouse penile nerves.

    PubMed

    Gonzalez-Cadavid, N F; Burnett, A L; Magee, T R; Zeller, C B; Vernet, D; Smith, N; Gitter, J; Rajfer, J

    2000-09-01

    Penile erection is mediated by nitric oxide (NO) synthesized by the neuronal nitric oxide synthase (nNOS). In the rat penis, the main nNOS mRNA variant, PnNOS, differs from cerebellar nNOS (CnNOS) by a 102 base pair insert encoding a 34-amino acid sequence. In the mouse, two nNOS mRNAs have been identified: nNOSalpha, encoding a 155-kDa protein, and an exon 2-deletion variant, nNOSbeta, encoding a 135-kDa protein that lacks a domain where a protein inhibitor of nNOS (PIN) binds. We wished to determine whether PnNOSalpha and beta are expressed in the rat penis and are located in the nerves and whether the beta form persists in the potent nNOS knock-out mouse (nNOS( big up tri, open big up tri, open)). A PnNOS antibody against the insert common to both PnNOSalpha and beta detected the expected 155-kDa protein in PnNOSalpha-transfected cells. This antibody, and the one common to PnNOS/CnNOS, showed (on Western blots) the 155- and 135-kDa nNOS variants in rat penile tissue during development and aging. PnNOSalpha mRNA and its subvariants were found as the main nNOS in the penile corpora, the cavernosal nerve, and the pelvic ganglia, with lower levels of PnNOSbeta mRNA. In tissue sections, PnNOS protein was immunodetected in the penile nerve endings in the rat and in the nNOS wild-type and nNOS( big up tri, open big up tri, open) mice. An antibody against the sequence encoded by exon 2 did not react (on Western blots) with the 135-kDa band, which confirms that this protein is the beta form. In conclusion, both PnNOSalpha and beta are expressed in the rat penis at all ages and are located in the nerves. The beta form may allow nitric oxide synthesis during erection to be partially insensitive to PIN. The residual expression of PnNOS, and possibly CnNOS, in the penis of the nNOS( big up tri, open big up tri, open) mouse occurs through transcription of the beta mRNA, and this may explain the retention of erectile function when the expression of nNOSalpha is disrupted. PMID

  10. Prevalence of Salmonella serotypes isolated in Spain from human and non human sources (1983-1987).

    PubMed

    Echeita, M A; Usera, M A

    1989-09-01

    Salmonella serotypes over a five year period were studied in order to know their prevalence in Spain. The Salmonella Reference Centre received a total of 17,612 strains from 1983-1987. The majority (16,133) were of human origin and only 1,479 strains were isolated from non-human sources. The serotyping yielded 100 different serotypes, Salmonella enterica serotype Enteritidis (8) being the commonest in both groups, 61.18% of human origin and 31.91% of non-human origin. Salmonella enterica serotype Typhimurium the commonest serotype in many countries, occupies second place in our results with the following percentages 11.87% and 9.67% respectively. Among the strains of human origin Salmonella enterica serotype Typhi occupies fourth place (3.24%). This is very low compared with the high number of clinically diagnosed typhoid fever cases declared in the country: over 5,000 cases per year.

  11. Isolation and characterization of a processed gene for human ceruloplasmin

    SciTech Connect

    Koschinsky, M.L.; Chow, B.K.C.; Schwartz, J.; Hamerton, J.L.; MacGillivray, R.T.A.

    1987-12-01

    A processed pseudogene for human ceruloplasmin has been isolated that contains DNA corresponding to the functional gene sequence encoding the carboxy-terminal 563 amino acid residues and the 3' untranslated region. The pseudogene appears to have arisen from a processed RNA species, since intervening sequences coincident with those of the functional gene have been removed, with the exception of a short segment of intronic sequence which denotes the 5' boundary of the pseudogene. The nucleotide sequence of the pseudogene is highly homologous (97% sequence identity) with that of the wild-type gene, suggesting that pseudogene formation was a relatively recent evolutionary event. In addition to single base substitutions, there is a large 213 base pair (bp) deletion in the pseudogene sequence which corresponds to the location of an intron-exon junction in the functional gene. A 4 bp duplication that occurs at amino acid residue 683 of the wild-type coding sequence results in a frameshift mutation and introduces a premature translational termination codon at this point. This is concordant with the inability to detect a human liver transcript corresponding to the pseudogene by nuclease S1 mapping analysis. The 3' end of the pseudogene is characterized by a 62 bp segment composed mainly of repeated TC dinucleotides. On the basis of genomic Southern blot analysis performed under high-stringency conditions, the pseudogene that the authors have identified seems to comprise the only sequence in the human genome that is closely related to the wild-type gene. Using somatic cell hybridization, they have mapped the pseudogene to human chromosome 8. This differs from the site of the wild-type ceruloplasmin locus, which has been assigned to chromosome 3.

  12. Penile Fracture: Our Experience in a Tertiary Care Hospital

    PubMed Central

    Mahapatra, Rajkumar Singha; Kundu, Anup Kumar

    2015-01-01

    Purpose Penile fracture is rare, but it is a urological emergency that always requires immediate attention. Moreover, penile fracture has been reported more frequently in recent years. It may have devastating physical, functional, and psychological consequences if not properly managed in time. Materials and Methods The objective of this study was to highlight the causes, clinical presentation, and outcomes of cases of penile fracture. This was a prospective observational study extending from November 2012 to November 2014. Each patient underwent a thorough clinical evaluation and received proper treatment. Results Twenty patients with penile fracture, aged 19 to 56 years (mean, 28 years) were evaluated in this study. Vaginal intercourse was the most common mechanism of injury. Most of the patients (95%) were diagnosed clinically with a proper history and clinical examination. Nineteen patients were treated surgically. The patients underwent six months of follow-up, and were evaluated with local examinations, questionnaires, and colour Doppler ultrasonography as necessary. Conclusions Although penile fracture is an under-reported urological emergency, its incidence is increasing. It is usually diagnosed based on a clinical examination, but ultrasonography can be very helpful in diagnosis. Especially in cases where treatment is delayed, surgery is preferable to conservative management, because it is associated with better outcomes and fewer long-term complications. PMID:26331126

  13. Receptors for kinins in the human isolated umbilical vein.

    PubMed

    Gobeil, F; Pheng, L H; Badini, I; Nguyen-Le, X K; Pizard, A; Rizzi, A; Blouin, D; Regoli, D

    1996-05-01

    1. The human umbilical vein has been found to contract in response to bradykinin (BK) and desArg9BK. 2. The rank order of potency of agonists, in the presence of the B1 receptor antagonist Lys[Leu8]desArg9BK, is as follows: [Hyp3, Tyr(Me)8]BK (pD2 8.88) = [Hyp3]BK (pD2 8.86) = LysBK (pD2 8.81) > or = BK (pD2 8.60) > [Aib7]BK (pD2 6.38) > desArg9BK and LysdesArg9BK (inactive). 3. Hoe 140 (pA2 8.42) inhibits the effects of BK while other B2 receptor peptide antagonists are very weak and WIN 64338 is practically inactive. 4. Venoconstrictor responses to desArg9BK of fresh tissues increase with time during the in vitro incubation and reach a maximum after 4-6 h. The activity of Hoe 140 (pA2 5.48) is negligible against B1 receptor agonists. 5. When measured in the presence of the selective B2 receptor antagonist Hoe 140 (400 nM), the order of potency of kinin related peptides on the B1 receptor is Lys[desArg9]BK (pD2 8.60) > desArg9BK (pD2 6.69). BK, LysBK, [Hyp3]BK and other B2 receptor agonists are inactive. 6. The B1 receptor antagonist, Lys[Leu8]desArg9BK (pA2 7.99), inhibits the response of the human vein to B1 receptor agonists (LysdesArg9BK or desArg9BK), but do not alter the effect of BK. 7. The results summarized in this paper indicate that the human isolated umbilical vein is a sensitive preparation containing both B1 and B2 receptors. The human B2 receptor shows some similarity with that of the rabbit (at least for agonist potencies) and differs from the B2 receptor of the guinea-pig. Compared to the rabbit B1 receptor, the human B1 receptor shows low sensitivity to peptides that lack the N-terminal Lys. PMID:8735629

  14. SNP genotyping of animal and human derived isolates of Mycobacterium avium subsp. paratuberculosis.

    PubMed

    Wynne, James W; Beller, Christie; Boyd, Victoria; Francis, Barry; Gwoźdź, Jacek; Carajias, Marios; Heine, Hans G; Wagner, Josef; Kirkwood, Carl D; Michalski, Wojtek P

    2014-08-27

    Mycobacterium avium subsp. paratuberculosis (MAP) is the aetiological agent of Johne's disease (JD), a chronic granulomatous enteritis that affects ruminants worldwide. While the ability of MAP to cause disease in animals is clear, the role of this bacterium in human inflammatory bowel diseases remains unresolved. Previous whole genome sequencing of MAP isolates derived from human and three animal hosts showed that human isolates were genetically similar and showed a close phylogenetic relationship to one bovine isolate. In contrast, other animal derived isolates were more genetically diverse. The present study aimed to investigate the frequency of this human strain across 52 wild-type MAP isolates, collected predominantly from Australia. A Luminex based SNP genotyping approach was utilised to genotype SNPs that had previously been shown to be specific to the human, bovine or ovine isolate types. Fourteen SNPs were initially evaluated across a reference panel of isolates with known genotypes. A subset of seven SNPs was chosen for analysis within the wild-type collection. Of the seven SNPs, three were found to be unique to paediatric human isolates. No wild-type isolates contain these SNP alleles. Interestingly, and in contrast to the paediatric isolates, three additional adult human isolates (derived from adult Crohn's disease patients) also did not contain these SNP alleles. Furthermore we identified two SNPs, which demonstrate extensive polymorphism within the animal-derived MAP isolates. One of which appears unique to ovine and a single camel isolate. From this study we suggest the existence of genetic heterogeneity between human derived MAP isolates, some of which are highly similar to those derived from bovine hosts, but others of which are more divergent.

  15. Optical power of the isolated human crystalline lens

    PubMed Central

    Borja, David; Manns, Fabrice; Ho, Arthur; Ziebarth, Noel; Rosen, Alexandre M.; Jain, Rakhi; Amelinckx, Adriana; Arrieta, Esdras; Augusteyn, Robert C.; Parel, Jean-Marie

    2009-01-01

    Purpose To characterize the age-dependence of isolated human crystalline lens power and quantify the contributions of the lens surfaces and refractive index gradient. Methods Experiments were performed on a total of 100 eyes from 73 donors (average post-mortem: 2.8±1.6 days) with an age range of 6 to 94 years. Lens power was measured with a modified commercial lensmeter or with an optical system based on the Scheiner principle. The radius of curvature and asphericity of the isolated lens surfaces were measured by shadowphotography. For each lens the contributions of the surfaces and the refractive index gradient to the measured lens power were calculated using an optical ray tracing software. The age dependency of these refractive powers was assessed. Results The total refractive power and surface refractive power both showed a biphasic age dependency. The total power decreased at a rate of −0.41D/year between age 6 and 58.1, and increased at a rate of 0.33D/year between age 58.1 and 82. The surface contribution decreased at a rate of −0.13D/year between age 6 and 55.2, and increased at a rate of 0.04D/year between age 55.2 and 94. The relative contribution of the surfaces increased by 0.17%/year. The equivalent refractive index also showed a biphasic age dependency with a decrease at a rate of −3.9×10−4/year from age 6 to 60.4 followed by a plateau. Conclusions The lens power decreases with age due mainly to a decrease in the contribution of the gradient. The use of a constant equivalent refractive index value to calculate lens power using the lens maker formula will underestimate the power of young lenses and overestimate the power of older lenses. PMID:18316704

  16. Identification of Putative Natriuretic Hormones Isolated from Human Urine.

    PubMed

    Kramer, Herbert J

    2015-01-01

    This brief review describes some representative methodological approaches to the isolation of putative endogenous inhibitors of epithelial sodium transport - i.e., as ouabain-like factors (OLF) that inhibit the sodium transport enzyme Na-K-ATPase or inhibit the epithelial sodium channel (ENaC). Gel chromatography and reverse-phase (RP)-high performance liquid chromatography (HPLC) of lyophilized and reconstituted 24 h-urine from salt-loaded healthy humans led to two active fractions, a hydrophilic OLF-1 and a lipophilic OLF-2, whose mass (Ms)-spectroscopic data indicate a Mr of 391 (1, 2). Further identification was attempted by Ms-, infrared (IR)-, ultraviolet (UV)-, and (1)H-NMR-spectroscopy. OLF-1 and OLF-2 may be closely related if not identical to (di)ascorbic acid or its salts such as vanadium (V)-V(v)-diascorbate with Mr 403 (3) and V(IV)-diascorbate. OLF-1 and V(v)-diascorbate are about 10-fold stronger inhibitors of Na-K-ATPase than OLF-2 and V(IV)-diascorbate, respectively. In conscious rats, i.v. infusion of OLF-1 and OLF-2 resulted in a strong natriuresis. In a similar study, Cain et al. (4) isolated a sodium transport inhibitor from the urine of uremic patients by gel chromatography and RP-HPLC. In uremic rats, a natriuretic response to the injection of the active material was found. Xanthurenic acid 8-O-β-d-glucoside (Mr 368) and xanthurenic acid 8-O-sulfate (Mr 284) were identified as endogenous inhibitors of sodium transport acting, e.g., by ENaC blockade. No definite relation to blood pressure, body fluid volume, or sodium balance has been reported for any of these above factors, and further studies to identify the natriuretic and/or ouabain-like compound(s) or hormone(s) will be needed. PMID:26052310

  17. [Deep dorsal penile vein thrombosis revealing Behcet's disease].

    PubMed

    Beddouche, Ali; Ouaziz, Hicham; Zougaghi, Sinane; Alaoui, Abdelilah; Dergamoun, Hamza; El Sayegh, Hachem; Iken, Ali; Benslimane, Lounis; Nouini, Yassine

    2016-01-01

    Deep dorsal penile vein thrombosis (DDPVT)is a rare and little known urologic emergency. It requires an early etiological and symptomatic approach to preserve erectile function and prevent recurrences. This study reports a case of dorsal penile vein thrombosis revealed by spontaneous priapism that didn't resolve adequately and confirmed by penile Doppler ultrasound. After management of priapism and DDPVT, the etiological investigation revealed Behcet's disease whose diagnosis was based on the association of a major criteria, such as oral aphthous ulcers with 3 minor criteria such as: genital aphthous ulcers, ocular involvement, and a positive skin pathergy test within 24h. The patient underwent etiological treatment with good clinical evolution and preservation of erectile function. PMID:27583081

  18. Radiation Dose-Volume Effects and the Penile Bulb

    SciTech Connect

    Roach, Mack; Nam, Jiho; Gagliardi, Giovanna; El Naqa, Issam; Deasy, Joseph O.; Marks, Lawrence B.

    2010-03-01

    The dose, volume, and clinical outcome data for penile bulb are reviewed for patients treated with external-beam radiotherapy. Most, but not all, studies find an association between impotence and dosimetric parameters (e.g., threshold doses) and clinical factors (e.g., age, comorbid diseases). According to the data available, it is prudent to keep the mean dose to 95% of the penile bulb volume to <50 Gy. It may also be prudent to limit the D70 and D90 to 70 Gy and 50 Gy, respectively, but coverage of the planning target volume should not be compromised. It is acknowledged that the penile bulb may not be the critical component of the erectile apparatus, but it seems to be a surrogate for yet to be determined structure(s) critical for erectile function for at least some techniques.

  19. Penile strangulation due to plastic bottle neck: a surgical emergency.

    PubMed

    Agarwal, Akshay Anand; Singh, Kul Ranjan; Kushwaha, Jitendra Kumar; Sonkar, Abhinav Arun

    2014-01-01

    Use of various metallic and non-metallic constricting objects on the external male genitalia for increasing sexual performance or because of autoerotic intentions is an unusual practice that can potentially lead to penile strangulation with severe consequences. Depending on the type of constricting material, emergency removal of such an object is a challenge. We report a case of a 45-year-old man who presented to our hospital with a hard plastic bottle neck at the base of his penis that led to penile strangulation. The constricting agent was successfully removed. The patient had an uneventful recovery. PMID:25427935

  20. Penile strangulation due to plastic bottle neck: a surgical emergency.

    PubMed

    Agarwal, Akshay Anand; Singh, Kul Ranjan; Kushwaha, Jitendra Kumar; Sonkar, Abhinav Arun

    2014-11-26

    Use of various metallic and non-metallic constricting objects on the external male genitalia for increasing sexual performance or because of autoerotic intentions is an unusual practice that can potentially lead to penile strangulation with severe consequences. Depending on the type of constricting material, emergency removal of such an object is a challenge. We report a case of a 45-year-old man who presented to our hospital with a hard plastic bottle neck at the base of his penis that led to penile strangulation. The constricting agent was successfully removed. The patient had an uneventful recovery.

  1. Penile cutaneous horn: An enigma-newer insights and perspectives.

    PubMed

    Karthikeyan, Kaliaperumal

    2015-01-01

    Cutaneous horn refers to unusually cohesive keratinized material and not a true pathologic diagnosis. Though cutaneous horn has been described at various sites, horn over the penis is very rare and represents the most unusual site. The role of chronic irritation, phimosis, surgical trauma and radiotherapy have been implicated in penile horn formation. Penile horns present as elongated, keratinous, white or yellowish projections that range from a few millimeters to centimeters in size arising from the glans penis. Histopathology of the keratotic mass reveals nothing but keratin. The underlying mass may vary from verruca vulgaris to squamous cell carcinoma. The treatment is based on the pathology. PMID:26392650

  2. Silicone-induced Penile Sclerosing Lipogranuloma: Magnetic Resonance Imaging Findings.

    PubMed

    Tsili, Athina C; Xiropotamou, Olga N; Nomikos, Michael; Argyropoulou, Maria I

    2016-01-01

    Sclerosing lipogranuloma is a rare benign disease, representing a peculiar granulomatous reaction of fatty tissue. The majority of cases are secondary to injection of exogenous foreign bodies, such as silicone, paraffin, mineral, or vegetable oils. To the best of our knowledge, we present the first case of a silicone-induced penile lipogranuloma in a 52-year-old man evaluated with a multiparametric magnetic resonance imaging (MRI) protocol, including diffusion-weighted imaging, magnetization transfer imaging, and dynamic contrast-enhanced MRI. MRI of the penis by combining both conventional and functional information represents an important imaging tool in the preoperative workup of silicone-induced penile lipogranuloma. PMID:26958433

  3. Possible function of the frenulum of prepuce in penile erection.

    PubMed

    Song, B; Cai, Z-M

    2012-02-01

    Fremulum of prepuce was the ruffle of foreskin while there was little about the function of fremulum. This study discusses the possible function of the frenulum of prepuce in penile erection. Twelve patients had premature ejaculation (PE) whose frenula were short. Two patients suffered unsatisfied intercourse whose frenula were damaged and departed 12 or 6 months earlier. We prolonged the short frenulum and reconstructed the ruptured frenulum. All patients reported satisfied sexual intercourse after 3-6 months. It is concluded that the frenulum is important in penile erection. PE might be treated by lengthening the frenulum.

  4. Prevotella colorans sp. nov., isolated from a human wound.

    PubMed

    Buhl, Michael; Willmann, Matthias; Liese, Jan; Autenrieth, Ingo B; Marschal, Matthias

    2016-08-01

    A strain of obligately anaerobic, Gram-stain-negative and non-spore-forming rod-shaped bacterium was isolated from a human wound and characterized both phenotypically and genotypically. The strain was moderately saccharolytic and proteolytic. Phylogenetic analysis was based on full-length 16S rRNA gene sequence analysis and revealed the strain to represent a member of the genus Prevotella, but to be different from the described species, with the closest relationship to Prevotella bergensis and Prevotella multisaccharivorax. The genomic DNA G+C content was 43.2 mol%. The most abundant cellular long-chain fatty acids were 3-OH iso-C17 : 0, anteiso-C15 : 0 and iso-C15 : 0. In view of phenotypical and biochemical characteristics as well as gene sequencing, strain A1336T is considered to represent a novel species within the genus Prevotella, for which the name Prevotella colorans sp. nov. is proposed. The type strain is A1336T (=DSM 100333T =CCUG 67421T =CCOS 902T). PMID:27150727

  5. Isolation and characterization of migratory human skin dendritic cells.

    PubMed Central

    Richters, C D; Hoekstra, M J; van Baare, J; Du Pont, J S; Hoefsmit, E C; Kamperdijk, E W

    1994-01-01

    A method is described to isolate and characterize human skin dendritic cells (DC). This method is based on the migratory capacities of these cells. The cells migrated 'spontaneously' out of split-skin explants into the medium during a 24-h culture period and contained up to 75% CD1a+ cells. After removal of co-migrated T cells and macrophages, the highly enriched (> 95% CD1a+) DC showed potent allo-antigen-presenting capacities. About 25% of the CD1a+ cells were also positive for the dermal DC marker CD1b, whereas only 15-20% of the cells contained Birbeck granules, the characteristic cell organelle of the epidermal Langerhans cell. Before culture, CD1a+ DC were observed on cryostat sections not only in the epidermis but also in the dermis. After culture, the number of CD1a+ cells in both epidermis and dermis had decreased. Not all the cells had migrated during the culture period; some CD1a+ cells could still be detected in the epidermis and dermis after culture. Thus, using this method, potent allo-stimulating CD1a+ cells, migrating from both epidermis and dermis, can be obtained without the use of enzymes. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:7955541

  6. The impact of diabetes mellitus on penile length in men undergoing inflatable penile prosthesis implantation

    PubMed Central

    Akın, Yiğit; Şahiner, İlker Fatih; Usta, Mustafa Faruk

    2013-01-01

    Objective: To evaluate the changing cavernosal length of patients with diabetes mellitus (DM) and organic erectile dysfunction (ED) who were treated with inflatable, three-piece penile prostheses, a current surgical treatment option in our clinic, over the course of 12 years. Materials and methods: Between April 2000 and December 2012, we retrospectively investigated data from patients who were diagnosed with organic ED and undergone penile prosthesis implantation (PPI). Of the 239 patients, 235 of them were included in the study. Four patients who were operated on for trans-sexuality were excluded from the study. All patients were divided into two groups as those with (Group 1) or without DM (Group 2). Data, including age, body mass index (BMI) in kg/m2, surgical history, comorbidities, International Index of Erectile Function (IIEF) questionnaire scores, combined intracavernous injection and stimulation (CIS) test results, length of corpus cavernosum while implanting the penile prosthesis, complications, operative times, mean hospital stay, and satisfaction of the patient and partner, were recorded. Kruskal-Wallis and Mann-Whitney U tests were used for statistical analysis. A p-value of <0.05 was considered to be statistically signifcant. Results: The mean age was 57.9±10.5 years. Study population consisted of patients with DM (n=65), hypertension (n=21), DM, and hypertension (n=28), hyperlipidemia (n=5), a history of previous radical pelvic surgery with (n=4) or without DM (n=51) or cases without any comorbidity (n=62). Mean length of the corpus cavernosum was 17.277±0.1509 cm in Group 1 and 17289±0.1598 cm in Group 2 (p<0.05). Additionally, the other parameters, including age, operative time, and the satisfaction of the patient and partner, were not different between these groups (p>0.05). Conclusion: The length of the corpus cavernosum and the destruction of cavernosal tissues do not depend only on DM. We conclude that these features may have multifactorial

  7. Identification of a novel herpesvirus associated with a penile proliferative lesion in a beluga (Delphinapterus leucas).

    PubMed

    Bellehumeur, Christian; Lair, Stéphane; Romero, Carlos H; Provost, Chantale; Nielsen, Ole; Gagnon, Carl A

    2015-01-01

    The carcass of an adult male beluga (Delphinapterus leucas) was found beach cast in 2008 on the shore of the St. Lawrence Estuary at Rivière-Ouelle, Quebec, Canada. The carcass was transported to the Faculté de médecine vétérinaire of the Université de Montréal for postmortem examination. Aspiration pneumonia was the probable cause of death. Necropsy revealed a focal papilloma-like penile lesion, characterized by focal mucosal thickening with disorganization of the epithelial layers and lymphoplasmacytic infiltration. A pan-herpesvirus nested PCR assay on frozen tissue from the penile lesion was positive. The PCR product sequencing revealed a partial herpesvirus DNA polymerase (DPOL) gene sequence of 600 nucleotides. Its nearest nucleotide identity was with the partial DPOL gene of an alphaherpesvirus, bovine herpesvirus 5 (79.5% identity). It also shared high identity with several other marine mammal herpesviruses (50.2 to 77.3% identity). This new herpesvirus was tentatively named beluga whale herpesvirus (BWHV). Virus isolation was unsuccessful. The pathogenic potential of BWHV is unknown, but the evaluation of archived tissues suggests that the virus is endemic in the St. Lawrence Estuary beluga population.

  8. Identification of a novel herpesvirus associated with a penile proliferative lesion in a beluga (Delphinapterus leucas).

    PubMed

    Bellehumeur, Christian; Lair, Stéphane; Romero, Carlos H; Provost, Chantale; Nielsen, Ole; Gagnon, Carl A

    2015-01-01

    The carcass of an adult male beluga (Delphinapterus leucas) was found beach cast in 2008 on the shore of the St. Lawrence Estuary at Rivière-Ouelle, Quebec, Canada. The carcass was transported to the Faculté de médecine vétérinaire of the Université de Montréal for postmortem examination. Aspiration pneumonia was the probable cause of death. Necropsy revealed a focal papilloma-like penile lesion, characterized by focal mucosal thickening with disorganization of the epithelial layers and lymphoplasmacytic infiltration. A pan-herpesvirus nested PCR assay on frozen tissue from the penile lesion was positive. The PCR product sequencing revealed a partial herpesvirus DNA polymerase (DPOL) gene sequence of 600 nucleotides. Its nearest nucleotide identity was with the partial DPOL gene of an alphaherpesvirus, bovine herpesvirus 5 (79.5% identity). It also shared high identity with several other marine mammal herpesviruses (50.2 to 77.3% identity). This new herpesvirus was tentatively named beluga whale herpesvirus (BWHV). Virus isolation was unsuccessful. The pathogenic potential of BWHV is unknown, but the evaluation of archived tissues suggests that the virus is endemic in the St. Lawrence Estuary beluga population. PMID:25375944

  9. Benign mast cell hyperplasia and atypical mast cell infiltrates in penile lichen planus in adult men.

    PubMed

    Regauer, Sigrid; Beham-Schmid, Christine

    2014-08-01

    Introduction. Lichen planus (LP) is a chronic cytokine-mediated disease of possible auto-immune etiology. 25% of men have anogenital manifestations. Erosive penile LP causes a scarring phimosis of the foreskin in uncircumcised men. Mast cells as potent immune modulators have been implicated in a number of autoimmune and chronic inflammatory diseases, but have not been investigated in LP. Material and Methods. Formalin-fixed tissues of 117 circumcision specimens of adult men affected by LP were evaluated for the extent of mast cell and lymphocyte infiltrates, characterized immunohistochemically with antibodies to CD 3, 4, 8, 20, 21, 25, 30, 117c and human mast cell tryptase. Specimens with dense mast cell infiltrates were analyzed for point mutations of the c-kit gene (D816V). Results. Unaffected skin and modified mucosa of foreskins contained ⟨5 mast cells/mm². The inflammatory infiltrate of LP-lesions displayed ⟨15 mast cells/mm² in 33/117 foreskins, 16-40 mast cells/mm² in 22/117 and ⟩40 mast cells/mm² (average 70, range 40-100) in 62/117 foreskins. Lesional mast cells of 29/117 (24%) foreskins showed aberrant CD25-expression and/or spindled morphology, with 11/29 men having erosive LP, 13/29 a lymphocytic vasculitis and 1/28 a systemic mastocytosis. Neither CD30-expression nor c-kit mutations were identified. Atypical mast cell infiltrates in LP correlated with high disease activity, erosive LP and presence of lymphocytic vasculitis Conclusions. Increased mast cells in penile LP, mostly representing a benign hyperplasia/activation syndrome, suggests them as targets for innovative therapy options for symptomatic LP-patients not responding to corticosteroid therapy. Presently, the biological implications of atypical mast cell infiltrates in penile LP are unknown. PMID:24402730

  10. Penile oxygen saturation in the flaccid and erect penis in men with and without erectile dysfunction.

    PubMed

    Padmanabhan, Priya; McCullough, Andrew R

    2007-01-01

    It is believed that a chronic state of corporal oxygen desaturation or hypoxemia secondary to the loss of nocturnal erections is a fundamental pathophysiological cause of erectile dysfunction (ED). Limited invasive blood gas measurements in human models have shown decreased oxygen tension in vasculogenic impotence. Normative data on flaccid and erect oxygen saturation (StO(2)) levels are lacking due to the invasive nature of blood gas determinations. Our objective was to determine StO(2) in the flaccid and erect penis in men with and without ED using a tissue oximeter. This FDA-approved instrument provides instantaneous, noninvasive, painless local tissue StO(2) measurements, which highly correlate to blood gas data. The study population included 171 men (18-90 years) who presented to one andrologist. They completed the Sexual Health Inventory for Men (SHIM) based on pharmacologically unassisted erectile function and had penile StO(2) measurements taken. 64 of these men had repeat measurements after PGE-1 induced erections. There are significant differences (P<.001) in corporal and glanular StO(2) in the flaccid (right corpora, 45.23%; left corpora, 52.50%) and erect state (right corpora, 76.58; left corpora, 80.42). Men with ED (right corpora, 45.04% vs 53.58%; P=.02; and left corpora, 50.95% vs 58.78%; P=.03) have significantly lower corporal penile StO(2). Future prospective data collection can correlate penile StO(2) in specific populations, such as diabetics and RRP patients. This may help further elucidate the relationship between corporal hypoxia and the development and progression of ED and possibly its treatment and prevention.

  11. AB023. Penile rehabilitation with phosphodiesterase type 5 inhibitors in men after nerve-sparing radical prostatectomy

    PubMed Central

    Jiann, Bang-Ping

    2016-01-01

    Post-radical prostatectomy (RP) erectile dysfunction (ED) remains a challenge for the urologist. Despite the improvements in surgical technique, ED occurs between 20% and 90% in patients treated with bilateral nerve-sparing RP. Patient factors, cancer selection, type of surgery, surgical techniques, and surgeon factors represent the key significant contributors to erectile function recovery. The aim of a penile rehabilitation program is to preserve the functional smooth-muscle content of the corpus cavernosum during the neuropraxia period. Phosphodiesterase type 5 (PDE5) inhibitors are commonly used in rehabilitation programs. In animal models, such an approach could promote erectile function recovery, improve smooth muscle-to-collagen penile ration, reduce penile apoptotic index, preserve penile endothelial function and promote neuroprotection during nerve damage. Despite the strong basic science support from animal studies, discordant results have been reached in humans. The previous randomized trials comparing chronic versus on-demand PDE-5 inhibitors use after RP may be affected by improper patients’ selection in that only men at low risk of postoperative ED were included. These patients would recover erectile function regardless of the type of PDE5 inhibitor administration because of their excellent baseline profile. Prospective, randomized trials have shown a significant benefit of daily PDE5-I administration as compared with placebo in terms of postoperative EF recovery. Patients with intermediate risk of ED after surgery are the best candidates for daily treatment with PDE5 inhibitor after bilateral nerve-sparing RP. The maximal effect of penile rehabilitation may be found in those men with a certain (but not high) degree of systemic and erectile impairment preoperatively. In conclusion, penile rehabilitation could achieve faster and better natural erectile function after RP and should be started as early as possible. Chronic use of PDE5-I may confer the

  12. Comparison of virulence factors and capsular types of Streptococcus agalactiae isolated from human and bovine infections.

    PubMed

    Emaneini, Mohammad; Khoramian, Babak; Jabalameli, Fereshteh; Abani, Samira; Dabiri, Hossein; Beigverdi, Reza

    2016-02-01

    Streptococcus agalactiae is a leading cause of human and bovine infections. A total of 194 S. agalactiae isolates, 55 isolates from bovines and 139 from humans, were analyzed for capsular types, virulence genes (scpB, hly, rib, bca and bac) and mobile genetic elements (IS1548 and GBSi1) using polymerase chain reaction (PCR) and multiplex PCR. Capsular type III was predominant (61%), followed by types V, II, Ib, and IV. The scpB, hly, bca and bac virulence genes were only found among human isolates. Twelve and 2 distinct virulence gene profiles were identified among human and bovine isolates respectively. The virulence gene profiles scpB- hly- IS1548- rib-bca (51%) and scpB- hly- IS1548- bca (19%) were only predominant among human isolates. The rib gene was the most common virulence gene in both human and bovine isolates. The study showed a high prevalence of virulence genes in S. agalactiae strains isolated from human infections, these result can support the idea that S. agalactiae isolated from humans and bovines are generally unrelated and probably belonged to separate populations.

  13. Complete Genome Sequence of Lactobacillus fermentum CECT 5716, a Probiotic Strain Isolated from Human Milk▿

    PubMed Central

    Jiménez, Esther; Langa, Susana; Martín, Virginia; Arroyo, Rebeca; Martín, Rocío; Fernández, Leónides; Rodríguez, Juan M.

    2010-01-01

    Lactobacillus fermentum is a heterofermentative lactic acid bacterium and is frequently isolated from mucosal surfaces of healthy humans. Lactobacillus fermentum CECT 5716 is a well-characterized probiotic strain isolated from human milk and, at present, is used in commercial infant formulas. Here, we report the complete and annotated genome sequence of this strain. PMID:20639335

  14. Description of Campylobacter fetus subsp. testudinum subsp. nov., isolated from humans and reptiles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A polyphasic study was undertaken to determine the taxonomic position of 13 Campylobacter fetus-like isolates from humans (n=8) and reptiles (n=5). Phenotypic characterization, Genusgenus-specific and sap insertion-PCR initially identified all human isolates as type A Campylobacter fetus. Phylogenet...

  15. Genome Sequence of Parascardovia denticolens IPLA 20019, Isolated from Human Breast Milk

    PubMed Central

    Gueimonde, Miguel; Bottacini, Francesca; van Sinderen, Douwe; Ventura, Marco; Margolles, Abelardo

    2012-01-01

    This work describes the draft genome of Parascardovia denticolens IPLA 20019, isolated from human milk. This species, usually isolated from caries lesions, is taxonomically related to the genus Bifidobacterium. The genetic information of IPLA 20019 enhances our understanding of the adaptation of this P. denticolens strain from human breast milk. PMID:22887674

  16. Detection of CC17 Enterococcus faecium in dogs and a comparison with human isolates.

    PubMed

    Kwon, K H; Moon, B Y; Hwang, S Y; Park, Y H

    2012-09-01

    Enterococcus faecium strains of clonal complex (CC) 17 were isolated from domestic dogs. The strains were more prevalent in infectious isolates than in colonized isolates, suggesting that strains of the CC17 lineage may have an advantage in causing infections in dogs. The pulsed field gel electrophoresis patterns of some dog and human isolates were over 90% similar. However, antimicrobial resistance patterns and virulence factors were not identical, which might reflect different use of antimicrobials in veterinary medicine or in host specificity.

  17. Characterization of Methicillin Resistant Staphylococcus aureus isolated from human and animal samples in Egypt.

    PubMed

    Bendary, M M; Solyman, S M; Azab, M M; Mahmoud, N F; Hanora, A M

    2016-02-29

    Staphylococcus aureus (S. aureus) has been one of the most problematic pathogens. Methicillin Resistant S. aureus (MRSA) has emerged as a major concern for both human and animal. Antibiotic resistance genes dissemination might be possible between human and animal bacteria. The aim of this study is to show phenotypic and genotypic diversity of human and animal MRSA isolates. Antibiogram typing and biofilm production were used as a primary phenotypic typing tool for the characterization of (40) animal and (38) human MRSA isolates. Genetic typing based on sequencing of 16S rRNA gene and virulence gene profiles were done. Antimicrobial resistance profiles of the animal isolates showed little evidence of widespread of resistance, although this was seen in many human isolates. The biofilm production was detected in higher percentage among animal isolates. Based on the genetic typing and multiple antibiotic resistance (MAR) index, the majority of animal isolates clustered into lineages that were not found in human isolates. Animal and human MRSA isolates showed diversity in antibiotic resistance and virulence gene profiles may be due to host adaptation or chances for contamination between the two hosts were not present in our study.

  18. Balanoposthitis and Penile Edema: Atypical Manifestations of Primary Syphilis.

    PubMed

    Rovira-López, Roger; García-Martínez, Pablo; Martín-Ezquerra, Gemma; Pujol, Ramon M; Aventín, Daniel López

    2015-09-01

    The typical finding in primary syphilis stage is a unique, painless chancre with indurated borders. We report a case of primary syphilis presenting as erosive and crusted balanoposthitis with an underlying chancre, penile edema, and bilateral inguinal lymphadenopathy in a heterosexual man.

  19. Supra and infralevator neurovascular pathways to the penile corpora cavernosa

    PubMed Central

    BENOIT, G.; DROUPY, S.; QUILLARD, J.; PARADIS, V.; GIULIANO, F.

    1999-01-01

    The aim of this study was to provide a comprehensive description of both penile innervation and vascularisation. Eighty-five male cadavers were examined through gross and microscopic anatomical analysis. The pelvic nerve plexus had both parasympathetic and sympathetic roots. It was distributed to the external urethral sphincter giving rise to cavernous nerves which anastomosed in 70% of the cases with the pudendal nerve in the penile root. Accessory pudendal arteries were present in the pelvis in 70% of the cases, anastomosing in 70% of the cases with the cavernous arteries that originated from the pudendal arteries. Transalbugineal anastomoses were always seen between the cavernous artery and the spongiosal arterial network. There were 2 venous pathways, 1 in the pelvis and 1 in the perineum with a common origin from the deep dorsal penile vein. It is concluded that there are 2 neurovascular pathways destined for the penis that are topographically distinct. One is located in the pelvis and the other in the perineum. We were unable to determine the functional balance between these 2 anastomosing pathways but experimental data have shown that they are both involved in penile erection. These 2 neurovascular pathways, above and below the levator ani, together with their anastomoses, form a neurovascular loop around the levator ani. PMID:10634698

  20. Bacterial biofilm in persistent penile prosthesis-associated infection.

    PubMed

    Nickel, J C; Heaton, J; Morales, A; Costerton, J W

    1986-03-01

    The ultrastructural microbiology of 2 cases of infection associated with rigid penile prostheses was studied. The persistence of these infections appeared to be related to the mode of growth of the bacteria in protected biofilms adherent to the inert surface of the prosthesis.

  1. Molecular typing of Brucella species isolates from livestock and human.

    PubMed

    Nagalingam, Mohandoss; Shome, Rajeswari; Balamurugan, Vinayagamurthy; Shome, Bibek Ranjan; NarayanaRao, Krishnamsetty; Vivekananda; Isloor, Shrikrishna; Prabhudas, Krishnamsetty

    2012-01-01

    Although host specificity has been observed in different species of Brucella, crossing the animal host boundary is likely to occur at any time. In this study, Bruce ladder PCR and abortus-melitensis-ovis-suis (AMOS) PCR assays were used to characterize 47 Brucella isolates from Indian origin in order to know exact species for understanding epidemiology of brucellosis. Out of them, 28, 14, and 5 isolates were found to be Brucella abortus, Brucella melitensis, and Brucella suis, respectively. Further analysis by AMOS PCR has identified that all the B. abortus isolates belong to any one of the biovar 1, 2, or 4; of the five B. suis isolates, three belong to biovar 1 and two belong to any one of the biovar 2, 3, 4, or 5. Although this multiplex Bruce ladder PCR is useful in differentiating all Brucella species, elaborate study is required to further characterize the isolates at exact biovar level.

  2. Antimicrobial susceptibility of animal and human isolates of Clostridium difficile by broth microdilution.

    PubMed

    Pirš, Tina; Avberšek, Jana; Zdovc, Irena; Krt, Brane; Andlovic, Alenka; Lejko-Zupanc, Tatjana; Rupnik, Maja; Ocepek, Matjaž

    2013-09-01

    A total of 188 human (n = 92) and animal (n = 96) isolates of Clostridium difficile of different PCR ribotypes were screened for susceptibility to 30 antimicrobials using broth microdilution. When comparing the prevalence of antimicrobial resistance, the isolates of animal origin were significantly more often resistant to oxacillin, gentamicin and trimethoprim/sulfamethoxazole (P<0.01). The most significant difference between the animal and human populations (P = 0.0006) was found in the level of imipenem resistance, with a prevalence of 53.3 % in isolates of human origin and 28.1 % in isolates of animal origin. Overall, the results show similar MICs for the majority of tested antimicrobials for isolates from human and animal sources, which were collected from the same geographical region and in the same time interval. This supports the hypothesis that C. difficile could be transmissible between human and animal hosts. Resistant isolates have been found in all animal species tested, including food and companion animals, and also among non-toxigenic isolates. The isolates of the most prevalent PCR ribotype 014/020 had low resistance rates for moxifloxacin, erythromycin, rifampicin and daptomycin, but a high resistance rate for imipenem. Multiresistant strains were found in animals and humans, belonging to PCR ribotypes 012, 017, 027, 045, 046, 078 and 150, and also to non-toxigenic strains of PCR ribotypes 010 and SLO 080.

  3. Yersinia enterocolitica: serotypes and biotypes isolated from humans and the environment in Quebec, Canada.

    PubMed Central

    Caprioli, T; Drapeau, A J; Kasatiya, S

    1978-01-01

    Thirty-one strains of Yersinia enterocolitica were isolated from food and surface water. During the period of January 1975 to June 1977, 157 strains from 143 human cases were also isolated. Among the different serotypes from nonhuman sources, serotypes 6,30 and 4,32 were the most common. Serotype 4,32 was present only in food and not in water. Serotype 3 was isolated only from humans. Among the different serotypes from human cases, 73.9% belonged to serotype 3. Only serotype 3 was isolated from children under 4 years old. The presence of other serotypes increased and that of serotype 3 decreased in frequency as the age progressed. No serotype 3 was isolated from human cases aged 50 years and more. PMID:670388

  4. The seasonality of human campylobacter infection and Campylobacter isolates from fresh, retail chicken in Wales.

    PubMed Central

    Meldrum, R. J.; Griffiths, J. K.; Smith, R. M. M.; Evans, M. R.

    2005-01-01

    Seasonal peaks in both human campylobacter infections and poultry isolates have been observed in several European countries but remain unexplained. We compared weekly data on human campylobacter infections with thermophilic Campylobacter isolation rates from fresh, retail chicken samples (n = 514) purchased weekly in Wales between January and December 2002. Human isolates (n = 2631) peaked between weeks 22 and 25 (early June) and chicken isolates (n = 364) between weeks 24 and 26 (late June). In the absence of a temporal association, we postulate that the seasonal rise in humans is not caused by a rise in isolation rates in poultry but that both are more likely to be associated with a common, but as yet unidentified, environmental source. PMID:15724710

  5. Penile rehabilitation after radical prostatectomy: does it work?

    PubMed Central

    Gandaglia, Giorgio; Suardi, Nazareno; Cucchiara, Vito; Bianchi, Marco; Shariat, Shahrokh F.; Roupret, Morgan; Salonia, Andrea; Montorsi, Francesco

    2015-01-01

    Context Erectile dysfunction (ED) represents one of the most common long-term side effects in patients with clinically localized prostate cancer (PCa) undergoing nerve-sparing radical prostatectomy (RP). Objective To analyze the role of penile rehabilitation in the recovery of erectile function (EF) after nerve-sparing RP. Evidence synthesis Penile rehabilitation is defined as the use of any intervention or combination with the goal not only to achieve erections sufficient for satisfactory sexual intercourses, but also to return EF to preoperative levels. The concept of rehabilitation is based on the implementation of protocols aimed at improving oxygenation, preserving endothelial structure, and preventing smooth muscle structural alterations. Nowadays, the most commonly adopted approaches for penile rehabilitation after nerve-sparing RP are represented by the administration of phosphodiesterase type-5 inhibitors (PDE5-Is), intracorporeal injection therapy, vacuum erection devices (VED), and the combination of these therapies. Several basic science studies support the rational for the adoption of penile rehabilitation protocols. Particularly, rehabilitation, set as early as possible, seems to be better than leaving the erectile tissues unassisted. On the other hand, results from solid prospective randomized trials finally assessing the long-term beneficial effects of PDE5-Is, intracavernosal injections, or VED on EF recovery after surgery are still lacking. Conclusions Although preclinical evidences support the rationale for penile rehabilitation after nerve-sparing RP, clinical studies reported conflicting results regarding its efficacy on long-term EF recovery. Nowadays, which is the optimal rehabilitation program still represents a matter of debate. PMID:26816818

  6. Penile Microbiota and Female Partner Bacterial Vaginosis in Rakai, Uganda

    PubMed Central

    Hungate, Bruce A.; Tobian, Aaron A. R.; Ravel, Jacques; Prodger, Jessica L.; Serwadda, David; Kigozi, Godfrey; Galiwango, Ronald M.; Nalugoda, Fred; Keim, Paul; Wawer, Maria J.; Price, Lance B.; Gray, Ronald H.

    2015-01-01

    ABSTRACT Bacterial vaginosis (BV) is a common vaginal bacterial imbalance associated with risk for HIV and poor gynecologic and obstetric outcomes. Male circumcision reduces BV-associated bacteria on the penis and decreases BV in female partners, but the link between penile microbiota and female partner BV is not well understood. We tested the hypothesis that having a female partner with BV increases BV-associated bacteria in uncircumcised men. We characterized penile microbiota composition and density (i.e., the quantity of bacteria per swab) by broad-coverage 16S rRNA gene-based sequencing and quantitative PCR (qPCR) in 165 uncircumcised men from Rakai, Uganda. Associations between penile community state types (CSTs) and female partner’s Nugent score were assessed. We found seven distinct penile CSTs of increasing density (CST1 to 7). CST1 to 3 and CST4 to 7 were the two major CST groups. CST4 to 7 had higher prevalence and abundance of BV-associated bacteria, such as Mobiluncus and Dialister, than CST1 to 3. Men with CST4 to 7 were significantly more likely to have a female partner with a high Nugent score (P = 0.03). Men with two or more extramarital partners were significantly more likely to have CST4 to 7 than men with only marital partners (CST4 to 7 prevalence ratio, 1.84; 95% confidence interval [CI], 1.16 to 2.92). Female partner Nugent BV is significantly associated with penile microbiota. Our data support the exchange of BV-associated bacteria through intercourse, which may explain BV recurrence and persistence. PMID:26081632

  7. Characterization of temperate phages infecting Clostridium difficile isolates of human and animal origins.

    PubMed

    Sekulovic, Ognjen; Garneau, Julian R; Néron, Audrey; Fortier, Louis-Charles

    2014-04-01

    Clostridium difficile is a Gram-positive pathogen infecting humans and animals. Recent studies suggest that animals could represent potential reservoirs of C. difficile that could then transfer to humans. Temperate phages contribute to the evolution of most bacteria, for example, by promoting the transduction of virulence, fitness, and antibiotic resistance genes. In C. difficile, little is known about their role, mainly because suitable propagating hosts and conditions are lacking. Here we report the isolation, propagation, and preliminary characterization of nine temperate phages from animal and human C. difficile isolates. Prophages were induced by UV light from 58 C. difficile isolates of animal and human origins. Using soft agar overlays with 27 different C. difficile test strains, we isolated and further propagated nine temperate phages: two from horse isolates (ΦCD481-1 and ΦCD481-2), three from dog isolates (ΦCD505, ΦCD506, and ΦCD508), and four from human isolates (ΦCD24-2, ΦCD111, ΦCD146, and ΦCD526). Two phages are members of the Siphoviridae family (ΦCD111 and ΦCD146), while the others are Myoviridae phages. Pulsed-field gel electrophoresis and restriction enzyme analyses showed that all of the phages had unique double-stranded DNA genomes of 30 to 60 kb. Phages induced from human C. difficile isolates, especially the members of the Siphoviridae family, had a broader host range than phages from animal C. difficile isolates. Nevertheless, most of the phages could infect both human and animal strains. Phage transduction of antibiotic resistance was recently reported in C. difficile. Our findings therefore call for further investigation of the potential risk of transduction between animal and human C. difficile isolates.

  8. Characterization of Clostridium difficile isolates from human fecal samples and retail meat from Pennsylvania.

    PubMed

    Varshney, Jyotika B; Very, Katherine J; Williams, Jen L; Hegarty, John P; Stewart, David B; Lumadue, Jeanne; Venkitanarayanan, Kumar; Jayarao, Bhushan M

    2014-10-01

    A study was conducted to determine the prevalence of Clostridium difficile and characterize C. difficile isolates from human stool and retail grocery meat samples. Human stool samples (n=317) were obtained from a clinical laboratory and meat samples (n=303) were collected from 8 retail grocery stores from October 2011 through September 2012 from Centre County of Pennsylvania and were examined for C. difficile. C. difficile was isolated from 16.7% of stool samples (n=317) and 6.9%, 11.5%, 14.5%, and 7.8% of beef (n=72), pork (n=78), turkey (n=76), and chicken (n=77) samples, respectively. Six different toxin gene profiles were detected in all human and meat isolates of C. difficile based on the presence or absence of toxin genes tcdA, tcdB, and cdtA and cdtB. Interestingly, 75.6% of the human C. difficile isolates lacked any deletion in the tcdC gene (139-bp), whereas a 39-bp deletion was observed in 61.3% of the C. difficile strains isolated from meat samples. C. difficile from meat samples were more susceptible to clindamycin, moxifloxacin, vancomycin, and metronidazole than C. difficile isolates from human samples. Twenty-five different ribotypes were identified in human and meat C. difficile isolates. In conclusion, significant genotypic and phenotypic differences were observed between human and meat isolates of C. difficile; however, a few C. difficile isolates from meat-in particular ribotypes 078, PA01, PA05, PA16, and PA22 with unique profiles (toxin gene, tcdC gene size and antimicrobial resistance profiles)-were similar to human C. difficile isolates.

  9. Prenatal exposure to phthalates is associated with decreased anogenital distance and penile size in male newborns.

    PubMed

    Bustamante-Montes, L P; Hernández-Valero, M A; Flores-Pimentel, D; García-Fábila, M; Amaya-Chávez, A; Barr, D B; Borja-Aburto, V H

    2013-08-01

    Reproductive effects from phthalate exposure have been documented mostly in animal studies. This study explored the association between prenatal exposure to phthalate metabolites, anogenital distance and penile measurements in male newborns in Toluca, State of Mexico. A total of 174 pregnant women provided urine samples for phthalate analysis during their last prenatal visit, and the 73 who gave birth to male infants were included in the study. The 73 male newborns were weighed and measured using standardized methods after delivery. After adjusting for creatinine and supine length at birth, significant inverse associations were observed between an index of prenatal exposure to total phthalate exposure and the distance from the anus to anterior base of the penis (β = -0.191 mm per 1 μg/l, P = 0.037), penile width (β = -0.0414, P = 0.050) and stretched length (β = -0.2137, P = 0.034); prenatal exposure to mono-2-ethylhexyl phthalate exposure was associated with a reduction in the stretched length of the penis (β = -0.2604, P = 0.050). Human exposure to phthalates is a public health concern, and the system most vulnerable to its potential effects seems to be the immature male reproductive tract.

  10. Prenatal exposure to phthalates is associated with decreased anogenital distance and penile size in male newborns

    PubMed Central

    Bustamante-Montes, L P; Hernández-Valero, M A; Flores-Pimentel, D; García-Fábila, M; Amaya-Chávez, A; Barr, D B; Borja-Aburto, V H

    2013-01-01

    Reproductive effects from phthalate exposure have been documented mostly in animal studies. This study explored the association between prenatal exposure to phthalate metabolites, anogenital distance and penile measurements in male newborns in Toluca, State of Mexico. A total of 174 pregnant women provided urine samples for phthalate analysis during their last prenatal visit, and the 73 who gave birth to male infants were included in the study. The 73 male newborns were weighed and measured using standardized methods after delivery. After adjusting for creatinine and supine length at birth, significant inverse associations were observed between an index of prenatal exposure to total phthalate exposure and the distance from the anus to anterior base of the penis (β = −0.191 mm per 1 µg/l, P = 0.037), penile width (β = −0.0414, P = 0.050) and stretched length (β = −0.2137, P = 0.034); prenatal exposure to mono-2-ethylhexyl phthalate exposure was associated with a reduction in the stretched length of the penis (β = −0.2604, P = 0.050). Human exposure to phthalates is a public health concern, and the system most vulnerable to its potential effects seems to be the immature male reproductive tract. PMID:24349678

  11. "Ihubacter massiliensis": a new bacterium isolated from the human gut.

    PubMed

    Ndongo, S; Lagier, J-C; Fournier, P-E; Raoult, D; Khelaifia, S

    2016-09-01

    We report here the main characteristics of "Ihubacter massiliensis," strain Marseille-P2843(T) (CSUR P2843), a new genus of the Clostridiales family isolated from a stool sample from a healthy 29-year-old woman. PMID:27579171

  12. Differential antimicrobial susceptibility between human and chicken isolates of vancomycin-resistant and sensitive Enterococcus faecium.

    PubMed

    Chen, Han Yuan; Hill, Robert L R; Kirk, Monica; Casewell, Mark W; Beighton, David

    2002-01-01

    To compare the differential antimicrobial susceptibilities of Enterococcus faecium from humans and whole chicken carcasses, MICs of 12 antimicrobial agents were determined for 54 clinical-isolates (31 vancomycin-resistant [VREF]) and 60 chicken-isolates (29 VREF). Chicken VREF were slightly but consistently more resistant to vancomycin, teicoplanin and avoparcin, compared with human VREF (P<0.01). MICs of LY333328 were human VREF were resistant to erythromycin and tylosin, compared with only 58.6% of chicken VREF (P<0.01). Streptogramins were active against all isolates except four chicken strains. MIC(90s) of amoxycillin and gentamicin for human E. faecium were 8-16-fold higher than chicken isolates. Chicken VREF were significantly more resistant to tetracycline but more susceptible to chloramphenicol than human VREF (P<0.001). PMID:11814766

  13. Prevalence of virulence genes in strains of Campylobacter jejuni isolated from human, bovine and broiler.

    PubMed

    González-Hein, Gisela; Huaracán, Bernardo; García, Patricia; Figueroa, Guillermo

    2013-12-01

    Campylobacter jejuni isolates of different origins (bovine, broiler meat, human) were screened by polymerase chain reaction for the presence of 4 genes cdtB, cst-II, ggt, and virB11, previously linked to virulence such as adherence, invasion, colonization, molecular mimicry, and cytotoxin production. In addition, the isolates were screened for the presence of the global gene regulator csrA linked to oxidative stress responses, biofilms formation, and cell adhesion. All the C. jejuni isolates were positive for cdtB gene. The csrA gene was detected in 100% and 92% of C. jejuni isolates from human and animal origin and the virB11 gene was detected in 7.3% and 3.6% isolates from chicken and human respectively. All isolates from bovine were negative for the virB11 gene. The isolates showed a wide variation for the presence of the remaining genes. Of the C. jejuni recovered from human 83.6%, and 32.7% were positive for cst-II, and ggt respectively. Out of the isolates from chicken 40% and 5.5% isolates revealed the presence of cst-II, and ggt, respectively. Finally of the C. jejuni isolates from bovine, 97.7% and 22.7% were positive for cst-II, and ggt respectively. We conclude that the genes of this study circulate among humans and animals. These results led us to hypothesize that the isolates associated with enteritis (cdtB positives) are not selected by environmental or host-specific factors. On the other hand, the high frequencies of csrA gene in C. jejuni show that this gene is important for the survival of C. jejuni in animals and humans.

  14. Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

    PubMed Central

    Kamaruddin, Mudyawati; Rahman, Md. Moshiur; Arayama, Shunsuke; Hidayati, Anggi P.N.; Syafruddin, Din; Asih, Puji B.S.; Yoshikawa, Hisao; Kawahara, Ei

    2014-01-01

    Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species. PMID:25352694

  15. Molecular characterization of various trichomonad species isolated from humans and related mammals in Indonesia.

    PubMed

    Kamaruddin, Mudyawati; Tokoro, Masaharu; Rahman, Md Moshiur; Arayama, Shunsuke; Hidayati, Anggi P N; Syafruddin, Din; Asih, Puji B S; Yoshikawa, Hisao; Kawahara, Ei

    2014-10-01

    Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species. PMID:25352694

  16. Molecular characterization of various trichomonad species isolated from humans and related mammals in Indonesia.

    PubMed

    Kamaruddin, Mudyawati; Tokoro, Masaharu; Rahman, Md Moshiur; Arayama, Shunsuke; Hidayati, Anggi P N; Syafruddin, Din; Asih, Puji B S; Yoshikawa, Hisao; Kawahara, Ei

    2014-10-01

    Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

  17. Draft Genome Sequence of the First Human Isolate of the Ruminant Pathogen Mycoplasma capricolum subsp. capricolum

    PubMed Central

    Fischer, Anne; Heller, Martin; Jores, Joerg; Sachse, Konrad; Mourier, Tobias; Hansen, Anders Johannes

    2015-01-01

    Mycoplasma capricolum subsp. capricolum is a well-known pathogen of small ruminants. A recent human case of septicemia involving this agent raised the question of its potential pathogenicity to humans. We present the first draft genome sequence of a human Mycoplasma capricolum subsp. capricolum isolate. PMID:26089408

  18. Diversity and antimicrobial susceptibility profiling of staphylococci isolated from bovine mastitis cases and close human contacts.

    PubMed

    Schmidt, T; Kock, M M; Ehlers, M M

    2015-09-01

    The objectives of this study were to examine the diversity of Staphylococcus spp. recovered from bovine intramammary infections and humans working in close contact with the animals and to evaluate the susceptibility of the staphylococcal isolates to different antimicrobials. A total of 3,387 milk samples and 79 human nasal swabs were collected from 13 sampling sites in the KwaZulu-Natal province of South Africa. In total, 146 Staph. aureus isolates and 102 coagulase-negative staphylococci (CNS) were recovered from clinical and subclinical milk samples. Staphylococcusaureus was isolated from 12 (15.2%) of the human nasal swabs and 95 representative CNS were recovered for further characterization. The CNS were identified using multiplex-PCR assays, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and tuf gene sequencing. Seven Staphylococcus spp. were identified among the CNS of bovine origin, with Staph.chromogenes (78.4%) predominating. The predominant CNS species recovered from the human nasal swabs was Staph.epidermidis (80%) followed by Staph.chromogenes (6.3%). The antimicrobial susceptibility of all staphylococcal isolates was evaluated using disk diffusion and was supplemented by screening for specific antimicrobial resistance genes. Ninety-eight (67.1%) Staph.aureus isolates of bovine origin were pansusceptible; 39 (26.7%) isolates were resistant to a single class, and 7 (4.8%) isolates were resistant to 2 classes of antimicrobials. Two Staph. aureus (1.4%) isolates were multidrug-resistant. Resistance to penicillin was common, with 28.8% of the bovine and 75% of the human Staph. aureus isolates exhibiting resistance. A similar observation was made with the CNS, where 37.3% of the bovine and 89.5% of the human isolates were resistant to penicillin. Multidrug-resistance was common among the human CNS, with 39% of the isolates exhibiting resistance to 3 or more classes of antimicrobials. The antimicrobial

  19. Diversity and antimicrobial susceptibility profiling of staphylococci isolated from bovine mastitis cases and close human contacts.

    PubMed

    Schmidt, T; Kock, M M; Ehlers, M M

    2015-09-01

    The objectives of this study were to examine the diversity of Staphylococcus spp. recovered from bovine intramammary infections and humans working in close contact with the animals and to evaluate the susceptibility of the staphylococcal isolates to different antimicrobials. A total of 3,387 milk samples and 79 human nasal swabs were collected from 13 sampling sites in the KwaZulu-Natal province of South Africa. In total, 146 Staph. aureus isolates and 102 coagulase-negative staphylococci (CNS) were recovered from clinical and subclinical milk samples. Staphylococcusaureus was isolated from 12 (15.2%) of the human nasal swabs and 95 representative CNS were recovered for further characterization. The CNS were identified using multiplex-PCR assays, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and tuf gene sequencing. Seven Staphylococcus spp. were identified among the CNS of bovine origin, with Staph.chromogenes (78.4%) predominating. The predominant CNS species recovered from the human nasal swabs was Staph.epidermidis (80%) followed by Staph.chromogenes (6.3%). The antimicrobial susceptibility of all staphylococcal isolates was evaluated using disk diffusion and was supplemented by screening for specific antimicrobial resistance genes. Ninety-eight (67.1%) Staph.aureus isolates of bovine origin were pansusceptible; 39 (26.7%) isolates were resistant to a single class, and 7 (4.8%) isolates were resistant to 2 classes of antimicrobials. Two Staph. aureus (1.4%) isolates were multidrug-resistant. Resistance to penicillin was common, with 28.8% of the bovine and 75% of the human Staph. aureus isolates exhibiting resistance. A similar observation was made with the CNS, where 37.3% of the bovine and 89.5% of the human isolates were resistant to penicillin. Multidrug-resistance was common among the human CNS, with 39% of the isolates exhibiting resistance to 3 or more classes of antimicrobials. The antimicrobial

  20. Tolerability and efficacy of newly developed penile injection of cross-linked dextran and polymethylmethacrylate mixture on penile enhancement: 6 months follow-up.

    PubMed

    Yang, D Y; Lee, W K; Kim, S C

    2013-05-01

    Cross-linked dextran and polymethylmethacrylate mixture (Lipen-10) is newly developed tissue filler. The purpose of this study was to evaluate tolerability and efficacy of Lipen-10 on penile enhancement. Twenty adult males were included in this study. Lipen-10 was injected into the subcutaneous tissue of the penile shaft. The penile girth and length were measured in the flaccid state, before and 1, 3 and 6 months after the injection. The circumference increased by 3.7±1.2 cm (50.8%, P<0.0001) at penile base, 4.2±0.9 cm (59.0%, P<0.001) at mid-shaft, and 3.8±1.0 cm (53.2%, P<0.0001) at distal shaft and the length increased by 2.3±1.4 cm (63.2%, P<0.001). There was, however, no significant difference between 3 and 6 months post-treatment in girth and length (P-values: 0.796, 0.498, 0.600 and 0.084 for penile base, mid- and distal-shaft and length, respectively). The complications were only one mild asymmetry of penile shape and one 5-mm-sized nodule in the injected site. There were no clinically significant adverse events in all subjects. Penile injection of Lipen-10 led to a significant increase in penile size, showed a good durability and was well-tolerated, without serious adverse events. These results suggest that penile injection of Lipen-10 may be a new effective method for penile enhancement. PMID:23171980

  1. Isolation and cryopreservation of human peripheral blood monocytes.

    PubMed

    Tsvetkov, T; Nickolov, C; Buckureshtliev, A; Mincheff, M; Tsoney, L; Terziev, R; Popov, D

    1986-12-01

    A modification of the Freundlich and Avdalovic method (J. Immunol. Methods 62, 31 (1983] is reported. Buffy coats, separated and pooled together, are used for isolation of monocytes (70% yield, 100% purity). Cell density of working suspension is increased up to 0.65 X 10(9) cells/75 cm2 surface by multiplication of the active fibronectin sites. For the purpose, cryoprecipitate is used instead of plasma for coating the glass-gelatin surface. Monocytes, isolated by that procedure, could be successfully cryopreserved with dimethyl sulfoxide cryoprotective solution.

  2. Draft Genome of Debaryomyces fabryi CBS 789T, Isolated from a Human Interdigital Mycotic Lesion.

    PubMed

    Tafer, Hakim; Sterflinger, Katja; Lopandic, Ksenija

    2016-01-01

    The yeast genus Debaryomyces comprises species isolated from various natural habitats, man-made environments, and clinical materials. Here, the draft genome of D. fabryi CBS 789(T), isolated from a human interdigital mycotic lesion, is presented. PMID:26847909

  3. Complete Genome Sequence of Bifidobacterium breve CECT 7263, a Strain Isolated from Human Milk

    PubMed Central

    Jiménez, Esther; Villar-Tajadura, M. Antonia; Marín, María; Fontecha, Javier; Requena, Teresa; Arroyo, Rebeca; Fernández, Leónides

    2012-01-01

    Bifidobacterium breve is an actinobacterium frequently isolated from colonic microbiota of breastfeeding babies. Here, we report the complete and annotated genome sequence of a B. breve strain isolated from human milk, B. breve CECT 7263. The genome sequence will provide new insights into the biology of this potential probiotic organism and will allow the characterization of genes related to beneficial properties. PMID:22740680

  4. Draft Genome of Debaryomyces fabryi CBS 789T, Isolated from a Human Interdigital Mycotic Lesion

    PubMed Central

    Tafer, Hakim; Sterflinger, Katja

    2016-01-01

    The yeast genus Debaryomyces comprises species isolated from various natural habitats, man-made environments, and clinical materials. Here, the draft genome of D. fabryi CBS 789T, isolated from a human interdigital mycotic lesion, is presented. PMID:26847909

  5. Human isolates of Listeria monocytogenes in Sweden during half a century (1958-2010).

    PubMed

    Lopez-Valladares, G; Tham, W; Parihar, V Singh; Helmersson, S; Andersson, B; Ivarsson, S; Johansson, C; Ringberg, H; Tjernberg, I; Henriques-Normark, B; Danielsson-Tham, M-L

    2014-11-01

    Isolates of Listeria monocytogenes (n = 932) isolated in Sweden during 1958-2010 from human patients with invasive listeriosis were characterized by serotyping and pulsed-field gel electrophoresis (PFGE) (AscI). Of the 932 isolates, 183 different PFGE types were identified, of which 83 were each represented by only one isolate. In all, 483 serovar 1/2a isolates were distributed over 114 PFGE types; 90 serovar 1/2b isolates gave 32 PFGE types; 21 serovar 1/2c isolates gave nine PFGE types; three serovar 3b isolates gave one PFGE type; and, 335 serovar 4b isolates gave 31 PFGE types. During the 1980s in Sweden, several serovar 4b cases were associated with the consumption of European raw soft cheese. However, as cheese-production hygiene has improved, the number of 4b cases has decreased. Since 1996, serovar 1/2a has been the dominant L. monocytogenes serovar in human listeriosis in Sweden. Therefore, based on current serovars and PFGE types, an association between human cases of listeriosis and the consumption of vacuum-packed gravad and cold-smoked salmon is suggested. PMID:24480252

  6. Characteristics of Yersinia enterocolitica and related species isolated from human, animal, and environmental sources.

    PubMed

    Shayegani, M; DeForge, I; McGlynn, D M; Root, T

    1981-09-01

    During a 4-year period, 4,448 human, animal, and environmental samples collected in New York State were tested for the presence of Yersinia enterocolitica or related species. A total of 339 isolates were identified as Yersinia and characterized according to source, species, biogroup, serogroup, and, in some instances, phage type. Four new biogroups of Y. intermedia were characterized. Of 149 human isolates, 120 (80.5%) were identified as Y. enterocolitica, and 29 were identified as either Y. intermedia (12.1%), Y. frederiksenii (5.4%), or Y. kristensenii (2.0%). Of the other 190 isolates, recovered from animals and the environment, 54 (28.4%) were Y. enterocolitica and 136 were either Y. intermedia (62.6%), Y. frederiksenii (4.7%), Y. kristensenii (3.7%), or an undescribed Yersinia species (0.5%). Two established human pathogenic strains of Y. enterocolitica were recovered: 59 isolates (37 from an outbreak) of "American strain" (serogroup O:8, biogroups Niléhn 2, Wauters 1, and Knapp and Thal 2, phage type Xz) and 11 isolates of "Canadian strain" (serogroup O:3, biogroups Niléhn 4, Wauters 4, and Knapp and Thal 1, phage types IXb). This was the first documented isolation of the Canadian strain in the United States. Isolates of other strains implicated in human disease (serogroups 0:4,33, 0:5, O:6,31, O:7,8, and O:8) were also recovered from both human and nonhuman sources.

  7. A strain of pathogenic Naegleria isolated from a human nasal swab.

    PubMed

    Chang, S L; Healy, G R; McCabe, L; Shumaker, J B; Schultz, M G

    1975-01-01

    The 161A strain of Naegleria isolated from a human nasal swab and reported earlier was further studied for its growth pattern, cyst-wall appearance, cytopathic effect on primary monkey kidney cells, and pathogenicity in mice. Results obtained in the study establish the pathogenic status of this strain. This constitutes the first isolation of a pathogenic Naegleria from a human nasal cavity. PMID:829906

  8. A zinc protein isolated from human parotid saliva.

    PubMed Central

    Henkin, R I; Lippoldt, R E; Bilstad, J; Edelhoch, H

    1975-01-01

    A zinc protein has been isolated and purified to apparent homogeneity from subjects with normal taste acuity by gel filtration and ion-exchange chromatography. The protein has a molecular weight of 37,000 and does not appear to have subunits. It is composed of 8% histidine residues and has 2 moles of zinc per mole of protein. Images PMID:1054831

  9. Role of Ultraosonography in Grading of Penile Fractures

    PubMed Central

    Bhagavan, B.C.; Sanjay, S.C.; Krishnappa, N.; Sahadev, Ramaiah; V., Satish

    2015-01-01

    Introduction: Penile fracture is an uncommon and less reported condition. Early diagnosis with corrective measures is the order of the day. Aim: To perform ultrasonography of penis and to understand its role in making correct diagnosis of fractures. To know how it will help surgeon in its management and in adopting a grading system. Materials and Methods: Emergency evaluation of 15 cases of penile fractures by ultrasonography before surgery. Entire penis was scanned all along its length and circumference. Tunica albugenia was seen as white covering of both corpora cavernosa and break in its continuity is seen as wedge shape defect. Hematoma on either sides of tunica was well appreciated. Results: After ultrasonography all patients underwent emergency surgery, the defect in corpora was well appreciated. Repair of tunica albugenia was done which confirmed our findings. Conclusion: Ultrasonography is the modality of choice for quick diagnosis, and no other radiological workup is required before surgery. PMID:26023616

  10. Genetic diversity and antibiotic resistance profiles of Campylobacter jejuni isolates from poultry and humans in Turkey.

    PubMed

    Abay, Secil; Kayman, Tuba; Otlu, Baris; Hizlisoy, Harun; Aydin, Fuat; Ertas, Nurhan

    2014-05-16

    In this study, the investigation of clonal relations between human and poultry Campylobacter jejuni isolates and the determination of susceptibilities of isolates to various antibiotics were aimed. A total of 200 C. jejuni isolates concurrently obtained from 100 chicken carcasses and 100 humans were genotyped by the Pulsed-Field Gel Electrophoresis (PFGE) and automated Repetitive Extragenic Palindromic PCR (Rep-PCR, DiversiLab system) methods and were tested for their susceptibility to six antibiotics with disk diffusion method. The minimum inhibitory concentration (MIC) values of ciprofloxacin (CI), enrofloxacin (EF) and erythromycin (EM) were evaluated by E-test. By using PFGE 174 of (87.0%) the isolates were able to be typed. The clonally related strains were placed in 35 different clusters and 115 different genotypes were obtained. All of the two hundred isolates could be typed by using Rep-PCR and were divided into 133 different genotypes. One hundred and fourteen clonally related isolates (57.0%) were included in 47 clusters. In disk diffusion test, while the susceptibility rates of AMC and S to human and chicken derived C. jejuni isolates were 84.0%-96.0% and 96.0%-98.0%, respectively, all isolates were susceptible to gentamicin. The resistance rates of human isolates to AMP, NA and TE were detected as 44.0%, 84.0% and 38.0% of the resistances of chicken isolates to these antibiotics were 34.0%, 95.0% and 56.0%, respectively. The MIC values of human and chicken isolates to CI, EF and EM were detected as 81.0-93.0%, 85.0-88.0% and 6.0-7.0%, respectively. The clonal proximity rates were detected between human and poultry origin C. jejuni isolates. The discriminatory power of PFGE and Rep-PCR was similar, with Simpson's diversity indexes of 0.993 and 0.995, respectively. Concordance of the two methods as determined by Adjusted Rand coefficient was 0.198 which showed the low congruence between Rep-PCR and PFGE. High rates of quinolone resistance were detected in

  11. Doramectin reduces sexual behavior and penile erection in male rats.

    PubMed

    Ferri, R; Todon E Silva, A F S; Cabral, D; Moreira, N; Spinosa, H S; Bernardi, M M

    2013-01-01

    Doramectin (DOR) is an antiparasitic drug that is widely used in domestic animals. In mammals, DOR acts as a γ-aminobutyric acid receptor agonist. This neurotransmitter plays an important role in the regulation of sexual behavior. The present study investigated the effects of two medically relevant doses of DOR on sexual behavior in male rats. We also examined whether previous sexual experience modulates responses to DOR. General activity was first observed in an open field 24, 48, and 72 h after administration of 0.1 and 0.3 mg/kg DOR to determine the dose and time effects of the drug. Apomorphine-induced penile erection and sexual behavior in inexperienced male rats were then analyzed. The effects of previous sexual experience on subsequent sexual behavior in DOR-treated rats (0.3 mg/kg, 24 h prior to the test) were also assessed. The standard therapeutic dose (0.2 mg/kg) did not modify general activity or penile erection. A slightly concentrated dose of 0.3 mg/kg, which is still within the therapeutic range, decreased apomorphine-induced penile erection, whereas 0.2 mg/kg did not modify this behavior. Compared with controls, sexual behavior in inexperienced male rats was impaired after 0.3 mg/kg DOR. Previous sexual experience had little impact on the effects of 0.3 mg/kg DOR. In conclusion, the 0.2 mg/kg dose of DOR did not affect motor behavior or apomorphine-induced penile erection. At a more slightly higher dose level, the appetitive and consummatory phases of sexual behavior in inexperienced male rats were impaired. Previous sexual experience was unable to reverse this sexual impairment, suggesting that previous sexual experience does not exert a positive effect in attenuating sexual impairment produced by DOR treatment.

  12. Physiology of penile erection and pathophysiology of erectile dysfunction.

    PubMed

    Dean, Robert C; Lue, Tom F

    2005-11-01

    This article reviews the physiology of penile erection, the components of erectile function, and the pathophysiology of erectile dysfunction. The molecular and clinical under-standing of erectile function continues to gain ground at a particularly fast rate. Advances in gene discovery have aided greatly in working knowledge of smooth muscle relaxation/contraction pathways. The understanding of the nitric oxide pathway has aided not only in the molecular understanding of the tumescence but also greatly in the therapy of erectile dysfunction.

  13. Subgenotype analysis of Cryptosporidium isolates from humans, cattle, and zoo ruminants in Portugal.

    PubMed

    Alves, Margarida; Xiao, Lihua; Sulaiman, Irshad; Lal, Altaf A; Matos, Olga; Antunes, Francisco

    2003-06-01

    Cryptosporidium parvum and Cryptosporidium hominis isolates from human immunodeficiency virus-infected patients, cattle, and wild ruminants were characterized by PCR and DNA sequencing analysis of the 60-kDa glycoprotein gene. Seven alleles were identified, three corresponding to C. hominis and four corresponding to C. parvum. One new allele was found (IId), and one (IIb) had only been found in Portugal. Isolates from cattle and wild ruminants clustered in two alleles. In contrast, human isolates clustered in seven alleles, showing extensive allelic diversity.

  14. Subgenotype Analysis of Cryptosporidium Isolates from Humans, Cattle, and Zoo Ruminants in Portugal

    PubMed Central

    Alves, Margarida; Xiao, Lihua; Sulaiman, Irshad; Lal, Altaf A.; Matos, Olga; Antunes, Francisco

    2003-01-01

    Cryptosporidium parvum and Cryptosporidium hominis isolates from human immunodeficiency virus-infected patients, cattle, and wild ruminants were characterized by PCR and DNA sequencing analysis of the 60-kDa glycoprotein gene. Seven alleles were identified, three corresponding to C. hominis and four corresponding to C. parvum. One new allele was found (IId), and one (IIb) had only been found in Portugal. Isolates from cattle and wild ruminants clustered in two alleles. In contrast, human isolates clustered in seven alleles, showing extensive allelic diversity. PMID:12791920

  15. Comparison of epidemiologically linked Campylobacter jejuni isolated from human and poultry sources.

    PubMed

    Lajhar, S A; Jennison, A V; Patel, B; Duffy, L L

    2015-12-01

    Campylobacter jejuni is responsible for most foodborne bacterial infections worldwide including Australia. The aim of this study was to investigate a combination of typing methods in the characterization of C. jejuni isolated from clinical diarrhoeal samples (n = 20) and chicken meat (n = 26) in order to identify the source of infection and rank isolates based on their relative risk to humans. Sequencing of the flaA short variable region demonstrated that 86% of clinical isolates had genotypes that were also found in chicken meat. A polymerase chain reaction binary typing system identified 27 different codes based on the presence or absence of genes that have been reported to be associated with various aspects of C. jejuni pathogenicity, indicating that not all isolates may be of equal risk to human health. The lipooligosaccharide (LOS) of the C. jejuni isolates was classified into six classes (A, B, C, E, F, H) with 10·4% remaining unclassified. The majority (72·7%) of clinical isolates possessed sialylated LOS classes. Sialylated LOS classes were also detected in chicken isolates (80·7%). Antimicrobial tests indicated a low level of resistance, with no phenotypic resistance found to most antibiotics tested. A combination of typing approaches was useful to assign isolates to a source of infection and assess their risk to humans.

  16. Penile cancer: a local case series and literature review

    PubMed Central

    Lau, Wei Da; Ong, Chin Hu; Lim, Tow Poh; Teo, Colin

    2015-01-01

    INTRODUCTION Penile cancer is an uncommon disease affecting only about one in 100,000 men worldwide in a year. The diagnosis of the condition is frequently delayed, and the disease and its treatment frequently result in significant morbidity in patients. METHODS We herein describe seven cases of penile tumours: six cases of squamous cell carcinomas and one case of B-cell lymphoma that presented to our hospital’s urology department between March 2011 and October 2012. We reviewed the literature to discuss the clinical presentation, natural history and current management of penile cancer. RESULTS The patients were followed up for 1–24 months. They were managed according to their disease stage and lymph node status. Four out of seven patients showed disease progression during the follow-up period. CONCLUSION The accurate staging of inguinal nodes in cases of low-risk disease is important to prescribe appropriate surgery for the inguinal nodes. Aggressive management of inguinal and pelvic lymph nodes remains the cornerstone in the treatment of high-risk disease cases. PMID:26668410

  17. Molecular Characterization of Staphylococcus sciuri Strains Isolated from Humans

    PubMed Central

    Couto, Isabel; Sanches, Ilda Santos; Sá-Leão, Raquel; de Lencastre, Hermínia

    2000-01-01

    We previously characterized over 100 Staphylococcus sciuri isolates, mainly of animal origin, and found that they all carried a genetic element (S. sciuri mecA) closely related to the mecA gene of methicillin-resistant Staphylococcus aureus (MRSA) strains. We also found a few isolates that carried a second copy of the gene, identical to MRSA mecA. In this work, we analyzed a collection of 28 S. sciuri strains isolated from both healthy and hospitalized individuals. This was a relatively heterogeneous group, as inferred from the different sources, places, and dates of isolation and as confirmed by pulsed-field gel electrophoresis analysis. All strains carried the S. sciuri mecA copy, sustaining our previous proposal that this element belongs to the genetic background of S. sciuri. Moreover, 46% of the strains also carried the MRSA mecA copy. Only these strains showed significant levels of resistance to beta-lactams. Strikingly, the majority of the strains carrying the additional MRSA mecA copy were obtained from healthy individuals in an antibiotic-free environment. Most of the 28 strains were resistant to penicillin, intermediately resistant to clindamycin, and susceptible to tetracycline, erythromycin, and gentamicin. Resistance to these last three antibiotics was found in some strains only. The findings reported in this work confirmed the role of S. sciuri in the evolution of the mechanism of resistance to methicillin in staphylococci and suggested that this species (like the pathogenic staphylococci) may accumulate resistance markers for several classes of antibiotics. PMID:10699009

  18. Characteristics and biotypes of Pasteurella multocida isolated from humans.

    PubMed

    Oberhofer, T R

    1981-03-01

    Fifty-two isolates of Pasteurella (48 strains of Pasteurella multocida and 4 strains of atypical Pasteurella) were identified by conventional and commercial test systems. All strains fermented glucose, sucrose, and fructose in purple broth base (Difco Laboratories) with bromocresol purple as indicator, although the atypical Pasteurella produced fermentation reactions that were barely perceptible. Eleven different biotypes were identified by fermentation reactions in maltose, mannitol, xylose, sorbitol, and trehalose media. There was a correlation of biotypes to cat bites, with 61% of cat bite isolates falling into biotype A and B. A correlation of biotype and dog bite isolates was not seen. The choice of medium used for fermentation tests was critical as evidenced by the inability of the organisms to grow in a second commercially purchased preparation of purple broth base. The reliability of commercial test systems in identifying Pasteurella was 81% for Oxi/Ferm (Roche Diagnostics, Div. Hoffmann-La Roche, Inc., Nutley, N.J.), 68% for API (Analytab Products, Plainview, N.Y.), and 11% for Minitek (BBL Microbiology Systems, Cockeysville, MD.).

  19. Social isolation disrupts hippocampal neurogenesis in young non-human primates.

    PubMed

    Cinini, Simone M; Barnabe, Gabriela F; Galvão-Coelho, Nicole; de Medeiros, Magda A; Perez-Mendes, Patrícia; Sousa, Maria B C; Covolan, Luciene; Mello, Luiz E

    2014-01-01

    Social relationships are crucial for the development and maintenance of normal behavior in non-human primates. Animals that are raised in isolation develop abnormal patterns of behavior that persist even when they are later reunited with their parents. In rodents, social isolation is a stressful event and is associated with a decrease in hippocampal neurogenesis but considerably less is known about the effects of social isolation in non-human primates during the transition from adolescence to adulthood. To investigate how social isolation affects young marmosets, these were isolated from other members of the colony for 1 or 3 weeks and evaluated for alterations in their behavior and hippocampal cell proliferation. We found that anxiety-related behaviors like scent-marking and locomotor activity increased after social isolation when compared to baseline levels. In agreement, grooming-an indicative of attenuation of tension-was reduced among isolated marmosets. These results were consistent with increased cortisol levels after 1 and 3 weeks of isolation. After social isolation (1 or 3 weeks), reduced proliferation of neural cells in the subgranular zone of dentate granule cell layer was identified and a smaller proportion of BrdU-positive cells underwent neuronal fate (doublecortin labeling). Our data is consistent with the notion that social deprivation during the transition from adolescence to adulthood leads to stress and produces anxiety-like behaviors that in turn might affect neurogenesis and contribute to the deleterious consequences of prolonged stressful conditions.

  20. A comparative study of strains of salmonella isolated from irrigation waters, vegetables and human infections.

    PubMed Central

    Garcia-Villanova Ruiz, B.; Cueto Espinar, A.; Bolaños Carmona, M. J.

    1987-01-01

    A total of 181 samples of irrigation water from the farmlands of Granada were examined for the presence of Salmonella spp. At the same time 849 samples of the crops from these farmlands and of vegetables sold in city market-places were studied. Sampling was done regularly over the period of study which ran from March 1981 to February 1983. Isolates from these sources were compared with 93 salmonellas isolated from human pathological material at various hospitals of the city of Granada from 1979-80, and again from 1981-3. The most commonly isolated serotypes of human origin were S. typhimurium and S. enteritidis. In irrigation waters and in crops, S. typhimurium, S. kapemba, S. london and S. blockley were found to be the most common. The results indicate a close relationship between the isolates from the irrigation waters and those from the vegetables, but their relationship to prevalent human infections is less clear. PMID:3595745

  1. Molecular characterization of Yersinia strains isolated from human and environmental sources.

    PubMed

    Marranzano, M; Agodi, A; Gulisano, M

    1993-01-01

    Thirty-seven strains of Yersinia isolated from human and environmental sources were investigated for virulence and molecular characteristics. Clinical isolates of Y. enterocolitica serotypes 0:3 and 0:9, harbouring the 45 MDa virulence plasmid were Ca++ dependent and responded positively to the CV-binding assay. The environmental Y. kristensenii strain, although harbouring a 45 MDa plasmid proved to be negative for both assays. All the environmental strains showed pyrazinamidase activity, while isolates from human sources proved negative. Two different plasmid profiles were disclosed: one Y. enterocolitica strain (serotype 0:7, 8) showed one plasmid of 67 MDa and one plasmid of 1.5 MDa. Eleven human isolates of Y. enterocolitica and one environmental strain of Y. krinstensenii showed only one plasmid of 45 MDa. Restriction endonuclease analysis of the 45 MDa plasmids supported the hypothesis that the environmental Y. krinstensenii plasmid is unrelated to the family of homologous virulence plasmids of Y. enterocolitica.

  2. Brucella isolated in humans and animals in Latin America from 1968 to 2006

    PubMed Central

    LUCERO, N. E.; AYALA, S. M.; ESCOBAR, G. I.; JACOB, N. R.

    2008-01-01

    SUMMARY We report a retrospective analysis of 1933 Brucella strains isolated from humans and animals in Latin American countries between 1968 and 1991 and in Argentina between 1994 and 2006. During the first period 50% of strains were from humans, mainly from Argentina, Mexico and Peru but, while B. suis was the main cause of infection in Argentina, B. melitensis was responsible for most infections in the other countries. In Argentina in the later years, B. melitensis and B. suis were observed more frequently than in the first period while isolation of B. abortus decreased. Of 145 B. melitensis human isolates, eight gave susceptibility patterns to dyes and penicillin and two were B. melitensis biovar 3, which has never been reported in animals. Forty-six percent of B. suis isolated were resistant to dyes which is an atypical feature in this species. PMID:17559694

  3. Clostridium difficile in Dutch animals: their presence, characteristics and similarities with human isolates.

    PubMed

    Koene, M G J; Mevius, D; Wagenaar, J A; Harmanus, C; Hensgens, M P M; Meetsma, A M; Putirulan, F F; van Bergen, M A P; Kuijper, E J

    2012-08-01

    The presence and characteristics of Clostridium difficile were investigated in 839 faecal samples from seven different animal species in the Netherlands. The number of positive samples ranged from 3.4% (cattle) to 25.0% (dogs). Twenty-two different PCR ribotypes were identified. Among 96 isolates, 53% harboured toxin genes. All C. difficile isolates from pigs, cattle and poultry were toxinogenic, whereas the majority of isolates from pet animals consisted of non-toxinogenic PCR ribotypes 010 and 039. Ribotype 012 was most prevalent in cattle and ribotype 078 in pigs. No predominant ribotypes were present in horse and poultry samples. Overall, PCR ribotypes 012, 014 and 078 were the most frequently recovered toxinogenic ribotypes from animal samples. Comparison with human isolates from the Dutch Reference Laboratory for C. difficile at Leiden University Medical Centre (LUMC) showed that these types were also recovered from human hospitalized patients in 2009/2010, encompassing 0.8%, 11.4% and 9.8% of all isolates, respectively. Application of multiple-locus variable-number tandem-repeat analysis indicated a genotypic relation of animal and human ribotype 078 strains, but a clear genotypic distinction for ribotypes 012 and 014. We conclude that toxinogenic C. difficile PCR ribotypes found in animals correspond to PCR ribotypes associated with human disease in hospitalized patients in the Netherlands. Contrary to PCR ribotype 078, significant genetic differences were observed between animal and human PCR ribotype 012 and 014 isolates.

  4. MLVA16 typing of Portuguese human and animal Brucella melitensis and Brucella abortus isolates.

    PubMed

    Ferreira, Ana Cristina; Chambel, Lélia; Tenreiro, Tania; Cardoso, Regina; Flor, Lídia; Dias, Isabel Travassos; Pacheco, Teresa; Garin-Bastuji, Bruno; Le Flèche, Philippe; Vergnaud, Gilles; Tenreiro, Rogério; de Sá, Maria Inácia Corrêa

    2012-01-01

    To investigate the epidemiological relationship of isolates from different Portuguese geographical regions and to assess the diversity among isolates, the MLVA16(Orsay) assay (panels 1, 2A and 2B) was performed with a collection of 126 Brucella melitensis (46 human and 80 animal isolates) and 157 B. abortus field isolates, seven vaccine strains and the representative reference strains of each species. The MLVA16(Orsay) showed a similar high discriminatory power (HGDI 0.972 and 0.902) for both species but panel 1 and 2A markers displayed higher diversity (HGDI 0.693) in B. abortus compared to B. melitensis isolates (HGDI 0.342). The B. melitensis population belong to the "Americas" (17%) and "East Mediterranean" (83%) groups. No isolate belonged to the "West Mediterranean" group. Eighty-five percent of the human isolates (39 in 46) fit in the "East-Mediterranean" group where a single lineage known as MLVA11 genotype 116 is responsible for the vast majority of Brucella infections in humans. B. abortus isolates formed a consistent group with bv1 and bv3 isolates in different clusters. Four MLVA11 genotypes were observed for the first time in isolates from S. Jorge and Terceira islands from Azores. From the collection of isolates analysed in this study we conclude that MLVA16(Orsay) provided a clear view of Brucella spp. population, confirming epidemiological linkage in outbreak investigations. In particular, it suggests recent and ongoing colonisation of Portugal with one B. melitensis lineage usually associated with East Mediterranean countries.

  5. MLVA16 Typing of Portuguese Human and Animal Brucella melitensis and Brucella abortus Isolates

    PubMed Central

    Ferreira, Ana Cristina; Chambel, Lélia; Tenreiro, Tania; Cardoso, Regina; Flor, Lídia; Dias, Isabel Travassos; Pacheco, Teresa; Garin-Bastuji, Bruno; Le Flèche, Philippe; Vergnaud, Gilles; Tenreiro, Rogério; de Sá, Maria Inácia Corrêa

    2012-01-01

    To investigate the epidemiological relationship of isolates from different Portuguese geographical regions and to assess the diversity among isolates, the MLVA16Orsay assay (panels 1, 2A and 2B) was performed with a collection of 126 Brucella melitensis (46 human and 80 animal isolates) and 157 B. abortus field isolates, seven vaccine strains and the representative reference strains of each species. The MLVA16Orsay showed a similar high discriminatory power (HGDI 0.972 and 0.902) for both species but panel 1 and 2A markers displayed higher diversity (HGDI 0.693) in B. abortus compared to B. melitensis isolates (HGDI 0.342). The B. melitensis population belong to the “Americas” (17%) and “East Mediterranean” (83%) groups. No isolate belonged to the “West Mediterranean” group. Eighty-five percent of the human isolates (39 in 46) fit in the “East-Mediterranean” group where a single lineage known as MLVA11 genotype 116 is responsible for the vast majority of Brucella infections in humans. B. abortus isolates formed a consistent group with bv1 and bv3 isolates in different clusters. Four MLVA11 genotypes were observed for the first time in isolates from S. Jorge and Terceira islands from Azores. From the collection of isolates analysed in this study we conclude that MLVA16Orsay provided a clear view of Brucella spp. population, confirming epidemiological linkage in outbreak investigations. In particular, it suggests recent and ongoing colonisation of Portugal with one B. melitensis lineage usually associated with East Mediterranean countries. PMID:22905141

  6. Comparison of Hemagglutination and Hemolytic Activity of Various Bacterial Clinical Isolates Against Different Human Blood Groups

    PubMed Central

    HRV, Rajkumar; Devaki, Ramakrishna

    2016-01-01

    Among the various pathogenic determinants shown by microorganisms hemagglutination and hemolysin production assume greater significance in terms of laboratory identification. This study evaluated the hemagglutination and hemolytic activity of various bacterial isolates against different blood groups. One hundred and fifty bacterial strains, isolated from clinical specimens like urine, pus, blood, and other body fluids were tested for their hemagglutinating and hemolytic activity against human A, B, AB, and O group red blood cells. Among the 150 isolates 81 were Escherichia coli, 18 were Klebsiella pneumoniae, 19 were Pseudomonas aeruginosa, 10 were Pseudomonas spp, six were Proteus mirabilis, and the rest 16 were Staphylococcus aureus. Nearly 85% of the isolates agglutinated A group cells followed by B and AB group (59.3% and 60.6% respectively). Least number of isolates agglutinated O group cells (38.0%). When the hemolytic activity was tested, out of these 150 isolates 79 (52.6%) hemolyzed A group cells, 61 (40.6%) hemolyzed AB group cells, 46 (30.6%) hemolyzed B group cells, and 57 (38.6%) isolates hemolyzed O group cells. Forty-six percent of the isolates exhibited both hemagglutinating and hemolytic property against A group cells, followed by B and AB group cells (28.6% and 21.3% respectively). Least number of isolates i.e., 32 (21.3%) showed both the properties against O group cells. The isolates showed wide variation in their hemagglutination and hemolytic properties against different combinations of human blood group cells. The study highlights the importance of selection of the type of cells especially when human RBCs are used for studying the hemagglutination and hemolytic activity of bacterial isolates because these two properties are considered as characteristic of pathogenic strains. PMID:27014523

  7. Isolating human DNA repair genes using rodent-cell mutants

    SciTech Connect

    Thompson, L.H.; Weber, C.A.; Brookman, K.W.; Salazar, E.P.; Stewart, S.A.; Mitchell, D.L.

    1987-03-23

    The DNA repair systems of rodent and human cells appear to be at least as complex genetically as those in lower eukaryotes and bacteria. The use of mutant lines of rodent cells as a means of identifying human repair genes by functional complementation offers a new approach toward studying the role of repair in mutagenesis and carcinogenesis. In each of six cases examined using hybrid cells, specific human chromosomes have been identified that correct CHO cell mutations affecting repair of damage from uv or ionizing radiations. This finding suggests that both the repair genes and proteins may be virtually interchangeable between rodent and human cells. Using cosmid vectors, human repair genes that map to chromosome 19 have cloned as functional sequences: ERCC2 and XRCC1. ERCC1 was found to have homology with the yeast excision repair gene RAD10. Transformants of repair-deficient cell lines carrying the corresponding human gene show efficient correction of repair capacity by all criteria examined. 39 refs., 1 fig., 1 tab.

  8. Comparative genotypic and pathogenic examination of Campylobacter concisus isolates from diarrheic and non-diarrheic humans

    PubMed Central

    2011-01-01

    Background Campylobacter concisus is an emerging enteric pathogen, yet it is commonly isolated from feces and the oral cavities of healthy individuals. This genetically complex species is comprised of several distinct genomospecies which may vary in pathogenic potential. Results We compared pathogenic and genotypic properties of C. concisus fecal isolates from diarrheic and healthy humans residing in the same geographic region. Analysis of amplified fragment length polymorphism (AFLP) profiles delineated two main clusters. Isolates assigned to AFLP cluster 1 belonged to genomospecies A (based on genomospecies-specific differences in the 23S rRNA gene) and were predominantly isolated from healthy individuals. This cluster also contained a reference oral strain. Isolates assigned to this cluster induced greater expression of epithelial IL-8 mRNA and more frequently contained genes coding for the zonnula occludins toxin and the S-layer RTX. Furthermore, isolates from healthy individuals induced greater apoptotic DNA fragmentation and increased metabolic activity than those from diarrheic individuals, and isolates assigned to genomospecies A (of which the majority were from healthy individuals) exhibited higher haemolytic activity compared to genomospecies B isolates. In contrast, AFLP cluster 2 was predominated by isolates belonging to genomospecies B and those from diarrheic individuals. Isolates from this cluster displayed greater mean epithelial invasion and translocation than cluster 1 isolates. Conclusion Two main genetically distinct clusters (i.e., genomospecies) were identified among C. concisus fecal isolates from healthy and diarrheic individuals. Strains within these clusters differed with respect to clinical presentation and pathogenic properties, supporting the hypothesis that pathogenic potential varies between genomospecies. ALFP cluster 2 isolates were predominantly from diarrheic patients, and exhibited higher levels of epithelial invasion and

  9. Dermabacter vaginalis sp. nov., isolated from human vaginal fluid.

    PubMed

    Chang, Dong-Ho; Rhee, Moon-Soo; Kim, Byoung-Chan

    2016-04-01

    A novel actinobacterial strain, AD1-86T, was isolated from the vaginal fluid of a Korean female and was characterized by a polyphasic approach. The strain was a facultatively anaerobic, Gram-stain-positive, non-spore-forming, non-motile, catalase-positive and oxidase-negative short rod. Colonies were creamy white, of low convexity and 1-2 mm in diameter after growth on DSM 92 agar plates at 37 °C for 2 days. The most closely related strains were Dermabacter hominis DSM 7083T and Helcobacillus massiliensis 6401990T (98.3 and 96.3 % 16S rRNA gene sequence similarity, respectively). The isolate grew optimally at 37 °C and pH 7 in the presence of 0.5% (w/v) NaCl. The cell-wall peptidoglycan contained meso-diaminopimelic acid and the cell-wall hydrolysates contained ribose, galactose and glucose. The DNA G+C content was 62.6 mol% and the mean DNA-DNA relatedness value of the isolate to D. hominis DSM 7083T was 31.1±3.0% (reciprocal: 48.2±5.3%). The major cellular fatty acids (>10%) were anteiso-C17:0, anteiso-C15:0 and iso-C16:0, and the menaquinones were MK-9, MK-8 and MK-7. The polar lipid profile of strain AD1-86T consisted of diphosphatidylglycerol, phosphatidylglycerol, two aminolipids and a glycolipid. Data from this polyphasic study indicate that strain AD1-86T represents a novel species of the genus Dermabacter, for which the name Dermabacter vaginalis sp. nov. is proposed; the type strain is AD1-86T (=KCTC 39585T=DSM 100050T). PMID:26867728

  10. Complete genome sequence of Streptococcus salivarius PS4, a strain isolated from human milk.

    PubMed

    Martín, Virginia; Maldonado-Barragán, Antonio; Jiménez, Esther; Ruas-Madiedo, Patricia; Fernández, Leónides; Rodríguez, Juan M

    2012-08-01

    Streptococcus salivarius is a commensal species commonly found in the human oropharyngeal tract. Some strains of this species have been developed for use as oral probiotics, while others have been associated with a variety of opportunistic human infections. Here, we report the complete sequence of strain PS4, which was isolated from breast milk of a healthy woman. PMID:22843595

  11. Complete Genome Sequence of Streptococcus salivarius PS4, a Strain Isolated from Human Milk

    PubMed Central

    Martín, Virginia; Maldonado-Barragán, Antonio; Jiménez, Esther; Ruas-Madiedo, Patricia; Fernández, Leónides

    2012-01-01

    Streptococcus salivarius is a commensal species commonly found in the human oropharyngeal tract. Some strains of this species have been developed for use as oral probiotics, while others have been associated with a variety of opportunistic human infections. Here, we report the complete sequence of strain PS4, which was isolated from breast milk of a healthy woman. PMID:22843595

  12. Equilibrium state and non-equilibrium steady state in an isolated human system

    NASA Astrophysics Data System (ADS)

    Zheng, Wen-Zhi; Liang, Yuan; Huang, Ji-Ping

    2014-02-01

    The principle of increasing entropy (PIE) is commonly considered as a universal physical law for natural systems. It also means that a non-equilibrium steady state (NESS) must not appear in any isolated natural systems. Here we experimentally investigate an isolated human social system with a clustering effect. We report that the PIE cannot always hold, and that NESSs can come to appear. Our study highlights the role of human adaptability in the PIE, and makes it possible to study human social systems by using some laws originating from traditional physics.

  13. Warty/basaloid penile intraepithelial neoplasia is more prevalent than differentiated penile intraepithelial neoplasia in nonendemic regions for penile cancer when compared with endemic areas: a comparative study between pathologic series from Paris and Paraguay.

    PubMed

    Soskin, Ana; Vieillefond, Anicke; Carlotti, Agnes; Plantier, Francoise; Chaux, Alcides; Ayala, Gustavo; Velazquez, Elsa F; Cubilla, Antonio L

    2012-02-01

    Penile squamous cell carcinoma shows an ample geographic variation in its prevalence with regions of low (North America, Europe, Japan, and Israel) and high (Africa, Asia, and South America) incidence. However, the geographic variation in the distribution of penile intraepithelial neoplasia is not well established. The aim of the present study was to compare the distribution of in situ and invasive lesions between geographic areas with low (France) and high (Paraguay) penile cancer incidence using a series of consecutive cases. The French series included 86 cases (57 in situ and 29 in situ + invasive squamous cell carcinoma), and the Paraguayan series, 117 cases (31 in situ and 86 in situ + invasive squamous cell carcinoma). Incidence of invasive squamous cell carcinoma in the overall samples was higher in the Paraguayan series (P < .00001). Comparing the Paraguayan and the French series, differentiated penile intraepithelial neoplasia was more prevalent in the former (65.0% versus 19.8%), whereas lesions showing warty and/or basaloid features predominated in the latter (35.0% versus 80.2%) to a significant level (P < .00001). This distinctive pattern of differential distribution was maintained when cases with associated invasive squamous cell carcinoma were excluded. The pattern of distribution of lichen sclerosus was also distinctive, with a significantly higher prevalence in the Paraguayan population when compared with the French series (32.5% versus 12.8%, P = .0015). In summary, there appears to be a distinctive distribution of penile precursor lesions depending on the geographic region in consideration. Penile intraepithelial neoplasia with warty and/or basaloid features predominated in low-incidence areas, whereas differentiated penile intraepithelial neoplasia was more prevalent in endemic regions for penile cancer. Further prospective studies in matched populations and from different geographic regions are needed to further clarify the reasons for this

  14. Tolerance of Bifidobacterium human isolates to bile, acid and oxygen.

    PubMed

    Andriantsoanirina, Valérie; Allano, Solène; Butel, Marie José; Aires, Julio

    2013-06-01

    Bifidobacteria are part of the human gastrointestinal microbiota and are used as probiotics in functional food products because of their health promoting properties. However, only few data are available on the phenotypic characteristics displayed by human bifidobacteria strain populations. In this study we compared the in vitro tolerance to acid, bile and oxygen of the largest number of independent human intestinal strains. Bile and acid tolerance varied among species and independent strains within a species: B. adolescentis strains were the most tolerant to bile followed by Bifidobacterium longum and B. breve; B. longum, B. breve and B. dentium showed the highest viability levels after exposure to acid pH. Oxygen tolerance was largely distributed among intestinal bifidobacteria: B. longum, B. breve and B. bifidum showed the highest oxygen tolerance. B. adolescentis showed the highest susceptibility to acid and oxygen stresses. The present study gave us the opportunity to update our knowledge about the phenotypic characteristics of human intestinal bifidobacteria. B. longum and B. breve harboured the best tolerance to oxygen, bile and acid stresses. Based on such biological characters, B. longum and B. breve species showed the highest interest in terms of potential selection of human probiotics.

  15. Characterization and biofilm forming ability of diarrhoeagenic enteroaggregative Escherichia coli isolates recovered from human infants and young animals.

    PubMed

    Vijay, Deepthi; Dhaka, Pankaj; Vergis, Jess; Negi, Mamta; Mohan, Vysakh; Kumar, Manesh; Doijad, Swapnil; Poharkar, Krupali; Kumar, Ashok; Malik, Satyaveer Singh; Barbuddhe, Sukhadeo Baliram; Rawool, Deepak B

    2015-02-01

    Enteroaggregative Escherichia coli (EAEC) is an important pathotype that causes infection in humans and animals. EAEC isolates (n=86) recovered from diarrhoeal cases in human infants (37) and young animals (49) were characterized as 'typical' and/or 'atypical' EAEC strains employing PCR for virulence associated genes (cvd432, aaiA, astA, pilS, irp2, ecp, pic, aggR, aafA, aggA, and agg3A). Besides, biofilm formation ability of human and animal EAEC isolates was assessed using microtiter plate assay. In addition, the transcriptional profile of biofilm associated genes (fis and ecp) was also evaluated and correlated with biofilm formation assay for few selected EAEC isolates of human and animal origins. Overall, a diverse virulence gene profile was observed for the EAEC isolates of human and animal origins as none of the EAEC isolates revealed the presence of all the genes that were targeted. Nine 'typical' EAEC isolates were identified (6 from humans and 3 from animals) while, the majority of the isolates were 'atypical' EAEC strains. Isolation and identification of three 'typical' EAEC isolates from animals (canines) appears to be the first report globally. Further, based on the observations of the biofilm formation assay, the study suggested that human EAEC isolates in particular were comparatively more biofilm producers than that of the animal EAEC isolates. The fis gene was highly expressed in majority of 'typical' EAEC isolates and the ecp gene in 'atypical' EAEC isolates.

  16. CRISPR Diversity in E. coli Isolates from Australian Animals, Humans and Environmental Waters

    PubMed Central

    Sheludchenko, Maxim S.; Huygens, Flavia; Stratton, Helen; Hargreaves, Megan

    2015-01-01

    Seventy four SNP genotypes and 54 E. coli genomes from kangaroo, Tasmanian devil, reptile, cattle, dog, horse, duck, bird, fish, rodent, human and environmental water sources were screened for the presence of the CRISPR 2.1 loci flanked by cas2 and iap genes. CRISPR 2.1 regions were found in 49% of the strains analysed. The majority of human E. coli isolates lacked the CRISPR 2.1 locus. We described 76 CRISPR 2.1 positive isolates originating from Australian animals and humans, which contained a total of 764 spacer sequences. CRISPR arrays demonstrated a long history of phage attacks especially in isolates from birds (up to 40 spacers). The most prevalent spacer (1.6%) was an ancient spacer found mainly in human, horse, duck, rodent, reptile and environmental water sources. The sequence of this spacer matched the intestinal P7 phage and the pO111 plasmid of E. coli. PMID:25946192

  17. Translational neuropharmacology: the use of human isolated gastrointestinal tissues.

    PubMed

    Sanger, G J; Broad, J; Kung, V; Knowles, C H

    2013-01-01

    Translational sciences increasingly emphasize the measurement of functions in native human tissues. However, such studies must confront variations in patient age, gender, genetic background and disease. Here, these are discussed with reference to neuromuscular and neurosecretory functions of the human gastrointestinal (GI) tract. Tissues are obtained after informed consent, in collaboration with surgeons (surgical techniques help minimize variables) and pathologists. Given the difficulties of directly recording from human myenteric neurones (embedded between muscle layers), enteric motor nerve functions are studied by measuring muscle contractions/relaxations evoked by electrical stimulation of intrinsic nerves; responses are regionally dependent, often involving cholinergic and nitrergic phenotypes. Enteric sensory functions can be studied by evoking the peristaltic reflex, involving enteric sensory and motor nerves, but this has rarely been achieved. As submucosal neurones are more accessible (after removing the mucosa), direct neuronal recordings are possible. Neurosecretory functions are studied by measuring changes in short-circuit current across the mucosa. For all experiments, basic questions must be addressed. Because tissues are from patients, what are the controls and the influence of disease? How long does it take before function fully recovers? What is the impact of age- and gender-related differences? What is the optimal sample size? Addressing these and other questions minimizes variability and raises the scientific credibility of human tissue research. Such studies also reduce animal use. Further, the many differences between animal and human GI functions also means that human tissue research must question the ethical validity of using strains of animals with unproved translational significance. PMID:22946540

  18. Genetic relatedness of Legionella longbeachae isolates from human and environmental sources in Australia

    SciTech Connect

    Lanser, J.A.; Doyle, R.; Sangster, N.; Steele, T.W. ); Adams, M. )

    1990-09-01

    The genetic relatedness of Legionella longbeachae isolated in Australia since 1987 was investigated by restriction fragment length polymorphism (RFLP) analysis and allozyme electrophoresis. Three radiolabeled probes were used in Southern hybridizations for the RFLP studies. They were Escherichia coli 16S and 23S rRNA and cloned fragments of L. longbeachae selected empirically from genomal banks in lambda and a cosmid. The legionellae included in the study comprised 11 Legionella longbeachae serogroup 1 organisms isolated form humans, 28 L. longbeachae serogroup 1 isolates from environmental sources, 3 L. longbeachae serogroup 2 environmental isolates. These were compared with the American Type Culture Collection reference strains of both serogroups and some other related Legionella species. Results of allozyme and RFLP analysis showed that all the isolates from humans and all but three of the environmental L. longbeachae serogroup 1 isolates were closely related. They were also closely related to L. longbeachae serogroup 1 ATCC 33462. There was wider variation among the three L. longbeachae serogroup 2 environmental isolates. One of these was closely related to L. longbeachae serogroup 2 ATCC 33484. RFLP studies with the rRNA probe provided the most discrimination among isolates but did not distinguish between the two serogroups.

  19. Contribution of Avian Salmonella enterica Isolates to Human Salmonellosis Cases in Constantine (Algeria).

    PubMed

    Elgroud, Rachid; Granier, Sophie A; Marault, Muriel; Kerouanton, Annaëlle; Lezzar, Abdesslem; Bouzitouna-Bentchouala, Chafia; Brisabois, Anne; Millemann, Yves

    2015-01-01

    An epidemiological investigation was carried out on one hundred Salmonella isolates from broiler farms, slaughterhouses, and human patients in the Constantine region of Algeria, in order to explore the contribution of avian strains to human salmonellosis cases in this region over the same period of time. The isolates were characterized by phenotypic as well as genotypic methods. A large variety of antimicrobial resistance profiles was found among human isolates, while only seven profiles were found among avian isolates. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR), Insertion Sequence 200-PCR (IS200-PCR), and Pulsed Field Gel Electrophoresis (PFGE) resulted in the allocation of the isolates to 16, 20, and 34 different profiles, respectively. The 3 genotyping methods led to complementary results by underlining the clonality of some serovars with the diffusion and persistence of a single clone in the Constantine area as well as stressing the polymorphism present in isolates belonging to other serovars, indicating the diversity of potential reservoirs of nontyphoidal Salmonella. Altogether, our results seem to indicate that nontyphoidal avian Salmonella may play an important role in human salmonellosis in the Constantine region.

  20. Contribution of Avian Salmonella enterica Isolates to Human Salmonellosis Cases in Constantine (Algeria)

    PubMed Central

    Elgroud, Rachid; Granier, Sophie A.; Marault, Muriel; Kerouanton, Annaëlle; Lezzar, Abdesslem; Bouzitouna-Bentchouala, Chafia; Brisabois, Anne; Millemann, Yves

    2015-01-01

    An epidemiological investigation was carried out on one hundred Salmonella isolates from broiler farms, slaughterhouses, and human patients in the Constantine region of Algeria, in order to explore the contribution of avian strains to human salmonellosis cases in this region over the same period of time. The isolates were characterized by phenotypic as well as genotypic methods. A large variety of antimicrobial resistance profiles was found among human isolates, while only seven profiles were found among avian isolates. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR), Insertion Sequence 200-PCR (IS200-PCR), and Pulsed Field Gel Electrophoresis (PFGE) resulted in the allocation of the isolates to 16, 20, and 34 different profiles, respectively. The 3 genotyping methods led to complementary results by underlining the clonality of some serovars with the diffusion and persistence of a single clone in the Constantine area as well as stressing the polymorphism present in isolates belonging to other serovars, indicating the diversity of potential reservoirs of nontyphoidal Salmonella. Altogether, our results seem to indicate that nontyphoidal avian Salmonella may play an important role in human salmonellosis in the Constantine region. PMID:26543858

  1. Molecular Characterization of Cryptosporidium Isolates Obtained from Humans in France

    PubMed Central

    Guyot, K.; Follet-Dumoulin, A.; Lelièvre, E.; Sarfati, C.; Rabodonirina, M.; Nevez, G.; Cailliez, J. C.; Camus, D.; Dei-Cas, E.

    2001-01-01

    Cryptosporidium parvum is usually considered the agent of human cryptosporidiosis. However, only in the last few years, molecular biology-based methods have allowed the identification of Cryptosporidium species and genotypes, and only a few data are available from France. In the present work, we collected samples of whole feces from 57 patients from France (11 immunocompetent patients, 35 human immunodeficiency virus [HIV]-infected patients, 11 immunocompromised but non-HIV-infected patients) in whom Cryptosporidium oocysts were recognized by clinical laboratories. A fragment of the Cryptosporidium 18S rRNA gene encompassing the hypervariable region was amplified by PCR and sequenced. The results revealed that the majority of the patients were infected with cattle (29 of 57) or human (18 of 57) genotypes of Cryptosporidium parvum. However, a number of immunocompromised patients were infected with C. meleagridis (3 of 57), C. felis (6 of 57), or a new genotype of C. muris (1 of 57). This is the first report of the last three species of Cryptosporidium in humans in France. These results indicate that immunocompromised individuals are susceptible to a wide range of Cryptosporidium species and genotypes. PMID:11574558

  2. The role of penile prosthetic surgery in the modern management of erectile dysfunction

    PubMed Central

    Jain, S; Bhojwani, A; Terry, T

    2000-01-01

    The management of erectile dysfunction looks set to be revolutionised with the introduction of effective oral therapies. There will remain, however, some men who do not respond to conservative measures. This article reviews the important role of penile prosthetic surgery as a treatment option in these patients.


Keywords: penile prosthesis; erectile dysfunction PMID:10622775

  3. Familial appearance of congenital penile curvature – case history of two brothers

    PubMed Central

    Matuszewski, Marcin; Krajka, Kazimierz; Rębała, Krzysztof

    2013-01-01

    The true prevalence of congenital penile curvature (CPC) is difficult to determine. Some study reports suggests that this problem may occur in as many as 10% of the male population [1]. However, a literature search of the Medline database revealed no reference concerning familial appearance of congenital penile curvature. For that reason we would like to present our case series. Two brothers aged 25 and 26 respectively were admitted to the department of urology due to congenital penile curvature. Each patient was assessed by a history, physical examination, auto-photography of the erect penis, and a thorough sexual history. Concomitant anomalies of penile layers were absent in both cases. The Yachia [2] and Essed-Schroeder [3] corporoplasty technique were applied respectively. In follow-up both brothers reported straight erections. A survey of the fetal penis at different stages of development shows some degree of curvature in a considerable number of embryos [4]. Penile curvature may thus be considered almost physiological in embryos between 35 and 45 mm in length. Thus, it has also been proposed that penile curvature is secondary to an arrest in normal penile development [5]. Therefore, some form of congenital local androgen deficiency may be responsible for inherited penile curvature. PMID:24579033

  4. Traumatic penile amputation in a 15-year-old boy presenting late in northwestern Nigeria.

    PubMed

    Musa, Muhammad Ujudud; Abdulmajid, Umar Farouk; Mashi, Sharfuddeen Abbas; Yunusa, Bashir

    2016-08-01

    Traumatic penile amputation is a serious urological emergency, although rare whenever it happens, there is a need to refer the patient early to urologist within 24 h, with the stump wrapped in saline; unfortunately, our patient presented late and as such could not benefit from penile reimplantation. PMID:27525085

  5. Characterization of hemolysins of Staphylococcus strains isolated from human and bovine, southern Iran.

    PubMed

    Moraveji, Z; Tabatabaei, M; Shirzad Aski, H; Khoshbakht, R

    2014-01-01

    The staphylococci are important pathogenic bacteria causing various infections in animals and human. Hemolysin is one of the virulence factors of coagulase-positive (CPS) and coagulase-negative staphylococci (CNS). The aims of the study were to characterize hemolysins of Staphylococcus spp. isolated from human and bovine origin, phenotypic- and genotypically. Characterization of hemolysin phenotypically based on hemolysis pattern of Staphylococcus spp. was done on the sheep, horse and rabbit blood agar plates. Genes encoding hemolysin were amplified with specific primers by using polymerase chain reaction (PCR) technique. Hemolytic activities phenotypically were determined in 60 and 90% of the total bovine and human isolates, respectively. All non hemolytic isolates were CNS (P≤0.05). In all isolates, hla and hld genes were determined by PCR amplification. None of the bovine and human isolates showed phenotypically and genotypically gamma hemolysin. The results from this study suggest that, in accordance with what is generally believed, some differences are apparent in hemolysin types among Staphylococcus strains of bovine and human origin. Furthermore, this study showed that CNS can be important as new pathogens.

  6. Characterization of hemolysins of Staphylococcus strains isolated from human and bovine, southern Iran

    PubMed Central

    Moraveji, Z; Tabatabaei, M; Shirzad Aski, H; Khoshbakht, R

    2014-01-01

    The staphylococci are important pathogenic bacteria causing various infections in animals and human. Hemolysin is one of the virulence factors of coagulase-positive (CPS) and coagulase-negative staphylococci (CNS). The aims of the study were to characterize hemolysins of Staphylococcus spp. isolated from human and bovine origin, phenotypic- and genotypically. Characterization of hemolysin phenotypically based on hemolysis pattern of Staphylococcus spp. was done on the sheep, horse and rabbit blood agar plates. Genes encoding hemolysin were amplified with specific primers by using polymerase chain reaction (PCR) technique. Hemolytic activities phenotypically were determined in 60 and 90% of the total bovine and human isolates, respectively. All non hemolytic isolates were CNS (P≤0.05). In all isolates, hla and hld genes were determined by PCR amplification. None of the bovine and human isolates showed phenotypically and genotypically gamma hemolysin. The results from this study suggest that, in accordance with what is generally believed, some differences are apparent in hemolysin types among Staphylococcus strains of bovine and human origin. Furthermore, this study showed that CNS can be important as new pathogens. PMID:27175125

  7. Multilocus Sequence Typing of Pathogenic Treponemes Isolated from Cloven-Hoofed Animals and Comparison to Treponemes Isolated from Humans

    PubMed Central

    Carter, Stuart D.; Birtles, Richard J.; Brown, Jennifer M.; Hart, C. Anthony; Evans, Nicholas J.

    2016-01-01

    ABSTRACT Treponema species are implicated in many diseases of humans and animals. Digital dermatitis (DD) treponemes are reported to cause severe lesions in cattle, sheep, pigs, goats, and wild elk, causing substantial global animal welfare issues and economic losses. The fastidiousness of these spirochetes has previously precluded studies investigating within-phylogroup genetic diversity. An archive of treponemes that we isolated enabled multilocus sequence typing to quantify the diversity and population structure of DD treponemes. Isolates (n = 121) were obtained from different animal hosts in nine countries on three continents. The analyses herein of currently isolated DD treponemes at seven housekeeping gene loci confirm the classification of the three previously designated phylogroups: the Treponema medium, Treponema phagedenis, and Treponema pedis phylogroups. Sequence analysis of seven DD treponeme housekeeping genes revealed a generally low level of diversity among the strains within each phylogroup, removing the need for the previously used “-like” suffix. Surprisingly, all isolates within each phylogroup clustered together, regardless of host or geographic origin, suggesting that the same sequence types (STs) can infect different animals. Some STs were derived from multiple animals from the same farm, highlighting probable within-farm transmissions. Several STs infected multiple hosts from similar geographic regions, identifying probable frequent between-host transmissions. Interestingly, T. pedis appears to be evolving more quickly than the T. medium or T. phagedenis DD treponeme phylogroup, by forming two unique ST complexes. The lack of phylogenetic discrimination between treponemes isolated from different hosts or geographic regions substantially contrasts with the data for other clinically relevant spirochetes. IMPORTANCE The recent expansion of the host range of digital dermatitis (DD) treponemes from cattle to sheep, goats, pigs, and wild elk

  8. Isolation and identification of Enterobacteriaceae from raw horsemeat intended for human consumption (Basashi).

    PubMed

    Furuhata, Katsunori; Ishizaki, Naoto; Sugi, Yutaka; Fukuyama, Masafumi

    2014-01-01

    The status of Enterobacteriaceae contamination was investigated in a total of 131 samples of raw horsemeat (Basashi) intended for human consumption purchased from a general meat shop or by mail-order from October 2012 to December 2013. The bacteria were isolated from 105 of the 112 samples (93.8%). Prominent differences in the isolation rate due to the place of manufacture/sale or by the cut of the meat were not observed. Moreover, in a comparison between domestic (92.6%) and imported (100%) samples, the isolation rate was slightly higher in the imported samples. When Enterobacteriaceae isolated from raw horsemeat was identified, it was highly diverse, with 14 species identified in total. From among these species, Hafnia alvei was the most common, with 33 strains (19.8%),followed by 27 strains (16.2%) of Klebsiella pneumoniae and 26 strains (15.6%) of Enterobacter cloacae, indicating that these three species were dominant. A trend was observed, with the dominant strain differing depending on the place of manufacture/sale or the cut of the meat. H. alvei was isolated at an especially high frequency from imported samples. An investigation was carried out regarding raw horsemeat intended for human consumption from Yamanashi Prefecture and Canada, regularly purchased from one store in Kanagawa Prefecture. Enterobacteriaceae were isolated during five of nine (55.6%) trials, in which the isolated bacteria were H. alvei, K. pneumoniae, etc. Moreover, they were isolated at a very high isolation rate of seven among 10 trials for the Canadian meat, and H. alvei was the most commonly isolated bacteria. Accordingly, when an investigation was carried out regarding the differences in the strain level in the six isolates of H. alvei periodically isolated from raw horsemeat from Canada by the pulsed-field gel electrophoresis (PFGE) pattern using a restriction enzyme, SfiI, there was a possibility that these were the same H-38 strain (November 2013) and H-64 strain (April 2014

  9. Molecular typing by pulsed-field gel electrophoresis of Spanish animal and human Listeria monocytogenes isolates.

    PubMed

    Vela, A I; Fernandez-Garayzabal, J F; Vazquez, J A; Latre, M V; Blanco, M M; Moreno, M A; de La Fuente, L; Marco, J; Franco, C; Cepeda, A; Rodriguez Moure, A A; Suarez, G; Dominguez, L

    2001-12-01

    A total of 153 strains of Listeria monocytogenes isolated from different sources (72 from sheep, 12 from cattle, 18 from feedstuffs, and 51 from humans) in Spain from 1989 to 2000 were characterized by pulsed-field gel electrophoresis. The strains of L. monocytogenes displayed 55 pulsotypes. The 84 animal, 51 human, and 18 feedstuff strains displayed 31, 29, and 7 different pulsotypes, respectively, indicating a great genetic diversity among the Spanish L. monocytogenes isolates studied. L. monocytogenes isolates from clinical samples and feedstuffs consumed by the diseased animals were analyzed in 21 flocks. In most cases, clinical strains from different animals of the same flock had identical pulsotypes, confirming the existence of a listeriosis outbreak. L. monocytogenes strains with pulsotypes identical to those of clinical strains were isolated from silage, potatoes, and maize stalks. This is the first study wherein potatoes and maize stalks are epidemiologically linked with clinical listeriosis.

  10. Mycobacterium bovis in Burkina Faso: Epidemiologic and Genetic Links between Human and Cattle Isolates

    PubMed Central

    Sanou, Adama; Tarnagda, Zekiba; Kanyala, Estelle; Zingué, Dezemon; Nouctara, Moumini; Ganamé, Zakaria; Combary, Adjima; Hien, Hervé; Dembele, Mathurin; Kabore, Antoinette; Meda, Nicolas; Van de Perre, Philippe; Neveu, Dorine

    2014-01-01

    Background In sub-Saharan Africa, bovine tuberculosis (bTB) is a potential hazard for animals and humans health. The goal of this study was to improve our understanding of bTB epidemiology in Burkina Faso and especially Mycobacterium bovis transmission within and between the bovine and human populations. Methodology/principal findings Twenty six M. bovis strains were isolated from 101 cattle carcasses with suspected bTB lesions during routine meat inspections at the Bobo Dioulasso and Ouagadougou slaughterhouses. In addition, 7 M. bovis strains were isolated from 576 patients with pulmonary tuberculosis. Spoligotyping, RDAf1 deletion and MIRU-VNTR typing were used for strains genotyping. The isolation of M. bovis strains was confirmed by spoligotyping and 12 spoligotype signatures were detected. Together, the spoligotyping and MIRU-VNTR data allowed grouping the 33 M. bovis isolates in seven clusters including isolates exclusively from cattle (5) or humans (1) or from both (1). Moreover, these data (genetic analyses and phenetic tree) showed that the M. bovis isolates belonged to the African 1 (Af1) clonal complex (81.8%) and the putative African 5 (Af5) clonal complex (18.2%), in agreement with the results of RDAf1 deletion typing. Conclusions/Significance This is the first detailed molecular characterization of M. bovis strains from humans and cattle in Burkina Faso. The distribution of the two Af1 and putative Af5 clonal complexes is comparable to what has been reported in neighbouring countries. Furthermore, the strain genetic profiles suggest that M. bovis circulates across the borders and that the Burkina Faso strains originate from different countries, but have a country-specific evolution. The genetic characterization suggests that, currently, M. bovis transmission occurs mainly between cattle, occasionally between cattle and humans and potentially between humans. This study emphasizes the bTB risk in cattle but also in humans and the difficulty to set up

  11. 5-HT4 receptors in isolated human corpus cavernosum?

    PubMed

    Hayes, E S; Adaikan, P G; Ratnam, S S; Ng, S C

    1999-08-01

    The novel serotonin subtype-4 (5-HT4) receptor agonist, SC53116 (SC), produced a limited relaxation of noradrenaline (NA) pre-contracted human corpus cavernosum (CC) smooth muscle in vitro. This effect was not significantly attenuated by the 5-HT4 antagonist SDZ250557 (SDZ). In the presence of (+/-) pindolol (1 microM) and methysergide (1 microM), employed to mask 5-HT1 and beta-adrenergic, and 5-HT2 receptors respectively, SC failed to relax NA pre-contracted CC strips to a greater extent than saline. Functional cAMP dependent relaxation pathways were demonstrated by a significant reduction in NA induced tone by prostaglandin E1 (PGE1) and isopropylnoradrenaline (IPNA), the action of the latter compound was effectively eliminated in the presence of (+/-) pindolol. Relaxation of NA induced tone caused by the nitric oxide donor nitro-glycerine (NTG) was significant and similar in the absence and presence of the 5-HT and beta-adrenergic antagonists. The results of this present study indicate that human corporal smooth muscle does not contain 5-HT4 receptors and that, although compounds like SC act to relax non-vascular smooth muscle via cAMP dependent mechanisms, 5-HT4 receptor agonists may be expected to be of limited utility in triggering cAMP dependent relaxation responses in human CC.

  12. Isolation and propagation of primary human and rodent embryonic neural progenitor cells and cortical neurons

    PubMed Central

    Darbinyan, Armine; Kaminski, Rafal; White, Martyn K; Darbinian, Nune; Khalili, Kamel

    2014-01-01

    Summary The research on human neural progenitor cells holds great potential for the understanding the molecular programs that control differentiation of cells of glial and neuronal lineages and pathogenetic mechanisms of neurological diseases. Stem cell technologies provide also opportunities for pharmaceutical industry to develop new approaches for regenerative medicine. Here we describe the protocol for isolation and maintenance of neural progenitor cells and cortical neurons using human fetal brain tissue. This protocol can be successfully adapted for preparation of rodent neural and oligodendrocyte progenitor cells. While several methods for isolation of neural and ologodendrocyte progenitors from rodent brain tissue have been described, including techniques which use gene transfer and magnetisc resonsnce beads, few methods are focused on derivation of human oligodendrocyte progenitor cells. Development of human culture provides the most physiologically relevent system for investigation of mechanisms which regulate function of oligodendrocyte, specifically of human origin. PMID:23975820

  13. Comparative study on human and bovine AT-SC isolation methods.

    PubMed

    Reshak, A H; Shahimin, M M; Buang, F

    2013-11-01

    Mammalian adipose tissue derived stem cells (AT-SC) have a tremendous potential in regenerative medicine for tissue engineering and somatic nuclear transfer (SNT). The isolation methods of human and bovine adipose tissue derived stem cells are compared in this paper to determine the feasibility and optimum method of isolation. The optimum isolation method will reduce the processing time, efforts and money as isolation is the first crucial and important step in stem cells research. Human abdominal subcutaneous adipose tissue and bovine abdominal subcutaneous adipose tissue are digested in three collagenase type 1 concentration 0.075%, 0.3% and 0.6% agitated at 1 h and 2 h under 37 °C in 5% CO2 incubator. The cultures are then morphologically characterised. Human adipose tissue stem cells are found to be best isolated using abdominal subcutaneous depot, using 0.075% collagenase type 1 agitated at 1 h under 37 °C in CO2 incubator. While bovine adipose tissue derived stem cells are best isolated using abdominal subcutaneous depot, using 0.6% collagenase type 1 agitated at 2 h under 37 °C in CO2 incubator.

  14. Bartonella Strains from Ground Squirrels Are Identical to Bartonella washoensis Isolated from a Human Patient

    PubMed Central

    Kosoy, Michael; Murray, Mike; Gilmore, Jr., Robert D.; Bai, Ying; Gage, Kenneth L.

    2003-01-01

    The most likely animal source of a human case of cardiac disease in Washoe County, Nev., was identified by comparison of DNA sequences of three genes (citrate synthase gltA, 60-kDa heat shock protein gene groEL, and 16S rRNA gene) of Bartonella washoensis cultured from the human patient in question and of Bartonella isolates obtained from the following Nevada rodents: Peromyscus maniculatus (17 isolates), Tamias minimus (11 isolates), Spermophilus lateralis (3 isolates), and Spermophilus beecheyi (7 isolates). Sequence analyses of gltA amplicons obtained from Bartonella from the rodents demonstrated considerable heterogeneity and resulted in the identification of 16 genetic variants that were clustered within three groups in phylogenetic analysis. Each of the three groups was associated with a rodent genus, Peromyscus, Tamias, or Spermophilus. The gltA, 16S rRNA gene, and groEL sequences of a Bartonella isolate obtained from a California ground squirrel (S. beecheyi) were completely identical to homologous sequences of B. washoensis, strongly suggesting that these animals were the source of infection in the human case. PMID:12574261

  15. Collection, isolation, and flow cytometric analysis of human endocervical samples.

    PubMed

    Juno, Jennifer A; Boily-Larouche, Genevieve; Lajoie, Julie; Fowke, Keith R

    2014-07-06

    Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female genital tract (FGT). Because heterosexual transmission now represents the dominant mechanism of HIV transmission, and given the continual spread of sexually transmitted infections (STIs), it is critical to understand the interplay between host and pathogen at the genital mucosa. The substantial gaps in knowledge around FGT immunity are partially due to the difficulty in successfully collecting and processing mucosal samples. In order to facilitate studies with sufficient sample size, collection techniques must be minimally invasive and efficient. To this end, a protocol for the collection of cervical cytobrush samples and subsequent isolation of cervical mononuclear cells (CMC) has been optimized. Using ex vivo flow cytometry-based immunophenotyping, it is possible to accurately and reliably quantify CMC lymphocyte/monocyte population frequencies and phenotypes. This technique can be coupled with the collection of cervical-vaginal lavage (CVL), which contains soluble immune mediators including cytokines, chemokines and anti-proteases, all of which can be used to determine the anti- or pro-inflammatory environment in the vagina.

  16. Human time perception in temporal isolation: effects of illumination intensity.

    PubMed

    Aschoff, J; Daan, S

    1997-11-01

    Living in isolation from time cues under relatively high and low light intensities for a total (on average) of 24 days, 18 subjects estimated the passage of time by "producing" short (10 to 120 seconds) and long (1h) intervals throughout the experiments. The 1h productions were independent of light intensity and highly positively correlated with the duration of wake times. The short-interval productions were markedly increased under high light intensity. In a subsample of 6 subjects, the interaction between effects of body temperature and light condition on 10-second production was analyzed. Productions were negatively correlated with body temperature. In both dim and bright light, productions decreased by a factor of 0.7 per degree C. In bright light, production was increased by a factor of 1.2 relative to dim light. This effect was not mediated by body temperature, which itself was on average slightly increased in bright light. Since subjective time is slowed by bright light, objective time seems to pass faster in bright light.

  17. Collection, isolation, and flow cytometric analysis of human endocervical samples.

    PubMed

    Juno, Jennifer A; Boily-Larouche, Genevieve; Lajoie, Julie; Fowke, Keith R

    2014-01-01

    Despite the public health importance of mucosal pathogens (including HIV), relatively little is known about mucosal immunity, particularly at the female genital tract (FGT). Because heterosexual transmission now represents the dominant mechanism of HIV transmission, and given the continual spread of sexually transmitted infections (STIs), it is critical to understand the interplay between host and pathogen at the genital mucosa. The substantial gaps in knowledge around FGT immunity are partially due to the difficulty in successfully collecting and processing mucosal samples. In order to facilitate studies with sufficient sample size, collection techniques must be minimally invasive and efficient. To this end, a protocol for the collection of cervical cytobrush samples and subsequent isolation of cervical mononuclear cells (CMC) has been optimized. Using ex vivo flow cytometry-based immunophenotyping, it is possible to accurately and reliably quantify CMC lymphocyte/monocyte population frequencies and phenotypes. This technique can be coupled with the collection of cervical-vaginal lavage (CVL), which contains soluble immune mediators including cytokines, chemokines and anti-proteases, all of which can be used to determine the anti- or pro-inflammatory environment in the vagina. PMID:25045942

  18. Prospective investigation of penile length with newborn male circumcision and second to fourth digit ratio

    PubMed Central

    Park, Jong Kwan; Doo, A. Ram; Kim, Joo Heung; Park, Hyung Sub; Do, Jung Mo; Choi, Hwang; Park, Seung Chol; Kim, Myung Ki; Jeong, Young Beom; Kim, Hyung Jim; Kim, Young Gon; Shin, Yu Seob

    2016-01-01

    Introduction: We prospectively investigated the relationship between newborn male circumcision (NMC) and second to fourth digit ratio with penile length. Methods: As participants for our study, we identified already circumcised young patients who visited our hospital for urological treatment. The age at which the circumcision had been done was assessed. The patients’ height and weight were measured. Second to fourth digit ratio was calculated by measuring the second and fourth digit lengths. The flaccid and erectile penile lengths were measured from the base of the penis to the tip of the glans in standing position. Results: A total of 248 patients were included in our study. In univariate analysis, height, second to fourth digit ratio, flaccid penile length, and age of circumcision were associated with erectile penile length. Among these variables, second to fourth digit ratio, flaccid penile length, and age of circumcision were significant predictive factors for erectile penile length in multivariate analysis. The subjects were divided into two groups, including 72 patients in the NMC group and 176 patients in the non-NMC group. No significant difference was found in height, weight, and second to fourth digit ratio between both groups. However, flaccid (p<0.001) and erectile (p=0.001) penile lengths were shorter in the NMC group than in the non-NMC group. Conclusions: Despite the small number of subjects, this study shows that NMC was associated with shorter penile length. Second to fourth digit ratio, flaccid penile length, and age of circumcision were also significant predictive factors for erectile penile length. Further multicentre studies with larger number of subjects and biochemical analyses are needed for potential clinical applicability. PMID:27695583

  19. Prospective investigation of penile length with newborn male circumcision and second to fourth digit ratio

    PubMed Central

    Park, Jong Kwan; Doo, A. Ram; Kim, Joo Heung; Park, Hyung Sub; Do, Jung Mo; Choi, Hwang; Park, Seung Chol; Kim, Myung Ki; Jeong, Young Beom; Kim, Hyung Jim; Kim, Young Gon; Shin, Yu Seob

    2016-01-01

    Introduction: We prospectively investigated the relationship between newborn male circumcision (NMC) and second to fourth digit ratio with penile length. Methods: As participants for our study, we identified already circumcised young patients who visited our hospital for urological treatment. The age at which the circumcision had been done was assessed. The patients’ height and weight were measured. Second to fourth digit ratio was calculated by measuring the second and fourth digit lengths. The flaccid and erectile penile lengths were measured from the base of the penis to the tip of the glans in standing position. Results: A total of 248 patients were included in our study. In univariate analysis, height, second to fourth digit ratio, flaccid penile length, and age of circumcision were associated with erectile penile length. Among these variables, second to fourth digit ratio, flaccid penile length, and age of circumcision were significant predictive factors for erectile penile length in multivariate analysis. The subjects were divided into two groups, including 72 patients in the NMC group and 176 patients in the non-NMC group. No significant difference was found in height, weight, and second to fourth digit ratio between both groups. However, flaccid (p<0.001) and erectile (p=0.001) penile lengths were shorter in the NMC group than in the non-NMC group. Conclusions: Despite the small number of subjects, this study shows that NMC was associated with shorter penile length. Second to fourth digit ratio, flaccid penile length, and age of circumcision were also significant predictive factors for erectile penile length. Further multicentre studies with larger number of subjects and biochemical analyses are needed for potential clinical applicability.

  20. Isolation of Human Amnion Epithelial Cells According to Current Good Manufacturing Procedures.

    PubMed

    Gramignoli, Roberto; Srinivasan, Raghuraman C; Kannisto, Kristina; Strom, Stephen C

    2016-05-12

    Different cell types can be isolated from human placental tissues, and some have been reported to retain phenotypic plasticity and characteristics that make them a promising source of cells for regenerative medicine. Among these are human amnion epithelial cells (hAECs). Adoption of current good manufacturing practices (cGMP) and enhanced quality control is essential when isolating hAECs in order to deliver a safe and effective cellular product for clinical purposes. This unit describes a detailed protocol for selective isolation of hAECs from human term placenta with little to no contamination by other cell types. A method for characterizing the heterogeneity of the hAEC suspension is also provided. The resulting cell product will be useful for clinical as well as basic research applications. © 2016 by John Wiley & Sons, Inc.

  1. Isolation and identification of bat viruses closely related to human, porcine and mink orthoreoviruses.

    PubMed

    Yang, Xing-Lou; Tan, Bing; Wang, Bo; Li, Wen; Wang, Ning; Luo, Chu-Ming; Wang, Mei-Niang; Zhang, Wei; Li, Bei; Peng, Cheng; Ge, Xing-Yi; Zhang, Li-Biao; Shi, Zheng-Li

    2015-12-01

    Bats have been identified as natural reservoirs of many viruses, including reoviruses. Recent studies have demonstrated the interspecies transmission of bat reoviruses to humans. In this study, we report the isolation and molecular characterization of six strains of mammalian orthoreovirus (MRV) from Hipposideros and Myotis spp. These isolates were grouped into MRV serotype 1, 2 or 3 based on the sequences of the S1 gene, which encodes the outer coat protein s1. Importantly, we found that three of six bat MRV strains shared high similarity with MRVs isolated from diseased minks, piglets or humans based on the S1 segment, suggesting that interspecies transmission has occurred between bats and humans or animals. Phylogenetic analyses based on the 10 segments showed that the genomic segments of these bat MRVs had different evolution lineages, suggesting that these bat MRVs may have arisen through reassortment of MRVs of different origins. PMID:26475793

  2. Isolation of Aeromonas salmonicida from Human Blood Sample: A Case Report.

    PubMed

    Tewari, Rachna; Dudeja, Mridu; Nandy, Shyamasree; Das, Ayan Kumar

    2014-02-01

    Aeromonas salmonicida belonging to the genus Aeromonas, is a common pathogen that causes furunculosis and septicaemia in variety of fishes. It infects cold blooded vertebrates living at low temperatures mainly salmonid fish hence named salmonicida. Untill recently Aeromanas salmonicida is considered to be a fish pathogen. A. salmonicida is considered to be non-pathogenic for humans as it cannot grow at 37ºC. "However, In our laboratory culture plates and broths were incubated twice at 37ºC and each time same type of colonies were isolated which were identified as A. samonicida by Vitek 2 compact system bioMerieux, Inc. (Durham, N.C.)". By far no report has been received regarding its isolation from humans biological sample. Here we present the first report of A. salmonicida isolated from the human blood. PMID:24701507

  3. Isolation of Human Amnion Epithelial Cells According to Current Good Manufacturing Procedures.

    PubMed

    Gramignoli, Roberto; Srinivasan, Raghuraman C; Kannisto, Kristina; Strom, Stephen C

    2016-01-01

    Different cell types can be isolated from human placental tissues, and some have been reported to retain phenotypic plasticity and characteristics that make them a promising source of cells for regenerative medicine. Among these are human amnion epithelial cells (hAECs). Adoption of current good manufacturing practices (cGMP) and enhanced quality control is essential when isolating hAECs in order to deliver a safe and effective cellular product for clinical purposes. This unit describes a detailed protocol for selective isolation of hAECs from human term placenta with little to no contamination by other cell types. A method for characterizing the heterogeneity of the hAEC suspension is also provided. The resulting cell product will be useful for clinical as well as basic research applications. © 2016 by John Wiley & Sons, Inc. PMID:27171794

  4. Isolation of Bacteroides from fish and human fecal samples for identification of unique molecular markers.

    PubMed

    Kabiri, Leila; Alum, Absar; Rock, Channah; McLain, Jean E; Abbaszadegan, Morteza

    2013-12-01

    Bacteroides molecular markers have been used to identify human fecal contamination in natural waters, but recent work in our laboratory confirmed cross-amplification of several human-specific Bacteroides spp. assays with fecal DNA from fish. For identification of unique molecular markers, Bacteroides from human (n = 4) and fish (n = 7) fecal samples were cultured and their identities were further confirmed using Rapid ID 32A API strips. The 16S rDNA from multiple isolates from each sample was PCR amplified, cloned, and sequenced to identify unique markers for development of more stringent human-specific assays. In human feces, Bacteroides vulgatus was the dominant species (75% of isolates), whereas in tilapia feces, Bacteroides eggerthii was dominant (66%). Bacteroides from grass carp, channel catfish, and blue catfish may include Bacteroides uniformis, Bacteroides ovatus, or Bacteroides stercoris. Phylogenic analyses of the 16S rRNA gene sequences showed distinct Bacteroides groupings from each fish species, while human sequences clustered with known B. vulgatus. None of the fish isolates showed significant similarity to Bacteroides sequences currently deposited in NCBI (National Center for Biotechnology Information). This study expands the current sequence database of cultured fish Bacteroides. Such data are essential for identification of unique molecular markers in human Bacteroides that can be utilized in differentiating fish and human fecal contamination in water samples.

  5. Epidemiology and public health significance of Cryptosporidium isolated from cattle, buffaloes, and humans in Egypt.

    PubMed

    Ibrahim, M A; Abdel-Ghany, A E; Abdel-Latef, G K; Abdel-Aziz, S A; Aboelhadid, S M

    2016-06-01

    The epidemiology and public health significance of Cryptosporidium species and genotypes were investigated in Beni-Suef Governorate, Egypt. A total of 610 animal fecal samples (480 from cattle and 130 from buffaloes) beside 290 stool samples from humans were collected in the period between January and December 2014. Based on the microscopic examination, the overall estimated prevalence of Cryptosporidium spp. in cattle, buffaloes, and humans was 10.2, 12.3, and 19 %, respectively. The highest detection rates were in calves less than 2 months of age (17.1 %) and diarrheic animals (13.0 %). Likewise in humans, the highest prevalence of Cryptosporidium was in infants (31.3 %) and diarrheic individuals (21.1 %). The gender distribution in humans denoted that Cryptosporidium was reported more frequently in males (21.7 %) than females (14.5 %). Based on the molecular characterization of Cryptosporidium, Cryptosporidium oocyst wall protein (COWP) and gp60 genes were successfully amplified in 36 out of 50 samples subjected to genotyping. Restriction fragment length polymorphism (RFLP) analysis of the COWP fragments revealed that Cryptosporidium parvum was the only species detected in cattle (12 isolates) and buffaloes (4 isolates), while in humans, the detected species were Cryptosporidium hominis (15 isolates) and C. parvum (5 isolates). Sequence analysis of the gp60 gene identified the subtype IIdA20G1 within C. parvum isolated from both animals and humans. The common occurrence of zoonotic subtypes of C. parvum in cattle and buffaloes highlights the potential role of these animals as significant reservoirs of infection to humans. Also, the presence of C. hominis and C. parvum in humans indicates that both anthroponotic and zoonotic pathways are expected.

  6. Epidemiology and public health significance of Cryptosporidium isolated from cattle, buffaloes, and humans in Egypt.

    PubMed

    Ibrahim, M A; Abdel-Ghany, A E; Abdel-Latef, G K; Abdel-Aziz, S A; Aboelhadid, S M

    2016-06-01

    The epidemiology and public health significance of Cryptosporidium species and genotypes were investigated in Beni-Suef Governorate, Egypt. A total of 610 animal fecal samples (480 from cattle and 130 from buffaloes) beside 290 stool samples from humans were collected in the period between January and December 2014. Based on the microscopic examination, the overall estimated prevalence of Cryptosporidium spp. in cattle, buffaloes, and humans was 10.2, 12.3, and 19 %, respectively. The highest detection rates were in calves less than 2 months of age (17.1 %) and diarrheic animals (13.0 %). Likewise in humans, the highest prevalence of Cryptosporidium was in infants (31.3 %) and diarrheic individuals (21.1 %). The gender distribution in humans denoted that Cryptosporidium was reported more frequently in males (21.7 %) than females (14.5 %). Based on the molecular characterization of Cryptosporidium, Cryptosporidium oocyst wall protein (COWP) and gp60 genes were successfully amplified in 36 out of 50 samples subjected to genotyping. Restriction fragment length polymorphism (RFLP) analysis of the COWP fragments revealed that Cryptosporidium parvum was the only species detected in cattle (12 isolates) and buffaloes (4 isolates), while in humans, the detected species were Cryptosporidium hominis (15 isolates) and C. parvum (5 isolates). Sequence analysis of the gp60 gene identified the subtype IIdA20G1 within C. parvum isolated from both animals and humans. The common occurrence of zoonotic subtypes of C. parvum in cattle and buffaloes highlights the potential role of these animals as significant reservoirs of infection to humans. Also, the presence of C. hominis and C. parvum in humans indicates that both anthroponotic and zoonotic pathways are expected. PMID:27044415

  7. Differences in biofilm formation and virulence factors between clinical and fecal enterococcal isolates of human and animal origin.

    PubMed

    Tsikrikonis, Giorgos; Maniatis, Antonios N; Labrou, Maria; Ntokou, Eleni; Michail, Giorgos; Daponte, Alexandros; Stathopoulos, Constantinos; Tsakris, Athanassios; Pournaras, Spyros

    2012-06-01

    The present study investigated the possible correlation between carriage of the virulence genes esp and fsrb, production of hemolysin and gelatinase and biofilm formation in human vs. animal enterococcal isolates. A collection of 219 enterococcal isolates recovered from clinical and fecal surveillance samples of hospitalized patients and 132 isolates from animal feces were studied. Isolates were tested for hemolysin and gelatinase phenotypically and for quantitative biofilm production by a microtitre method. Genes esp and fsrb were detected by PCR. Human Enterococcus faecium and Enterococcus faecalis isolates from both surveillance and clinical samples produced biofilm significantly more often than animal isolates (P < 0.0001 for both species). The quantity of biofilm did not differ significantly between human and animal isolates, while was significantly higher in esp-positive compared with esp-negative human E. faecium isolates (P < 0.0001). The frequency of esp gene carriage was significantly higher in human compared with animal E. faecium and E. faecalis isolates (P < 0.0001). The gene fsrb was detected significantly more often in animal than human E. faecium isolates (P 0.004). Hemolysin production was significantly more common in human clinical compared with animal E. faecalis isolates (P < 0.0001). Similar proportions of animal and human E. faecalis produced gelatinase, which was significantly correlated with the presence of fsrb gene (P < 0.0001) in both human clinical and animal E. faecalis isolates. The hemolysin trait did not exhibit any correlation with the presence of esp and fsrb genes, but appeared to be linked with enhanced quantity of biofilm production in both human clinical and animal E. faecalis isolates. Production of gelatinase was associated with the proportion and the degree of biofilm production mainly in animal E. faecalis isolates.

  8. Genetic stability of pneumococcal isolates during 35 days of human experimental carriage

    PubMed Central

    Gladstone, R.A.; Gritzfeld, J.F.; Coupland, P.; Gordon, S.B.; Bentley, S.D.

    2015-01-01

    Background Pneumococcal carriage is a reservoir for transmission and a precursor to pneumococcal disease. The experimental human pneumococcal carriage model provides a useful tool to aid vaccine licensure through the measurement of vaccine efficacy against carriage (VEcol). Documentation of the genetic stability of the experimental human pneumococcal carriage model is important to further strengthen confidence in its safety and conclusions, enabling it to further facilitate vaccine licensure through providing evidence of VEcol. Methods 229 isolates were sequenced from 10 volunteers in whom experimental human pneumococcal carriage was established, sampled over a period of 35 days. Multiple isolates from within a single volunteer at a single time provided a deep resolution for detecting variation. HiSeq data from the isolates were mapped against a PacBio reference of the inoculum to call variable sites. Results The observed variation between experimental carriage isolates was minimal with the maximum SNP distance between any isolate and the reference being 3 SNPs. Conclusion The low-level variation described provides evidence for the stability of the experimental human pneumococcal carriage model over 35 days, which can be reliably and confidently used to measure VEcol and aid future progression of pneumococcal vaccination. PMID:26006086

  9. Comparison of Campylobacter jejuni pulsotypes isolated from humans and poultry in Split and Dalmatia County, Croatia.

    PubMed

    Kovačić, Ana; Carev, Merica; Tripković, Ingrid; Srečec, Siniša; Siško-Kraljević, Katarina

    2015-01-01

    Consumption of poultry is considered to be an important source of human infection with Campylobacter. In the period from 2008 to 2010, 50 isolates of Campylobacter jejuni from human faeces were analysed and compared with 61 isolates from poultry by pulsed field gel electrophoresis using SmaI and KpnI. Based on the analysis of SmaI macrorestriction profiles, 86 isolates (77.5 %) were assigned to 15 S clusters: 31 (62 %) from humans and 55 from poultry (90.2 %). Altogether 21 isolates (19 %) exhibited macrorestriction profiles common to both humans and poultry after restriction with SmaI and KpnI. A total of five identical pulsotypes were isolated from both poultry and patients and one of them appeared in eight different locations in the time interval of one year. These results indicate that poultry could be an important source of Campylobacter infection in Split and Dalmatia County which is the biggest County in Croatia and the most important tourist destination.

  10. Genotypic characterization of Staphylococcus aureus isolated from bovines, humans, and food in Indonesia

    PubMed Central

    Tato, Syarifudin; Sugiyono, Ngalijan; Ariyanti, Dwi; Prabawati, Feny

    2011-01-01

    The present study determined the genetic relationships between 41 Staphyloccocus (S.) aureus isolates from bovines, humans, and food using a single enzyme amplified fragment length polymorphism (AFLP) technique. We evaluated the prevalence of staphylococcal enterotoxin (SE) genes and other virulence gene determinants by PCR. The identification of S. aureus was based on culturing and biochemical tests, and by amplifying a specific section of the 23S rRNA gene. PCR amplification of the SE genes (sea, seb, sec, see, seg, seh, and sei) singly or in combination was observed. Most isolates of bovine origin harbored hla (84%) and cap5 (74%), while most isolates from humans harbored hla (73%), cap8 (91%), and fnbA (100%). Strains from food sources were positive for hla (100%), cap5 (100%), and cap8 (64%) unlike isolates from humans or bovines. A single enzyme AFLP analysis revealed a correlation between AFLP clusters of some strains and the source of the isolates The genotypic results of the present study might help to better understand the distribution of prevalent S. aureus clones among humans, bovines, and food and will help control S. aureus infections in Indonesia. PMID:22127020

  11. Fusobacterium hwasookii sp. nov., Isolated from a Human Periodontitis Lesion.

    PubMed

    Cho, Eugene; Park, Soon-Nang; Lim, Yun Kyong; Shin, Yeseul; Paek, Jayoung; Hwang, Cheol Ho; Chang, Young-Hyo; Kook, Joong-Ki

    2015-02-01

    In this study, we classified the five strains (ChDC F128(T), ChDC F145, ChDC F174, ChDC F206, and ChDC F300) as a novel species of genus Fusobacterium by DNA-DNA hybridization and multi-locus phylogenetic analysis (MLPA), based on a single sequence (24,715 bp) of 22 concatenated housekeeping genes, with morphological and chemotaxonomic characteristics. DNA-DNA hybridization data showed that the values of genomic relatedness between ChDC F128(T) and each of the other novel strains were ranged from 79.0 to 82.6 %, while those of genomic relatedness between ChDC F128(T) and type strain of each of subspecies of F. nucleatum or Fusobacterium periodonticum were ranged from 40.9 to 54.4 %. MLPA revealed that the 5 strains were clustered as one group and clearly discriminated with F. nucleatum and F. periodonticum with 100 % bootstrap value. The DNA G+C content of the five novel strains were ranged from 26.9 to 27.0 mol%. The cellular fatty acid analysis of clinical isolates and type strains revealed C14:0, C16:0, and cis-9 C16:1 as the major fatty acids. The cell wall peptidoglycan of the 5 strains was comprised of meso-lanthionine. These results show that the 5 strains are novel species and belong to the genus Fusobacterium. Strain ChDC F128(T) (=KCOM 1249(T) = KCTC 5108(T) = JCM 30218(T)) is suggested to be the type strain of a novel species of genus Fusobacterium, for which the name Fusobacterium hwasookii sp. nov. is proposed.

  12. Fusobacterium hwasookii sp. nov., Isolated from a Human Periodontitis Lesion.

    PubMed

    Cho, Eugene; Park, Soon-Nang; Lim, Yun Kyong; Shin, Yeseul; Paek, Jayoung; Hwang, Cheol Ho; Chang, Young-Hyo; Kook, Joong-Ki

    2015-02-01

    In this study, we classified the five strains (ChDC F128(T), ChDC F145, ChDC F174, ChDC F206, and ChDC F300) as a novel species of genus Fusobacterium by DNA-DNA hybridization and multi-locus phylogenetic analysis (MLPA), based on a single sequence (24,715 bp) of 22 concatenated housekeeping genes, with morphological and chemotaxonomic characteristics. DNA-DNA hybridization data showed that the values of genomic relatedness between ChDC F128(T) and each of the other novel strains were ranged from 79.0 to 82.6 %, while those of genomic relatedness between ChDC F128(T) and type strain of each of subspecies of F. nucleatum or Fusobacterium periodonticum were ranged from 40.9 to 54.4 %. MLPA revealed that the 5 strains were clustered as one group and clearly discriminated with F. nucleatum and F. periodonticum with 100 % bootstrap value. The DNA G+C content of the five novel strains were ranged from 26.9 to 27.0 mol%. The cellular fatty acid analysis of clinical isolates and type strains revealed C14:0, C16:0, and cis-9 C16:1 as the major fatty acids. The cell wall peptidoglycan of the 5 strains was comprised of meso-lanthionine. These results show that the 5 strains are novel species and belong to the genus Fusobacterium. Strain ChDC F128(T) (=KCOM 1249(T) = KCTC 5108(T) = JCM 30218(T)) is suggested to be the type strain of a novel species of genus Fusobacterium, for which the name Fusobacterium hwasookii sp. nov. is proposed. PMID:25257648

  13. Orandi flap for penile urethral stricture: Polishing the gold standard

    PubMed Central

    Goel, Apul; Kumar, Manoj; Singh, Manmeet

    2015-01-01

    Introduction: We describe the combined use of the Orandi flap and the scrotal skin advancement flap to reduce complications for pendulous urethral stricture in men >40 years old. Methods: Over the last 40 months, 10 men underwent urethroplasty for pendulous stricture by the modified Orandi urethroplasty. In this, additionally, a midline hairless scrotal skin flap of the size of the ventral skin defect on the pendulous portion was raised based on the dartos fascia. This flap was mobilized so that it reached the pendulous portion without tension and covered the penile defect. The catheter was removed after 4 weeks. Patients were followed every 3 months using uroflowmetry and the American Urological Association (AUA) symptom score. Results: The mean age was 55.5. Of the 10 patients, the etiology was post-catheterization in 5 and idiopathic in the remaining 5. Three men also had stricture extending into the bulbous urethra (repaired using buccal graft). The mean additional time needed for the flap coverage was 36.2 minutes (range: 30–45). The median follow-up was 12 months (range: 3–40). The mean postoperative symptom score was 5.2 and the mean flow rate was 20.1 mL/sec. In 2 men, the meatus got retracted to the distal penile part (probably due to downward traction by scrotal skin). No patient complained of disfigurement. Two men reported recurrence (1 each in bulbous and penile urethra). The limitations are small number of patients and the observational nature of this study. Conclusions: The intermediate-term results show that the modified Orandi urethroplasty is an acceptable treatment option with acceptable cosmetic results. PMID:26085873

  14. Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles.

    PubMed

    Yang, Ruifeng; Xu, Xiaowei

    2016-01-01

    The hair follicle undergoes lifelong cycling and growth. Previous studies have been focused on epithelial stem cells in the hair follicles. Neural crest stem cells (NCSCs) are pluripotent cells that can persist in adult tissues. We have previously demonstrated that human NCSCs can be isolated from hair follicles. Here, we present a protocol to isolate NCSCs from human hair follicles based on their specific surface-marker expression of CD271/HNK1 or CD271/CD49D (alpha4 integrin). NCSCs can be expanded in the culture as neural spheres or attached cells.

  15. Isolation of dengue virus with a human promonocyte cell line.

    PubMed

    Liu, W T; Chen, C L; Lee, S S; Chan, C C; Lo, F L; Ko, Y C

    1991-05-01

    In October-November, 1988 there was an outbreak of dengue fever in the Kaoshiung area of southern Taiwan. We collected 100 serum samples from 96 patients at the onset of their fever for virus cultures and identification. A human promonocyte cell line (HL-CZ) established in our laboratory was used and proved to be susceptible for dengue virus propagation. Type 1 dengue virus in the HL-CZ cell culture was identified by immunofluorescence tests using monoclonal antibodies, and also by hemagglutination tests with goose red blood cells. The density of the virus particles, as measured by sucrose gradient ultracentrifugation, ranged from 1.186 to 1.224 g/ml. The virus yield from this cell culture is comparable with that from the C6/36 mosquito cell line. There was a significant correlation between the antibody responses tested with Western dot blots and hemagglutination inhibition techniques.

  16. [Isolation and properties of 2 kallikreins from human saliva].

    PubMed

    Vovchuk, S V; Barabash, R D; Levitsky, A P

    1975-01-01

    Two kallikreins (K-I and K-II) were purified from mixed, parotid and submandibular human saliva. The kallikreins were separated by chromatography on CM-cellulose. The pH optima of activity were at pH 9.3 and pH 9.6-9.8; Km for BAEE was 1.10-3M and 4.10-3M, respectively. The esterase activity of K-I and K-II was inhibited by trasylol-like inhibitors, while the plant and synthetic inhibitors of trypsin were uneffective. In dog, rabbit and rat the hypotensive effect of K-II and its action upon the permeability of rabbit skin was 4-5 fold higher than the effects of K-I. Ki and K-II did not alter the arterial blood pressure in guinea pig. PMID:3025

  17. A material isolated from human hands that attracts female mosquitoes.

    PubMed

    Schreck, C E; Smith, N; Carlson, D A; Price, G D; Haile, D; Godwin, D R

    1982-02-01

    The residue left on glass surfaces by human hands was found to be attractive to femaleAedes aegypti (L.) andAnopheles quadrimaculatus Say mosquitoes. The material lost half of its activity in 1 hr. A solvent wash technique was developed to recover and concentrate the residuum from handled glass beads. The residuum could be recovered effectively with absolute ethanol and less effectively with several other solvents. More mosquitoes were attracted to heated than to unheated residuum, an indication of its volatility. Also, attraction of the residuum decreased with decreasing concentration or dose. Concentrated residuum collections, stored under refrigeration and tested for longevity, showed no appreciable loss of attractiveness up to 60 days of storage.

  18. Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program.

    PubMed

    Walker, Douglas G; Whetzel, Alexis M; Serrano, Geidy; Sue, Lucia I; Lue, Lih-Fen; Beach, Thomas G

    2016-09-01

    Many factors affect the integrity of messenger RNA from human autopsy tissues including postmortem interval (PMI) between death and tissue preservation and the pre-mortem agonal and disease states. In this communication, we describe RNA isolation and characterization of 389 samples from 18 different tissues from elderly donors who were participants in a rapid whole-body autopsy program located in Sun City, Arizona ( www.brainandbodydonationprogram.org ). Most tissues were collected within a PMI of 2-6 h (median 3.15 h; N = 455), but for this study, tissue from cases with longer PMIs (1.25-29.25 h) were included. RNA quality was assessed by RNA integrity number (RIN) and total yield (ng RNA/mg tissue). RIN correlated with PMI for heart (r = -0.531, p = 0.009) and liver (r = -558, p = 0.0017), while RNA yield correlated with PMI for colon (r = -485, p = 0.016) and skin (r = -0.460, p = 0.031). RNAs with the lowest integrity were from skin and cervix where 22.7 and 31.4 % of samples respectively failed to produce intact RNA; by contrast all samples from esophagus, lymph node, jejunum, lung, stomach, submandibular gland and kidney produced RNA with measurable RINs. Expression levels in heart RNA of 4 common housekeeping normalization genes showed significant correlations of Ct values with RIN, but only one gene, glyceraldehyde-3 phosphate dehydrogenase, showed a correlation of Ct with PMI. There were no correlations between RIN values obtained for liver, adrenal, cervix, esophagus and lymph node and those obtained from corresponding brain samples. We show that high quality RNA can be produced from most human autopsy tissues, though with significant differences between tissues and donors. The RNA stability and yield did not depend solely on PMI; other undetermined factors are involved, but these do not include the age of the donor.

  19. Isolated 'idiopathic' micropenis: hidden genetic defects?

    PubMed

    Paris, F; De Ferran, K; Bhangoo, A; Ten, S; Lahlou, N; Audran, F; Servant, N; Poulat, F; Philibert, P; Sultan, C

    2011-12-01

    Micropenis is defined as a stretched penile length of less than 2-2.5SD for age. Aetiologies include hypogonadotropic hypogonadism, testicular dysgenesis, defects in testosterone synthesis, androgen resistance [5α-reductase (5αR) deficiency or partial androgen insensitivity] and other rare causes like growth hormone GH deficiency. Often, the cause remains unknown. The aim of this study was to determine whether isolated micropenis with normal plasma testosterone could hide a molecular defect in the androgen pathway. Twenty-six boys with isolated micropenis were included in this study. All of them had 46,XY karyotype, normal luteinizing hormone and follicle-stimulating hormone and a normal plasma testosterone response to human chorionic gonadotropin testing. Androgen receptor (AR), 5αR and steroidogenic factor 1 (SF1) genes were sequenced. A mutation in the AR gene was found in two patients, and a new mutation in the SF1 gene was found in one patient who was the only one to have a low level of inhibin B (InhB). This is the first report of isolated micropenis as a revealing symptom of AR and SF1 mutations. Anti-Mullerian hormone and InhB should thus be evaluated in patients with isolated micropenis, even when plasma testosterone is in the normal range. Detection of gene mutations is helpful for diagnosis, treatment and genetic counselling for probands.

  20. Study of penile circulation before and after radiation in patients with prostate cancer and its effect on impotence

    SciTech Connect

    Mittal, B.

    1985-06-01

    Decrease in penile blood flow has been implicated as the cause of erectile impotence in patients receiving pelvic irradiation. To determine any changes in the penile circulation secondary to pelvic irradiation, the authors measured the penile blood flow before and 6-9 months following completion of irradiation in six patients with prostate cancer. None of these patients had hormonal manipulation. The non-invasive techniques of Penile Brachial Index (PBI) and Penile Flow Index (PFI) were used to study penile circulation. Two patients developed impotence 2 to 4 1/2 months following completion of irradiation. There was no significant change in penile blood flow following irradiation in any of the six patients studied. The etiology of post-irradiation impotence is probably multifactorial and it may be an oversimplification to attribute it to a single organic cause.

  1. Relaxation of human isolated mesenteric arteries by vasopressin and desmopressin.

    PubMed Central

    Martínez, M C; Vila, J M; Aldasoro, M; Medina, P; Flor, B; Lluch, S

    1994-01-01

    1. The effects of vasopressin and deamino-8-D-arginine vasopressin (DDAVP, desmopressin) were studied in artery rings (0.8-1 mm in external diameter) obtained from portions of human omentum during the course of abdominal operations (27 patients). 2. In arterial rings under resting tension, vasopressin produced concentration-dependent, endothelium-independent contractions with an EC50 of 0.59 +/- 0.12 nM. The V1 antagonist d(CH2)5Tyr(Me)AVP (1 microM) and the mixed V1-V2 antagonist desGly-d(CH2)5D-Tyr(Et)ValAVP (0.01 microM) displaced the control curve to vasopressin to the right in a parallel manner without differences in the maximal responses. In the presence of indomethacin (1 microM) the contractile response to vasopressin was significantly increased (P < 0.01). 3. In precontracted arterial rings, previously treated with the V1 antagonist, d(CH2)5Tyr(Me)AVP (1 microM), vasopressin produced endothelium-dependent relaxation. This relaxation was reduced significantly (P < 0.05) by indomethacin (1 microM) and unaffected by the V1-V2 receptor antagonist desGly-d(CH2)5D-Tyr(Et)ValAVP (1 microM) or by NG-nitro-L-arginine methyl ester (L-NAME, 0.1 mM). 4. The selective V2 receptor agonist, DDAVP, caused endothelium-independent, concentration-dependent relaxations in precontracted arterial rings that were inhibited by the mixed V1-V2 receptor antagonist, but not by the V1 receptor antagonist or by pretreatment with indomethacin or L-NAME. 5. Results from this study suggest that vasopressin is primarily a constrictor of human mesenteric arteries by V1 receptor stimulation; vasopressin causes dilatation only during V1 receptor blockade. The relaxation appears to be mediated by the release of vasodilator prostaglandins from the endothelial cell layer and is independent of V2 receptor stimulation or release of nitric oxide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7834191

  2. Morphine attenuates cholinergic nerve activity in human isolated colonic muscle.

    PubMed Central

    Burleigh, D. E.; Trout, S. J.

    1986-01-01

    The action of morphine on cholinergic nerves in human sigmoid taenia coli muscle strips (taenia) was investigated using a radiolabelling technique. Basal release of tritiated material from taenia was increased by electrical field stimulation (EFS). This increase was tetrodotoxin (3.14 microM)-sensitive and calcium-dependent. Analysis of basal and stimulated release of tritiated material indicated that evoked release (i.e. stimulated minus basal) is almost entirely due to an increase in [3H]-acetylcholine ([3H]-ACh) output. Evoked release of [3H]-ACh was dependent on the current strength and could be greatly reduced by exposing taenia to hemicholinium (34.8, 87.0 microM) before and during incubation with [3H]-choline (4 microCi ml-1, 15 Ci mmol-1). Spontaneous activity, muscle tone and the motor response of taenia to EFS were unaffected by morphine. Evoked, but not basal, release of tritiated material was inhibited by morphine (1.32-13.20 microM) in a concentration-dependent manner. The inhibition of release was frequency-dependent and naloxone (0.28 microM)-sensitive. The possible relationship between the effects of morphine on cholinergic nerves in taenia muscle and its actions in vivo are discussed. PMID:2873856

  3. Campylobacter species isolated from poultry and humans, and their analysis using PFGE in southern Brazil.

    PubMed

    Silva, Daiani Teixeira da; Tejada, Talita Schneid; Blum-Menezes, Dulcinea; Dias, Priscila Alves; Timm, Cláudio Dias

    2016-01-18

    Campylobacteriosis is a bacterial disease transmitted to humans through ingestion of contaminated food. Six hundred samples were collected, 200 from human stool samples, 200 from poultry products and 200 from poultry feces in Southern Brazil, and then inoculated on blood agar plates. A total of 58% of the poultry feces, 17% of the poultry meat, and 2% the of human stools tested positive for Campylobacter. Positive Campylobacter colonies were identified as Campylobacter jejuni or Campylobactercoli by multiplex PCR. Campylobacter isolates were analyzed using PFGE to compare different profiles according to the source. This study demonstrated that there are different Campylobacter clones distributed in different aviaries in Southern Brazil. In addition, PFGE molecular profiles suggested that broilers can be a source of contamination of poultry products. However, the human isolate studied did not show any relationship with other strains examined.

  4. Virulence-associated gene pattern of porcine and human Yersinia enterocolitica biotype 4 isolates.

    PubMed

    Schneeberger, M; Brodard, I; Overesch, G

    2015-04-01

    Yersinia enterocolitica 4/O:3 is the most important human pathogenic bioserotype in Europe and the predominant pathogenic bioserotype in slaughter pigs. Although many studies on the virulence of Y. enterocolitica strains have showed a broad spectrum of detectable factors in pigs and humans, an analysis based on a strict comparative approach and serving to verify the virulence capability of porcine Y. enterocolitica as a source for human yersiniosis is lacking. Therefore, in the present study, strains of biotype (BT) 4 isolated from Swiss slaughter pig tonsils and feces and isolates from human clinical cases were compared in terms of their spectrum of virulence-associated genes (yadA, virF, ail, inv, rovA, ymoA, ystA, ystB and myfA). An analysis of the associated antimicrobial susceptibility pattern completed the characterization. All analyzed BT 4 strains showed a nearly similar pattern, comprising the known fundamental virulence-associated genes yadA, virF, ail, inv, rovA, ymoA, ystA and myfA. Only ystB was not detectable among all analyzed isolates. Importantly, neither the source of the isolates (porcine tonsils and feces, humans) nor the serotype (ST) had any influence on the gene pattern. From these findings, it can be concluded that the presence of the full complement of virulence genes necessary for human infection is common among porcine BT 4 strains. Swiss porcine BT 4 strains not only showed antimicrobial susceptibility to chloramphenicol, cefotaxime, ceftazidime, ciprofloxacin, colistin, florfenicol, gentamicin, kanamycin, nalidixic acid, sulfamethoxazole, streptomycin, tetracycline and trimethoprim but also showed 100% antibiotic resistance to ampicillin. The human BT 4 strains revealed comparable results. However, in addition to 100% antibiotic resistance to ampicillin, 2 strains were resistant to chloramphenicol and nalidixic acid. Additionally, 1 of these strains was resistant to sulfamethoxazole. The results demonstrated that Y. enterocolitica BT 4

  5. Antibacterial susceptibility of enterobacteriaceae isolated from raw horsemeat isolated for human consumption (basashi).

    PubMed

    Furuhata, Katsunori; Ishizaki, Naoto; Fukuyama, Masafumi

    2015-01-01

    Drug susceptibility testing was carried out using 14 antibiotics in order to identify trends in the antibiotic tolerance of 142 strains of Enterobacteriaceae isolated from horsemeat commercially available for raw consumption (basashi). A comparison of the sensitivity to the 14 antibiotics using the 90% MIC (minimum inhibitory concentration) values (MIC90) showed the strongest tolerance to ampicillin (ABPC) at a concentration of > 128 μg/mL, followed by that to fosfomycin (FOM) at a concentration of 128 μg/mL. When the sensitivity to these antibiotics was examined for each individual genus of tested bacteria, Hafnia spp. exhibited relative tolerance to ceftazidime (CAZ) and ceftriaxone (CTRX) at a concentration of 4 μg/mL and 2 μg/mL, respectively, which was high in comparison to that observed for the other strains. Furthermore, Raoultella spp. and Serratia spp. were found to be highly resistant to tetracycline (TC) at a concentration of 128 μg/mL and 64 μg/mL, respectively. Of the 142 strains of test bacteria, 140 (98.6%) demonstrated resistance to ABPC, with the exception of Hafnia alvei and Klebsiella pneumonia. In addition, a total of eight strains (5.6%), seven Serratia marcescens strains and one Raoultella terrigena strain, were found to be resistant to TC. Furthermore, one strain of Citrobacter freundii exhibited resistance to nalidixic acid (NA), while another displayed resistance to ofloxacin (OFLX) (0.7% each), and one strain (0.7%) each of Enterobacter cloacae, Serratia marcescens and Citrobacter youngae demonstrated resistance to fosfomycin (FOM), streptomycin (SM) and kanamycin (KM), respectively. A single strain of C. freundii was found to be resistant to three antibiotics, ABPC, NA and OFLX. Resistance to two antibiotics was confirmed in 11 strains, including seven strains of S. marcescens and one strain of R. terrigena (a total of eight strains) resistant to ABPC and TC, and one strain each of C. youngae, S. marcescens and E. cloacae resistant

  6. Antibacterial susceptibility of enterobacteriaceae isolated from raw horsemeat isolated for human consumption (basashi).

    PubMed

    Furuhata, Katsunori; Ishizaki, Naoto; Fukuyama, Masafumi

    2015-01-01

    Drug susceptibility testing was carried out using 14 antibiotics in order to identify trends in the antibiotic tolerance of 142 strains of Enterobacteriaceae isolated from horsemeat commercially available for raw consumption (basashi). A comparison of the sensitivity to the 14 antibiotics using the 90% MIC (minimum inhibitory concentration) values (MIC90) showed the strongest tolerance to ampicillin (ABPC) at a concentration of > 128 μg/mL, followed by that to fosfomycin (FOM) at a concentration of 128 μg/mL. When the sensitivity to these antibiotics was examined for each individual genus of tested bacteria, Hafnia spp. exhibited relative tolerance to ceftazidime (CAZ) and ceftriaxone (CTRX) at a concentration of 4 μg/mL and 2 μg/mL, respectively, which was high in comparison to that observed for the other strains. Furthermore, Raoultella spp. and Serratia spp. were found to be highly resistant to tetracycline (TC) at a concentration of 128 μg/mL and 64 μg/mL, respectively. Of the 142 strains of test bacteria, 140 (98.6%) demonstrated resistance to ABPC, with the exception of Hafnia alvei and Klebsiella pneumonia. In addition, a total of eight strains (5.6%), seven Serratia marcescens strains and one Raoultella terrigena strain, were found to be resistant to TC. Furthermore, one strain of Citrobacter freundii exhibited resistance to nalidixic acid (NA), while another displayed resistance to ofloxacin (OFLX) (0.7% each), and one strain (0.7%) each of Enterobacter cloacae, Serratia marcescens and Citrobacter youngae demonstrated resistance to fosfomycin (FOM), streptomycin (SM) and kanamycin (KM), respectively. A single strain of C. freundii was found to be resistant to three antibiotics, ABPC, NA and OFLX. Resistance to two antibiotics was confirmed in 11 strains, including seven strains of S. marcescens and one strain of R. terrigena (a total of eight strains) resistant to ABPC and TC, and one strain each of C. youngae, S. marcescens and E. cloacae resistant

  7. Detection and quantification of human adenovirus genomes in Acanthamoeba isolated from swimming pools.

    PubMed

    Staggemeier, Rodrigo; Arantes, Thalita; Caumo, Karin S; Rott, Marilise B; Spilki, Fernando R

    2016-01-01

    Acanthamoeba is the most common free-living environmental amoeba, it may serve as an important vehicle for various microorganisms living in the same environment, such as viruses, being pathogenic to humans. This study aimed to detect and quantify human adenoviruses (HAdV) in Acanthamoebas isolated from water samples collected from swimming pools in the city of Porto Alegre, Southern Brazil. Free-living amoebae of the genus Acanthamoeba were isolated from water samples, and isolates (n=16) were used to investigate the occurrence of HAdVs. HAdV detection was performed by quantitative real-time polymerase chain reaction (qPCR). HAdVs were detected in 62.5% (10/16) of Acanthamoeba isolates, ranging from 3.24x103 to 5.14x105 DNA copies per milliliter of isolate. HAdV viral loads found in this study are not negligible, especially because HAdV infections are associated with several human diseases, including gastroenteritis, respiratory distress, and ocular diseases. These findings reinforce the concept that Acanthamoeba may act as a reservoir and promote HAdV transmission through water. PMID:27142544

  8. Isolation of blood-vessel-derived multipotent precursors from human skeletal muscle.

    PubMed

    Chen, William C W; Saparov, Arman; Corselli, Mirko; Crisan, Mihaela; Zheng, Bo; Péault, Bruno; Huard, Johnny

    2014-01-01

    Since the discovery of mesenchymal stem/stromal cells (MSCs), the native identity and localization of MSCs have been obscured by their retrospective isolation in culture. Recently, using fluorescence-activated cell sorting (FACS), we and other researchers prospectively identified and purified three subpopulations of multipotent precursor cells associated with the vasculature of human skeletal muscle. These three cell populations: myogenic endothelial cells (MECs), pericytes (PCs), and adventitial cells (ACs), are localized respectively to the three structural layers of blood vessels: intima, media, and adventitia. All of these human blood-vessel-derived stem cell (hBVSC) populations not only express classic MSC markers but also possess mesodermal developmental potentials similar to typical MSCs. Previously, MECs, PCs, and ACs have been isolated through distinct protocols and subsequently characterized in separate studies. The current isolation protocol, through modifications to the isolation process and adjustments in the selective cell surface markers, allows us to simultaneously purify all three hBVSC subpopulations by FACS from a single human muscle biopsy. This new method will not only streamline the isolation of multiple BVSC subpopulations but also facilitate future clinical applications of hBVSCs for distinct therapeutic purposes. PMID:25177794

  9. Phalloplasty: A panacea for 46,XY disorder of sex development conditions with penile deficiency?

    PubMed

    Callens, Nina; Hoebeke, Piet

    2014-01-01

    In cases of severe penile inadequacy, such as in pathological conditions involving penile amputation (e.g. penile cancer), or in 46,XY disorders of sex development with severe undervirilization or maldevelopment of the penis (e.g. idiopathic micropenis, cloacal exstrophy), standard (surgical) penile lengthening techniques do not provide patients with a phallus suitable for sexual intercourse. Genital dissatisfaction can lead to low self-esteem and psychosexual dysfunction. Therefore, phalloplasty, the gold standard in transgender surgery, may provide a possibility to achieve a satisfactory genital appearance and sexual function. Small series have reported cosmetically acceptable and erogenous sensate neophalli with incorporation of a neourethra to allow voiding in a standing position and with enough bulk to allow penile prosthesis insertion for pleasurable intercourse. Although early results seem promising, further publication of series with large numbers and longer follow-up is needed to evaluate to what extent phalloplasty improves physical and sexual outcomes. Complications are of particular concern because of associated scarring and loss of sensitive tissue. Without full preoperative workups assessing patients' expectations and reasons for undergoing surgery, they may still struggle with self/penile image and with psychological barriers for engaging in sexual activity. Recommendations for the psychosocial management of boys and men with penile deficiency are suggested.

  10. Molecular typing of Leptospira spp. strains isolated from field mice confirms a link to human leptospirosis.

    PubMed

    Li, S J; Wang, D M; Zhang, C C; Li, X W; Yang, H M; Tian, K C; Wei, X Y; Liu, Y; Tang, G P; Jiang, X G; Yan, J

    2013-11-01

    In recent years, human leptospirosis has been reported in Jinping and Liping counties, Guizhou province, but the leptospires have never been isolated. To track the source of infection and understand the aetiological characteristics, we performed surveillance for field mice carriage of leptospirosis in 2011. Four strains of leptospire were isolated from Apodemus agrarius. PCR confirmed the four isolates as pathogenic. Multiple-locus variable-number tandem repeat analysis (MLVA) showed that the four strains were closely related to serovar Lai strain 56601 belonging to serogroup Icterohaemorrhagiae, which is consistent with the antibody detection results from local patients. Furthermore, the diversity of leptospiral isolates from different hosts and regions was demonstrated with MLVA. Our results suggest that A. agrarius may be the main carrier of Leptospira in Jinping and Liping counties, and the serogroup Icterohaemorrhagiae serovar may be the epidemic serogroup of Leptospira. This will contribute to the control and prevention of leptospirosis in these localities. PMID:23406882

  11. Genotoxicity to human cells induced by air particulates isolated during the Kuwait oil fires.

    PubMed

    Kelsey, K T; Xia, F; Bodell, W J; Spengler, J D; Christiani, D C; Dockery, D W; Liber, H L

    1994-01-01

    In an effort to examine the potential of exposure to soot from the 1991 oil fires in the Kuwait desert for inducing genetic effects we studied the in vitro genotoxicity of this material. Air particulates isolated near the Kuwait oil fires were studied using three assays. Dose-dependent increases were observed for both sister chromatid exchanges in human peripheral blood lymphocytes and mutation at the hprt locus in the metabolically competent human lymphoblast cell line AHH-1. Similar magnitudes of response were seen using these two assays when testing a standard air particulate sample which had been isolated from the Washington, DC, area. Using the 32P-postlabeling assay, no increase in DNA adduct formation was observed in AHH-1 cells treated with particulates isolated from sampling in Kuwait.

  12. Isolation and Expansion of Mesenchymal Stem/Stromal Cells Derived from Human Placenta Tissue

    PubMed Central

    Pelekanos, Rebecca A.; Sardesai, Varda S.; Futrega, Kathryn; Lott, William B.; Kuhn, Michael; Doran, Michael R.

    2016-01-01

    Mesenchymal stem/stromal cells (MSC) are promising candidates for use in cell-based therapies. In most cases, therapeutic response appears to be cell-dose dependent. Human term placenta is rich in MSC and is a physically large tissue that is generally discarded following birth. Placenta is an ideal starting material for the large-scale manufacture of multiple cell doses of allogeneic MSC. The placenta is a fetomaternal organ from which either fetal or maternal tissue can be isolated. This article describes the placental anatomy and procedure to dissect apart the decidua (maternal), chorionic villi (fetal), and chorionic plate (fetal) tissue. The protocol then outlines how to isolate MSC from each dissected tissue region, and provides representative analysis of expanded MSC derived from the respective tissue types. These methods are intended for pre-clinical MSC isolation, but have also been adapted for clinical manufacture of placental MSC for human therapeutic use. PMID:27340821

  13. Isolation and clinical sample typing of human leptospirosis cases in Argentina.

    PubMed

    Chiani, Yosena; Jacob, Paulina; Varni, Vanina; Landolt, Noelia; Schmeling, María Fernanda; Pujato, Nazarena; Caimi, Karina; Vanasco, Bibiana

    2016-01-01

    Leptospira typing is carried out using isolated strains. Because of difficulties in obtaining them, direct identification of infective Leptospira in clinical samples is a high priority. Multilocus sequence typing (MLST) proved highly discriminatory for seven pathogenic species of Leptospira, allowing isolate characterization and robust assignment to species, in addition to phylogenetic evidence for the relatedness between species. In this study we characterized Leptospira strains circulating in Argentina, using typing methods applied to human clinical samples and isolates. Phylogenetic studies based on 16S ribosomal RNA gene sequences enabled typing of 8 isolates (6 Leptospira interrogans, one Leptospira wolffii and one Leptospira broomii) and 58 out of 85 (68.2%) clinical samples (55 L. interrogans, 2 Leptospira meyeri, and one Leptospira kirschneri). MLST results for the L. interrogans isolates indicated that five were probably Canicola serogroup (ST37) and one was probably Icterohaemorrhagiae serogroup (ST17). Eleven clinical samples (21.6%), provided MLST interpretable data: five were probably Pyrogenes serogroup (ST13), four Sejroe (ST20), one Autumnalis (ST22) and one Canicola (ST37). To the best of our knowledge this study is the first report of the use of an MLST typing scheme with seven loci to identify Leptospira directly from clinical samples in Argentina. The use of clinical samples presents the advantage of the possibility of knowing the infecting strain without resorting to isolates. This study also allowed, for the first time, the characterization of isolates of intermediate pathogenicity species (L. wolffii and L. broomii) from symptomatic patients. PMID:26658064

  14. Isolation and clinical sample typing of human leptospirosis cases in Argentina.

    PubMed

    Chiani, Yosena; Jacob, Paulina; Varni, Vanina; Landolt, Noelia; Schmeling, María Fernanda; Pujato, Nazarena; Caimi, Karina; Vanasco, Bibiana

    2016-01-01

    Leptospira typing is carried out using isolated strains. Because of difficulties in obtaining them, direct identification of infective Leptospira in clinical samples is a high priority. Multilocus sequence typing (MLST) proved highly discriminatory for seven pathogenic species of Leptospira, allowing isolate characterization and robust assignment to species, in addition to phylogenetic evidence for the relatedness between species. In this study we characterized Leptospira strains circulating in Argentina, using typing methods applied to human clinical samples and isolates. Phylogenetic studies based on 16S ribosomal RNA gene sequences enabled typing of 8 isolates (6 Leptospira interrogans, one Leptospira wolffii and one Leptospira broomii) and 58 out of 85 (68.2%) clinical samples (55 L. interrogans, 2 Leptospira meyeri, and one Leptospira kirschneri). MLST results for the L. interrogans isolates indicated that five were probably Canicola serogroup (ST37) and one was probably Icterohaemorrhagiae serogroup (ST17). Eleven clinical samples (21.6%), provided MLST interpretable data: five were probably Pyrogenes serogroup (ST13), four Sejroe (ST20), one Autumnalis (ST22) and one Canicola (ST37). To the best of our knowledge this study is the first report of the use of an MLST typing scheme with seven loci to identify Leptospira directly from clinical samples in Argentina. The use of clinical samples presents the advantage of the possibility of knowing the infecting strain without resorting to isolates. This study also allowed, for the first time, the characterization of isolates of intermediate pathogenicity species (L. wolffii and L. broomii) from symptomatic patients.

  15. Antibiotic susceptibility profile of bacilli isolated from the skin of healthy humans.

    PubMed

    Tarale, Prashant; Gawande, Sonali; Jambhulkar, Vinay

    2015-01-01

    In the present work, twelve bacilli were isolated from four different regions of human skin from Bela population of Nagpur district, India. The isolated bacilli were identified by their morphological, cultural and biochemical characteristics. Seven isolates were Gram negative rods, out of which five were belong to genus Pseudomonas. Three among the five Gram positive isolates were identified as Dermabactor and the remaining two Bacillus. Their antimicrobial susceptibility profile was determined by Kirby-Bauer disc diffusion method. The isolates showed resistance to several currently used broad-spectrum antibiotics. The Dermabactor genus was resistant to vancomycin, although it was earlier reported to be susceptible. Imipenem was found to be the most effective antibiotic for Pseudomonas while nalidixic acid, ampicillin and tetracycline were ineffective. Isolates of Bacillus displayed resistance to the extended spectrum antibiotics cephalosporin and ceftazidime. Imipenem, carbenicillin and ticarcillin were found to be the most effective antibiotics as all the investigated isolates were susceptible to them. Antibiotic resistance may be due to the overuse or misuse of antibiotics during the treatment, or following constant exposure to antibiotic-containing cosmetic formulations.

  16. Draft Genome Sequence of Lactobacillus hominis Strain CRBIP 24.179T, Isolated from Human Intestine.

    PubMed

    Cousin, Sylvie; Creno, Sophie; Ma, Laurence; Clermont, Dominique; Loux, Valentin; Bizet, Chantal; Bouchier, Christiane

    2013-01-01

    We report the draft genome sequence of the strain Lactobacillus hominis CRBIP 24.179(T), isolated from a human clinical sample. The total length of the 28 contigs is about 1.9 Mb, with a G+C content of 37% and 1,983 coding sequences.

  17. Complete Genome Sequences of Four Different Bordetella sp. Isolates Causing Human Respiratory Infections

    PubMed Central

    Peng, Yanhui; Loparev, Vladimir; Batra, Dhwani; Bowden, Katherine E.; Cassiday, Pamela K.; Davis, Jamie K.; Johnson, Taccara; Juieng, Phalasy; Miner, Christine E.; Rowe, Lori; Sheth, Mili; Tondella, M. Lucia; Williams, Margaret M.

    2016-01-01

    Species of the genus Bordetella associate with various animal hosts, frequently causing respiratory disease. Bordetella pertussis is the primary agent of whooping cough and other Bordetella species can cause similar cough illness. Here, we report four complete genome sequences from isolates of different Bordetella species recovered from human respiratory infections. PMID:27795250

  18. The Psychology of Isolated and Confined Environments: Understanding Human Behavior in Antarctica.

    ERIC Educational Resources Information Center

    Palinkas, Lawrence A.

    2003-01-01

    Reviews lessons learned from research in Antarctica with relevance to understanding human behavior in other isolated and confined environments. Outlines four distinct characteristics of psychosocial adaptation to such environments and discusses some of the benefits for individuals seeking challenging experiences. (Contains references.) (SLD)

  19. Ribotype and serotype diversity of Escherichia coli isolates from poultry and human sources

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Escherichia coli isolates from the poultry and human sources were analyzed to evaluate the genetic relatedness and serotype. Ribotype was determined using the RiboPrinter® Microbial Characterization System following DNA digestion with the restriction enzyme EcoRI. A total of 41 ribogroups were iso...

  20. Solely Penile Skin for Neovaginal Construction in Sex Reassignment Surgery

    PubMed Central

    Sigurjonsson, Hannes; Rinder, Johan; Lindqvist, Ebba K.; Farnebo, Filip

    2016-01-01

    Background: Gender reassignment surgery due to transsexualism (International Classification of Diseases, Tenth Revision: F64.0) is a procedure becoming increasingly common worldwide as a result of a significant increase in diagnostic incidence. Several methods have been described for this complex surgery, but no internationally agreed upon gold standard exists, in particular with regard to which methods allow for creating a sufficient neovaginal depth. Methods: We use a 2-stage technique using solely penile skin for creating a neovaginal cavity and present the long-term outcome in terms of measured neovaginal depth. Eighty patients were included. Patients’ neovaginal depth was measured in a standardized fashion 6 months or more after initial surgery. Results were compared with published data on female anatomy. Results: The average neovaginal depth achieved was 10.2 cm. Having had a postoperative complication and noncompliance to neovaginal dilatation were both negatively correlated with neovaginal depth, whereas higher body mass index was not. Most patients received a neovaginal depth sufficient for penetrative intercourse and within the range for biological women. Conclusions: Using solely penile skin for the vaginal lining is a satisfactory surgical method to achieve adequate vaginal depth, provided that the postoperative dilatation regimen is followed. This holds true regardless of age or body mass index. PMID:27482506

  1. Penile Fracture: Experience from a Third World Country

    PubMed Central

    Bali, Rajandeep Singh; Mushtaque, Majid; Nabi, Shakeeb; Thakur, Sajad Ahmad; Bhat, Rouf Ahmad

    2013-01-01

    Aim. To ascertain the clinical presentation, commonest age group affected, and treatment of patients diagnosed to have penis fracture. Materials and Methods. We performed a retrospective study carried at a tertiary care hospital from January 2005 to January 2011. All the 36 patients diagnosed to have penile fracture were enrolled in the study group. The diagnosis was made based on the clinical findings in the patients. All, except two patients, were managed by a standard surgical procedure, same for all the patients, on the day of presentation to the hospital. All the data pertaining to the presentation, management, and followup of these patients were studied and scrutinized thoroughly. Results. Thirty-four patients were operated while 2 refused surgery. Most of our patients were between 16 and 30 years (55.6%) of age. The commonest presenting complaints were penile swelling and detumescence during sexual intercourse or an erection. All except two of our patients were managed with immediate surgical repair which had excellent results even in the presence of associated urethral injury. Conclusion. Fracture of the penis is a surgical emergency which can be best managed by immediate surgical repair with excellent results even in the presence of urethral injury. PMID:23956740

  2. Targets of Wnt/ß-Catenin Transcription in Penile Carcinoma

    PubMed Central

    Henrique, Rui; Millar, Michael; Hamblin, Ruth; Davda, Reena; Aare, Kristina; Masters, John R.; Thomson, Calum; Muneer, Asif; Patel, Hitendra R. H.; Ahmed, Aamir

    2015-01-01

    Penile squamous cell carcinoma (PeCa) is a rare malignancy and little is known regarding the molecular mechanisms involved in carcinogenesis of PeCa. The Wnt signaling pathway, with the transcription activator ß-catenin as a major transducer, is a key cellular pathway during development and in disease, particularly cancer. We have used PeCa tissue arrays and multi-fluorophore labelled, quantitative, immunohistochemistry to interrogate the expression of WNT4, a Wnt ligand, and three targets of Wnt-ß-catenin transcription activation, namely, MMP7, cyclinD1 (CD1) and c-MYC in 141 penile tissue cores from 101 unique samples. The expression of all Wnt signaling proteins tested was increased by 1.6 to 3 fold in PeCa samples compared to control tissue (normal or cancer adjacent) samples (p<0.01). Expression of all proteins, except CD1, showed a significant decrease in grade II compared to grade I tumors. High magnification, deconvolved confocal images were used to measure differences in co-localization between the four proteins. Significant (p<0.04-0.0001) differences were observed for various permutations of the combinations of proteins and state of the tissue (control, tumor grades I and II). Wnt signaling may play an important role in PeCa and proteins of the Wnt signaling network could be useful targets for diagnosis and prognostic stratification of disease. PMID:25901368

  3. Role of hydrogen sulfide in the physiology of penile erection.

    PubMed

    Qiu, Xuefeng; Villalta, Jackie; Lin, Guiting; Lue, Tom F

    2012-01-01

    Hydrogen sulfide (H(2)S), which is a well-known toxic gas, has recently been recognized as a biological messenger that plays an important role in physiological and pathophysiological conditions. Relatively high levels of H(2)S have been discovered in mammalian tissues. It is mainly synthesized by 2 enzymes, including cystathionine β-synthase and cystathionine γ-lysase, which utilize L-cysteine as substrate to produce H(2)S. H(2)S has been demonstrated to exhibit potent vasodilator activity both in vitro and in vivo by relaxing vascular smooth muscle. Recently, H(2)S has been discovered in penile tissue with smooth muscle relaxant effects. Furthermore, other effects of H(2)S could play a role in the physiology of erection. Understanding H(2)S in the physiology of erection might provide alternative erectile dysfunction strategies for those patients with poor or no response to type 5 phosphodiesterase inhibitors. This review intends to present the H(2)S pathway in penile tissue and the potential role of H(2)S in the physiology of erections.

  4. Biofilm-Forming Abilities of Shiga Toxin-Producing Escherichia coli Isolates Associated with Human Infections

    PubMed Central

    Vogeleer, Philippe; Tremblay, Yannick D. N.; Jubelin, Grégory; Jacques, Mario

    2015-01-01

    Forming biofilms may be a survival strategy of Shiga toxin-producing Escherichia coli to enable it to persist in the environment and the food industry. Here, we evaluate and characterize the biofilm-forming ability of 39 isolates of Shiga toxin-producing Escherichia coli isolates recovered from human infection and belonging to seropathotypes A, B, or C. The presence and/or production of biofilm factors such as curli, cellulose, autotransporter, and fimbriae were investigated. The polymeric matrix of these biofilms was analyzed by confocal microscopy and by enzymatic digestion. Cell viability and matrix integrity were examined after sanitizer treatments. Isolates of the seropathotype A (O157:H7 and O157:NM), which have the highest relative incidence of human infection, had a greater ability to form biofilms than isolates of seropathotype B or C. Seropathotype A isolates were unique in their ability to produce cellulose and poly-N-acetylglucosamine. The integrity of the biofilms was dependent on proteins. Two autotransporter genes, ehaB and espP, and two fimbrial genes, z1538 and lpf2, were identified as potential genetic determinants for biofilm formation. Interestingly, the ability of several isolates from seropathotype A to form biofilms was associated with their ability to agglutinate yeast in a mannose-independent manner. We consider this an unidentified biofilm-associated factor produced by those isolates. Treatment with sanitizers reduced the viability of Shiga toxin-producing Escherichia coli but did not completely remove the biofilm matrix. Overall, our data indicate that biofilm formation could contribute to the persistence of Shiga toxin-producing Escherichia coli and specifically seropathotype A isolates in the environment. PMID:26712549

  5. Probiotic attributes of Lactobacillus strains isolated from food and of human origin.

    PubMed

    Gaudana, Sandeep B; Dhanani, Akhilesh S; Bagchi, Tamishraha

    2010-06-01

    Lactobacilli isolated from various sources were identified on the basis of 16S-23S rRNA gene intergenic region amplification and subsequent sequencing of the smaller intergenic region. An in vitro analysis of probiotic properties including binding, ability to tolerate different concentrations of bile, survival in acidic buffer and antimicrobial activity of four different isolates and two standard strains (Lactobacillus plantarum American Type Culture Collection (ATCC) 8014 and L. rhamnosus GG (LGG)) was carried out. The ability of each isolate to stimulate Caco-2 cells, human peripheral blood mononuclear cells (PBMC) and THP-1 cells resulting in immunomodulation of these cells was analysed. Isolates L. rhamnosus CS25 and L. delbrueckii M and standard strain ATCC 8014 showed broad antimicrobial activity, and isolates CS25 (percentage of survival 6.9 % at pH 2.5, 5.1 % at pH 2.0) and L. plantarum CS23 (5.7 % at pH 2.5, 4.9 % at pH 2.0) have shown good tolerance to acidic pH. Isolate CS23 showed a good survival (14 %) after 2 h incubation in de Man, Rogosa and Sharpe (MRS) medium containing 3 % bile salts. Isolates CS23, CS25 and L. fermentum ASt1 could stimulate Caco-2 cells, human PBMC and THP-1 cells for a strong and varied immunomodulatory response in these cells. Though LGG showed poor antimicrobial activity as well as bile and acid tolerance, it was found to be the best binding strain tested. Child faecal isolate CS23 from the present study showed high binding ability (seventeen bacteria/Caco-2), high tolerance to acidic pH and bile salts and significant immunomodulation; therefore it is a good potential probiotic candidate.

  6. Draft Genome Sequences of 40 Salmonella enterica Serovar Typhimurium Strains Isolated from Humans and Food in Brazil

    PubMed Central

    Almeida, Fernanda; Medeiros, Marta Inês Cazentini; Rodrigues, Dália Prazeres; Payne, Justin; Timme, Ruth E.

    2016-01-01

    Salmonellosis is an important health problem worldwide and Salmonella enterica serovar Typhimurium is one of the most common isolated serovars. Here, we reported the draft genomes of 40 S. Typhimurium strains isolated from humans and food in Brazil. These draft genomes will improve phylogenetic analysis and will help enhance our understanding of strains of this serovar isolated in Brazil. PMID:27660768

  7. Draft Genome Sequences of 40 Salmonella enterica Serovar Typhimurium Strains Isolated from Humans and Food in Brazil.

    PubMed

    Almeida, Fernanda; Medeiros, Marta Inês Cazentini; Rodrigues, Dália Prazeres; Payne, Justin; Timme, Ruth E; Allard, Marc W; Falcão, Juliana Pfrimer

    2016-01-01

    Salmonellosis is an important health problem worldwide and Salmonella enterica serovar Typhimurium is one of the most common isolated serovars. Here, we reported the draft genomes of 40 S Typhimurium strains isolated from humans and food in Brazil. These draft genomes will improve phylogenetic analysis and will help enhance our understanding of strains of this serovar isolated in Brazil. PMID:27660768

  8. A penal problem: the increasing incidence of implantation of penile foreign bodies.

    PubMed

    Flynn, Ryan M; Mostafa, Hesham I; Khan, Omar A; Haselhuhn, Gregory D; Jain, Samay

    2014-12-01

    Our objective is to describe a novel presentation of subcutaneous penile insertion of foreign bodies. This is a practice performed globally and mostly has been reported outside of the United States. We present three cases of incarcerated males that implanted sculpted dominos into the penile subcutaneous tissue. The patients presented with erosion of the foreign bodies through the skin without evidence of infection. We believe that insertion of foreign bodies into penile subcutaneous tissue by incarcerated American males for sexual enhancement is more widespread than previously reported. Erosion is a novel presentation.

  9. Cryptococcus gattii in the United States: Genotypic Diversity of Human and Veterinary Isolates

    PubMed Central

    Lockhart, Shawn R.; Iqbal, Naureen; Harris, Julie R.; Grossman, Nina T.; DeBess, Emilio; Wohrle, Ron; Marsden-Haug, Nicola; Vugia, Duc J.

    2013-01-01

    Background Cryptococcusgattii infections are being reported in the United States (US) with increasing frequency. Initially, US reports were primarily associated with an ongoing C. gattii outbreak in the Pacific Northwest (PNW) states of Washington and Oregon, starting in 2004. However, reports of C. gattii infections in patients from other US states have been increasing since 2009. Whether this is due to increasing frequency of disease, greater recognition within the clinical community, or both is currently unknown. Methodology/Principal Findings During 2005–2013, a total of 273 C. gattii isolates from human and veterinary sources in 16 US states were collected. Of these, 214 (78%) were from the Pacific Northwest (PNW) and comprised primarily the clonal C. gattii genotypes VGIIa (64%), VGIIc (21%) and VGIIb (9%). The 59 isolates from outside the PNW were predominantly molecular types VGIII (44%) and VGI (41%). Genotyping using multilocus sequence typing revealed small clusters, including a cluster of VGI isolates from the southeastern US, and an unrelated cluster of VGI isolates and a large cluster of VGIII isolates from California. Most of the isolates were mating type MATα, including all of the VGII isolates, but one VGI and three VGIII isolates were mating type MATa. Conclusions/Significance We provide the most comprehensive report to date of genotypic diversity of US C. gattii isolates both inside and outside of the PNW. C. gattii may have multiple endemic regions in the US, including a previously-unrecognized endemic region in the southeast. Regional clusters exist both in California and the Southeastern US. VGII strains associated with the PNW outbreak do not appear to have spread substantially beyond the PNW. PMID:24019979

  10. Antimicrobial susceptibility and molecular typing of salmonella agona isolated from humans and other sources.

    PubMed

    Kuang, Dai; Zhang, Jianmin; Meng, Jianghong; Yang, Xiaowei; Jin, Huiming; Shi, Weimin; Luo, Kaijian; Tao, Yan; Pan, Haijian; Xu, Xuebin; Ren, Tao

    2014-11-01

    Salmonella enterica serotype Agona (Salmonella Agona) has been among the top 10 serotypes that cause human diarrheal diseases in China. A total of 95 Salmonella Agona (67 from humans, and 28 from animals, food of animal origins, and environmental sources) recovered in Shanghai, China from 2005 to 2011 were subjected to antimicrobial susceptibility testing and molecular subtyping using pulsed-field gel electrophoresis (PFGE). Approximately 68.4% of the Salmonella Agona isolates were pansusceptible to 15 antimicrobial agents tested, and 4 isolates (4.21%) were resistant to at least 3 antimicrobials. PFGE analysis resulted in 41 unique patterns, of which 4 major PFGE patterns (X3, X4, X5, and X6) were grouped together at 96.1% similarity. Isolates of the four patterns included those from food (pork, beef, and chicken) and humans. Our findings showed that the same clones of Salmonella Agona were recovered from human patients and food, and that food of animal origin was potentially a major vehicle of Salmonella Agona in human salmonellosis in Shanghai.

  11. Phenotypic and genotypic characterization of Vagococcus fluvialis, including strains isolated from human sources.

    PubMed Central

    Teixeira, L M; Carvalho, M G; Merquior, V L; Steigerwalt, A G; Brenner, D J; Facklam, R R

    1997-01-01

    This study presents phenotypic and genotypic data for seven isolates of Vagococcus fluvialis, including four strains recovered from human clinical sources, one strain isolated from an environmental source, and two strains isolated from pigs. On the basis of phenotypic characteristics, most isolates were initially classified as "unidentified enterococci," because they resembled atypical arginine-negative enterococcal species. All seven strains as well as the type strain of V. fluvialis reacted with the AccuProbe Enterococcus genetic probe. The seven isolates had virtually indistinguishable whole-cell protein profiles that were similar to that of the V. fluvialis type strain and distinct from those of Enterococcus and Lactococcus species. DNA-DNA reassociation experiments confirmed that the strains were V. fluvialis. They were 71% or more related to the V. fluvialis type strain under optimum and stringent conditions, with 2.5% or less divergence within related sequences. All strains were susceptible to ampicillin, cefotaxime, trimethoprim-sulfamethoxazole, and vancomycin and were resistant to clindamycin, lomefloxacin, and ofloxacin. Strain-to-strain variation was observed in relation to susceptibilities to 18 other antimicrobial agents. Chromosomal DNA was analyzed by pulsed-field gel electrophoresis (PFGE) after digestion with SmaI. Distinctive PFGE patterns were generated, suggesting the nonclonal nature of V. fluvialis strains. Although the number of strains was small, this report provides molecular characterization of V. fluvialis and the first evidence of a possible connection of this species with human infections. PMID:9350732

  12. Antigenic typing of Brazilian rabies virus samples isolated from animals and humans, 1989-2000.

    PubMed

    Favoretto, Silvana Regina; Carrieri, Maria Luiza; Cunha, Elenice Maria S; Aguiar, Elizabeth A C; Silva, Luzia Helena Q; Sodre, Miriam M; Souza, Maria Conceição A M; Kotait, Ivanete

    2002-01-01

    Animal and human rabies samples isolated between 1989 and 2000 were typified by means of a monoclonal antibody panel against the viral nucleoprotein. The panel had been previously established to study the molecular epidemiology of rabies virus in the Americas. Samples were isolated in the Diagnostic Laboratory of the Pasteur Institute and in other rabies diagnostic centers in Brazil. In addition to the fixed virus samples CVS-31/96-IP, preserved in mouse brain, and PV-BHK/97, preserved in cell culture, a total of 330 rabies virus samples were isolated from dogs, cats, cattle, horses, bats, sheep, goat, swine, foxes, marmosets, coati and humans. Six antigenic variants that were compatible with the pre-established monoclonal antibodies panel were defined: numbers 2 (dog), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (vampire bat from Venezuela), 6 (Lasiurus cinereus) and Lab (reacted to all used antibodies). Six unknown profiles, not compatible with the panel, were also found. Samples isolated from insectivore bats showed the greatest variability and the most commonly isolated variant was variant-3 (Desmodus rotundus). These findings may be related to the existence of multiple independent transmission cycles, involving different bat species. PMID:12048546

  13. Relatedness of Mycobacterium avium subspecies hominissuis clinical isolates of human and porcine origins assessed by MLVA.

    PubMed

    Leão, Célia; Canto, Ana; Machado, Diana; Sanches, Ilda Santos; Couto, Isabel; Viveiros, Miguel; Inácio, João; Botelho, Ana

    2014-09-17

    Mycobacterium avium subsp. hominissuis (MAH) is an important opportunistic pathogen, infecting humans and animals, notably pigs. Several methods have been used to characterize MAH strains. RFLP and PFGE typing techniques have been used as standard methods but are technically demanding. In contrast, the analysis of VNTR loci is a simpler, affordable and highly reliable PCR-based technique, allowing a numerical and reproductive digitalization of typing data. In this study, the analysis of Mycobacterium avium tandem repeats (MATRs) loci was adapted to evaluate the genetic diversity of epidemiological unrelated MAH clinical strains of human (n=28) and porcine (n=69) origins, collected from diverse geographical regions across mainland Portugal. These MAH isolates were found to be genetically diverse and genotypes are randomly distributed across the country. Some of the human strains shared identical VNTR profiles with porcine isolates. Our study shows that the VNTR genotyping using selected MATR loci is a useful analysis technique for assessing the genetic diversity of MAH isolates from Portugal. This typing method could be successfully applied in other countries toward the implementation of a worldwide open-access database of MATR-VNTR profiles of MAH isolates, allowing a better assessment of the global epidemiology traits of this important pathogenic species.

  14. Molecular Epidemiological and Antibiotic Susceptibility Characterization of Brucella Isolates from Humans in Sicily, Italy▿

    PubMed Central

    Marianelli, Cinzia; Graziani, Caterina; Santangelo, Carmela; Xibilia, Maria Teresa; Imbriani, Alida; Amato, Rosa; Neri, Domenico; Cuccia, Mario; Rinnone, Sebastiano; Di Marco, Vincenzo; Ciuchini, Franco

    2007-01-01

    Brucellosis is a serious problem in Sicily. Brucella melitensis was identified as the species most frequently isolated in humans in Italy. No data, however, are available about the molecular epidemiological characterization of Brucella isolates from humans. We have conducted this study to molecularly characterize clinical isolates of Brucella spp. and to evaluate their antimicrobial susceptibilities. Twenty Brucella isolates were studied. Differential growth characteristics and DNA polymorphisms such as the restriction patterns of the PCR-amplified omp2a and omp2b genes, rpoB nucleotide sequencing, and multiple-locus variable-number tandem repeat analysis of 16 loci (MLVA-16) were used to characterize the strains. In vitro antibiotic susceptibility was determined by the E-test method on two different agar media, and the results were compared. All isolates were identified as B. melitensis biovar 3. rpoB nucleotide sequence analysis allowed the identification of two different genotypes of B. melitensis biovar 3. On the other hand, the MLVA-16 typing assay recognized 17 distinct genotypes. All isolates were sensitive to all tested antibiotics (rifampin, doxycycline, ciprofloxacin, ceftriaxone, and trimethoprim-sulfamethoxazole), and the Mueller-Hinton agar plate is recommended for antibiotic susceptibility testing by the E-test method. Our findings identify B. melitensis biovar 3 as the etiological agent isolated in Sicily and encourage the use of both molecular methods, and in particular of the MLVA-16 assay, in epidemiological trace-back analysis. This study represents the first epidemiological data from molecular typing of Brucella strains circulating in Italy and, in particular, in eastern Sicily. PMID:17634297

  15. Genotypic and phenotypic variation among Staphylococcus saprophyticus from human and animal isolates

    PubMed Central

    2010-01-01

    Background The main aim of this study was to examine the genotypic and phenotypic diversity of Staphylococcus saprophyticus isolates from human and animal origin. Findings In total, 236 clinical isolates and 15 animal isolates of S. saprophyticus were characterized in respect of the occurrence of 9 potential virulence genes and four surface properties. All strains were PCR positive for the regulatory genes agr, sar >it>A and rot as well as for the surface proteins UafA and Aas. Nearly 90% of the clinical isolates were found to possess the gene for the surface-associated lipase Ssp and 10% for the collagen binding MSCRAMM SdrI. All animal isolates were negative forsdrI. Lipolytic activity could be detected in 66% of the clinical and 46% of the animal isolates. Adherence to collagen type I was shown of 20% of the clinical strains and 6% of the strains of animal origin. Most S. saprophyticus strains showed hydrophobic properties and only few could agglutinate sheep erythrocytes. Conclusions We described a broad analysis of animal and human S. saprophyticus isolates regarding virulence genes and phenotypic properties such as lipase activity, hydrophobicity, and adherence. While S. saprophyticus strains from animal sources have prerequisites for colonization of the urinary tract like the D-serine-deaminase, out findings suggested that they need to acquire new genes e.g. MSCRAMMS for adherence like sdrI and to modulate their existing properties e.g. increasing the lipase activity or reducing hydrophobicity. These apparently important new genes or properties for virulence have to be further analyzed. PMID:20537181

  16. Non-typhoidal Salmonella Typhimurium ST313 isolates that cause bacteremia in humans stimulate less inflammasome activation than ST19 isolates associated with gastroenteritis.

    PubMed

    Carden, Sarah; Okoro, Chinyere; Dougan, Gordon; Monack, Denise

    2015-06-01

    Salmonella is an enteric pathogen that causes a range of diseases in humans. Non-typhoidal Salmonella (NTS) serovars such as Salmonella enterica serovar Typhimurium generally cause a self-limiting gastroenteritis whereas typhoidal serovars cause a systemic disease, typhoid fever. However, S. Typhimurium isolates within the multi-locus sequence type ST313 have emerged in sub-Saharan Africa as a major cause of bacteremia in humans. The S. Typhimurium ST313 lineage is phylogenetically distinct from classical S. Typhimurium lineages, such as ST19, that cause zoonotic gastroenteritis worldwide. Previous studies have shown that the ST313 lineage has undergone genome degradation when compared to the ST19 lineage, similar to that observed for typhoidal serovars. Currently, little is known about phenotypic differences between ST313 isolates and other NTS isolates. We find that representative ST313 isolates invade non-phagocytic cells less efficiently than the classical ST19 isolates that are more commonly associated with gastroenteritis. In addition, ST313 isolates induce less Caspase-1-dependent macrophage death and IL-1β release than ST19 isolates. ST313 isolates also express relatively lower levels of mRNA of the genes encoding the SPI-1 effector sopE2 and the flagellin, fliC, providing possible explanations for the decrease in invasion and inflammasome activation. The ST313 isolates have invasion and inflammatory phenotypes that are intermediate; more invasive and inflammatory than Salmonella enterica serovar Typhi and less than ST19 isolates associated with gastroenteritis. This suggests that both phenotypically and at the genomic level ST313 isolates are evolving signatures that facilitate a systemic lifestyle in humans.

  17. Non-typhoidal Salmonella Typhimurium ST313 isolates that cause bacteremia in humans stimulate less inflammasome activation than ST19 isolates associated with gastroenteritis

    PubMed Central

    Carden, Sarah; Okoro, Chinyere; Dougan, Gordon; Monack, Denise

    2014-01-01

    Salmonella is an enteric pathogen that causes a range of diseases in humans. Non-typhoidal Salmonella (NTS) serovars such as Salmonella enterica serovar Typhimurium generally cause a self-limiting gastroenteritis whereas typhoidal serovars cause a systemic disease, typhoid fever. However, S. Typhimurium isolates within the multi-locus sequence type ST313 have emerged in sub-Saharan Africa as a major cause of bacteremia in humans. The S. Typhimurium ST313 lineage is phylogenetically distinct from classical S. Typhimurium lineages, such as ST19, that cause zoonotic gastroenteritis worldwide. Previous studies have shown that the ST313 lineage has undergone genome degradation when compared to the ST19 lineage, similar to that observed for typhoidal serovars. Currently, little is known about phenotypic differences between ST313 isolates and other NTS isolates. We find that representative ST313 isolates invade non-phagocytic cells less efficiently than the classical ST19 isolates that are more commonly associated with gastroenteritis. In addition, ST313 isolates induce less Caspase-1-dependent macrophage death and IL-1β release than ST19 isolates. ST313 isolates also express relatively lower levels of mRNA of the genes encoding the SPI-1 effector sopE2 and the flagellin, fliC, providing possible explanations for the decrease in invasion and inflammasome activation. The ST313 isolates have invasion and inflammatory phenotypes that are intermediate; more invasive and inflammatory than Salmonella enterica serovar Typhi and less than ST19 isolates associated with gastroenteritis. This suggests that both phenotypically and at the genomic level ST313 isolates are evolving signatures that facilitate a systemic lifestyle in humans. PMID:25808600

  18. Failure to attain stretched penile length after intracavernosal injection of a vasodilator agent is predictive of veno-occlusive dysfunction on penile duplex Doppler ultrasonography.

    PubMed

    Yafi, F A; Libby, R P; McCaslin, I R; Sangkum, P; Sikka, S C; Hellstrom, W J G

    2015-09-01

    Penile duplex Doppler ultrasound (PDDU) assesses the etiology of erectile dysfunction. Peak systolic velocity (PSV), end-diastolic velocity (EDV), and resistive index (RI) are common PDDU parameters. We assessed whether stretched penile length (SPL) in the flaccid state and measured penile length at peak erection after intracavernosal injection (ICI) of a vasodilator during PDDU correlated with the etiology of erectile dysfunction. We performed a retrospective review of 93 patients who underwent PDDU for erectile dysfunction. Normal and stretched penile length were measured, both at a flaccid state prior to ICI and at peak erection during PDDU. Collected data included patient demographics, vascular, and anatomic parameters. The mean age was 52 years. SPL was equivalent to peak penile length after ICI in 60 patients (65%, group 1) and did not match in 33 (35%, group 2). There were no significant differences between the two groups in terms of flaccid, stretched, and post-ICI erect penile lengths, IIEF score, PSV, percent rigidity or tumescence, and vasodilator dose used. Patients in group 2 had less of a change in penile length from flaccid to erect state (36% vs. 44%, p = 0.02), higher EDV (12.0 vs. 8.5, p = 0.041), lower RI (0.6 vs. 1.0, p = 0.046), and more veno-occlusive dysfunction (82% vs. 53%, p = 0.001). On multivariate analysis, failure to reach maximum SPL at peak ICI erection (OR 2.255, CI 1.191-4.271, p = 0.0126), EDV (OR 1.281, CI 1.115-1.471, p < 0.001) and RI (OR 0.694, CI 0.573-0.723, p = 0.009) predicted veno-occlusive dysfunction. Failure to reach maximal SPL during PDDU using ICI with a vasodilator agent predicted veno-occlusive dysfunction, which is independent of both penile rigidity and tumescence. This measurement could serve as another diagnostic tool for predicting veno-occlusive dysfunction when PDDU is not readily available. Limitations include the subjective nature of penile measurements and different PGE1 doses used.

  19. Isolation of Hofbauer Cells from Human Term Placentas with High Yield and Purity

    PubMed Central

    Tang, Zhonghua; Tadesse, Serkalem; Norwitz, Errol; Mor, Gil; Abrahams, Vikki M.; Guller, Seth

    2011-01-01

    Problem Placental villus macrophages (i.e. Hofbauer cells, HBCs) were identified more than 100 years ago. Alterations in their numbers and characteristics are associated with several complications of pregnancy. Although HBCs have previously been isolated and cultured, there is no consensus methodology to obtain these cells with high yield and purity for in vitro studies. Method of Study HBCs were isolated from human term placentas using protocols in which cytotrophoblasts (CTs) and fibroblasts (FIBs), other major villous cell types, were isolated in parallel. Enzymatic digestion, Percoll gradients, and immunoselection were used to isolate the three cell types. Purity was assessed by morphology, flow cytometry, and in phagocytosis assays. Results HBCs were isolated with 98–99% purity and a yield of 130–200 ×106 cells/80 to 100 g of tissue. HBCs exhibited a pleiomorphic and vacuolated appearance for at least 5 days in culture medium with and without serum. High levels of phagocytosis in HBCs, but not in CTs, or FIBs, confirmed macrophage function in HBCs. Phagocytotic activity was maintained across several days in culture. Conclusion HBCs were isolated from term placenta with high yield and purity using protocols in which CTs and FIBs were also obtained. This methodology will foster future studies which examine the role of HBCs in regulating villus function. PMID:21545365

  20. Detecting Genetic Isolation in Human Populations: A Study of European Language Minorities

    PubMed Central

    Capocasa, Marco; Battaggia, Cinzia; Anagnostou, Paolo; Montinaro, Francesco; Boschi, Ilaria; Ferri, Gianmarco; Alù, Milena; Coia, Valentina; Crivellaro, Federica; Bisol, Giovanni Destro

    2013-01-01

    The identification of isolation signatures is fundamental to better understand the genetic structure of human populations and to test the relations between cultural factors and genetic variation. However, with current approaches, it is not possible to distinguish between the consequences of long-term isolation and the effects of reduced sample size, selection and differential gene flow. To overcome these limitations, we have integrated the analysis of classical genetic diversity measures with a Bayesian method to estimate gene flow and have carried out simulations based on the coalescent. Combining these approaches, we first tested whether the relatively short history of cultural and geographical isolation of four “linguistic islands” of the Eastern Alps (Lessinia, Sauris, Sappada and Timau) had left detectable signatures in their genetic structure. We then compared our findings to previous studies of European population isolates. Finally, we explored the importance of demographic and cultural factors in shaping genetic diversity among the groups under study. A combination of small initial effective size and continued genetic isolation from surrounding populations seems to provide a coherent explanation for the diversity observed among Sauris, Sappada and Timau, which was found to be substantially greater than in other groups of European isolated populations. Simulations of micro-evolutionary scenarios indicate that ethnicity might have been important in increasing genetic diversity among these culturally related and spatially close populations. PMID:23418562

  1. Detecting genetic isolation in human populations: a study of European language minorities.

    PubMed

    Capocasa, Marco; Battaggia, Cinzia; Anagnostou, Paolo; Montinaro, Francesco; Boschi, Ilaria; Ferri, Gianmarco; Alù, Milena; Coia, Valentina; Crivellaro, Federica; Destro Bisol, Giovanni

    2013-01-01

    The identification of isolation signatures is fundamental to better understand the genetic structure of human populations and to test the relations between cultural factors and genetic variation. However, with current approaches, it is not possible to distinguish between the consequences of long-term isolation and the effects of reduced sample size, selection and differential gene flow. To overcome these limitations, we have integrated the analysis of classical genetic diversity measures with a bayesian method to estimate gene flow and have carried out simulations based on the coalescent. Combining these approaches, we first tested whether the relatively short history of cultural and geographical isolation of four "linguistic islands" of the Eastern Alps (Lessinia, Sauris, Sappada and Timau) had left detectable signatures in their genetic structure. We then compared our findings to previous studies of European population isolates. Finally, we explored the importance of demographic and cultural factors in shaping genetic diversity among the groups under study. A combination of small initial effective size and continued genetic isolation from surrounding populations seems to provide a coherent explanation for the diversity observed among Sauris, Sappada and Timau, which was found to be substantially greater than in other groups of European isolated populations. Simulations of micro-evolutionary scenarios indicate that ethnicity might have been important in increasing genetic diversity among these culturally related and spatially close populations.

  2. Genomic Variability of Serial Human Isolates of Salmonella enterica Serovar Typhimurium Associated with Prolonged Carriage.

    PubMed

    Octavia, Sophie; Wang, Qinning; Tanaka, Mark M; Sintchenko, Vitali; Lan, Ruiting

    2015-11-01

    Salmonella enterica serovar Typhimurium is an important foodborne human pathogen that often causes self-limiting but severe gastroenteritis. Prolonged excretion of S. Typhimurium after the infection can lead to secondary transmissions. However, little is known about within-host genomic variation in bacteria associated with asymptomatic shedding. Genomes of 35 longitudinal isolates of S. Typhimurium recovered from 11 patients (children and adults) with culture-confirmed gastroenteritis were sequenced. There were three or four isolates obtained from each patient. Single nucleotide polymorphisms (SNPs) were analyzed in these isolates, which were recovered between 1 and 279 days after the initial diagnosis. Limited genomic variation (5 SNPs or fewer) was associated with short- and long-term carriage of S. Typhimurium. None of the isolates was shown to be due to reinfection. SNPs occurred randomly, and the majority of the SNPs were nonsynonymous. Two nonsense mutations were observed. A nonsense mutation in flhC rendered the isolate nonmotile, whereas the significance of a nonsense mutation in yihV is unknown. The estimated mutation rate is 1.49 × 10(-6) substitution per site per year. S. Typhimurium isolates excreted in stools following acute gastroenteritis in children and adults demonstrated limited genomic variability over time, regardless of the duration of carriage. These findings have important implications for the detection of possible transmission events suspected by public health genomic surveillance of S. Typhimurium infections.

  3. Temporal Variabilities in Genetic Patterns and Antibiotic Resistance Profiles of Enterococci Isolated from Human Feces.

    PubMed

    Nishiyama, Masateru; Shimauchi, Hidetaka; Suzuki, Yoshihiro

    2016-06-25

    Temporal variabilities in the genetic patterns and antibiotic resistance profiles of enterococci were monitored over a 7-month period. Enterococcus faecalis isolates (103 strains) collected from feces showed only one genetic pattern and antibiotic resistance profile within 0 d and 30 d. In contrast, after 60 d and 90 d, the genetic patterns and antibiotic resistance profiles of all E. faecalis isolates (8 strains) clearly differed within 30 d. These results indicate that the genetic patterns and antibiotic resistance profiles of E. faecalis in human feces changed to completely dissimilar patterns between 1 and 2 months.

  4. Temporal Variabilities in Genetic Patterns and Antibiotic Resistance Profiles of Enterococci Isolated from Human Feces

    PubMed Central

    Nishiyama, Masateru; Shimauchi, Hidetaka; Suzuki, Yoshihiro

    2016-01-01

    Temporal variabilities in the genetic patterns and antibiotic resistance profiles of enterococci were monitored over a 7-month period. Enterococcus faecalis isolates (103 strains) collected from feces showed only one genetic pattern and antibiotic resistance profile within 0 d and 30 d. In contrast, after 60 d and 90 d, the genetic patterns and antibiotic resistance profiles of all E. faecalis isolates (8 strains) clearly differed within 30 d. These results indicate that the genetic patterns and antibiotic resistance profiles of E. faecalis in human feces changed to completely dissimilar patterns between 1 and 2 months. PMID:27265342

  5. 76 FR 51374 - Direct Discovery of HLA Associated Influenza Epitopes Isolated From Human Cells for Vaccine and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-18

    ... Isolated From Human Cells for Vaccine and Therapeutic Evaluation and Development (U01) AGENCY: Food and... processed and presented on soluble HLA (human leucocyte antigen) expressed by human cells. Initial studies... produced from influenza infected human lung cell lines. There is a growing interest in developing...

  6. Anti-caspase-3 preconditioning increases proinsulin secretion and deteriorates posttransplant function of isolated human islets.

    PubMed

    Brandhorst, Daniel; Brandhorst, Heide; Maataoui, Vidya; Maataoui, Adel; Johnson, Paul R V

    2013-06-01

    Human islet isolation is associated with adverse conditions inducing apoptosis and necrosis. The aim of the present study was to assess whether antiapoptotic preconditioning can improve in vitro and posttransplant function of isolated human islets. A dose-finding study demonstrated that 200 μmol/L of the caspase-3 inhibitor Ac-DEVD-CMK was most efficient to reduce the expression of activated caspase-3 in isolated human islets exposed to severe heat shock. Ac-DEVD-CMK-pretreated or sham-treated islets were transplanted into immunocompetent or immunodeficient diabetic mice and subjected to static glucose incubation to measure insulin and proinsulin secretion. Antiapoptotic pretreatment significantly deteriorated graft function resulting in elevated nonfasting serum glucose when compared to sham-treated islets transplanted into diabetic nude mice (p < 0.01) and into immunocompetent mice (p < 0.05). Ac-DEVD-CMK pretreatment did not significantly change basal and glucose-stimulated insulin release compared to sham-treated human islets but increased the proinsulin release at high glucose concentrations (20 mM) thus reducing the insulin-to-proinsulin ratio in preconditioned islets (p < 0.05). This study demonstrates that the caspase-3 inhibitor Ac-DEVD-CMK interferes with proinsulin conversion in preconditioned islets reducing their potency to cure diabetic mice. The mechanism behind this phenomenon is unclear so far but may be related to the ketone CMK linked to the Ac-DEVD molecule. Further studies are required to identify biocompatible caspase inhibitors suitable for islet preconditioning.

  7. Structure activity characterization of Bordetella petrii lipid A, from environment to human isolates.

    PubMed

    Basheer, Soorej M; Bouchez, Valerie; Novikov, Alexey; Augusto, Luis A; Guiso, Nicole; Caroff, Martine

    2016-01-01

    Bordetella petrii, a facultative anaerobic species, is the only known member of the Bordetella genus with environmental origin. However it was also recently isolated from humans. The structures of the B. petrii lipid A moieties of the endotoxins were characterized here for the first time for an environmental strain and compared to that of human isolates. Characterization was achieved using chemical analyses, gas chromatography-mass spectrometry, and Matrix Assisted Laser Desorption Ionisation mass spectrometry. The analyses revealed that the different lipid A structures contain a common bisphosphorylated β-(1→6)-linked d-glucosamine disaccharide with hydroxytetradecanoic acid in amide as well at the C-3' in ester linkages. Similar to Bordetella pertussis and Bordetella bronchiseptica lipids A, the hydroxytetradecanoic acid at the C-2' position was substituted by tetradecanoic acid. Unlike B. pertussis, the hydroxytetradecanoic acid at the C-2 position was substituted with either 12:0 or 14:0 and/or their 2-OH forms. Depending on the environmental or human origin the structures differed in the length and degree of fatty acid acylation and impacted the IL-6 and TNF-α inflammatory responses tested. In one isolate we showed the presence at the C-3 position of the short-chain 10:0(3-OH), which according to our previous analyses is more characteristic of the human pathogens in the genus like B. pertussis and Bordetella parapertussis. PMID:26164553

  8. Isolation of Human Neural Stem Cells from the Amniotic Fluid with Diagnosed Neural Tube Defects.

    PubMed

    Chang, Yu-Jen; Su, Hong-Lin; Hsu, Lee-Feng; Huang, Po-Jui; Wang, Tzu-Hao; Cheng, Fu-Chou; Hsu, Li-Wen; Tsai, Ming-Song; Chen, Chih-Ping; Chang, Yao-Lung; Chao, An-Shine; Hwang, Shiaw-Min

    2015-08-01

    Human neural stem cells (NSCs) are particularly valuable for the study of neurogenesis process and have a therapeutic potential in treating neurodegenerative disorders. However, current progress in the use of human NSCs is limited due to the available NSC sources and the complicated isolation and culture techniques. In this study, we describe an efficient method to isolate and propagate human NSCs from the amniotic fluid with diagnosed neural tube defects (NTDs), specifically, anencephaly. These amniotic fluid-derived NSCs (AF-NSCs) formed neurospheres and underwent long-term expansion in vitro. In addition, these cells showed normal karyotypes and telomerase activity and expressed NSC-specific markers, including Nestin, Sox2, Musashi-1, and the ATP-binding cassette G2 (ABCG2). AF-NSCs displayed typical morphological patterns and expressed specific markers that were consistent with neurons, astrocytes, oligodendrocytes, and dopaminergic neurons after proper induction conditions. Furthermore, grafted AF-NSCs improved the physiological functions in a rat stroke model. The ability to isolate and bank human NSCs from this novel source provides a unique opportunity for translational studies of neurological disorders. PMID:25923707

  9. Structure activity characterization of Bordetella petrii lipid A, from environment to human isolates.

    PubMed

    Basheer, Soorej M; Bouchez, Valerie; Novikov, Alexey; Augusto, Luis A; Guiso, Nicole; Caroff, Martine

    2016-01-01

    Bordetella petrii, a facultative anaerobic species, is the only known member of the Bordetella genus with environmental origin. However it was also recently isolated from humans. The structures of the B. petrii lipid A moieties of the endotoxins were characterized here for the first time for an environmental strain and compared to that of human isolates. Characterization was achieved using chemical analyses, gas chromatography-mass spectrometry, and Matrix Assisted Laser Desorption Ionisation mass spectrometry. The analyses revealed that the different lipid A structures contain a common bisphosphorylated β-(1→6)-linked d-glucosamine disaccharide with hydroxytetradecanoic acid in amide as well at the C-3' in ester linkages. Similar to Bordetella pertussis and Bordetella bronchiseptica lipids A, the hydroxytetradecanoic acid at the C-2' position was substituted by tetradecanoic acid. Unlike B. pertussis, the hydroxytetradecanoic acid at the C-2 position was substituted with either 12:0 or 14:0 and/or their 2-OH forms. Depending on the environmental or human origin the structures differed in the length and degree of fatty acid acylation and impacted the IL-6 and TNF-α inflammatory responses tested. In one isolate we showed the presence at the C-3 position of the short-chain 10:0(3-OH), which according to our previous analyses is more characteristic of the human pathogens in the genus like B. pertussis and Bordetella parapertussis.

  10. Cyclic AMP-dependent phosphorylation of neuronal nitric oxide synthase mediates penile erection

    PubMed Central

    Hurt, K. Joseph; Sezen, Sena F.; Lagoda, Gwen F.; Musicki, Biljana; Rameau, Gerald A.; Snyder, Solomon H.; Burnett, Arthur L.

    2012-01-01

    Nitric oxide (NO) generated by neuronal NO synthase (nNOS) initiates penile erection, but has not been thought to participate in the sustained erection required for normal sexual performance. We now show that cAMP-dependent phosphorylation of nNOS mediates erectile physiology, including sustained erection. nNOS is phosphorylated by cAMP-dependent protein kinase (PKA) at serine(S)1412. Electrical stimulation of the penile innervation increases S1412 phosphorylation that is blocked by PKA inhibitors but not by PI3-kinase/Akt inhibitors. Stimulation of cAMP formation by forskolin also activates nNOS phosphorylation. Sustained penile erection elicited by either intracavernous forskolin injection, or augmented by forskolin during cavernous nerve electrical stimulation, is prevented by the NOS inhibitor l-NAME or in nNOS-deleted mice. Thus, nNOS mediates both initiation and maintenance of penile erection, implying unique approaches for treating erectile dysfunction. PMID:23012472

  11. Cyclic AMP-dependent phosphorylation of neuronal nitric oxide synthase mediates penile erection.

    PubMed

    Hurt, K Joseph; Sezen, Sena F; Lagoda, Gwen F; Musicki, Biljana; Rameau, Gerald A; Snyder, Solomon H; Burnett, Arthur L

    2012-10-01

    Nitric oxide (NO) generated by neuronal NO synthase (nNOS) initiates penile erection, but has not been thought to participate in the sustained erection required for normal sexual performance. We now show that cAMP-dependent phosphorylation of nNOS mediates erectile physiology, including sustained erection. nNOS is phosphorylated by cAMP-dependent protein kinase (PKA) at serine(S)1412. Electrical stimulation of the penile innervation increases S1412 phosphorylation that is blocked by PKA inhibitors but not by PI3-kinase/Akt inhibitors. Stimulation of cAMP formation by forskolin also activates nNOS phosphorylation. Sustained penile erection elicited by either intracavernous forskolin injection, or augmented by forskolin during cavernous nerve electrical stimulation, is prevented by the NOS inhibitor L-NAME or in nNOS-deleted mice. Thus, nNOS mediates both initiation and maintenance of penile erection, implying unique approaches for treating erectile dysfunction.

  12. Valproic Acid Prevents Penile Fibrosis and Erectile Dysfunction in Cavernous Nerve Injured Rats

    PubMed Central

    Hannan, Johanna L.; Kutlu, Omer; Stopak, Bernard L.; Liu, Xiaopu; Castiglione, Fabio; Hedlund, Petter; Burnett, Arthur L.; Bivalacqua, Trinity J.

    2014-01-01

    Introduction Bilateral cavernous nerve injury (BCNI) causes profound penile changes such as apoptosis and fibrosis leading to erectile dysfunction (ED). Histone deacetylase (HDAC) has been implicated in chronic fibrotic diseases. Aims This study will characterize the molecular changes in penile HDAC after BCNI and determine if HDAC inhibition can prevent BCNI-induced ED and penile fibrosis. Methods Five groups of rats (8–10 wks, n=10/group) were utilized: 1) sham, 2&3) BCNI 14 and 30 days following injury, and 4&5) BCNI treated with HDAC inhibitor valproic acid (VPA 250mg/kg; 14 and 30 days). All groups underwent cavernous nerve stimulation (CNS) to determine intracavernosal pressure (ICP). Penile HDAC3, HDAC4, fibronectin, and transforming growth factor-β1 (TGF-β1) protein expression (Western blot) were assessed. Trichrome staining and the fractional area of fibrosis were determined in penes from each group. Cavernous smooth muscle content was assessed by immunofluorescence to alpha smooth muscle actin (α-SMA) antibodies. Main Outcome Measures ICP; HDAC3, HDAC4, fibronectin and TGF-β1 protein expression; penile fibrosis; penile α-SMA content. Results There was a voltage-dependent decline (p<0.05) in ICP to CNS 14 and 30 days after BCNI. Penile HDAC3, HDAC4, and fibronectin were significantly increased (P<0.05) 14 days after BCNI. There was a slight increase in TGF-β1 protein expression after BCNI. Histological analysis showed increased (P<0.05) corporal fibrosis after BCNI at both time points. VPA treatment decreased (P<0.05) penile HDAC3, HDAC4, and fibronectin protein expression as well as corporal fibrosis. There was no change in penile α-SMA between all groups. Furthermore, VPA-treated BCNI rats had improved erectile responses to CNS (P<0.05). Conclusion HDAC-induced pathological signaling in response to BCNI contributes to penile vascular dysfunction after BCNI. Pharmacological inhibition of HDAC prevents penile fibrosis, normalizes fibronectin

  13. Isolation of human erythrocyte acetylcholinesterase using phase separation with Triton X-114 and monoclonal immunosorbent chromatography.

    PubMed

    Bjerrum, O J; Selmer, J; Hangaard, J; Larsen, F

    1985-01-01

    A generally applicable approach to the preparative isolation of amphiphilic membrane proteins that follow the Triton X-114 phase during a temperature-dependent phase separation is described. The phase separations were performed direct on whole blood and a 650-fold purification of human erythrocyte membrane acetylcholinesterase (AchE) was obtained. Thus, 0.2 mg enzyme was isolated per 1 liter of blood, with a specific activity of 13 IU/mg, the major contaminants being glycophorin and hemoglobin. The protein material was isolated from the detergent phase by Cu2+ chelate chromatography. This material was used to raise monoclonal anti-AchE antibodies which, when applied to immunosorbent chromatography of washed Triton X-100-lysed erythrocytes in one step, allowed a 246,000-fold purification of AchE with a yield of 88% and a specific activity of 3800 IU/mg.

  14. Flow-Cytometric Isolation of Human Antibodies from a Nonimmune Saccharomyces cerevisiae Surface Display Library

    SciTech Connect

    Feldhaus, Michael ); Siegel, Robert W. ); Opresko, Lee ); Coleman, James R. ); Feldhaus, Jane M. ); Yeung, Yik A.; Cochran, Jennifer R.; Heinzelman, Peter; Colby, David; Swers, Jeffrey; Graff, Christilyn; Wiley, H Steven ); Wittrup, K D.

    2003-02-28

    A nonimmune library of 109 human antibody scFv fragments has been cloned and expressed on the surface of yeast, and nanomolar-affinity scFvs routinely obtained by magnetic bead screening and flow cytometric sorting. The yeast library can be amplified 1010-fold without measurable loss of clonal diversity, enabling effectively indefinite expansion of the library. The expression, stability, and antigen binding properties of more than 50 isolated scFv clones were assessed directly on the yeast cell surface by immunofluorescent labeling and flow cytometry, obviating separate subcloning, expression, and purification steps and thereby expediting the isolation of novel affinity reagents. The ability to use multiplex library screening demonstrates the utility of this approach for high throughput antibody isolation for proteomics applications.

  15. Genotypic diversity and virulent factors of Staphylococcus epidermidis isolated from human breast milk.

    PubMed

    Begović, Jelena; Jovčić, Branko; Papić-Obradović, Milena; Veljović, Katarina; Lukić, Jovanka; Kojić, Milan; Topisirović, Ljubiša

    2013-02-22

    Staphylococcus epidermidis strains were isolated from the expressed human breast milk (EHM) of 14 healthy donor mothers. Genetic diversity was evaluated using RAPD-PCR REP-PCR and pulse-field gel electrophoresis (PFGE). PFGE allowed the best discrimination of the isolates, since it provided for the greatest diversity of the analyzed genomes. Among the S. epidermidis strains, resistance to gentamicin, tetracycline, erythromycin, clindamycin or vancomycin was detected, whilst four isolates were multiresistant. The results from our study demonstrate that staphylococci from EHM could be reservoirs of resistance genes, since we showed that tetK could be transferred from EHM staphylococci to Gram-negative Escherichia coli. Most of the staphylococcal strains displayed excellent proteolytic and lipolytic activities. Additionally, the presence of ica genes, which was related to their ability to form a biofilm on tissue culture plates, and the presence of virulence factors including autolysin/adhesin AtLE, point to their pathogenic potential.

  16. Xenogenic transfer of isolated murine mitochondria into human rho0 cells can improve respiratory function.

    PubMed

    Katrangi, Eyad; D'Souza, Gerard; Boddapati, Sarathi V; Kulawiec, Mariola; Singh, Keshav K; Bigger, Brian; Weissig, Volkmar

    2007-12-01

    Mitochondrial DNA mutations are the direct cause of several physiological disorders and are also associated with the aging process. The modest progress made over the past two decades towards manipulating the mitochondrial genome and understanding its function within living mammalian cells means that cures for mitochondrial DNA mutations are still elusive. Here, we report that transformed mammalian cells internalize exogenous isolated mitochondria upon simple co-incubation. We first demonstrate the physical presence of internalized mitochondria within recipient cells using fluorescence microscopy. Second, we show that xenogenic transfer of murine mitochondria into human cells lacking functional mitochondria can functionally restore respiration in cells lacking mtDNA. Third, utilizing the natural competence of isolated mitochondria to take up linear DNA molecules, we demonstrate the feasibility of using cellular internalization of isolated exogenous mitochondria as a potential tool for studying mitochondrial genetics in living mammalian cells. PMID:18069915

  17. Mutagenicity towards Salmonella typhimurium of some known genotoxic agents, activated by isolated hepatocytes of monkey (Macaca fascicularis). Comparison with isolated human hepatocytes.

    PubMed

    Neis, J M; Roelofs, H M; van Gemert, P J; Henderson, P T

    1986-06-01

    This paper describes some striking differences between isolated human and monkey hepatocytes in their capacity to activate some known genotoxic agents into products mutagenic towards Salmonella typhimurium. Isolated monkey hepatocytes, in contrast to human hepatocytes, appeared to activate benzidine (BZ), N-acetylbenzidine (MABZ), N,N'-diacetylbenzidine (DABZ), 2-aminofluorene (2-AF) and 2-acetylaminofluorene (2-AAF) poorly. With monkey hepatocytes BZ was slightly more mutagenic than DABZ, whereas with human hepatocytes DABZ was more active than BZ. N-Nitrosodimethylamine (DMN) and N-nitrosodiethylamine (DEN) were also found to be poorly mutagenic when activated by monkey hepatocytes, unlike the human hepatocytes. However, the polycyclic arylhydrocarbons benzo[a]pyrene (B[a]P) and 7,12-dimethylbenzanthracene (7,12-DMBA) were highly active in the presence of monkey hepatocytes, unlike the human hepatocytes. A metabolic study showed that monkey liver preparations seem to possess a higher monooxygenase activity towards B[a]P than human liver preparations.

  18. Penile entrapment in a plastic bottle - a case for using an oscillating splint saw.

    PubMed

    May, Matthias; Gunia, Sven; Helke, Christian; Kheyri, Reza; Hoschke, Bernd

    2006-01-01

    Penile entrapment is a rare but serious urological emergency, which can easily lead to stangulation and infarction. We report a case of penile entrapment in a polyethylene terephthalate (PET) bottle in a 49-year-old male. Attempts to cut the bottle with a scalpel or a glass saw were ineffective. Finally, the bottle neck was cut longitudinally with an oscillating saw intended for cutting plaster casts.

  19. Penile enhancement using a porcine small intestinal submucosa graft in a rat model.

    PubMed

    Leungwattanakij, S; Pummangura, N; Ratana-Olarn, K

    2006-01-01

    Several biodegradable materials have been experimented for penile enhancement, but none show the potential for clinical use. This study was designed to use porcine small intestinal submucosa (SIS) augmenting the normal tunica albuginea to increase the functional girth of the rat penis. In all, 20 adult male Sprague-Dawley rats constituted the study population. The animals were divided into two groups: group 1 consisted of the control (n=10) and group 2 (n=10) consisted of rats that underwent penile enhancement by a longitudinal I-shaped incision of the tunica albuginea on both sides, and the dissection of the plane between tunica albuginea and cavernosal tissue was carried out (n=10). The incision was then patched with a 3 x 10 mm2 piece of SIS, using a 6/0 nylon suture material. The penile length and mid-circumference were then measured using a Vernier Caliper before and 2 months after surgery. All rat penises underwent histological examination using Masson's trichome and Verhoff's van Giesen's stain for collagen and elastic fibers. The penile length, mid-circumference and degree of fibrosis score were expressed as mean+/-s.e. (standard error) and analyzed using a Wilcoxon rank-sum test. A statistical significance was accepted at P-value < or =0.05. Our results showed similar preoperative penile length and circumference in both groups. However, 2 months after the surgery, the mean penile circumference of the SIS group has grown significantly larger than the control group, while the mean penile length remained unchanged. The histological study of the rat penises revealed minimal amounts of fibrosis under the graft, and the elastic fibers of the graft showed orientation in a circular manner. In conclusion, SIS appears promising for material use in a penile enhancement. PMID:16049525

  20. Reference standard of penile size and prevalence of buried penis in Japanese newborn male infants.

    PubMed

    Matsuo, Nobutake; Ishii, Tomohiro; Takayama, John I; Miwa, Masayuki; Hasegawa, Tomonobu

    2014-01-01

    The present study set forth the reference values for penile size and determined the prevalence of buried penis in Japanese full-term newborns. The stretched penile length was measured and the presence of buried penis was assessed at 1-7 days of age in 547 Japanese full-term newborn infants born between 2008 and 2012 in Tokyo. The stretched penile lengths were compared at 1-12 hours and 1-7 days of age in 63 infants and by two observers in 73 infants to estimate postnatal changes and interobserver variation, respectively. The mean stretched penile length was 3.06 cm (SD, 0.26; 95% confidence interval [CI], 3.04-3.08) and the mean ratio of penile length to body length was 6.24 × 100(-1) (SD, 0.55 × 100(-1)), both of which were significantly smaller than those in Caucasian newborn infants. Buried penis was identified in 20 of 547 infants (3.7%; 95% CI, 2.1-5.2%). The first measurements of penile length at 1-12 hours were significantly smaller than the next measurements at 1-7 days (95% CI of the difference, 0.22-0.34). The 95% CI for the limits of agreement in the penile lengths measured by the two observers was -0.58 to -0.40 for the lower limit and 0.33 to 0.51 for the upper limit. These findings indicate that the penile length should be assessed after 24 hours of age by the reference standard of the same ethnicity for identifying micropenis and that buried penis is not uncommon in Japanese full-term newborns.

  1. Current status of penile rehabilitation after radical prostatectomy

    PubMed Central

    Kim, Jae Heon

    2015-01-01

    Although disease-free survival remains the primary goal of prostate cancer treatment, erectile dysfunction (ED) remains a common complication that affects the quality of life. Even though several preventive and therapeutic strategies are available for ED after radical prostatectomy (RP), no specific recommendations have been made on the optimal rehabilitation or treatment strategy. Several treatment options are available, including phosphodiesterase-5 inhibitors, vacuum erection devices, intracavernosal or intraurethral prostaglandin injections, and penile prostheses. Urologists must consider more effective ways to establish optimal treatments for ED after RP. ED is an important issue among patients with prostate cancer, and many patients hope for early ED recovery after surgery. This review highlights the currently available treatment options for ED after RP and discusses the limitations of each. PMID:25685296

  2. Pityriasis versicolor on penile shaft in a renal transplant recipient.

    PubMed

    Ryu, Han-Won; Cho, Jae-We; Lee, Kyu-Suk

    2012-08-01

    Pityriasis versicolor is a superficial infection of the stratum corneum, which is caused by the Malassezia species. Tge Malassezia species consist of 12 subspecies, including M. furfur, M. pachydermatis, M. symphodialis and M. globasa. The Malassezia species are classified as a normal flora, particularly in the sebum rich areas of the skin, and they convert from saprophytic yeast to parasitic mycelial morpholgic form to cause clinical disease. But majorities of their distributions are in the upper back, the neck, the thighs, and the forearm, and not in the penis. It is well known that the renal transplant patients, who take immunosuppressive agents, have impairment in the protective cell mediated immunity. Thus, they are more susceptible to infectious diseases, such as a fungal infection. Therefore, clinical manifestations show higher incidence of disease, but they mostly occur in an expected distribution. We here report a case of pityriasis versicolor in a renal transplant recipient on penile shaft, which is an unusual area.

  3. Genotypes and Mouse Virulence of Toxoplasma gondii Isolates from Animals and Humans in China

    PubMed Central

    Liu, Daohua; Huo, Xingxing; Gao, Jiangmei; Song, Xiaorong; Xu, Xiucai; Huang, Kaiquan; Liu, Wenqi; Wang, Yong; Lu, Fangli; Lun, Zhao-Rong; Luo, Qingli; Wang, Xuelong; Shen, Jilong

    2013-01-01

    Background Recent population structure studies of T. gondii revealed that a few major clonal lineages predominated in different geographical regions. T. gondii in South America is genetically and biologically divergent, whereas this parasite is remarkably clonal in North America and Europe with a few major lineages including Types I, II and III. Information on genotypes and mouse virulence of T. gondii isolates from China is scarce and insufficient to investigate its population structure, evolution, and transmission. Methodology/Principal Findings Genotyping of 23 T. gondii isolates from different hosts using 10 markers for PCR-restriction fragment length polymorphism analyses (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) revealed five genotypes; among them three genotypes were atypical and two were archetypal. Fifteen strains belong to the Chinese 1 lineage, which has been previously reported as a widespread lineage from swine, cats, and humans in China. Two human isolates fall into the type I and II lineages and the remaining isolates belong to two new atypical genotypes (ToxoDB#204 and #205) which has never been reported in China. Our results show that these genotypes of T. gondii isolates are intermediately or highly virulent in mice except for the strain TgCtwh6, which maintained parasitemia in mice for 35 days post infection although it possesses the uniform genotype of Chinese 1. Additionally, phylogenetic network analyses of all isolates of genotype Chinese 1 are identical, and there is no variation based on the sequence data generated for four introns (EF1, HP2, UPRT1 and UPRT7) and two dense granule proteins (GRA6 and GRA7). Conclusion/Significance A limited genetic diversity was found and genotype Chinese 1 (ToxoDB#9) is dominantly circulating in mainland China. The results will provide a useful profile for deep insight to the population structure, epidemiology and biological characteristics of T. gondii in China. PMID:23308233

  4. Carboxybetaine Modified Interface for Electrochemical Glycoprofiling of Antibodies Isolated from Human Serum

    PubMed Central

    2015-01-01

    Impedimetric lectin biosensors capable of recognizing two different carbohydrates (galactose and sialic acid) in glycans attached to antibodies isolated from human serum were prepared. The first step entailed the modification of a gold surface by a self-assembled monolayer (SAM) deposited from a solution containing a carboxybetaine-terminated thiol applied to the subsequent covalent immobilization of lectins and to resist nonspecific protein adsorption. In the next step, Sambucus nigra agglutinin (SNA) or Ricinus communis agglutinin (RCA) was covalently attached to the SAM, and the whole process of building a bioreceptive layer was optimized and characterized using a diverse range of techniques including electrochemical impedance spectroscopy, cyclic voltammetry, quartz crystal microbalance, contact angle measurements, zeta-potential assays, X-ray photoelectron spectroscopy, and atomic force microscopy. In addition, the application of the SNA-based lectin biosensor in the glycoprofiling of antibodies isolated from the human sera of healthy individuals and of patients suffering from rheumatoid arthritis (RA) was successfully validated using an SNA-based lectin microarray. The results showed that the SNA lectin, in particular, is capable of discriminating between the antibodies isolated from healthy individuals and those from RA patients based on changes in the amount of sialic acid present in the antibodies. In addition, the results obtained by the application of RCA and SNA biosensors indicate that the abundance of galactose and sialic acid in antibodies isolated from healthy individuals is age-related. PMID:26048139

  5. Characterization of quinolone resistance in Salmonella spp. isolates from food products and human samples in Brazil

    PubMed Central

    Pribul, Bruno Rocha; Festivo, Marcia Lima; de Souza, Miliane Moreira Soares; dos Prazeres Rodrigues, Dalia

    2016-01-01

    Non-typhoidal salmonellosis is an important zoonotic disease caused by Salmonella enterica. The aim of this study was to investigate the prevalence of plasmid-mediated quinolone resistance in Salmonella spp. and its association with fluoroquinolone susceptibility in Brazil. A total of 129 NTS isolates (samples from human origin, food from animal origin, environmental, and animal) grouped as from animal (n = 62) and human (n = 67) food were evaluated between 2009 and 2013. These isolates were investigated through serotyping, antimicrobial susceptibility testing, and the presence of plasmid-mediated quinolone resistance (PMQR) genes (qnr, aac(6′)-Ib) and associated integron genes (integrase, and conserved integron region). Resistance to quinolones and/or fluoroquinolones, from first to third generations, was observed. Fifteen isolates were positive for the presence of qnr genes (8 qnrS, 6 qnrB, and 1 qnrD) and twenty three of aac(6′)-Ib. The conserved integron region was detected in 67 isolates as variable regions, from ±600 to >1000 pb. The spread of NTS involving PMQR carriers is of serious concern and should be carefully monitored. PMID:26887245

  6. Characterization of quinolone resistance in Salmonella spp. isolates from food products and human samples in Brazil.

    PubMed

    Pribul, Bruno Rocha; Festivo, Marcia Lima; de Souza, Miliane Moreira Soares; Rodrigues, Dalia dos Prazeres

    2016-01-01

    Non-typhoidal salmonellosis is an important zoonotic disease caused by Salmonella enterica. The aim of this study was to investigate the prevalence of plasmid-mediated quinolone resistance in Salmonella spp. and its association with fluoroquinolone susceptibility in Brazil. A total of 129 NTS isolates (samples from human origin, food from animal origin, environmental, and animal) grouped as from animal (n=62) and human (n=67) food were evaluated between 2009 and 2013. These isolates were investigated through serotyping, antimicrobial susceptibility testing, and the presence of plasmid-mediated quinolone resistance (PMQR) genes (qnr, aac(6')-Ib) and associated integron genes (integrase, and conserved integron region). Resistance to quinolones and/or fluoroquinolones, from first to third generations, was observed. Fifteen isolates were positive for the presence of qnr genes (8 qnrS, 6 qnrB, and 1 qnrD) and twenty three of aac(6')-Ib. The conserved integron region was detected in 67 isolates as variable regions, from ±600 to >1000pb. The spread of NTS involving PMQR carriers is of serious concern and should be carefully monitored.

  7. Virulence characterization of non-O157 Shiga toxin-producing Escherichia coli isolates from food, humans and animals.

    PubMed

    Shen, Jinling; Rump, Lydia; Ju, Wenting; Shao, Jingdong; Zhao, Shaohua; Brown, Eric; Meng, Jianghong

    2015-09-01

    A total of 359 non-O157 STEC isolates from food, humans and animals were examined for serotypes, Shiga toxin subtypes and intimin subtypes. Isolates solely harboring stx2 from the three sources were selected for Vero cell cytotoxicity test. stx subtypes in eae negative isolates were more diverse than in eae positive isolates primarily carrying stx2a. Four eae subtypes (eaeβ,eaeε1,eaeγ1 and eaeγ2/θ) were observed and correlated with serotypes and flagella. Food isolates showed more diverse serotypes, virulence factors and cell cytotoxicities than human isolates. Some isolates from produce belonged to serotypes that have been implicated in human diseases, carried stx2a or/and stx2dact and exhibited high cell cytotoxicity similar to human isolates. This indicates that foods can be contaminated with potentially pathogenic STEC isolates that may cause human diseases. Given the increased produce consumption and growing burden of foodborne outbreaks due to produce, produce safety should be given great importance.

  8. Prevalence and pathogenic potential of campylobacter isolates from free-living, human-commensal american crows.

    PubMed

    Weis, Allison M; Miller, Woutrina A; Byrne, Barbara A; Chouicha, Nadira; Boyce, Walter M; Townsend, Andrea K

    2014-03-01

    Recent studies have suggested a potential role for wild birds in zoonotic transmission of Campylobacter jejuni, the leading cause of gastroenteritis in humans worldwide. In this study, we detected Campylobacter spp. in 66.9% (85/127) of free-ranging American crows (Corvus brachyrhyncos) sampled in the Sacramento Valley of California in 2012 and 2013. Biochemical testing and sequence analysis of 16S rRNA revealed that 93% of isolates (n = 70) were C. jejuni, with cytolethal distending toxin (CDT) and flagellin A genes detected by PCR in 20% and 46% of the C. jejuni isolates (n = 59), respectively. The high prevalence of C. jejuni, coupled with the occurrence of known virulence markers CDT and flagellin A, demonstrates that crows shed Campylobacter spp. in their feces that are potentially pathogenic to humans. Crows are abundant in urban, suburban, and agricultural settings, and thus further study to determine their role in zoonotic transmission of Campylobacter will inform public health.

  9. Biotransformation of luteoloside by a newly isolated human intestinal bacterium using UHPLC-Q-TOF/MS.

    PubMed

    Tao, Jin-hua; Wang, Dong-geng; Yang, Chi; Huang, Jin-hua; Qiu, Wen-qian; Zhao, Xi

    2015-06-01

    To explore the metabolic pathways and metabolites of luteoloside yielded by the isolated human intestinal bacteria from healthy human feces and characterize the β-d-glucosidase activity of the specific strain which catalyzed the breakdown of luteoloside, a preculture bacterial GAM broth and luteoloside were mixed incubated together for 48h. UHPLC-Q-TOF/MS was used for analysis of the metabolites of luteoloside in the corresponding supernatant fractions from fermentation. Aliquots of the reactive solutions were collected at different times and were measured with a microplate reader at 405nm to evaluate the enzymatic activity. Three metabolites (acetylated luteoloside, luteolin and deoxygenated luteolin) were detected in the fractions isolated from the bacterial samples. The variation of β-d-glucosidase activity inside the bacterium was in coincidence with the changes in luteolin generation or luteoloside degradation in different time periods.

  10. Prevalence and Pathogenic Potential of Campylobacter Isolates from Free-Living, Human-Commensal American Crows

    PubMed Central

    Weis, Allison M.; Miller, Woutrina A.; Byrne, Barbara A.; Chouicha, Nadira; Boyce, Walter M.

    2014-01-01

    Recent studies have suggested a potential role for wild birds in zoonotic transmission of Campylobacter jejuni, the leading cause of gastroenteritis in humans worldwide. In this study, we detected Campylobacter spp. in 66.9% (85/127) of free-ranging American crows (Corvus brachyrhyncos) sampled in the Sacramento Valley of California in 2012 and 2013. Biochemical testing and sequence analysis of 16S rRNA revealed that 93% of isolates (n = 70) were C. jejuni, with cytolethal distending toxin (CDT) and flagellin A genes detected by PCR in 20% and 46% of the C. jejuni isolates (n = 59), respectively. The high prevalence of C. jejuni, coupled with the occurrence of known virulence markers CDT and flagellin A, demonstrates that crows shed Campylobacter spp. in their feces that are potentially pathogenic to humans. Crows are abundant in urban, suburban, and agricultural settings, and thus further study to determine their role in zoonotic transmission of Campylobacter will inform public health. PMID:24375131

  11. Relationship between plasma cholesterol levels and cholesterol esterification in isolated human mononuclear cells

    SciTech Connect

    Dallongeville, J.; Davignon, J.; Lussier-Cacan, S. )

    1990-01-01

    The authors studied the relationship between plasma lipoprotein concentrations and cholesterol esterification in freshly isolated human mononuclear cells from 27 normolipidemic and 32 hyperlipidemic individuals. Cells were either incubated for 5 hours with radiolabeled oleate immediately after isolation or were preincubated for 18 hours in the presence of exogenous cholesterol, and then incubated with ({sup 14}C)sodium-oleate-albumin complex. In the absence of exogenous cholesterol, control and hypercholesterolemic subjects had similarly low values of intracellular cholesterol esterification. In the presence of exogenous cholesterol, both hypertriglyceridemic and hypercholesterolemic subjects had higher cholesterol esterification than controls. There was a significant correlation between the rate of cholesterol esterification and plasma total cholesterol. These results suggest that plasma cholesterol levels may regulate mononuclear cell intra-cellular cholesterol esterification in humans.

  12. Distribution and genetic variability among Campylobacter spp. isolates from different animal species and humans in Switzerland.

    PubMed

    Keller, J; Wieland, B; Wittwer, M; Stephan, R; Perreten, V

    2007-01-01

    In Switzerland, a national database with 1028 Campylobacter isolates from poultry, pigs, cats, dogs, cattle, humans, zoo animals and water has been created. The database contains the genetic fingerprint and background information of each Campylobacter isolate. Dominant species could be identified in the different sources with a majority of Campylobacter jejuni in poultry (73%), humans (79%), cattle (95%), zoo animals (40%) and water (100%), of Campylobacter coli in pigs (72%), and of Campylobacter upsaliensis/helveticus in cats and dogs (55%). The comparison of three genotyping methods, amplified fragment length polymorphism (AFLP), pulsed field gel electrophoresis and restriction fragment length polymorphism, revealed that AFLP allows discrimination between the different Campylobacter species and is the most appropriate method to distinguish specific strains within the same species. Genotyping analysis demonstrated that the Campylobacter population is heterogeneous among the different sources and that no dominant clone is spread in the country. Genotyping and the resulting database are useful tools to trace back future Campylobacter infections.

  13. A comparative study of two methods for the isolation of human leucocytes for DNA extraction.

    PubMed

    Lim, L H; Ton, S H; Cheong, S K

    1990-06-01

    The 'Dextran' and the 'Buffy-coat' methods for isolation of human leucocytes for DNA extraction were compared on the basis of DNA yield from the same amounts (10 ml) of blood. Human leucocytes from a total of 11 samples were isolated using both methods for each sample after which DNA was extracted. Extracted DNA samples were treated with ribonucleases and proteinase K after which the yields were quantitated by measuring absorbance at 260 nm. The 'Buffy-coat' method yielded a mean concentration of DNA of 476.7 micrograms/ml (range: 212 to 700 micrograms/ml) while the 'Dextran' method yielded 188.4 micrograms/ml (range: 64 to 340 micrograms/ml). The difference was confirmed by subjecting the extracted DNA samples to agarose gel electrophoresis.

  14. Isolation and Characterization of Broad and Ultrapotent Human Monoclonal Antibodies with Therapeutic Activity against Chikungunya Virus.

    PubMed

    Smith, Scott A; Silva, Laurie A; Fox, Julie M; Flyak, Andrew I; Kose, Nurgun; Sapparapu, Gopal; Khomandiak, Solomiia; Khomadiak, Solomiia; Ashbrook, Alison W; Kahle, Kristen M; Fong, Rachel H; Swayne, Sherri; Doranz, Benjamin J; McGee, Charles E; Heise, Mark T; Pal, Pankaj; Brien, James D; Austin, S Kyle; Diamond, Michael S; Dermody, Terence S; Crowe, James E

    2015-07-01

    Chikungunya virus (CHIKV) is a mosquito-transmitted RNA virus that causes acute febrile infection associated with polyarthralgia in humans. Mechanisms of protective immunity against CHIKV are poorly understood, and no effective therapeutics or vaccines are available. We isolated and characterized human monoclonal antibodies (mAbs) that neutralize CHIKV infectivity. Among the 30 mAbs isolated, 13 had broad and ultrapotent neutralizing activity (IC50 < 10 ng/ml), and all of these mapped to domain A of the E2 envelope protein. Potent inhibitory mAbs blocked post-attachment steps required for CHIKV membrane fusion, and several were protective in a lethal challenge model in immunocompromised mice, even when administered at late time points after infection. These highly protective mAbs could be considered for prevention or treatment of CHIKV infection, and their epitope location in domain A of E2 could be targeted for rational structure-based vaccine development.

  15. Isolation and characterization of broad and ultrapotent human monoclonal antibodies with therapeutic activity against chikungunya virus

    PubMed Central

    Smith, Scott A.; Silva, Laurie A.; Fox, Julie M.; Flyak, Andrew; Kose, Nurgun; Sapparapu, Gopal; Khomadiak, Solomiia; Ashbrook, Alison W.; Kahle, Kristen M.; Fong, Rachel H.; Swayne, Sherri; Doranz, Benjamin J.; McGee, Charles E.; Heise, Mark T.; Pal, Pankaj; Brien, James D.; Austin, S. Kyle; Diamond, Michael S.; Dermody, Terence S.; Crowe, James E.

    2015-01-01

    SUMMARY Chikungunya virus (CHIKV) is a mosquito-transmitted RNA virus that causes acute febrile infection associated with polyarthralgia in humans. Mechanisms of protective immunity against CHIKV are poorly understood, and no effective therapeutics or vaccines are available. We isolated and characterized human monoclonal antibodies (mAbs) that neutralize CHIKV infectivity. Among the 30 mAbs isolated, 13 had broad and ultrapotent neutralizing activity (IC50 < 10 ng/mL), and all of these mapped to domain A of the E2 envelope protein. Potent inhibitory mAbs blocked post-attachment steps required for CHIKV membrane fusion, and several were protective in a lethal challenge model in immunocompromised mice, even when administered at late time points after infection. These highly protective mAbs could be considered for prevention or treatment of CHIKV infection, and their epitope location in domain A of E2 could be targeted for rational structure-based vaccine development. PMID:26159721

  16. Radioisotope penile plethysmography: A technique for evaluating corpora cavernosal blood flow during early tumescence

    SciTech Connect

    Schwartz, A.N.; Graham, M.M.; Ferency, G.F.; Miura, R.S.

    1989-04-01

    Radioisotope penile plethysmography is a nuclear medicine technique which assists in the evaluation of patients with erectile dysfunction. This technique attempts to noninvasively quantitate penile corpora cavernosal blood flow during early penile tumescence using technetium-99m-labeled red blood cells. Penile images and counts were acquired in a steady-state blood-pool phase prior to and after the administration of intracorporal papaverine. Penile counts, images, and time-activity curves were computer analyzed in order to determine peak corporal flow and volume changes. Peak corporal flow rates were compared to arterial integrity (determined by angiography) and venosinusoidal corporal leak (determined by cavernosometry). Peak corporal flow correlated well with arterial integrity (r = 0.91) but did not correlate with venosinusoidal leak parameters (r = 0.01). This report focuses on the methodology and the assumptions which form the foundation of this technique. The strong correlation of peak corporal flow and angiography suggests that radioisotope penile plethysmography could prove useful in the evaluation of arterial inflow disorders in patients with erectile dysfunction.

  17. Electrophysiological actions of the dopamine agonist apomorphine in the paraventricular nucleus during penile erection.

    PubMed

    Richards, Natalie; Wayman, Chris; Allers, Kelly A

    2009-11-20

    The ability to achieve and maintain penile erection is necessary for successful copulation. Studies have demonstrated that dopamine receptor stimulation in the paraventricular nucleus (PVN) of the hypothalamus induces penile erection in rodents, and the dopamine agonist apomorphine has been used to treat erectile dysfunction. The aim of this study was to determine the electrophysiological characteristics of PVN neuronal firing activity in anaesthetised rodents during apomorphine-induced erection. Our findings can be placed in two categories; those effects that occur immediately upon apomorphine administration and continue for up to several minutes prior to penile erection, deemed 'pre-erectile', and those effects that were only observed during penile erection and seminal emission. In the pre-erectile period, apomorphine acts on two different populations of PVN neurons to increase or decrease firing rates and increases alpha1 frequency band power in local field potentials. Decreased delta and increased theta frequency power in PVN local field potentials occur only during penile erection and seminal emission. These studies provide further understanding of the coordinated neuronal activity that occurs in the PVN during apomorphine-induced penile erection.

  18. Methicillin (Oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans.

    PubMed

    Lee, John Hwa

    2003-11-01

    From May 2001 to April 2003, various types of specimens from cattle, pigs, and chickens were collected and examined for the presence of methicillin (oxacillin)-resistant Staphylococcus aureus (MRSA). S. aureus was isolated and positively identified by using Gram staining, colony morphology, tests for coagulase and urease activities, and an API Staph Ident system. Among 1,913 specimens collected from the animals, 421 contained S. aureus; of these, 28 contained S. aureus resistant to concentrations of oxacillin higher than 2 micro g/ml. Isolates from 15 of the 28 specimens were positive by PCR for the mecA gene. Of the 15 mecA-positive MRSA isolates, 12 were from dairy cows and 3 were from chickens. Antimicrobial susceptibility tests of mecA-positive MRSA strains were performed by the disk diffusion method. All isolates were resistant to members of the penicillin family, such as ampicillin, oxacillin, and penicillin. All isolates were also susceptible to amikacin, vancomycin, and trimethoprim-sulfamethoxazole. To determine molecular epidemiological relatedness of these 15 animal MRSA isolates to isolates from humans, random amplified polymorphic DNA (RAPD) patterns were generated by arbitrarily primed PCR. The RAPD patterns of six of the isolates from animals were identical to the patterns of certain isolates from humans. The antibiotypes of the six animal isolates revealed types similar to those of the human isolates. These data suggested that the genomes of the six animal MRSA isolates were very closely related to those of some human MRSA isolates and were a possible source of human infections caused by consuming contaminated food products made from these animals. PMID:14602604

  19. Microcin determinants are associated with B2 phylogroup of human fecal Escherichia coli isolates.

    PubMed

    Micenková, Lenka; Bosák, Juraj; Štaudová, Barbora; Kohoutová, Darina; Čejková, Darina; Woznicová, Vladana; Vrba, Martin; Ševčíková, Alena; Bureš, Jan; Šmajs, David

    2016-06-01

    Escherichia coli strains are classified into four main phylogenetic groups (A, B1, B2, and D) and strains of these phylogroups differ in a number of characteristics. This study tested whether human fecal E. coli isolates belonging to different phylogroups differ in prevalence of bacteriocinogenic isolates and prevalence of individual bacteriocinogenic determinants. A set of 1283 fecal E. coli isolates from patients with different diseases was tested for the presence of DNA regions allowing classification into E. coli phylogroups and for the ability to produce bacteriocins (23 colicins and 7 microcins). Of the isolates tested, the most common was phylogroup B2 (38.3%) followed by phylogroups A (28.3%), D (26.3%) and B1 (7.2%). Altogether, 695 bacteriocin producers were identified representing 54.2% of all tested isolates. The highest prevalence of bacteriocin producers was found in group B2 (60.3%) and the lowest in group B1 (44.6%). Determinants encoding colicins E1, Ia, and microcin mV were most common in phylogroup A, determinants encoding microcins mM and mH47 were most common in phylogroup B2, and determinant encoding mB17 was most common in phylogroup D. The highest prevalence of bacteriocinogeny was found in phylogroup B2, suggesting that bacteriocinogeny and especially the synthesis of microcins was associated with virulent and resident E. coli strains.

  20. Antimicrobial Susceptibility Profiles of Human Campylobacter jejuni Isolates and Association with Phylogenetic Lineages

    PubMed Central

    Cha, Wonhee; Mosci, Rebekah; Wengert, Samantha L.; Singh, Pallavi; Newton, Duane W.; Salimnia, Hossein; Lephart, Paul; Khalife, Walid; Mansfield, Linda S.; Rudrik, James T.; Manning, Shannon D.

    2016-01-01

    Campylobacter jejuni is a zoonotic pathogen and the most common bacterial cause of human gastroenteritis worldwide. With the increase of antibiotic resistance to fluoroquinolones and macrolides, the drugs of choice for treatment, C. jejuni was recently classified as a serious antimicrobial resistant threat. Here, we characterized 94 C. jejuni isolates collected from patients at four Michigan hospitals in 2011 and 2012 to determine the frequency of resistance and association with phylogenetic lineages. The prevalence of resistance to fluoroquinolones (19.1%) and macrolides (2.1%) in this subset of C. jejuni isolates from Michigan was similar to national reports. High frequencies of fluoroquinolone-resistant C. jejuni isolates, however, were recovered from patients with a history of foreign travel. A high proportion of these resistant isolates were classified as multilocus sequence type (ST)-464, a fluoroquinolone-resistant lineage that recently emerged in Europe. A significantly higher prevalence of tetracycline-resistant C. jejuni was also found in Michigan and resistant isolates were more likely to represent ST-982, which has been previously recovered from ruminants and the environment in the U.S. Notably, patients with tetracycline-resistant C. jejuni infections were more likely to have contact with cattle. These outcomes prompt the need to monitor the dissemination and diversification of imported fluoroquinolone-resistant C. jejuni strains and to investigate the molecular epidemiology of C. jejuni recovered from cattle and farm environments to guide mitigation strategies. PMID:27199922

  1. Antimicrobial Susceptibility Profiles of Human Campylobacter jejuni Isolates and Association with Phylogenetic Lineages.

    PubMed

    Cha, Wonhee; Mosci, Rebekah; Wengert, Samantha L; Singh, Pallavi; Newton, Duane W; Salimnia, Hossein; Lephart, Paul; Khalife, Walid; Mansfield, Linda S; Rudrik, James T; Manning, Shannon D

    2016-01-01

    Campylobacter jejuni is a zoonotic pathogen and the most common bacterial cause of human gastroenteritis worldwide. With the increase of antibiotic resistance to fluoroquinolones and macrolides, the drugs of choice for treatment, C. jejuni was recently classified as a serious antimicrobial resistant threat. Here, we characterized 94 C. jejuni isolates collected from patients at four Michigan hospitals in 2011 and 2012 to determine the frequency of resistance and association with phylogenetic lineages. The prevalence of resistance to fluoroquinolones (19.1%) and macrolides (2.1%) in this subset of C. jejuni isolates from Michigan was similar to national reports. High frequencies of fluoroquinolone-resistant C. jejuni isolates, however, were recovered from patients with a history of foreign travel. A high proportion of these resistant isolates were classified as multilocus sequence type (ST)-464, a fluoroquinolone-resistant lineage that recently emerged in Europe. A significantly higher prevalence of tetracycline-resistant C. jejuni was also found in Michigan and resistant isolates were more likely to represent ST-982, which has been previously recovered from ruminants and the environment in the U.S. Notably, patients with tetracycline-resistant C. jejuni infections were more likely to have contact with cattle. These outcomes prompt the need to monitor the dissemination and diversification of imported fluoroquinolone-resistant C. jejuni strains and to investigate the molecular epidemiology of C. jejuni recovered from cattle and farm environments to guide mitigation strategies. PMID:27199922

  2. Cannabidiol induced a contrasting pro-apoptotic effect between freshly isolated and precultured human monocytes

    SciTech Connect

    Wu, Hsin-Ying; Chang, An-Chi; Wang, Chia-Chi; Kuo, Fu-Hua; Lee, Chi-Ya; Liu, Der-Zen; Jan, Tong-Rong

    2010-08-01

    It has been documented that cannabidiol (CBD) induced apoptosis in a variety of transformed cells, including lymphocytic and monocytic leukemias. In contrast, a differential sensitivity between normal lymphocytes and monocytes to CBD-mediated apoptosis has been reported. The present study investigated the pro-apoptotic effect of CBD on human peripheral monocytes that were either freshly isolated or precultured for 72 h. CBD markedly enhanced apoptosis of freshly isolated monocytes in a time- and concentration-dependent manner, whereas precultured monocytes were insensitive. By comparison, both cells were sensitive to doxorubicin-induced apoptosis. CBD significantly diminished the cellular thiols and glutathione in freshly isolated monocytes. The apoptosis induced by CBD was abrogated in the presence of N-acetyl-{sub L}-cysteine, a precursor of glutathione. In addition, precultured monocytes contained a significantly greater level of glutathione and heme oxygenase-1 (HO-1) compared to the freshly isolated cells. The HO-1 competitive inhibitor zinc protoporphyrin partially but significantly restored the sensitivity of precultured monocytes to CBD-mediated apoptosis. Collectively, our results demonstrated a contrasting pro-apoptotic effect of CBD between precultured and freshly isolated monocytes, which was closely associated with the cellular level of glutathione and the antioxidative capability of the cells.

  3. Isolation of a novel human papillomavirus (type 51) from a cervical condyloma

    SciTech Connect

    Nuovo, G.J.; Crum, C.P.; Levine, R.U.; Silverstein, S.J. ); De Villiers, E.M. )

    1988-04-01

    The authors cloned the DNA from a novel human papillomavirus (HPV) present in a cervical condyloma. When DNA from this isolate was hybridized at high stringency with HPV types 1 through 50 (HPV-1 through HPV-50), it showed weak homology with HPV-6 and -16 and stronger homology with HPV-26. A detailed restriction endonuclease map was prepared which showed marked differences from the maps for other HPVs that have been isolated from the female genital tract. Reassociation kinetic analysis revealed that HPV-26 and this new isolate were less than 10% homologous; hence, the new isolate is a noel strain of HPV. The approximate positions of the open reading frames of the new strain were surmised by hybridization with probes derived from individual open reading frames of HPV-16. In an analysis of 175 genital biopsies from patients with abnormal Papanicolaou smears, sequences hybridizing under highly stringent conditions to probes from this novel HPV type were found in 4.2, 6.1, and 2.4% of biopsies containing normal squamous epithelium, condylomata, and intraepithelial neoplasia, respectively. In addition, sequences homologous to probes from this novel isolate were detected in one of five cervical carcinomas examined.

  4. Modeling of Hepatocytes Proliferation Isolated from Proximal and Distal Zones from Human Hepatocellular Carcinoma Lesion

    PubMed Central

    Montalbano, Mauro; Curcurù, Giuseppe; Shirafkan, Ali; Vento, Renza; Rastellini, Cristiana; Cicalese, Luca

    2016-01-01

    Isolation of hepatocytes from cirrhotic human livers and subsequent primary culture are important new tools for laboratory research and cell-based therapeutics in the study of hepatocellular carcinoma (HCC). Using such techniques, we have previously identified different subpopulations of human hepatocytes and among them one is showing a progressive transformation of hepatocytes in HCC-like cells. We have hypothesized that increasing the distance from the neoplastic lesion might affect hepatocyte function and transformation capacity. However, limited information is available in comparing the growth and proliferation of human hepatocytes obtained from different areas of the same cirrhotic liver in relation to their distance from the HCC lesion. In this study, hepatocytes from 10 patients with cirrhosis and HCC undergoing surgical resections from specimens obtained at a proximal (CP) and distal (CD) distance from the HCC lesion were isolated and placed in primary culture. CP hepatocytes (CP-Hep) were isolated between 1 to 3 cm (leaving at least 1cm margin to avoid cancer cells and/or satellite lesions), while CD hepatocytes (CD-Hep) were isolated from more than 5 cm or from the contralateral-lobe. A statistical model was built to analyze the proliferation rates of these cells and we evaluated expression of HCC markers (Glypican-3 (GPC3), αSmooth Muscle Actin (α-SMA) and PCNA). We observed a significant difference in proliferation and in-vitro growth showing that CP-Hep had a proliferation pattern and rate significantly different than CD-Hep. Based on these data, this model can provide information to predict growth of human hepatocytes in primary culture in relation to their pre-cancerous state with significant differences in the HCC markers expression. This model provides an important innovative tool for in-vitro analysis of HCC. PMID:27074018

  5. Isolation, Characterization, Cryopreservation of Human Amniotic Stem Cells and Differentiation to Osteogenic and Adipogenic Cells

    PubMed Central

    Gholizadeh-Ghaleh Aziz, Shiva; Pashaei-Asl, Fatima; Fardyazar, Zahra; Pashaiasl, Maryam

    2016-01-01

    Human stem cells and progenitor cells can be used to treat cancer and replace dysfunctional cells within a tissue or organ. The objective of this study was to identify the appropriate cells type in regenerative medicine and targeted therapy. As an alternative to embryonic and bone marrow stem cells, we examined human amniotic fluid stem cells (hAFSCs), one of the potential source of multipotent stem cells isolated from both cell pellet (using single-stage method), and supernatant of human amniotic fluid. Source of isolation and unique property of the cells emphasize that these cells are one of the promising new tools in therapeutic field. Double sources for isolation and availability of the left over samples in diagnostic laboratory at the same time have less legal and ethical concerns compared with embryonic stem cell studies. Cells were isolated, cultured for 18th passage for 6 months and characterized using qPCR and flow cytometry. Cells showed good proliferative ability in culture condition. The cells successfully differentiated into the adipogenic and osteogenic lineages. Based on these findings, amniotic fluid can be considered as an appropriate and convenient source of human amniotic fluid stem cells. These cells provide potential tools for therapeutic applications in the field of regenerative medicine. To get a better understanding of crosstalk between Oct4/NANOG with osteogenesis and adipogenesis, we used network analysis based on Common Targets algorithm and Common Regulators algorithm as well as subnetwork discovery based on gene set enrichment. Network analysis highlighted the possible role of MIR 302A and MIR let-7g. We demonstrated the high expression of MIR 302A and low expression of MIR let7g in hAFSCs by qPCR. PMID:27434028

  6. Cryptolepine, isolated from Sida acuta, sensitizes human gastric adenocarcinoma cells to TRAIL-induced apoptosis.

    PubMed

    Ahmed, Firoj; Toume, Kazufumi; Ohtsuki, Takashi; Rahman, Mahmudur; Sadhu, Samir Kumar; Ishibashi, Masami

    2011-01-01

    Bioassay guided separation of Sida acuta whole plants led to the isolation of an alkaloid, cryptolepine (1), along with two kaempferol glycosides (2-3). Compound 1 showed strong activity in overcoming TRAIL-resistance in human gastric adenocarcinoma (AGS) cells at 1.25, 2.5 and 5 μm. Combined treatment of 1 and TRAIL sensitized AGS cells to TRAIL-induced apoptosis at the aforementioned concentrations.

  7. Comparative genomic analysis of human Chlamydia pneumoniae isolates from respiratory, brain and cardiac tissues.

    PubMed

    Roulis, Eileen; Bachmann, Nathan L; Myers, Garry S A; Huston, Wilhelmina; Summersgill, James; Hudson, Alan; Dreses-Werringloer, Ute; Polkinghorne, Adam; Timms, Peter

    2015-12-01

    Chlamydia pneumoniae is an obligate intracellular bacterium implicated in a wide range of human diseases including atherosclerosis and Alzheimer's disease. Efforts to understand the relationships between C. pneumoniae detected in these diseases have been hindered by the availability of sequence data for non-respiratory strains. In this study, we sequenced the whole genomes for C. pneumoniae isolates from atherosclerosis and Alzheimer's disease, and compared these to previously published C. pneumoniae genomes. Phylogenetic analyses of these new C. pneumoniae strains indicate two sub-groups within human C. pneumoniae, and suggest that both recombination and mutation events have driven the evolution of human C. pneumoniae. Further fine-detailed analyses of these new C. pneumoniae sequences show several genetically variable loci. This suggests that similar strains of C. pneumoniae are found in the brain, lungs and cardiovascular system and that only minor genetic differences may contribute to the adaptation of particular strains in human disease.

  8. Plasticity of interneuronal networks of the functionally isolated human spinal cord

    PubMed Central

    Harkema, Susan J.

    2009-01-01

    The loss of walking after human spinal cord injury has been attributed to the dominance of supraspinal over spinal mechanisms. The evidence for central pattern generation in humans is limited due to the inability to conclusively isolate the circuitry from descending and afferent input. However, studying individuals following spinal cord injury with no detectable influence on spinal networks from supraspinal centers can provide insight to their interaction with afferent input. The focus of this article is on the interaction of sensory input with human spinal networks in the generation of locomotor patterns. The functionally isolated human spinal cord has the capacity to generate locomotor patterns with appropriate afferent input. Locomotor Training is a rehabilitative strategy that has evolved from animal and humans studies focused on the neural plasticity of the spinal cord and has been successful for many people with acute and chronic incomplete spinal cord injury. However, even those individuals with clinically complete spinal cord injury that generate appropriate locomotor patterns during stepping with assistance on a treadmill with body weight support cannot sustain overground walking. This suggests that although a significant control of locomotion can occur at the level of spinal interneuronal networks the level of sustainable excitability of these circuits is still compromised. Future studies should focus on approaches to increase the central state of excitability and may include neural repair strategies, pharmacological interventions or epidural stimulation in combination with Locomotor Training. PMID:18042493

  9. [Isolation of a gamma heavy chain fragment from normal human serum].

    PubMed

    Irurzun, P L; Miranda, M P

    1976-01-01

    Several components of catodic electrophoretic migration in serum and urine are present in normal individuals and in rabbit serum. There also exists in man and in some animals, serum fractions of low molecular weight. These types of serum components may be or may not be related with the IgG. In a previous study we have isolated two components in the slow catodic electrophoretic area of the normal human serum (NHS). One of them was identified as an IgG subclass and the other component presented a clear line of precipitation to gamma heavy chain specific immuno-serum. This latter component was found in the post gamma-globulin area crossing the IgG arc in the I.E. analysis. Its molecular weight was variable from 3700 to 9500. In this paper a differential analysis of the gamma fragment isolated for us, is made and its relationship with Fc subfragments of pepsin-digested IgG is studied. In order to obtain this comparative study, the electrophoresis, gel diffusion immunoelectrophoresis gel chromatography and analytic ultracentrifugation techniques are employed. The post-gammaglobulin fraction has been isolated from total normal human serum without previous manipulation, or with the gammaglobulin fraction precipitated with saturated ammonium sulphate, in Sephadex G-200 chromatography. These two fractions present similar immunelectrophoretical characteristics. The constant sedimentation is 0.90 S and the approximated molecular weight is 7000. Since the pepsin digestion of IgG produced Fc subfragments of low molecular weight, we have isolated and submitted this immunoglobulin to peptic digestion. The G-75 Sephadex filtration shows an isolated post-gamma-globulin of I.E. sedimentation constant and molecular weight whose characteristics are similar to the isolated serum post-gammaglobulin fraction. The antigenical analysis in I.D. shows a total identity between the pepsin digested post-gammaglobulin and the fragment obtained for us from the human serum to an anti-heavy gamma

  10. The molecular structure and lubricating activity of lubricin isolated from bovine and human synovial fluids.

    PubMed Central

    Swann, D A; Silver, F H; Slayter, H S; Stafford, W; Shore, E

    1985-01-01

    Lubricin was isolated from bovine ankle, metacarpophalangeal and knee and human knee synovial fluids. The lubricins isolated from the bovine joint fluids had the same amino acid and carbohydrate compositions, but differences were observed in the relative molecular masses. The Mr values of bovine metacarpophalangeal and ankle lubricin determined by light-scattering measurements were about 200 000, whereas values of 132 000 and 143 000 were obtained for the bovine knee lubricin. The human knee lubricin had a similar carbohydrate composition to bovine knee lubricin except for the higher glucosamine content, and the amino acid composition differed slightly. The human sample had a lower glutamic acid content and a leucine/isoleucine ratio of 2:1 compared with 1:1 in the bovine. The Mr value of the human knee lubricin (166 000) was also lower than that of the bovine metacarpophalangeal and ankle samples. The Mr value of the bovine knee lubricin determined by sedimentation-equilibrium measurements was 171 000. The length measurements determined by electron microscopy and also the sedimentation measurements showed considerable polydispersity and indicate that the degree of extension of lubricin molecules can vary. Friction measurements showed that the human knee synovial-fluid lubricin had equivalent lubricating ability in a test system in vitro to that observed for lubricin isolated from normal bovine synovial fluids. The lubricating ability of lubricin was concentration-dependent, and each lubricin sample was able to act as a lubricant in vitro in an equivalent manner to whole synovial fluid at concentrations that are thought to occur in vivo. PMID:3977823

  11. Isolation of contractile cardiomyocytes from human pluripotent stem-cell-derived cardiomyogenic cultures using a human NCX1-EGFP reporter.

    PubMed

    Ovchinnikov, Dmitry A; Hidalgo, Alejandro; Yang, Seung-Kwon; Zhang, Xinli; Hudson, James; Mazzone, Stuart B; Chen, Chen; Cooper-White, Justin J; Wolvetang, Ernst J

    2015-01-01

    The prospective isolation of defined contractile human pluripotent stem cell (hPSC)-derived cardiomyocytes is advantageous for regenerative medicine and drug screening applications. Currently, enrichment of cardiomyocyte populations from such cultures can be achieved by combinations of cell surface markers or the labor-intensive genetic modification of cardiac developmental genes, such as NKX2.5 or MYH6, with fluorescent reporters. To create a facile, portable method for the isolation of contractile cardiomyocytes from cardiomyogenic hPSC cultures, we employed a highly conserved cardiac enhancer sequence in the SLC8A1 (NCX1) gene to generate a lentivirally deliverable, antibiotic-selectable NCX1cp-EGFP reporter. We show that human embryonic stem cells (and induced pluripotent stem cells) transduced with the NCX1cp-EGFP reporter cassette exhibit enhanced green fluorescent protein (EGFP) expression in cardiac progenitors from 5 days into the directed cardiac hPSC differentiation protocol, with all reporter-positive cells transitioning to spontaneously contracting foci 3 days later. In subsequent stages of cardiomyocyte maturation, NCX1cp-EGFP expression was exclusively limited to contractile cells expressing high levels of cardiac troponin T (CTNT), MLC2a/v, and α-actinin proteins, and was not present in CD90/THY1(+) cardiac stromal cells or CD31/PECAM(+) endothelial cells. Flow-assisted cytometrically sorted EGFP(+) fractions of differentiated cultures were highly enriched in both early (NKX2.5 and TBX5) and late (CTNT/TNNI2, MYH6, MYH7, NPPA, and MYL2) cardiomyocyte markers, with a significant proportion of cells displaying a ventricular-like action potential pattern in patch-clamp recordings. We conclude that the use of the cardiac-specific promoter of the human SLC8A1(NCX1) gene is an effective strategy to isolate contractile cardiac cells and their progenitors from hPSC-derived cardiomyogenic cultures. PMID:25075536

  12. The isolation and culture of endothelial colony-forming cells from human and rat lungs.

    PubMed

    Alphonse, Rajesh S; Vadivel, Arul; Zhong, Shumei; Zong, Shumei; McConaghy, Suzanne; Ohls, Robin; Yoder, Mervin C; Thébaud, Bernard

    2015-11-01

    Blood vessels are crucial for the normal development, lifelong repair and homeostasis of tissues. Recently, vascular progenitor cell-driven 'postnatal vasculogenesis' has been suggested as an important mechanism that contributes to new blood vessel formation and organ repair. Among several described progenitor cell types that contribute to blood vessel formation, endothelial colony-forming cells (ECFCs) have received widespread attention as lineage-specific 'true' vascular progenitors. Here we describe a protocol for the isolation of pulmonary microvascular ECFCs from human and rat lung tissue. Our technique takes advantage of an earlier protocol for the isolation of circulating ECFCs from the mononuclear cellular fraction of peripheral blood. We adapted the earlier protocol to isolate resident ECFCs from the distal lung tissue. After enzymatic dispersion of rat or human lung samples into a cellular suspension, CD31-expressing cells are positively selected using magnetic-activated cell sorting and plated in endothelial-specific growth conditions. The colonies arising after 1-2 weeks in culture are carefully separated and expanded to yield pure ECFC cultures after a further 2-3 weeks. The resulting cells demonstrate the defining characteristics of ECFCs such as (i) 'cobblestone' morphology of cultured cell monolayers; (ii) acetylated low-density lipoprotein uptake and Ulex europaeus lectin binding; (iii) tube-like network formation in Matrigel; (iv) expression of endothelial cell-specific surface markers and the absence of hematopoietic or myeloid surface antigens; (v) self-renewal potential displayed by the most proliferative cells; and (vi) contribution to de novo vessel formation in an in vivo mouse implant model. Assuming typical initial cell adhesion and proliferation rates, the entire procedure can be completed within 4 weeks. Isolation and culture of lung vascular ECFCs will allow assessment of the functional state of these cells in experimental and human

  13. [Genotyping of Giardia intestinalis strains isolated from humans in Aydin, Turkey].

    PubMed

    Ertuğ, Sema; Ertabaklar, Hatice; Özlem Çalışkan, Serçin; Malatyalı, Erdoğan; Bozdoğan, Bülent

    2016-01-01

    Giardia intestinalis which is a flagellate, intestinal protozoon of humans and a variety of mammalian species, shows worldwide distribution. To date, eight genotypes of the parasite have been identified. Among these genotypes, assemblage A and B have zoonotic characteristics with low host specificity, thus they are responsible for the human infections. The aim of this study was to identify G.intestinalis genotypes in Aydin, located in Aegean region of Turkey. A total of 40 stool samples that were found positive for G.intestinalis by direct microscopic examination, from Adnan Menderes University, Research and Training Hospital, Parasitology Laboratory from January 2011 to December 2014 were included in the study. DNA isolation from stool samples performed with commercial kit (QIAamp DNA Stool Mini Kit, Qiagen, Germany) followed by polymerase chain reaction (PCR) for G.intestinalis 16S rRNA and beta-giardin genes and then the amplicons were sequenced. Out of 40 isolates 11 (27.5%) were positive with 16S rRNA PCR and 10 (25%) were positive with beta-giardin PCR. Of 21 sequenced amplicons, 10 (47.6%) of them showed 98%-100% similarity with reference sequences and their genotypes could be identified. The distribution of genotypes were as follows: cluster A1 (n: 3), cluster A2 (n: 3), cluster A3 (n: 2) and assemblage B (n: 2). In the light of our results the isolates detected in humans might be zoonotic origin. In accordance with the previous reports in Turkey, assemblage A (8/10) was more common than assemblage B (2/10). In the present study, 10 (25%) out of 40 isolates could be genotyped and sequencing of beta-giardin gene yielded more effective results than sequencing of 16S rRNA for the determination of assemblages. The present study indicated that, there is a need for prospective studies with extended number of cases allowing the comparison of the two genes used for G.intestinalis genotyping.

  14. The isolation and culture of endothelial colony-forming cells from human and rat lungs.

    PubMed

    Alphonse, Rajesh S; Vadivel, Arul; Zhong, Shumei; Zong, Shumei; McConaghy, Suzanne; Ohls, Robin; Yoder, Mervin C; Thébaud, Bernard

    2015-11-01

    Blood vessels are crucial for the normal development, lifelong repair and homeostasis of tissues. Recently, vascular progenitor cell-driven 'postnatal vasculogenesis' has been suggested as an important mechanism that contributes to new blood vessel formation and organ repair. Among several described progenitor cell types that contribute to blood vessel formation, endothelial colony-forming cells (ECFCs) have received widespread attention as lineage-specific 'true' vascular progenitors. Here we describe a protocol for the isolation of pulmonary microvascular ECFCs from human and rat lung tissue. Our technique takes advantage of an earlier protocol for the isolation of circulating ECFCs from the mononuclear cellular fraction of peripheral blood. We adapted the earlier protocol to isolate resident ECFCs from the distal lung tissue. After enzymatic dispersion of rat or human lung samples into a cellular suspension, CD31-expressing cells are positively selected using magnetic-activated cell sorting and plated in endothelial-specific growth conditions. The colonies arising after 1-2 weeks in culture are carefully separated and expanded to yield pure ECFC cultures after a further 2-3 weeks. The resulting cells demonstrate the defining characteristics of ECFCs such as (i) 'cobblestone' morphology of cultured cell monolayers; (ii) acetylated low-density lipoprotein uptake and Ulex europaeus lectin binding; (iii) tube-like network formation in Matrigel; (iv) expression of endothelial cell-specific surface markers and the absence of hematopoietic or myeloid surface antigens; (v) self-renewal potential displayed by the most proliferative cells; and (vi) contribution to de novo vessel formation in an in vivo mouse implant model. Assuming typical initial cell adhesion and proliferation rates, the entire procedure can be completed within 4 weeks. Isolation and culture of lung vascular ECFCs will allow assessment of the functional state of these cells in experimental and human

  15. Adherence to human lung microvascular endothelial cells (HMVEC-L) of Plasmodium vivax isolates from Colombia

    PubMed Central

    2013-01-01

    Background For years Plasmodium vivax has been considered the cause of benign malaria. Nevertheless, it has been observed that this parasite can produce a severe disease comparable to Plasmodium falciparum. It has been suggested that some physiopathogenic processes might be shared by these two species, such as cytoadherence. Recently, it has been demonstrated that P. vivax-infected erythrocytes (Pv-iEs) have the capacity to adhere to endothelial cells, in which intercellular adhesion molecule-1 (ICAM-1) seems to be involved in this process. Methods Adherence capacity of 21 Colombian isolates, from patients with P. vivax mono-infection to a microvascular line of human lung endothelium (HMVEC-L) was assessed in static conditions and binding was evaluated at basal levels or in tumor necrosis factor (TNF) stimulated cells. The adherence specificity for the ICAM-1 receptor was determined through inhibition with an anti-CD54 monoclonal antibody. Results The majority of P. vivax isolates, 13 out of 21 (61.9%), adhered to the HMVEC-L cells, but P. vivax adherence was at least seven times lower when compared to the four P. falciparum isolates. Moreover, HMVEC-L stimulation with TNF led to an increase of 1.6-fold in P. vivax cytoadhesion, similar to P. falciparum isolates (1.8-fold) at comparable conditions. Also, blockage of ICAM-1 receptor with specific antibodies showed a significant 50% adherence reduction. Conclusions Plasmodium vivax isolates found in Colombia are also capable of adhering specifically in vitro to lung endothelial cells, via ICAM-1 cell receptor, both at basal state and after cell stimulation with TNF. Collectively, these findings reinforce the concept of cytoadherence for P. vivax, but here, to a different endothelial cell line and using geographical distinct isolates, thus contributing to understanding P. vivax biology. PMID:24080027

  16. Isolation and chromosomal mapping of the human immunoglobulin-associated B29 gene (IGB)

    SciTech Connect

    Wood, W.J. Jr.; Thompson, A.A.; Korenberg, J.; Xianing Chen; May, W.; Wall, R.; Denny, C.T. )

    1993-04-01

    The B29 gene encodes a B-cell-specific membrane protein in the immunoglobulin antigen receptor complex. B29 is a crucial member of this receptor complex and is believed to function as an effector of signal transduction in a manner analogous to that of the CD3 components of the T cell antigen receptor. The authors have isolated a full-length human B29 cDNA clone by using a murine B29 cDNA probe. They show that there is an extremely high degree of evolutionary conservation between the human and mouse proteins, particularly in the transmembrane and intracytoplasmic regions, where the identity is 96%. In addition, the intracytoplasmic region in both proteins contains an identical peptide motif that is present in a number of molecules involved in lymphocyte activation. Genomic Southern blot analysis of human cell lines hybridized with both murine and human B29 cDNAs gives patterns consistent with a single-copy gene occupying a small region of the genomic sequence. Using human B29 cosmid DNA, they have localized the B29 gene to human chromosome 17q23 via fluorescence in situ hybridization. B29 is the first gene localized to this area of the genome. Interestingly, a subset of human B cell chronic lymphocytic leukemias (CLL) has translocations in this locus on chromosome 17. 18 refs., 4 figs.

  17. Invasion characteristics of a Plasmodium knowlesi line newly isolated from a human

    PubMed Central

    Amir, Amirah; Russell, Bruce; Liew, Jonathan Wee Kent; Moon, Robert W.; Fong, Mun Yik; Vythilingam, Indra; Subramaniam, Vellayan; Snounou, Georges; Lau, Yee Ling

    2016-01-01

    Plasmodium knowlesi is extensively used as an important malaria model and is now recognized as an important cause of human malaria in Malaysia. The strains of P. knowlesi currently used for research were isolated many decades ago, raising concerns that they might no longer be representative of contemporary parasite populations. We derived a new P. knowlesi line (University Malaya line, UM01), from a patient admitted in Kuala Lumpur, Malaysia, and compared it with a human-adapted laboratory line (A1-H.1) derived from the P. knowlesi H strain. The UM01 and A1-H.1 lines readily invade human and macaque (Macaca fascicularis) normocytes with a preference for reticulocytes. Whereas invasion of human red blood cells was dependent on the presence of the Duffy antigen/receptor for chemokines (DARC) for both parasite lines, this was not the case for macaque red blood cells. Nonetheless, differences in invasion efficiency, gametocyte production and the length of the asexual cycle were noted between the two lines. It would be judicious to isolate and characterise numerous P. knowlesi lines for use in future experimental investigations of this zoonotic species. PMID:27097521

  18. Isolation and Characterization of Human Adult Epithelial Stem Cells from the Periodontal Ligament.

    PubMed

    Athanassiou-Papaefthymiou, M; Papagerakis, P; Papagerakis, S

    2015-11-01

    We report a novel method for the isolation of adult human epithelial stem cells (hEpiSCs) from the epithelial component of the periodontal ligament-the human epithelial cell rests of Malassez (hERM). hEpiSC-rich integrin-α6(+ve) hERM cells derived by fluorometry can be clonally expanded, can grow organoids, and express the markers of pluripotency (OCT4, NANOG, SOX2), polycomb protein RING1B, and the hEpiSC supermarker LGR5. They maintain the growth profile of their originating hERM in vitro. Subcutaneous cotransplantation with mesenchymal stem cells from the dental pulp on poly-l-lactic acid scaffolds in nude mice gave rise to perfect heterotopic ossicles in vivo with ultrastructure of dentin, enamel, cementum, and bone. These remarkable fully mineralized ossicles underscore the importance of epithelial-mesenchymal crosstalk in tissue regeneration using human progenitor stem cells, which may have already committed to lineage despite maintaining hallmarks of pluripotency. In addition, we report the clonal expansion and isolation of human LGR5(+ve) cells from the hERM in xeno-free culture conditions. The genetic profile of LGR5(+ve) cells includes both markers of pluripotency and genes important for secretory epithelial and dental epithelial cell differentiation, giving us a first insight into periodontal ligament-derived hEpiSCs. PMID:26392003

  19. Invasion characteristics of a Plasmodium knowlesi line newly isolated from a human.

    PubMed

    Amir, Amirah; Russell, Bruce; Liew, Jonathan Wee Kent; Moon, Robert W; Fong, Mun Yik; Vythilingam, Indra; Subramaniam, Vellayan; Snounou, Georges; Lau, Yee Ling

    2016-01-01

    Plasmodium knowlesi is extensively used as an important malaria model and is now recognized as an important cause of human malaria in Malaysia. The strains of P. knowlesi currently used for research were isolated many decades ago, raising concerns that they might no longer be representative of contemporary parasite populations. We derived a new P. knowlesi line (University Malaya line, UM01), from a patient admitted in Kuala Lumpur, Malaysia, and compared it with a human-adapted laboratory line (A1-H.1) derived from the P. knowlesi H strain. The UM01 and A1-H.1 lines readily invade human and macaque (Macaca fascicularis) normocytes with a preference for reticulocytes. Whereas invasion of human red blood cells was dependent on the presence of the Duffy antigen/receptor for chemokines (DARC) for both parasite lines, this was not the case for macaque red blood cells. Nonetheless, differences in invasion efficiency, gametocyte production and the length of the asexual cycle were noted between the two lines. It would be judicious to isolate and characterise numerous P. knowlesi lines for use in future experimental investigations of this zoonotic species. PMID:27097521

  20. Intermedilysin, a novel cytotoxin specific for human cells secreted by Streptococcus intermedius UNS46 isolated from a human liver abscess.

    PubMed Central

    Nagamune, H; Ohnishi, C; Katsuura, A; Fushitani, K; Whiley, R A; Tsuji, A; Matsuda, Y

    1996-01-01

    A novel cytotoxin (intermedilysin) specific for human cells was identified as a cytolytic factor of Streptococcus intermedius UNS46 isolated from a human liver abscess. Intermedilysin caused human cell death with membrane blebs. Intermedilysin was purified from UNS46 culture medium by means of gel filtration and hydrophobic chromatography. The purified toxin was resolved into major and minor bands of 54 and 53 kDa, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These proteins reacted with an antibody against intermedilysin. Five internal peptide fragments of intermedilysin were sequenced and found to have 42 to 71% homology with the thiol-activated cytotoxin pneumolysin. However, the action of intermedilysin differed from that of thiol-activated cytotoxins, especially in terms of a lack of activation by dithiothreitol and resistance to treatments with N-ethylmaleimide and 5,5'-dithio-bis-(2-nitrobenzoic acid), although cholesterol inhibited the toxin activity. Intermedilysin was potently hemolytic on human erythrocytes but was 100-fold less effective on chimpanzee and cynomolgus monkey erythrocytes. Intermedilysin was not hemolytic in nine other animal species tested. Since human erythrocytes treated with trypsin were far less sensitive to intermedilysin than were the intact cells, a cell membrane protein(s) may participate in the intermedilysin action. These data demonstrated that intermedilysin is distinguishable from all known bacterial cytolysins. PMID:8757839

  1. Detection of Salmonella enterica in pigs at slaughter and comparison with human isolates in Italy.

    PubMed

    Bonardi, Silvia; Alpigiani, Irene; Bruini, Ilaria; Barilli, Elena; Brindani, Franco; Morganti, Marina; Cavallini, Pierugo; Bolzoni, Luca; Pongolini, Stefano

    2016-02-01

    In 2013-2014, 201 pigs belonging to 67 batches were tested for Salmonella in their mesenteric lymph nodes (MLN) in one abattoir of Northern Italy. For each batch, faecal material was collected at lairage by swabbing the pen floor for approximately 1600 cm(2). The aim of this study was to investigate the prevalence of Salmonella in MLN of pigs at slaughter, to assess Salmonella contamination at lairage and to evaluate the effect of lairage duration on its prevalence. Serotyping, XbaI PFGE typing and antimicrobial testing of the isolates were performed. Pig and human Salmonella isolates of the same region of Italy were compared to evaluate possible correlations. Salmonella enterica was isolated from 19.9% of the MLN and 49.3% of the environmental faecal samples. Nine different serovars were identified among 75 S. enterica isolates. In MLN Salmonella Derby was the most common (52.5%), followed by S. enterica 4,[5],12:i:- (17.5%) and Salmonella Rissen (10.0%). In faecal samples S. Derby was prevalent (51.4%), followed by S. enterica 4,[5], 12:i:- (20.0%) and Salmonella Brandenburg (14.3%). Lairage holding varied between 1 and ≥ 12 h (median value: 2.5h). In pigs held for 1-3h, 14.1% were positive for Salmonella in MLN but the prevalence reached 31.8% when they were held for ≥ 12 h. The contamination of MLN was statistically different (p=0.0045) between the two groups, thus confirming the role of long-lasting lairage in Salmonella contamination of pigs. XbaI PFGE typing detected 36 PFGE types. Twenty-three PFGE types were identified among the 40 MLN isolates and 22 PFGE types among the 35 faecal isolates. A total of 11 PFGE types were shared between the MLN of pigs and the lairage environment. Among S. Derby, 6 shared PFGE types between MLN and faeces were found and among S. enterica 4,[5],12:i:- one PFGE type was common between MLN and the faecal samples. Shared profiles between human and swine isolates of S. Derby, S. enterica 4,[5],12:i:-, S. Rissen, Salmonella

  2. Phocaeicola abscessus gen. nov., sp. nov., an anaerobic bacterium isolated from a human brain abscess sample.

    PubMed

    Al Masalma, Mouhamad; Raoult, Didier; Roux, Véronique

    2009-09-01

    A strictly anaerobic bacterial strain, 7401987T, was isolated from a human brain abscess sample. Cells were Gram-negative, non-spore-forming, coccoid to rod-shaped and motile by flagella in a lophotrichous arrangement. The isolate was asaccharolytic and the major cellular fatty acids were anteiso-C15:0 (28.2%), C16:0 (18.0%), iso-C15:0 (12.3%) and iso-C17:0 3-OH (11.7%). 16S rRNA gene sequence comparisons showed that the isolate was distantly related to members of the genera Bacteroides (<83.6% similarity), Parabacteroides (<79.9% similarity), Tannerella (<79.8% similarity), Dysgonomonas (<79.6% similarity), Porphyromonas (<79.3% similarity) and Prevotella (<78.9% similarity). The low 16S rRNA gene sequence similarity values and physiological and biochemical characteristics differentiated strain 7401987T from all known species and indicate that our isolate represents a novel species in a new genus within the phylum Bacteroidetes. The name Phocaeicola abscessus gen. nov., sp. nov. is proposed; the type strain of Phocaeicola abscessus is 7401987T (=CCUG 55929T=CSUR P22T=DSM 21584T). PMID:19620382

  3. Trichosporon isolation from human ungueal infections: is there a pathogenic role?*

    PubMed Central

    de Magalhães, Alba Regina; Nishikawa, Marília Martins; de Mondino, Silvia Suzana Bona; de Macedo, Heloisa Werneck; da Rocha, Elisabeth Martins da Silva; Baptista, Andrea Regina de Souza

    2016-01-01

    BACKGROUND Although dermatophytes are considered the major cause of onychomycosis, many reports have incriminated non-dermatophyte moulds and yeasts in the disease’s etiology. Successive Trichosporon isolation from onychomycosis has led to the genus being suspected as a nail primary pathogen. OBJECTIVE To determine the prevalence of Trichosporon isolation in onychomycosis patients who attended a mycology diagnostic service in Rio de Janeiro, Brazil, between January 2003 and December 2006. The study also includes a worldwide review on Trichosporon isolation prevalence in ungueal disease, emphasizing T. ovoides. METHODS This retrospective study was conducted with the support of staff from the Mycology Laboratory at the Dermatological Service of Rio de Janeiro’s Santa Casa da Misericórdia (MLDS). RESULTS Mycological analysis provided positive results equaling 47/5036 (0.93%) for Trichosporon spp.; obtained mainly as a single agent (72.35%), and from mixed cultures (27.65%; X2= 6.397; p= 0.018). The great majority belongs to the T. ovoides species (91.5%; n=43), obtained as a single isolate (74.41%; n= 32/43; X2 = 7.023; p= 0.014). CONCLUSIONS Although T. ovoides is classically associated as an etiologic agent of white piedra, this study highlights its potential as a human nail disease pathogen. Our study opens doors for future epidemiologic and virulence factors aimed at determining whether T. ovoides is an important causative agent of onychomycosis in Brazil. PMID:27192516

  4. Bone resorption by isolated human osteoclasts in vitro: effects of calcitonin.

    PubMed

    Murrills, R J; Shane, E; Lindsay, R; Dempster, D W

    1989-04-01

    Human osteoclasts were isolated from 12- to 17-week-old fetal tissue and from transiliac crest bone biopsies for an in vitro study of their biology. A hypodermic needle was used to flush either the fetal long bones or the trabeculae of the iliac crest bone biopsy with tissue culture medium and the resulting cell suspension sedimented briefly either onto the surface of plastic tissue culture dishes, for time-lapse microcinematography, or onto slices of devitalized bovine cortical bone for quantitative assay of bone resorption. The osteoclasts were motile, tartrate-resistant acid phosphatase positive and capable of excavating pits in slices of devitalized bovine cortical bone. Human calcitonin, at doses of 1 ng/ml and 1 microgram/ml, caused a 70% inhibition of bone resorption by human fetal osteoclasts over a 24 h period but had no apparent effect on the morphology or motility of either fetal or adult osteoclasts. PMID:2728929

  5. A Methodology for Concomitant Isolation of Intimal and Adventitial Endothelial Cells from the Human Thoracic Aorta

    PubMed Central

    Leclercq, Anne; Veillat, Véronique; Loriot, Sandrine; Spuul, Pirjo; Madonna, Francesco; Roques, Xavier; Génot, Elisabeth

    2015-01-01

    Background Aortic diseases are diverse and involve a multiplicity of biological systems in the vascular wall. Aortic dissection, which is usually preceded by aortic aneurysm, is a leading cause of morbidity and mortality in modern societies. Although the endothelium is now known to play an important role in vascular diseases, its contribution to aneurysmal aortic lesions remains largely unknown. The aim of this study was to define a reliable methodology for the isolation of aortic intimal and adventitial endothelial cells in order to throw light on issues relevant to endothelial cell biology in aneurysmal diseases. Methodology/Principal Findings We set up protocols to isolate endothelial cells from both the intima and the adventitia of human aneurysmal aortic vessel segments. Throughout the procedure, analysis of cell morphology and endothelial markers allowed us to select an endothelial fraction which after two rounds of expansion yielded a population of >90% pure endothelial cells. These cells have the features and functionalities of freshly isolated cells and can be used for biochemical studies. The technique was successfully used for aortic vessel segments of 20 patients and 3 healthy donors. Conclusions/Significance This simple and highly reproducible method allows the simultaneous preparation of reasonably pure primary cultures of intimal and adventitial human endothelial cells, thus providing a reliable source for investigating their biology and involvement in both thoracic aneurysms and other aortic diseases. PMID:26599408

  6. Genotype analysis of faecal and blood isolates of Salmonella dublin from humans in England and Wales.

    PubMed Central

    Chowdry, N.; Threlfall, E. J.; Rowe, B.; Stanley, J.

    1993-01-01

    An analysis of genotype was made for representative strains of Salmonella dublin. The collection consisted primarily of strains isolated from humans in England and Wales, and were of both intestinal and extra-intestinal origin. Three genetic elements were characterized by DNA hybridization. They were the spvBC genes, extrachromosomal virulence determinants, the salmonella-specific insertion sequence IS200, and the 16S ribosomal RNA genes, a phylogenetic marker. Two clones of S. dublin (SdRI and SdRII) which shared an identical IS200 profile, were identified on the basis of restriction fragment length polymorphism at the 16S rRNA locus. With one exception, all strains harboured a 52 MDa plasmid which contained a conserved 3.7 kbp Hind III fragment homologous to the spvBC mouse-virulence genes of S. typhimurium. However, a single plasmid-free strain of SdRI, isolated from a patient with septicaemia exhibited no spc homology. In SdRI there was no observable genotype distinction between strains causing gastroenteritis or bacteraemia. In contrast, none of the strains of SdRII were from cases of bacteraemia, and all human isolates of this clone were from cases of gastroenteritis. Images Fig. 1 Fig. 2 PMID:8097167

  7. Comparison of human hepatocytes isolated from livers accepted or discarded for orthotopic transplantation.

    PubMed

    Groothuis, G M; Sandker, G W; Pruim, J; Weert, B; Slooff, M J; Meijer, D K

    1995-12-01

    The aim of this study was to compare human hepatocytes isolated from livers accepted and from livers discarded for transplantation with respect to viability and drug transport function. In addition, the influence of age of the donor and preservation time of the liver on cell viability was determined. Cell viability was assessed by trypan blue exclusion, MTT reduction, morphological integrity and ATP content, and drug transport function by uptake and excretion of taurocholic acid. Hepatocytes could be isolated successfully from livers accepted as well as from livers discarded for transplantation, with a median yield of 5.0 x 10(6) cells/g (range 0.1 to 42.4) and 0.7 x 10(6) cells/g (range 0.0 to 22.7), respectively (not significantly different). These cells were not significantly different with respect to viability and transport rate of taurocholate. Neither the age of the donor nor the duration of liver preservation (6-43 hr in University of Wisconsin solution) significantly influenced cell yield and viability. It is concluded that because of this overlap in cell viability, hepatocytes isolated both from accepted and from discarded livers can in principle be used to investigate drug transport functions in the human liver. PMID:20650173

  8. Genetic variability of vancomycin-resistant Enterococcus faecium and Enterococcus faecalis isolates from humans, chickens, and pigs in Malaysia.

    PubMed

    Getachew, Yitbarek; Hassan, Latiffah; Zakaria, Zunita; Abdul Aziz, Saleha

    2013-08-01

    Vancomycin-resistant enterococci (VRE) have been reported to be present in humans, chickens, and pigs in Malaysia. In the present study, representative samples of VRE isolated from these populations were examined for similarities and differences by using the multilocus sequence typing (MLST) method. Housekeeping genes of Enterococcus faecium (n = 14) and Enterococcus faecalis (n = 11) isolates were sequenced and analyzed using the MLST databases eBURST and goeBURST. We found five sequence types (STs) of E. faecium and six STs of E. faecalis existing in Malaysia. Enterococcus faecium isolates belonging to ST203, ST17, ST55, ST79, and ST29 were identified, and E. faecium ST203 was the most common among humans. The MLST profiles of E. faecium from humans in this study were similar to the globally reported nosocomial-related strain lineage belonging to clonal complex 17 (CC17). Isolates from chickens and pigs have few similarities to those from humans, except for one isolate from a chicken, which was identified as ST203. E. faecalis isolates were more diverse and were identified as ST4, ST6, ST87, ST108, ST274, and ST244, which were grouped as specific to the three hosts. E. faecalis, belonging to the high-risk CC2 and CC87, were detected among isolates from humans. In conclusion, even though one isolate from a chicken was found clonal to that of humans, the MLST analysis of E. faecium and E. faecalis supports the findings of others who suggest VRE to be predominantly host specific and that clinically important strains are found mainly among humans. The infrequent detection of a human VRE clone in a chicken may in fact suggest a reverse transmission of VRE from humans to animals.

  9. Oral treatment with herbal formula B401 alleviates penile toxicity in aging mice with manganism.

    PubMed

    Hsu, Chih-Hsiang; Lin, Ching-Lung; Wang, Sheue-Er; Sheu, Shuenn-Jyi; Chien, Chiang-Ting; Wu, Chung-Hsin

    2015-01-01

    The present study aims to elucidate the roles of nitric oxide synthase activity, oxidative stress, inflammation, and apoptosis in penile toxicity of aging mice associated with excess manganese (Mn) treatment and to investigate the effect of oral treatment with the herbal formula B401 in this respect. ICR strain mice were divided into two groups: the vehicle (sham group) and the B401 (50 mg/kg) group. The mice were orally treated for 5 days; then a high single dose of MnCl2 (100 mg/kg) was given by intraperitoneal injection to the mice. One day after MnCl2 treatment, corpora cavernosal tissues of both Mn-treated mice and their controls were simultaneously sampled to examine their immunohistochemical staining and Western blot analysis. Nitric oxide (NO) production, levels of neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS), expression levels of factors governing angiogenesis (vascular endothelial growth factor), oxidative stress (catalase, superoxide dismutase 2,4-hydroxynonenal), inflammation (tumor necrosis factor alpha), apoptosis (B-cell lymphoma 2 [Bcl-2], Bcl-2-associated X protein [Bax], cleaved poly(adenosine diphosphate-ribose) polymerase [c-PARP], cytochrome C, caspase-12, and caspase-3) were evaluated in penile corpus cavernosum of the mice. We found that penile toxicity in the mice was enhanced under excess Mn treatment through reduction of NOS activity and increase in oxidative stress, inflammation, and apoptosis in the penile cavernous tissue. Furthermore, the penile toxicity in mice with manganism was alleviated by oral B401 treatment through enhancement of both nitric oxide synthesis and angiogenesis, with simultaneous reduction of oxidative stress, inflammation, and apoptosis in penile corpus cavernosum. We suggest that the herbal formula B401 may serve as a potential dietotherapeutic supplement for penile toxicity or dysfunction in aging males.

  10. Oral treatment with herbal formula B401 alleviates penile toxicity in aging mice with manganism

    PubMed Central

    Hsu, Chih-Hsiang; Lin, Ching-Lung; Wang, Sheue-Er; Sheu, Shuenn-Jyi; Chien, Chiang-Ting; Wu, Chung-Hsin

    2015-01-01

    The present study aims to elucidate the roles of nitric oxide synthase activity, oxidative stress, inflammation, and apoptosis in penile toxicity of aging mice associated with excess manganese (Mn) treatment and to investigate the effect of oral treatment with the herbal formula B401 in this respect. ICR strain mice were divided into two groups: the vehicle (sham group) and the B401 (50 mg/kg) group. The mice were orally treated for 5 days; then a high single dose of MnCl2 (100 mg/kg) was given by intraperitoneal injection to the mice. One day after MnCl2 treatment, corpora cavernosal tissues of both Mn-treated mice and their controls were simultaneously sampled to examine their immunohistochemical staining and Western blot analysis. Nitric oxide (NO) production, levels of neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS), expression levels of factors governing angiogenesis (vascular endothelial growth factor), oxidative stress (catalase, superoxide dismutase 2,4-hydroxynonenal), inflammation (tumor necrosis factor alpha), apoptosis (B-cell lymphoma 2 [Bcl-2], Bcl-2-associated X protein [Bax], cleaved poly(adenosine diphosphate-ribose) polymerase [c-PARP], cytochrome C, caspase-12, and caspase-3) were evaluated in penile corpus cavernosum of the mice. We found that penile toxicity in the mice was enhanced under excess Mn treatment through reduction of NOS activity and increase in oxidative stress, inflammation, and apoptosis in the penile cavernous tissue. Furthermore, the penile toxicity in mice with manganism was alleviated by oral B401 treatment through enhancement of both nitric oxide synthesis and angiogenesis, with simultaneous reduction of oxidative stress, inflammation, and apoptosis in penile corpus cavernosum. We suggest that the herbal formula B401 may serve as a potential dietotherapeutic supplement for penile toxicity or dysfunction in aging males. PMID:26064043

  11. Isolation, Characterization, and Differentiation of Progenitor Cells from Human Adult Adrenal Medulla

    PubMed Central

    Santana, Magda M.; Chung, Kuei-Fang; Vukicevic, Vladimir; Rosmaninho-Salgado, Joana; Kanczkowski, Waldemar; Cortez, Vera; Hackmann, Karl; Bastos, Carlos A.; Mota, Alfredo; Schrock, Evelin; Bornstein, Stefan R.; Cavadas, Cláudia

    2012-01-01

    Chromaffin cells, sympathetic neurons of the dorsal ganglia, and the intermediate small intensely fluorescent cells derive from a common neural crest progenitor cell. Contrary to the closely related sympathetic nervous system, within the adult adrenal medulla a subpopulation of undifferentiated progenitor cells persists, and recently, we established a method to isolate and differentiate these progenitor cells from adult bovine adrenals. However, no studies have elucidated the existence of adrenal progenitor cells within the human adrenal medulla. Here we describe the isolation, characterization, and differentiation of chromaffin progenitor cells obtained from adult human adrenals. Human chromaffin progenitor cells were cultured in low-attachment conditions for 10–12 days as free-floating spheres in the presence of fibroblast growth factor-2 (FGF-2) and epidermal growth factor. These primary human chromosphere cultures were characterized by the expression of several progenitor markers, including nestin, CD133, Notch1, nerve growth factor receptor, Snai2, Sox9, Sox10, Phox2b, and Ascl1 on the molecular level and of Sox9 on the immunohistochemical level. In opposition, phenylethanolamine N-methyltransferase (PNMT), a marker for differentiated chromaffin cells, significantly decreased after 12 days in culture. Moreover, when plated on poly-l-lysine/laminin-coated slides in the presence of FGF-2, human chromaffin progenitor cells were able to differentiate into two distinct neuron-like cell types, tyrosine hydroxylase (TH)+/β-3-tubulin+ cells and TH−/β-3-tubulin+ cells, and into chromaffin cells (TH+/PNMT+). This study demonstrates the presence of progenitor cells in the human adrenal medulla and reveals their potential use in regenerative medicine, especially in the treatment of neuroendocrine and neurodegenerative diseases. PMID:23197690

  12. Isolation, characterization, and differentiation of progenitor cells from human adult adrenal medulla.

    PubMed

    Santana, Magda M; Chung, Kuei-Fang; Vukicevic, Vladimir; Rosmaninho-Salgado, Joana; Kanczkowski, Waldemar; Cortez, Vera; Hackmann, Klaus; Bastos, Carlos A; Mota, Alfredo; Schrock, Evelin; Bornstein, Stefan R; Cavadas, Cláudia; Ehrhart-Bornstein, Monika

    2012-11-01

    Chromaffin cells, sympathetic neurons of the dorsal ganglia, and the intermediate small intensely fluorescent cells derive from a common neural crest progenitor cell. Contrary to the closely related sympathetic nervous system, within the adult adrenal medulla a subpopulation of undifferentiated progenitor cells persists, and recently, we established a method to isolate and differentiate these progenitor cells from adult bovine adrenals. However, no studies have elucidated the existence of adrenal progenitor cells within the human adrenal medulla. Here we describe the isolation, characterization, and differentiation of chromaffin progenitor cells obtained from adult human adrenals. Human chromaffin progenitor cells were cultured in low-attachment conditions for 10-12 days as free-floating spheres in the presence of fibroblast growth factor-2 (FGF-2) and epidermal growth factor. These primary human chromosphere cultures were characterized by the expression of several progenitor markers, including nestin, CD133, Notch1, nerve growth factor receptor, Snai2, Sox9, Sox10, Phox2b, and Ascl1 on the molecular level and of Sox9 on the immunohistochemical level. In opposition, phenylethanolamine N-methyltransferase (PNMT), a marker for differentiated chromaffin cells, significantly decreased after 12 days in culture. Moreover, when plated on poly-l-lysine/laminin-coated slides in the presence of FGF-2, human chromaffin progenitor cells were able to differentiate into two distinct neuron-like cell types, tyrosine hydroxylase (TH)(+)/β-3-tubulin(+) cells and TH(-)/β-3-tubulin(+) cells, and into chromaffin cells (TH(+)/PNMT(+)). This study demonstrates the presence of progenitor cells in the human adrenal medulla and reveals their potential use in regenerative medicine, especially in the treatment of neuroendocrine and neurodegenerative diseases. PMID:23197690

  13. Complete Genome Sequences of Two Methicillin-Sensitive Staphylococcus aureus Isolates Representing a Population Subset Highly Prevalent in Human Colonization

    PubMed Central

    Weber, Robert E.; Layer, Franziska; Fuchs, Stephan; Bender, Jennifer K.; Fiedler, Stefan; Werner, Guido

    2016-01-01

    Here, we report the high-quality draft genome sequences of two methicillin-susceptible Staphylococcus aureus isolates, 08-02119 and 08-02300. Belonging to sequence type 582 (ST582) and ST7, both isolates are representatives of clonal lineages often associated with asymptomatic colonization of humans. PMID:27469954

  14. Complete Genome Sequences of Two Methicillin-Sensitive Staphylococcus aureus Isolates Representing a Population Subset Highly Prevalent in Human Colonization.

    PubMed

    Weber, Robert E; Layer, Franziska; Fuchs, Stephan; Bender, Jennifer K; Fiedler, Stefan; Werner, Guido; Strommenger, Birgit

    2016-01-01

    Here, we report the high-quality draft genome sequences of two methicillin-susceptible Staphylococcus aureus isolates, 08-02119 and 08-02300. Belonging to sequence type 582 (ST582) and ST7, both isolates are representatives of clonal lineages often associated with asymptomatic colonization of humans. PMID:27469954

  15. Complete Genome Sequences of Two Methicillin-Sensitive Staphylococcus aureus Isolates Representing a Population Subset Highly Prevalent in Human Colonization.

    PubMed

    Weber, Robert E; Layer, Franziska; Fuchs, Stephan; Bender, Jennifer K; Fiedler, Stefan; Werner, Guido; Strommenger, Birgit

    2016-01-01

    Here, we report the high-quality draft genome sequences of two methicillin-susceptible Staphylococcus aureus isolates, 08-02119 and 08-02300. Belonging to sequence type 582 (ST582) and ST7, both isolates are representatives of clonal lineages often associated with asymptomatic colonization of humans.

  16. Complete Genome Sequences of Two Novel Isolates of Human Parainfluenza Virus 1 Associated with Acute Respiratory Infection

    PubMed Central

    Denson, J. L.; Kennedy, J. L.; Dehority, W. N.; Eickman, M. M.; Schwalm, K. S.; Stoner, A. N.; Kincaid, J. C.; Abramo, T. J.; Thompson, T. M.; Ulloa, E. M.; Burchiel, S. W.; Young, S. A.

    2016-01-01

    Using target capture of viral nucleic acid and next-generation sequencing, we generated the complete genomes of two novel human parainfluenza virus 1 isolates. Isolates AR001 (accession no. KX570602) and NM001 (accession no. KX639498) were collected 3 months apart from pediatric patients with acute respiratory infection from Arkansas and New Mexico, respectively. PMID:27738046

  17. Full genome sequences and molecular characterization of tick-borne encephalitis virus strains isolated from human patients.

    PubMed

    Formanová, Petra; Černý, Jiří; Bolfíková, Barbora Černá; Valdés, James J; Kozlova, Irina; Dzhioev, Yuri; Růžek, Daniel

    2015-02-01

    Tick-borne encephalitis virus (TBEV) causes tick-borne encephalitis (TBE), one of the most important human neuroinfections across Eurasia. Up to date, only three full genome sequences of human European TBEV isolates are available, mostly due to difficulties with isolation of the virus from human patients. Here we present full genome characterization of an additional five low-passage TBEV strains isolated from human patients with severe forms of TBE. These strains were isolated in 1953 within Central Bohemia in the former Czechoslovakia, and belong to the historically oldest human TBEV isolates in Europe. We demonstrate here that all analyzed isolates are distantly phylogenetically related, indicating that the emergence of TBE in Central Europe was not caused by one predominant strain, but rather a pool of distantly related TBEV strains. Nucleotide identity between individual sequenced TBEV strains ranged from 97.5% to 99.6% and all strains shared large deletions in the 3' non-coding region, which has been recently suggested to be an important determinant of virulence. The number of unique amino acid substitutions varied from 3 to 9 in individual isolates, but no characteristic amino acid substitution typical exclusively for all human TBEV isolates was identified when compared to the isolates from ticks. We did, however, correlate that the exploration of the TBEV envelope glycoprotein by specific antibodies were in close proximity to these unique amino acid substitutions. Taken together, we report here the largest number of patient-derived European TBEV full genome sequences to date and provide a platform for further studies on evolution of TBEV since the first emergence of human TBE in Europe. PMID:25311899

  18. Molecular Characterization of Echinococcus granulosus from Hydatid Cysts Isolated from Human and Animals in Golestan Province, North of Iran

    PubMed Central

    Gholami, Sh; Sosari, M; Fakhar, M; Sharif, M; Daryani, A; Hashemi, MB; Vahadi, M

    2012-01-01

    Background The aim of the present study was to determine the molecular characteristics of Echinococcus granulosus from paraffin-embedded tissues of hydatid cysts isolated from human and protoscoleces of hydatid cysts from sheep, cattle and camel isolates using PCR- RFLP of ITS1- rDNA analysis in Golestan Province, northern Iran. Methods E. granulosus isolates from human patients infected with hydatid cyst and protoscoleces from hydatid cysts of sheep, cattle and camel isolates were collected from different hospitals and the abattoir throughout the Golestan Province. In all, 60 E. granulosus genomic DNA were extracted and examined by PCR - ITS1 of rDNA and amplified using BD1 / 4S and EGF1 / EGR2 primers, followed by RFLP using Alu1, Msp1 and TaqI restriction enzymes. Results The PCR-ITS1 products obtained from sheep, cattle and human isolates were similar to sheep strain (1000 bp and 391 bp). Majority of the camel samples yielded 295 bp DNA bands. RFLP -ITS1 of E. granulosus with Taq1 in human, sheep and cattle isolates showed similar patterns in the number and size of DNA. RFLP methods in camel isolates showed a different genotype, using Taq1, whereas no DNA bands were observed using Alu1 in camel and human isolates. Therefore, two clearly distinguishable banding patterns of E. granulosus were obtained with the three enzymes, which separating human, sheep and cattle isolates from the camel origin. Conclusion The results indicate the possible of transmission of the G1 and G6 genotypes of E. granulosus between livestock animals and human in Golestan Province. PMID:23323086

  19. Isolation and characterization of cytotoxic effector cells and antibody producing cells from human intestine.

    PubMed

    MacDermott, R P

    1985-01-01

    We have examined the ability of intestinal and peripheral blood mononuclear cells isolated from patients with inflammatory bowel disease to mediate killing against cell line targets in spontaneous, antibody-dependent, lectin-induced, and interferon-induced cell-mediated cytotoxicity assays, as well as responsiveness in the allogeneic mixed leukocyte reaction, and effector capabilities in cell-mediated lympholysis. IMC were poor mediators of spontaneous or antibody-dependent cellular cytotoxicity with cell line cells as targets (in comparison to normal PBMC, but were capable of killing antibody coated chicken red blood cells. Although IMC were capable of responding to allogeneic cell surface antigens in the mixed leukocyte reaction, they did not exhibit effector function in cell-mediated lympholysis. Mitogenic lectins induced cell-mediated cytotoxicity by isolated intestinal mononuclear cells from controls and patients. HFIF induces cytotoxicity by control but not inflammatory bowel disease intestinal cells. Pokeweed mitogen was the lectin which induced the greatest amount of killing against human cell line targets. We therefore speculate that exogenous agents, or endogenous factors released during viral infection, could play a role in inducing cell mediated cytotoxic damage to the intestine in inflammatory bowel disease patients. In addition, the functional differences between IMC and PBMC indicate that intestinal MNC may have unique cell capabilities which must be better understood prior to the delineation of immunopathologic events in solid organ tissues. We have also examined the secretion of IgA, IgM, and IgG by isolated human IMC, human bone marrow MNC from rib specimens, and PBMC from patients with CD, UC, SLE, or Henoch-Schoenlein purpura (HSP). Control IMC exhibited high spontaneous secretion of IgA, while intestinal MNC from UC and CD patients exhibited only modest increases in IgA secretion. PBMC from patients with CD, UC, SLE, or HSP exhibited markedly

  20. Human alpha-L-iduronidase: cDNA isolation and expression.

    PubMed Central

    Scott, H S; Anson, D S; Orsborn, A M; Nelson, P V; Clements, P R; Morris, C P; Hopwood, J J

    1991-01-01

    alpha-L-Iduronidase (IDUA; EC 3.2.1.76) is a lysosomal hydrolase in the metabolic pathway responsible for the degradation of the glycosaminoglycans heparan sulfate and dermatan sulfate. A deficiency of IDUA in humans leads to the accumulation of these glycosaminoglycans and results in the lysosomal storage disorder mucopolysaccharidosis type I. We have isolated and sequenced cDNA clones containing part of the human IDUA coding region and used PCR from reverse-transcribed RNA to obtain the full IDUA sequence. Analysis of the predicted 653-amino acid precursor protein shows that IDUA has a 26-amino acid signal peptide that is cleaved immediately prior to the amino terminus of the 74-kDa polypeptide present in human liver IDUA. The protein sequence contains six potential N-glycosylation sites. Northern blot analysis with IDUA cDNA detected only a single 2.3-kilobase mRNA species in human placental RNA; however, PCR analysis of fibroblast, liver, kidney, and placental RNA showed the existence of alternatively spliced mRNA from the IDUA gene. Southern blot analysis failed to detect major deletions or gene rearrangements in any of the 40 mucopolysaccharidosis type I patients studied. Expression of a full-length IDUA cDNA construct in Chinese hamster ovary cells produced human IDUA protein at a level 13-fold higher than, and with a specific activity comparable to, IDUA present in normal human fibroblasts. Images PMID:1946389

  1. Isolation and mapping of 68 RFLP markers on human chromosome 6

    SciTech Connect

    Saito, Susumo SRL Inc., Tokyo ); Okui, Keiko; Tokino, Takashi; Nakamura, Yusuke ); Oshimura, Mitsuo )

    1992-01-01

    The authors have isolated 68 new RFLP markers on human chromosome 6. Of these, 64 were localized on chromosomal bands by the fluorescent in-situ hybridization (FISH) method, 25 on the short arm and 39 on the long arm. Their distribution was uneven; the makers were localized predominantly in regions of R-positive banding. Eleven markers defined VNTR loci. This expanded collection of DNA markers will contribute to high-resolution linkage mapping of genes causing inherited disorders and will provide useful reagents for isolation of putative tumor-suppressor genes on chromosome 6 that appear to be involved in malignancies. Furthermore, the new markers will be guideposts for detailed linkage and physical maps of this chromosome.

  2. Isolation and refined regional mapping of expressed sequences from human chromosome 21

    SciTech Connect

    Kao, F.T.; Yu, J.; Patterson, D.

    1994-10-01

    To increase candidate genes from human chromosome 21 for the analysis of Down syndrome and other genetic diseases localized on this chromosome, we have isolated and studied 9 cDNA clones encoded by chromosome 21. For isolating cDNAs, single-copy microclones from a chromosome 21 microdissection library were used in direct screening of various cDNA libraries. Seven of the cDNA clones have been regionally mapped on chromosome 21 using a comprehensive hybrid mapping panel comprising 24 cell hybrids that divide the chromosome into 33 subregions. These cDNA clones with refined mapping positions should be useful for identification and cloning of genes responsible for the specific component phenotypes of Down syndrome and other diseases on chromosome 21, including progressive myoclonus epilepsy in 21q22.3. 12 refs., 2 figs., 1 tab.

  3. The isolation and structure of C4, the fourth component of human complement.

    PubMed Central

    Gigli, I; von Zabern, I; Porter, R R

    1977-01-01

    The fourth component of complement, C4, was isolated from human serum in good yield, and in confirmation of previous reports was shown to be formed from three peptide chains, alpha, beta and gamma, with apparent mol.wts. 90 000, 80 000 and 30 000 respectively. Preparative methods are described for the isolation of the three peptide chains and their amino acid analyses reported. Component C4 contains 7.0% carbohydrate, alpha-chain 8.6% and the beta-chain 5.6%. The N-terminal amino acid sequences are given for 12 residues of the alpha-chain, eight of the beta-chain and 19 of the gamma-chain. Images PLATE 1 PMID:921758

  4. Isolation of Klebsiella terrigena from human feces: biochemical reactions, capsule types, and antibiotic sensitivity.

    PubMed

    Podschun, R

    1991-04-01

    Colonization of the human intestinal tract by a newly proposed species, K. terrigena, was investigated. 5377 different stool specimens from healthy persons (food handlers) yielded 50 isolates (0.9%). Biochemically, low frequencies in the degradation of urea, dulcitol, and utilization of citrate at 37 degrees C were found when compared to K. pneumoniae. At 30 degrees C, urea hydrolysis was observed twice as often as at 37 degrees C. Apart from ampicillin, K. terrigena was susceptible to 12 other antimicrobial drugs tested. Multiple drug resistance was rare, few isolates being resistant against 2-4 antibiotic agents. Capsule typing revealed 30 different serotypes, K 70 and K 14 were the most frequent. Six strains expressed capsule types K 2 and K 5, which have been reported to be associated with virulence in K. pneumoniae. A possible pathogenic role of K. terrigena is discussed.

  5. Isolation and characterization of human cerebellum cDNAs containing polymorphic CAG trinucleotide repeats

    SciTech Connect

    Igarashi, S.; Onodera, O.; Tanaka, H.

    1994-09-01

    It has been discovered that neurologic diseases such as X linked spinal and bulbar muscular atrophy, Huntington`s disease, spinocerebellar ataxia type 1 (SCA1), and dentatorubral-pallidoluysian atrophy (DRPLA) are caused by unstable expansions of CAG repeats, which shed a light on a new mechanism of human hereditary diseases. The genetic anticipation, a common genetic feature in these diseases, can be explained by the trinucleotide repeat expansions, and an inverse correlation between the ages of onset and the numbers of trinucleotide repeats is demonstrated in these diseases. Furthermore, there have been diseases such as spinocerebellar ataxia 2 (SCA2) and Machado-Joseph disease showing similar genetic anticipation, which suggests that their causative mutations are unstable expansions of trinucleotide repeats. To identify candidate genes for neurodegenerative diseases which are expressed in human cerebellum and contain CAG repeats, we screened a human cerebellum cDNA library with an oligonucleotide (CAG){sub 10}, labelled with [{gamma}{sup 32}P]ATP. Out of 78 clones we have isolated, 43 clones were partially sequenced and 31 clones were shown to contain CAG or CTG tinucleotide repeats. From homology searches, 12 of the 59 clones were identified to contain known sequences including human MAR/SAR DNA binding protein, human glial fibrillary acidic protein, human myelin transcription factor 1, human neuronal growth protein 43 and human myocyte-specific enhancer 2. From 6 clones out of the 43 novel genes, we were able to develop primer pairs flanking CAG repeats and determined chromosomal localizations with human and rodent hybrid mapping panels. These CAG repeats were shown to be polymorphic and mapped to 1, 15, 17 and 18. These novel cDNAs will be useful as candidate genes for hereditary neurologic diseases showing genetic anticipation.

  6. Salmonella L-forms: formation in human bile in vitro and isolation culture from patients' gallbladder samples by a non-high osmotic isolation technique.

    PubMed

    Wang, D N; Wu, W J; Wang, T; Pan, Y Z; Tang, K L; She, X L; Ding, W J; Wang, H

    2015-05-01

    Bacterial L-forms have always been considered as osmotic-pressure-sensitive cell-wall-deficient bacteria and isolation culture of L-forms must use media with high osmotic pressure. However, isolation culture of stable L-forms formed in humans and animals is very difficult because they have adapted to the physiological osmotic pressure condition of the host. We use a non-high osmotic isolation technique to isolate stable L-forms of Salmonella Typhi and Salmonella Paratyphi A from bile-inducer cultures in vitro and from patients' gallbladder specimens. Multiplex PCR assay for Salmonella-specific genes and nucleotide sequencing are used to identify the Salmonella L-forms in stable L-form isolates. Using this method, we confirmed that Salmonella Paratyphi A and Salmonella Typhi cannot be isolated from bile-inducer cultures cultured for 6 h or 48 h, but the L-forms can be isolated from 1 h to 45 days. In the 524 gallbladder samples, the positive rate for bacterial forms was 19.7% and the positive rate for Salmonella spp. was 0.6% by routine bacteriological methods. The positive rate for bacterial L-forms was 75.4% using non-high osmotic isolation culture. In the L-form isolates, the positive rate of Salmonella invA gene was 3.1%. In these invA-positive L-form isolates, four were positive for the invA and flic-d genes of Salmonella Typhi, and ten were positive for the invA and flic-a genes of Salmonella Paratyphi A.

  7. Prevalence and Characterization of Methicillin-Resistant Staphylococcus aureus Isolates from Retail Meat and Humans in Georgia

    PubMed Central

    Davis, Johnnie A.; Barrett, John B.

    2013-01-01

    There is increasing interest in the presence of Staphylococcus aureus, specifically methicillin-resistant S. aureus (MRSA), on retail meat products. In this study, staphylococci were isolated from retail pork and retail beef in Georgia, and MRSA from the products was compared to human MRSA from the same geographic area using broth microdilution antimicrobial susceptibility testing, multilocus sequence typing (MLST), spa typing, SCCmec typing, and pulsed-field gel electrophoresis (PFGE). S. aureus was isolated from 45% (45/100) of pork products and 63% (63/100) of beef products; mecA was detected in S. aureus from both pork (3/100; 3%) and beef (4/100; 4%). Fifty percent (50/100) of human S. aureus also contained mecA. Multidrug resistance was detected among MRSA from all sources. All MRSA (n = 57) was SCCmec type IV, and nine different spa types were present among the isolates (t002, t008, t012, t024, t179, t337, t548, t681, and t1062). Four sequence types (ST5, ST8, ST9, and ST30) were detected using MLST; the majority of MRSA isolates belonged to ST8, followed by ST5. One retail beef MRSA isolate belonged to ST8, while the remaining three were ST5. In retail pork MRSA, ST5, ST9, and ST30 were observed. The majority of human MRSA isolates belonged to ST8. Thirty-seven MRSA isolates, one of which was a retail beef MRSA isolate, were pvl+. Using PFGE, MLST, and spa typing, three retail beef MRSA isolates were found to be identical in PFGE pattern, ST, and spa type to two human clonal MRSA isolates (USA100 and USA300). One additional retail beef MRSA isolate had a PFGE pattern similar to that of a human MRSA isolate, whereas none of the retail pork MRSA isolates had PFGE patterns similar to those of human MRSA isolates. These data suggest that the retail beef samples were contaminated by a human source, possibly during processing of the meat, and may present a source of MRSA for consumers and others who handle raw meat. PMID:23363837

  8. Genetic Relatedness Among Escherichia coli Pathotypes Isolated from Food Products for Human Consumption in Cartagena, Colombia.

    PubMed

    Amézquita-Montes, Zorangel; Tamborski, Maria; Kopsombut, Usa G; Zhang, Chengxian; Arzuza, Octavio S; Gómez-Duarte, Oscar G

    2015-05-01

    Foodborne pathogens are a leading cause of mild-to-severe gastrointestinal illnesses worldwide. Escherichia coli pathotypes have been known to cause gastrointestinal illnesses in children less than 5 years old in Colombia. However, insufficient information is available on the prevalence of E. coli contamination of food products and the kind of E. coli food product reservoirs. The two objectives of this study were designed to address this issue. The first objective was to ascertain coliform, E. coli, and pathogenic E. coli contamination of food products readily available for human consumption in Cartagena, Colombia. The second objective was to evaluate the relationship between pathogenic E. coli isolated from food products and those isolated from cases of diarrhea in children. Food product samples consisting of pasteurized milk, unpasteurized fruit juice, ground beef, cheese, and vegetables were obtained at four retail stores. The food samples were cultured in liquid media and tested for the presence of coliforms and E. coli. E. coli isolates were tested by polymerase chain reaction for the presence of pathogenic E. coli. Coliforms, E. coli, and E. coli intestinal pathotypes contamination were detected in 88.4%, 53%, and 2.1% of food product samples, respectively. Ground beef and cheese were the only food samples contaminated with E. coli intestinal pathotypes including enteropathogenic (EPEC), Shiga toxin-producing (STEC), and enterotoxigenic E. coli (ETEC). Closed multilocus sequencing typing relationships between diarrheagenic E. coli isolates from food products and from individuals with diarrhea suggest that food products readily available at public markets in Cartagena can transmit ETEC and possibly EPEC and STEC. We demonstrated that a high proportion of food products for human consumption available at public markets in Cartagena are contaminated with coliforms, E. coli, and E. coli intestinal pathogens. Furthermore, food products containing E. coli intestinal

  9. Genetic Relatedness Among Escherichia coli Pathotypes Isolated from Food Products for Human Consumption in Cartagena, Colombia

    PubMed Central

    Amézquita-Montes, Zorangel; Tamborski, Maria; Kopsombut, Usa G.; Zhang, Chengxian; Arzuza, Octavio S.

    2015-01-01

    Abstract Foodborne pathogens are a leading cause of mild-to-severe gastrointestinal illnesses worldwide. Escherichia coli pathotypes have been known to cause gastrointestinal illnesses in children less than 5 years old in Colombia. However, insufficient information is available on the prevalence of E. coli contamination of food products and the kind of E. coli food product reservoirs. The two objectives of this study were designed to address this issue. The first objective was to ascertain coliform, E. coli, and pathogenic E. coli contamination of food products readily available for human consumption in Cartagena, Colombia. The second objective was to evaluate the relationship between pathogenic E. coli isolated from food products and those isolated from cases of diarrhea in children. Food product samples consisting of pasteurized milk, unpasteurized fruit juice, ground beef, cheese, and vegetables were obtained at four retail stores. The food samples were cultured in liquid media and tested for the presence of coliforms and E. coli. E. coli isolates were tested by polymerase chain reaction for the presence of pathogenic E. coli. Coliforms, E. coli, and E. coli intestinal pathotypes contamination were detected in 88.4%, 53%, and 2.1% of food product samples, respectively. Ground beef and cheese were the only food samples contaminated with E. coli intestinal pathotypes including enteropathogenic (EPEC), Shiga toxin–producing (STEC), and enterotoxigenic E. coli (ETEC). Closed multilocus sequencing typing relationships between diarrheagenic E. coli isolates from food products and from individuals with diarrhea suggest that food products readily available at public markets in Cartagena can transmit ETEC and possibly EPEC and STEC. We demonstrated that a high proportion of food products for human consumption available at public markets in Cartagena are contaminated with coliforms, E. coli, and E. coli intestinal pathogens. Furthermore, food products containing E. coli

  10. Isolation of Cellular Lipid Droplets: Two Purification Techniques Starting from Yeast Cells and Human Placentas

    PubMed Central

    Dalhaimer, Paul

    2014-01-01

    Lipid droplets are dynamic organelles that can be found in most eukaryotic and certain prokaryotic cells. Structurally, the droplets consist of a core of neutral lipids surrounded by a phospholipid monolayer. One of the most useful techniques in determining the cellular roles of droplets has been proteomic identification of bound proteins, which can be isolated along with the droplets. Here, two methods are described to isolate lipid droplets and their bound proteins from two wide-ranging eukaryotes: fission yeast and human placental villous cells. Although both techniques have differences, the main method - density gradient centrifugation - is shared by both preparations. This shows the wide applicability of the presented droplet isolation techniques. In the first protocol, yeast cells are converted into spheroplasts by enzymatic digestion of their cell walls. The resulting spheroplasts are then gently lysed in a loose-fitting homogenizer. Ficoll is added to the lysate to provide a density gradient, and the mixture is centrifuged three times. After the first spin, the lipid droplets are localized to the white-colored floating layer of the centrifuge tubes along with the endoplasmic reticulum (ER), the plasma membrane, and vacuoles. Two subsequent spins are used to remove these other three organelles. The result is a layer that has only droplets and bound proteins. In the second protocol, placental villous cells are isolated from human term placentas by enzymatic digestion with trypsin and DNase I. The cells are homogenized in a loose-fitting homogenizer. Low-speed and medium-speed centrifugation steps are used to remove unbroken cells, cellular debris, nuclei, and mitochondria. Sucrose is added to the homogenate to provide a density gradient and the mixture is centrifuged to separate the lipid droplets from the other cellular fractions. The purity of the lipid droplets in both protocols is confirmed by Western Blot analysis. The droplet fractions from both preps

  11. Genetic Relatedness Among Escherichia coli Pathotypes Isolated from Food Products for Human Consumption in Cartagena, Colombia.

    PubMed

    Amézquita-Montes, Zorangel; Tamborski, Maria; Kopsombut, Usa G; Zhang, Chengxian; Arzuza, Octavio S; Gómez-Duarte, Oscar G

    2015-05-01

    Foodborne pathogens are a leading cause of mild-to-severe gastrointestinal illnesses worldwide. Escherichia coli pathotypes have been known to cause gastrointestinal illnesses in children less than 5 years old in Colombia. However, insufficient information is available on the prevalence of E. coli contamination of food products and the kind of E. coli food product reservoirs. The two objectives of this study were designed to address this issue. The first objective was to ascertain coliform, E. coli, and pathogenic E. coli contamination of food products readily available for human consumption in Cartagena, Colombia. The second objective was to evaluate the relationship between pathogenic E. coli isolated from food products and those isolated from cases of diarrhea in children. Food product samples consisting of pasteurized milk, unpasteurized fruit juice, ground beef, cheese, and vegetables were obtained at four retail stores. The food samples were cultured in liquid media and tested for the presence of coliforms and E. coli. E. coli isolates were tested by polymerase chain reaction for the presence of pathogenic E. coli. Coliforms, E. coli, and E. coli intestinal pathotypes contamination were detected in 88.4%, 53%, and 2.1% of food product samples, respectively. Ground beef and cheese were the only food samples contaminated with E. coli intestinal pathotypes including enteropathogenic (EPEC), Shiga toxin-producing (STEC), and enterotoxigenic E. coli (ETEC). Closed multilocus sequencing typing relationships between diarrheagenic E. coli isolates from food products and from individuals with diarrhea suggest that food products readily available at public markets in Cartagena can transmit ETEC and possibly EPEC and STEC. We demonstrated that a high proportion of food products for human consumption available at public markets in Cartagena are contaminated with coliforms, E. coli, and E. coli intestinal pathogens. Furthermore, food products containing E. coli intestinal

  12. Genetic features of human and bovine Escherichia coli O157:H7 strains isolated in Argentina.

    PubMed

    Pianciola, L; D'Astek, B A; Mazzeo, M; Chinen, I; Masana, M; Rivas, M

    2016-02-01

    Shiga toxin-producing Escherichia coli (STEC) are important food-borne pathogens associated with human diseases. In Argentina, O157:H7 is the dominant serotype in hemolytic uremic syndrome (HUS) cases. Previously, we have described the almost exclusive circulation of human E. coli O157 strains belonging to the hypervirulent clade 8 in Neuquén Province. The aim of the present study was to investigate, by a broad molecular characterization, if this particular distribution of E. coli O157 clades in Neuquén is similar to the situation in other regions of the country and if it may be originated in a similar profile in cattle, its main reservoir. Two-hundred and eighty O157 strains (54 bovine and 226 human) isolated between 2006 and 2008 in different regions of Argentina were studied. All strains harbored rfbO157, fliCH7, eae, and ehxA genes. The predominant genotype was stx2a/stx2c in human (76.1%) and bovine (55.5%) strains. All human isolates tested by Lineage-Specific Polymorphism Assay (LSPA-6), were lineage I/II; among bovine strains, 94.1% belonged to lineage I/II and 5.9% to lineage I. No LSPA-6 lineage II isolates were detected. Single nucleotide polymorphism (SNP) analysis has revealed the existence of nine clade phylogenetic groups. In our clinical strains collection, 87.6% belonged to the hypervirulent clade 8, and 12.4% were classified as clade 4/5. In bovine isolates, 59.3% strains were clade 8, 33.3% clade 4/5 and 7.4% clade 3. More than 80% of human strains showed the presence of 6 of the 7 virulence determinants described in the TW14359 O157 strain associated with the raw spinach outbreak in the U.S. in 2006. More than 80% of bovine strains showed the presence of 3 of these factors. The q933 allele, which has been related to high toxin production, was present in 98.2% of clinical strains and 75.9% of the bovine isolates. The molecular characterization of human STEC O157 strains allows us to conclude that the particular situation previously described

  13. Genetic features of human and bovine Escherichia coli O157:H7 strains isolated in Argentina.

    PubMed

    Pianciola, L; D'Astek, B A; Mazzeo, M; Chinen, I; Masana, M; Rivas, M

    2016-02-01

    Shiga toxin-producing Escherichia coli (STEC) are important food-borne pathogens associated with human diseases. In Argentina, O157:H7 is the dominant serotype in hemolytic uremic syndrome (HUS) cases. Previously, we have described the almost exclusive circulation of human E. coli O157 strains belonging to the hypervirulent clade 8 in Neuquén Province. The aim of the present study was to investigate, by a broad molecular characterization, if this particular distribution of E. coli O157 clades in Neuquén is similar to the situation in other regions of the country and if it may be originated in a similar profile in cattle, its main reservoir. Two-hundred and eighty O157 strains (54 bovine and 226 human) isolated between 2006 and 2008 in different regions of Argentina were studied. All strains harbored rfbO157, fliCH7, eae, and ehxA genes. The predominant genotype was stx2a/stx2c in human (76.1%) and bovine (55.5%) strains. All human isolates tested by Lineage-Specific Polymorphism Assay (LSPA-6), were lineage I/II; among bovine strains, 94.1% belonged to lineage I/II and 5.9% to lineage I. No LSPA-6 lineage II isolates were detected. Single nucleotide polymorphism (SNP) analysis has revealed the existence of nine clade phylogenetic groups. In our clinical strains collection, 87.6% belonged to the hypervirulent clade 8, and 12.4% were classified as clade 4/5. In bovine isolates, 59.3% strains were clade 8, 33.3% clade 4/5 and 7.4% clade 3. More than 80% of human strains showed the presence of 6 of the 7 virulence determinants described in the TW14359 O157 strain associated with the raw spinach outbreak in the U.S. in 2006. More than 80% of bovine strains showed the presence of 3 of these factors. The q933 allele, which has been related to high toxin production, was present in 98.2% of clinical strains and 75.9% of the bovine isolates. The molecular characterization of human STEC O157 strains allows us to conclude that the particular situation previously described

  14. Screening of Bifidobacterium strains isolated from human faeces for antagonistic activities against potentially bacterial pathogens.

    PubMed

    Bevilacqua, Lorella; Ovidi, Monia; Di Mattia, Elena; Trovatelli, Luigi Daniele; Canganella, Francesco

    2003-01-01

    As probiotic bacteria, strains belonging to the genus Bifidobacterium colonise the gastro-intestinal tract of humans and animals at the time of birth, and they are found in young as well as in adult individuals in great numbers. Moreover, they can interact with the development of enteric infections by the production of antimicrobial metabolites. In this work 281 strains of bifidobacteria were anaerobically isolated from human faecal samples, supplied by volunteers of different ages (youngs, adults, elders), and preliminarly described by microscopic observation. All strains were screened by the fructose 6-phosphate phosphoketolase (F6PPK) test in order to confirm their classification within the genus Bifidobacterium. Selected strains were used to evaluate their antagonistic activities against Escherichia coli, Salmonella thyphimurium, Staphylococcus lentus, Enterococcus faecalis, Acinetobacter calcoaceticus, Sphingomonas paucimobilis, Listeria monocytogenes, Yersinia enterocolitica, Bacillus cereus, Clostridium sporogenes. Experiments were performed in vitro by different methods based on the observation of growth inhibition in Petri dishes. The strains that showed the highest inhibiting activities were compared by SDS-PAGE for total cell proteins, using type strains of human origin as references. Representative isolates were metabolically characterised by the BIOLOG system; a specific database was created with strains obtained from our collection and a statistical evaluation for metabolic patterns was carried out.

  15. Multiple isolates and characteristics of human T-cell leukemia virus type II.

    PubMed Central

    Hall, W W; Takahashi, H; Liu, C; Kaplan, M H; Scheewind, O; Ijichi, S; Nagashima, K; Gallo, R C

    1992-01-01

    Human T-cell leukemia (or lymphotropic) virus type II (HTLV-II) was isolated from eight HTLV-seropositive patients, six of whom were also infected with human immunodeficiency virus, by cocultivation of peripheral blood mononuclear cells (PBMCs) with BJAB, a continuous B-cell line. Restriction endonuclease mapping of the proviruses demonstrated consistent differences among isolates, and two distinct physical map patterns were observed. The results suggest the existence of two closely related molecular subtypes of HTLV-II, which are tentatively designated HTLV-IIa and HTLV-IIb. This finding was supported by preliminary nucleotide sequence analysis of the env gene region encoding the transmembrane glycoprotein gp21, which showed consistent differences between the two proposed virus subtypes. Exploitation of differences in restriction endonuclease sites allowed polymerase chain reaction amplification to detect and differentiate the two subtypes in fresh PBMCs of HTLV-seropositive intravenous drug abusers (IVDAs). The results of these studies confirm that HTLV-II infection is the prominent HTLV infection in seropositive IVDAs and also show that infection with both subtypes occurs. The finding of genetic heterogeneity in the HTLV-II group of viruses may have important implications for studies on its role in human disease and will be useful in characterizing the viruses present in newly discovered endemic foci in New World indigenous populations. Images PMID:1347796

  16. In vivo pathogenic properties of two clonal human immunodeficiency virus type 1 isolates.

    PubMed Central

    Jamieson, B D; Pang, S; Aldrovandi, G M; Zha, J; Zack, J A

    1995-01-01

    We have investigated the in vivo pathogenic properties of two molecularly cloned strains of human immunodeficiency virus type 1 (HIV-1), HIV-1NL4-3 and HIV-1JR-CSF, in human fetal thymus/liver implants in severe combined immunodeficient mice. Studies comparing their in vivo replication kinetics and abilities to induce CD4+ thymocyte depletion were performed. HIV-1NL4-3 replicated in vivo with faster kinetics and induced greater levels of CD4+ thymocyte depletion than did HIV-1JR-CSF. These results demonstrate that different viral isolates have different pathogenic properties in this system. In the SCID-hu model, this pathogenesis most likely occurs in the absence of an immune response. Therefore, we investigated whether the absence of immune selection resulted in extensive genetic variation and the generation of viral quasispecies. To this end, DNA corresponding to the fourth variable domain region of the viral envelope gp120 protein recovered from biopsy samples at 6 weeks postinfection was sequenced. Little genetic variation was noted in either HIV-1JR-CSF- or HIV-1NL4-3-infected implants. The mutation levels demonstrated in both viral strains were more reflective of the acute rather than the chronic phase of HIV-1 infection in humans. These results suggest that the SCID-hu mouse model can be used to study the in vivo pathogenicity of different HIV-1 isolates in the absence of host immune selective pressures. PMID:7666526

  17. Isolation of the human Xp21 glycerol kinase gene by positional cloning.

    PubMed

    Walker, A P; Muscatelli, F; Monaco, A P

    1993-02-01

    The gene for human glycerol kinase deficiency (GK) maps in Xp21.3 in a critical region of about 50-250 kb located distal to the Duchenne muscular dystrophy gene (DMD) by analysis of patient deletions and YAC contigs. We have used a genomic exon amplification strategy to isolate potential exons from two cosmids which mapped to this interval. The genomic exons were used to isolate six overlapping cDNA clones from human fetal liver which encode the X-linked glycerol kinase gene. The cDNA clones map to cosmids, YAC clones and deletions in patients which define the GK critical region and also hybridize to several autosomal fragments and one Xq fragment in genomic DNA. The GK gene is expressed most in human liver with three transcript sizes of 1.85, 2.7, and 3.7 kb. Sequence analysis of 1.5 kb of several overlapping liver cDNA clones predicted a protein with approximately 63% similarity to the E. coli and B. subtilis glycerol kinase genes. The liver cDNA clones have sequence identity with four genomic exons and the 3' untranslated region from an Xp21.3 cosmid thus indicating that this is the expressed GK gene which when deleted in patients gives rises to GK deficiency. PMID:8499898

  18. Isolation and characterization of lymphatic endothelial cells from human glossal lymphangioma.

    PubMed

    You, Lanying; Wu, Mingfu; Chen, Ying; Xu, Gang; Wei, Juncheng; Li, Qiong; Song, Anping; Zhao, Liangpin; Li, Shuang; Han, Zhiqiang; Zhou, Jianfeng; Lu, Yunpin; Wang, Shixuan; Ma, Ding; Meng, Li

    2010-01-01

    Abnormal lymphangiogenesis is associated with several diseases such as tumor metastasis and lymphangioma. Human lymphangioma originated from the transformation of lymphatic endothelium is a benign malformation of lymphatic vessels and its pathogenesis has up to date not been illuminated and its cell model has also not been established. An optimized method was used to isolate lymphatic endothelial cells from human glossal lymphangioma (GL-LECs) and GL-LECs were further primarily cultured and expanded. GL-LECs were of typical cobblestone appearance when they reached confluence. The weible-palade body was observed in the GL-LECs cytoplasm. Almost all GL-LECs were strongly positive for specific lymphatic markers FLT-4, LYVE-1 and prox-1 by immunocytochemistry. Furthermore, three-dimension tube-like capillaries of GL-LECs resembled the lymphatic system in vivo, and the GL-LECs spheroids sprouted radically out to form three-dimensional buds when embedded in the cultured BME. These results indicated that high purity GL-LECs were successfully isolated and expanded. They had the abilities of tube formation and differentiation in vitro, which provide a favorable cell model for further uncovering the pathogenesis of human lymphangiomas.

  19. Genome sequence of Oceanobacillus picturae strain S1, an halophilic bacterium first isolated in human gut.

    PubMed

    Lagier, Jean-Christophe; Khelaifia, Saber; Azhar, Esam Ibraheem; Croce, Olivier; Bibi, Fehmida; Jiman-Fatani, Asif Ahmad; Yasir, Muhammad; Helaby, Huda Ben; Robert, Catherine; Fournier, Pierre-Edouard; Raoult, Didier

    2015-01-01

    Oceanobacillus picturae is a strain of a moderately halophilic bacterium, first isolated from a mural painting. We demonstrate, for the first time, the culture of human Oceanobacillus picturae, strain S1(T), whose genome is described here, from a stool sample collected from a 25-year-old Saoudian healthy individual. We used a slightly modified standard culture medium adding 100 g/L of NaCl. We provide a short description of this strain including its MALDI-TOF spectrum, the main identification tool currently used in clinical microbiology. The 3,675,175 bp long genome exhibits a G + C content of 39.15 % and contains 3666 protein-coding and 157 RNA genes. The draft genome sequence of Oceanobacillus picturae has a similar size to the Oceanobacillus kimchii (respectively 3.67 Mb versus 3.83 Mb). The G + C content was higher compared with Oceanobacillus kimchii (respectively 39.15 % and 35.2 %). Oceanobacillus picturae shared almost identical number of genes (3823 genes versus 3879 genes), with a similar ratio of genes per Mb (1041 genes/Mb versus 1012 genes/Mb). The genome sequencing of Oceanobacillus picturae strain S1 isolated for the first time in a human, will be added to the 778 genome projects from the gastrointestinal tract listed by the international consortium Human Microbiome Project.

  20. Characterization of the 5-hydroxytryptamine receptor which mediates contraction of the human isolated basilar artery.

    PubMed

    Parsons, A A; Whalley, E T

    1989-01-01

    This paper reports part of a study which investigated the identity of the receptor involved in 5-hydroxytryptamine (5-HT) mediated contraction of the human basilar artery in vitro. 5-HT and a variety of 5-HT receptor agonists contracted human isolated basilar artery with a rank order of agonist potency: 5-carboxamidotryptamine greater than 5-HT greater than GR43175 much much greater than 2-methyl-5-HT. The maximum response produced by these agonists differed. The contractile responses to both 5-HT and GR43175 were resistant to antagonism by the 5-HT2 antagonist ketanserin and the 5-HT3 antagonist GR38032, indicating that neither 5-HT nor GR43175 activate 5-HT2 and 5-HT3 receptors in this tissue. In striking contrast, methiothepin (an antagonist at 5-HT1-like receptors) proved a potent antagonist of the contractile actions of both 5-HT and GR43175. Methiothepin did not antagonize the contractile response to the thromboxane-A2 mimetic U-46619. It is concluded that 5-HT and GR43175 contract the human isolated basilar artery by activating 5-HT1-like receptors. It is suggested that the antimigraine action of GR43175 may reflect its ability to constrict certain cranial arteries via 5-HT1-like receptor activation. PMID:2544283

  1. Isolation and characterization of cDNA clones for human erythrocyte. beta. -spectrin

    SciTech Connect

    Prchal, J.T.; Morley, B.J.; Yoon, S.H.; Coetzer, T.L.; Palek, J.; Conboy, J.G.; Kan, Y.W.

    1987-11-01

    Spectrin is an important structural component of the membrane skeleton that underlies and supports the erythrocyte plasma membrane. It is composed of nonidentical ..cap alpha.. (M/sub r/ 240,000) and ..beta.. (M/sub r/ 220,000) subunits, each of which contains multiple homologous 106-amino acid segments. The authors report here the isolation and characterization of a human erythroid-specific ..beta..-spectrin cDNA clone that encodes parts of the ..beta..-9 through ..beta..-12 repeat segments. This cDNA was used as a hybridization probe to assign the ..beta..-spectrin gene to human chromosome 14 and to begin molecular analysis of the gene and its mRNA transcripts. RNA transfer blot analysis showed that the reticulocyte ..beta..-spectrin mRNA is 7.8 kilobases in length. Southern blot analysis of genomic DNA revealed the presence of restriction fragment length polymorphisms (RFLPs) within the ..beta..-spectrin gene locus. The isolation of human spectrin cDNA probes and the identification of closely linked RFLPs will facilitate analysis of mutant spectrin genes causing congenital hemolytic anemias associated with quantitative and qualitative spectrin abnormalities.

  2. Comparison of virulence factors and biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections.

    PubMed

    Khoramian, Babak; Jabalameli, Fereshteh; Niasari-Naslaji, Amir; Taherikalani, Morovat; Emaneini, Mohammad

    2015-11-01

    The aim of this study was to find different prevalence of genes involved in the biofilm formation process and to assess the phenotypic and genotypic markers of biofilm formation among Staphylococcus aureus strains isolated from human and bovine infections. In this study, 215 S. aureus strains were collected from human and dairy cow's infections. The biofilm forming capacity of the strains was evaluated using a colorimetric microtiter plate assay. The genes encoding microbial surface components, recognizing adhesive matrix molecules (MSCRAMMs) (ebpS, eno, fib, fnbA, fnbB, cna and bap), and the intracellular adhesion (ica) genes (icaA, and icaD) were targeted by polymerase chain reaction (PCR)-based method. Approximately 70% of the isolates produced biofilm. Among these, 59.3% were producers of weakly adherent biofilms while 34.8% and 5.8% produced moderate and strong biofilms, respectively. The most prevalent gene was icaD found in 88.4% of the isolates, followed by icaA, fib and eno found in 87.9%, 75.8% and 75.3% of the isolates, respectively. The bap gene was not detected in any of the isolates. The prevalence of ebpS and fnbA genes among bovine isolates were significantly higher than those in human isolates, whilst the prevalence of cna gene was significantly higher in the human isolates. In this study, a high prevalence of biofilm production was found among S. aureus strains isolated from human and bovine infections. Most biofilm producing isolates were positive for MSCRAMM, icaA, and icaD genes.

  3. Featured Article: Isolation, characterization, and cultivation of human hepatocytes and non-parenchymal liver cells

    PubMed Central

    Pfeiffer, Elisa; Kegel, Victoria; Zeilinger, Katrin; Hengstler, Jan G; Nüssler, Andreas K; Seehofer, Daniel

    2015-01-01

    Primary human hepatocytes (PHH) are considered to be the gold standard for in vitro testing of xenobiotic metabolism and hepatotoxicity. However, PHH cultivation in 2D mono-cultures leads to dedifferentiation and a loss of function. It is well known that hepatic non-parenchymal cells (NPC), such as Kupffer cells (KC), liver endothelial cells (LEC), and hepatic stellate cells (HSC), play a central role in the maintenance of PHH functions. The aims of the present study were to establish a protocol for the simultaneous isolation of human PHH and NPC from the same tissue specimen and to test their suitability for in vitro co-culture. Human PHH and NPC were isolated from tissue obtained by partial liver resection by a two-step EDTA/collagenase perfusion technique. The obtained cell fractions were purified by Percoll density gradient centrifugation. KC, LEC, and HSC contained in the NPC fraction were separated using specific adherence properties and magnetic activated cell sorting (MACS®). Identified NPC revealed a yield of 1.9 × 106 KC, 2.7 × 105 LEC and 4.7 × 105 HSC per gram liver tissue, showing viabilities >90%. Characterization of these NPC showed that all populations went through an activation process, which influenced the cell fate. The activation of KC strongly depended on the tissue quality and donor anamnesis. KC became activated in culture in association with a loss of viability within 4–5 days. LEC lost specific features during culture, while HSC went through a transformation process into myofibroblasts. The testing of different culture conditions for HSC demonstrated that they can attenuate, but not prevent dedifferentiation in vitro. In conclusion, the method described allows the isolation and separation of PHH and NPC in high quality and quantity from the same donor. PMID:25394621

  4. Featured Article: Isolation, characterization, and cultivation of human hepatocytes and non-parenchymal liver cells.

    PubMed

    Pfeiffer, Elisa; Kegel, Victoria; Zeilinger, Katrin; Hengstler, Jan G; Nüssler, Andreas K; Seehofer, Daniel; Damm, Georg

    2015-05-01

    Primary human hepatocytes (PHH) are considered to be the gold standard for in vitro testing of xenobiotic metabolism and hepatotoxicity. However, PHH cultivation in 2D mono-cultures leads to dedifferentiation and a loss of function. It is well known that hepatic non-parenchymal cells (NPC), such as Kupffer cells (KC), liver endothelial cells (LEC), and hepatic stellate cells (HSC), play a central role in the maintenance of PHH functions. The aims of the present study were to establish a protocol for the simultaneous isolation of human PHH and NPC from the same tissue specimen and to test their suitability for in vitro co-culture. Human PHH and NPC were isolated from tissue obtained by partial liver resection by a two-step EDTA/collagenase perfusion technique. The obtained cell fractions were purified by Percoll density gradient centrifugation. KC, LEC, and HSC contained in the NPC fraction were separated using specific adherence properties and magnetic activated cell sorting (MACS®). Identified NPC revealed a yield of 1.9 × 10(6) KC, 2.7 × 10(5) LEC and 4.7 × 10(5) HSC per gram liver tissue, showing viabilities >90%. Characterization of these NPC showed that all populations went through an activation process, which influenced the cell fate. The activation of KC strongly depended on the tissue quality and donor anamnesis. KC became activated in culture in association with a loss of viability within 4-5 days. LEC lost specific features during culture, while HSC went through a transformation process into myofibroblasts. The testing of different culture conditions for HSC demonstrated that they can attenuate, but not prevent dedifferentiation in vitro. In conclusion, the method described allows the isolation and separation of PHH and NPC in high quality and quantity from the same donor.

  5. Clonal relationship between human and avian ciprofloxacin-resistant Escherichia coli isolates in North-Eastern Algeria.

    PubMed

    Agabou, A; Lezzar, N; Ouchenane, Z; Khemissi, S; Satta, D; Sotto, A; Lavigne, J-P; Pantel, A

    2016-02-01

    The objectives of this study were to determine rates, patterns, and mechanisms of antibiotic resistance, and to assess connections between chicken commensal, human commensal, and pathogenic ciprofloxacin-resistant Escherichia coli isolates. All E. coli isolates collected from chickens, their farmers, and patients in the Constantine region (North-east Algeria) were analyzed for bla and plasmid-mediated quinolone resistance (PMQR) gene contents, phylogroups, Rep-PCR profiles, and multilocus sequence types. A high prevalence of resistance to fluoroquinolones (51.4 % to ciprofloxacin) was recorded in avian isolates. Of these, 22.2 % carried the aac(6')-Ib-cr gene, whereas lower resistance levels to these antibiotics were recorded in chicken farmers' isolates. None of the commensal isolates harbored the qnr, qepA, or oqxAB genes. One human pathogenic isolate was ertapenem-resistant and harbored the bla OXA-48 gene, 84 showed an extended-spectrum β-lactamase phenotype, with bla CTX-M-15 gene prevalent in 87.2 % of them. Seventy isolates were resistant to fluoroquinolones, with aac(6')-Ib-cr present in 72.8 %, qnrB in 5.7 %, and qnrS in 10 %. Three Rep-PCR profiles were common to chicken commensal and human pathogenic isolates (phylogroups D and B1; ST21, ST48, and ST471 respectively); one was found in both chicken and chicken-farmer commensal strains (D; ST108), while another profile was identified in a chicken-farmer commensal strain and a human pathogenic one (B1; ST19). These findings suggest clonal and epidemiologic links between chicken and human ciprofloxacin-resistant E. coli isolates and the important role that poultry may play in the epidemiology of human E. coli infections in the Constantine region.

  6. Isolation, expression, and chromosomal localization of the human mitochondrial capsule selenoprotein gene (MCSP)

    SciTech Connect

    Aho, Hanne; Schwemmer, M.; Tessmann, D.; Murphy, D.

    1996-03-01

    The mitochondrial capsule selenoprotein (MCS) (HGMW-approved symbol MCSP) is one of three proteins that are important for the maintenance and stabilization of the crescent structure of the sperm mitochondria. We describe here the isolation of a cDNA, the exon-intron organization, the expression, and the chromosomal localization of the human MCS gene. Nucleotide sequence analysis of the human and mouse MCS cDNAs reveals that the 5{prime}- and 3{prime}-untranslated sequences are more conserved (71%) than the coding sequences (59%). The open reading frame encodes a 116-amino-acid protein and lacks the UGA codons, which have been reported to encode the selenocysteines in the N-terminal of the deduced mouse protein. The deduced human protein shows a low degree of amino acid sequence identity to the mouse protein. The deduced human protein shows a low degree of amino acid sequence identity to the mouse protein (39%). The most striking homology lies in the dicysteine motifs. Northern and Southern zooblot analyses reveal that the MCS gene in human, baboon, and bovine is more conserved than its counterparts in mouse and rat. The single intron in the human MCS gene is approximately 6 kb and interrupts the 5{prime}-untranslated region at a position equivalent to that in the mouse and rat genes. Northern blot and in situ hybridization experiments demonstrate that the expression of the human MCS gene is restricted to haploid spermatids. The human gene was assigned to q21 of chromosome 1. 30 refs., 9 figs.

  7. Isolation and characterization of the human G sub s. alpha. gene

    SciTech Connect

    Kozasa, Tohru; Itoh, Hiroshi; Tsukamoto, Toshihiko; Kaziro, Yoshito )

    1988-04-01

    The gene for G{sub s}{alpha} (the {alpha} subunit of the guanine nucleotide-binding protein G{sub s}) was isolated from human genomic libraries using rat G{sub s}{alpha} cDNA as a probe. Comparison of the nucleotide sequence of the human gene with that of the rat cDNA revealed that the human G{sub s}{alpha} gene spans {approx}20 kilobases and is composed of 13 exons and 12 introns. Genomic Southern blot analysis suggests that the human haploid genome contains a single G{sub s}{alpha} gene. Previous reports indicated the presence of multiple species of G{sub s}{alpha} cDNA. The structure of the human G{sub s}{alpha} gene suggests that four types of G{sub s}{alpha} mRNAs may be generated from a single G{sub s}{alpha} gene by alternate use of exon 3 and/or of two 3{prime} splice sites of intron 3, where an unusual splice junction sequence (TG) instead of the consensus (AG) is used. S1 nuclease mapping analysis of human G{sub s}{alpha} mRNA identified multiple transcriptional initiation sites. The promoter region of the human G{sub s}{alpha} gene has extremely high G+C content (85%). It contains 4 GC boxes, but no typical TATA or CAAT box sequence. In the 5{prime} flanking region, there are several blocks of sequences that are similar to the sequences of the 5{prime} flanking region of the human cKi-ras2 gene.

  8. The science of vacuum erectile device in penile rehabilitation after radical prostatectomy

    PubMed Central

    Lin, Haocheng

    2013-01-01

    Introduction Radical prostatectomy (RP) is a standard surgical treatment for clinically localized prostate cancer. Erectile dysfunction (ED) and penile shrinkage are common complications. Vacuum Erectile Device (VED) therapy uses negative pressure to distend the corporal sinusoids and to increase blood inflow into the penis. It is the second most commonly used method for penile rehabilitation after RP. However, the underlying mechanisms are still unclear. This paper is designed to review the scientific evidences of VED therapy after RP and discuss the possible mechanisms. Methods We reviewed published papers of post-prostatectomy penile rehabilitation using VED. We analyzed the scientific evidences of VED therapy and discussed the possible underlying mechanisms. Results There are existing clinical evidences for VED therapy to improve ED and preserve penile size. Emerging basic scientific evidence is available and further study is still needed to understand the mechanisms at the molecular level. Conclusions Current clinical evidences support the safety, tolerability, effectiveness and benefits of early VED therapy after RP. The available basic scientific evidences demonstrate that VED therapy for penile rehabilitation is achieved by increasing arterial inflow, anti-apoptotic, anti-fibrotic and anti-hypoxia mechanisms. PMID:26816725

  9. Factors predicting outcomes of penile rehabilitation with udenafil 50 mg following radical prostatectomy.

    PubMed

    Kim, T-H; Ha, Y-S; Choi, S H; Yoo, E S; Kim, B W; Yun, S-J; Kim, W-J; Kwon, Y S; Kwon, T G

    2016-01-01

    Udenafil is a selective phosphodiesterase type 5 inhibitor made available in recent years for the treatment of erectile dysfunction. Herein, we evaluated independent predictors of potency recovery in radical prostatectomy (RP) patients who underwent penile rehabilitation with udenafil 50 mg. One hundred and forty-three men who underwent RP were enrolled in a penile rehabilitation program using udenafil 50 mg every other day. The rate of regained potency in the study group was significantly higher compared with the recovery rate seen in patients who were not part of the penile rehabilitation program (41.3% vs 13.0%; P<0.001). On the multivariate Cox analyses, preoperative International Index of Erectile Function-5 scores (hazard ratio (HR), 1.049; P=0.040), alcohol consumption (HR, 2.043; P=0.020) and Gleason biopsy score (HR, 0.368; P=0.024) were independent preoperative predictors for potency recovery. Among post-RP variables, the use of robotic procedures (HR, 2.287; P=0.030) and pathologic stage (HR, 0.506; P=0.038) were significantly associated with potency recovery. This study identified predictive factors for the recovery of potency in patients undergoing penile rehabilitation with udenafil following RP. Our results could provide physicians with useful information for counseling RP patients and selecting optimal candidates for penile rehabilitation.

  10. Mechanisms predisposing penile fracture and long-term outcomes on erectile and voiding functions.

    PubMed

    Reis, Leonardo O; Cartapatti, Marcelo; Marmiroli, Rafael; de Oliveira Júnior, Eduardo Jeronimo; Saade, Ricardo Destro; Fregonesi, Adriano

    2014-01-01

    Purpose. To determine the mechanisms predisposing penile fracture as well as the rate of long-term penile deformity and erectile and voiding functions. Methods. All fractures were repaired on an emergency basis via subcoronal incision and absorbable suture with simultaneous repair of eventual urethral lesion. Patients' status before fracture and voiding and erectile functions at long term were assessed by periodic follow-up and phone call. Detailed history included cause, symptoms, and single-question self-report of erectile and voiding functions. Results. Among the 44 suspicious cases, 42 (95.4%) were confirmed, mean age was 34.5 years (range: 18-60), mean follow-up 59.3 months (range 9-155). Half presented the classical triad of audible crack, detumescence, and pain. Heterosexual intercourse was the most common cause (28 patients, 66.7%), followed by penile manipulation (6 patients, 14.3%), and homosexual intercourse (4 patients, 9.5%). "Woman on top" was the most common heterosexual position (n = 14, 50%), followed by "doggy style" (n = 8, 28.6%). Four patients (9.5%) maintained the cause unclear. Six (14.3%) patients had urethral injury and two (4.8%) had erectile dysfunction, treated by penile prosthesis and PDE-5i. No patient showed urethral fistula, voiding deterioration, penile nodule/curve or pain. Conclusions. "Woman on top" was the potentially riskiest sexual position (50%). Immediate surgical treatment warrants long-term very low morbidity. PMID:24822062

  11. Epidermoid cyst of the coronal sulcus mimicking penile cancer: a case report

    PubMed Central

    2014-01-01

    Introduction Epidermoid cysts represent common benign tumors occurring anywhere in the body but very rarely in the penis. Only a few cases of penile localization have been reported in the literature so far, most of them being congenital and/or idiopathic, usually presenting in children as slow-growing, solitary, well-delimited cystic lesions. Here, we describe the case of a patient with a penile epidermoid cyst presenting as an ulcerated lesion of the coronal sulcus, thus mimicking penile cancer. Case presentation A 36-year-old Caucasian man presented with a three-month history of a rapidly growing asymptomatic ulcerated lesion in the ventral portion of the penile coronal sulcus. At surgical exploration, the area under the ulcerated lesion had a well-demarcated cystic shape; following its wide excision, an intraoperative histological examination revealed an epidermoid cyst. No recurrence had occurred at nine years of follow-up. Conclusions Rare benign tumors of the penis, like the described epidermoid cyst, may mimic cancer. Nevertheless, penile ulcerated lesions should always be surgically explored as wide excision and intraoperative histological examination remain the only means of obtaining a precise disease definition and, consequently, administering the appropriate treatment. PMID:24906506

  12. A delayed foreskin-sparing approach to the management of penile fractures in uncircumcised Jamaican men

    PubMed Central

    Wong, Dean P.; Morrison, Belinda F.; Mayhew, Richard G.; Reid, Gareth A.; Aiken, William D.

    2015-01-01

    Introduction The traditional surgical approach to penile fracture is to perform a circumferential subcoronal degloving incision emergently to repair the injury. This approach necessitates circumcision to avoid foreskin complications. We present four men who had a delayed foreskin-sparing approach and discuss its advantages. Presentation of case Four of five uncircumcised patients who had suspected penile fractures secondary to coital injury, and without suspicion of concomitant urethral injury, had a delayed exploration, seven days after injury, utilizing an incision directly over the palpable haematoma, at the location of the tunical defect, thereby resulting in foreskin preservation. Two of 5 patients had repair under general anaesthesia, one under local anaesthesia and surgery was cancelled in another because upon reassessment at seven days he had normal erections and a normal penile examination. At follow up, all men had good functional and cosmetic outcomes. Discussion Uncircumcised patients with penile fractures, without suspicion of urethral injury, may undergo a delayed repair without prophylactic circumcision since there is minimal risk of foreskin complications. Delayed repair decreases the incidence of negative explorations by fostering a conservative approach in mimicking conditions such as superficial vein lacerations. It also enables the use of local anaesthesia in an elective ambulatory setting. Conclusion Delayed repair of penile fractures results in foreskin preservation, facilitates elective ambulatory care under local anaesthesia and decreases the incidence of negative surgical explorations. PMID:26551556

  13. Ultrasound-guided dorsal penile nerve block for ED paraphimosis reduction.

    PubMed

    Flores, Stefan; Herring, Andrew A

    2015-06-01

    Adequate anesthesia for emergency department management of painful penile conditions such as paraphimosis or priapism is often both technically challenging and inconsistent using traditional landmark-based techniques of the dorsal penile block (DPB). The pudendal nerves branch to form the paired dorsal nerves of the penis providing sensory innervation to the skin of both the dorsal and ventral aspects of the penis. "Blind" DPB techniques tend to rely on subtle tactile feedback from the needle and visual landmark approximation to identify the appropriate subpubic fascial compartment for injection. The landmark-based DPB is not standardized with options including “10 o'clock and 2 o'clock” infrapubic injections with or without ventral infiltration or a ring block. Given the lack of standardization and inherent technical imprecision with the landmark-based DPB, large volumes of local anesthetic (up to 50 mL) are sometimes required to achieve a clinically adequate block. In addition, inadvertent injection into the corpora cavernosa may occur. More recently, an ultrasound-guided approach has been developed. Using ultrasound, the dorsal penile nerves can be precisely targeted in the fascial compartment just deep to Buck fascia, potentially increasing block success rate and reducing the need for large local anesthetic volumes. Herein, we report the first adult case of an ultrasound-guided dorsal penile nerve block performed in the emergency department for the reduction of a paraphimosis and review the relevant penile anatomy and technical details of the procedure. PMID:25605058

  14. An unusual course after injection of industrial silicone for penile augmentation

    PubMed Central

    Shin, Yu Seob; You, Jae Hyung; Choi, Hwang; Zhang, Li Tao; Zhao, Chen; Choi, In Sung; Park, Jong Kwan

    2015-01-01

    A 48-year-old male patient had an injection of industrial silicone under the penile skin for augmentation by non-medical practitioners a week before. There was complete necrosis of the dorsal part of the penile skin and soft tissue. In a penile magnetic resonance image, big masses of silicone under the penile skin were found and a part of the silicone was partially exposed. Debridement of the necrotic tissue was done. As the right side of the tunica albuginea was thin-walled, a silicone-induced infection developed. Because of this, the wet dressing was done daily without closing the wound for the next 23 days. Finally, both scrotal skins were drawn and sutured to the dorsal glandular skin after the total penile skin was completely removed and sutured with T-style anastomosis. The ventral flap was anastomosed to the ventral glandular skin with the end-to-end technique with inverted V incision at 1 cm proximal from the sutured margin. Flaps survived completely without skin necrosis or dehiscence. PMID:25844103

  15. Effect of penile size on nocturnal erections: evaluation with NPTR testing with men having micropenis.

    PubMed

    Yaman, O; Soygür, T; Akand, M; Tokatli, Z

    2005-01-01

    There has been conflicting opinions in the literature regarding sexual function in hypogonadal men with micropenis. In this study we aimed to evaluate erectile function in hypogonadal men with micropenis by nocturnal penile tumescence and rigidity testing (NPTR) and compared the results with young potent normal penile sized men. A total of 15 men (ages 17-30 y) defined having a micropenis with a stretched penile length of less than 9.3 cm were constituted the study group. Mean stretched penile length was 6.8+/-1.6 cm (range 3.6-7.8 cm). Karyotype analysis showed 46XY in all cases. Control group included 22 potent and normal penile sized men (23-29 y). All subjects completed three sessions of consecutive nights using the RigiScan Plus device. Comparison of the results of NPTR of control group with study group revealed that number and duration of erectile episodes (P < 0.001), duration of tip rigidity > 60% (P < 0.01), TAU tip and TAU base (P = 0.001), and RAU base (P = 0.01) were found to be significantly lower in men with micropenis. In conclusion, our study showed that men with micropenis are associated with decreased nocturnal erectile activity.

  16. Semi-rigid penile prosthesis as a salvage management of idiopathic ischemic stuttering priapism

    PubMed Central

    Faddan, Amr A; Aksenov, Alexey V; Naumann, Carsten M; Jünemann, Klaus P; Osmonov, Daniar K

    2015-01-01

    Introduction Priapism is the persistent erection resulting from dysfunction of the mechanisms that regulate penile swelling, stiffness, and sagging. It is a full or partial erection that persists for a period more than 4 hours beyond sexual stimulation and/or orgasm or is unrelated to sexual stimulation. Ischemic priapism should be managed in a step-by-step fashion. Objective To demonstrate step-by-step management of stuttering refractory ischemic priapism. We report a case of stuttering refractory ischemic priapism. Moreover, we reviewed different approaches to priapism management in the literature. Case presentation A 53-year-old male presented with a painful erection of 29 hours’ duration, probably caused by consumption of alcohol. The penile blood gas showed a pH of 7.08, PCO2 of 75 mmHg and PO2 of 39 mmHg. Aspiration was followed by irrigation of an α-adrenergic, Winter and T-shunt operations were preformed, and finally a semi-rigid penile prosthesis was implanted to overcome the refractory stuttering ischemic priapism. Conclusion In case of stuttering refractory ischemic priapism, immediate implantation of a penile prosthesis is a simple and effective procedure that manages both the acute episode and the inevitable erectile dysfunction that would otherwise occur, while preserving penile length. PMID:26380229

  17. Characterization of Fosfomycin Resistant Extended-Spectrum β-Lactamase-Producing Escherichia coli Isolates from Human and Pig in Taiwan

    PubMed Central

    Tseng, Sung-Pin; Wang, Sheng-Fan; Kuo, Cheng-Yu; Huang, Jun-Wei; Hung, Wei-Chun; Ke, Guan-Ming; Lu, Po-Liang

    2015-01-01

    To investigate the efficacy of fosfomycin against extended-spectrum β-lactamases (ESBL) producing Escherichia coli in Taiwan and the resistance mechanisms and characterization of human and pig isolates, we analyzed 145 ESBL-producing isolates collected from two hospitals (n = 123) and five farms (n = 22) in Taiwan from February to May, 2013. Antimicrobial susceptibilities were determined. Clonal relatedness was determined by PFGE and multi-locus sequence typing. ESBLs, ampC, and fosfomycin resistant genes were detected by PCR, and their flanking regions were determined by PCR mapping and sequencing. The fosfomycin resistant mechanisms, including modification of the antibiotic target (MurA), functionless transporters (GlpT and UhpT) and their regulating genes such as uhpA, cyaA, and ptsI, and antibiotic inactivation by enzymes (FosA and FosC), were examined. The size and replicon type of plasmids carrying fosfomycin resistant genes were analyzed. Our results revealed the susceptibility rates of fosfomycin were 94% for human ESBL-producing E. coli isolates and 77% for pig isolates. The PFGE analysis revealed 79 pulsotypes. No pulsotype was found existing in both human and pig isolates. Three pulsotypes were distributed among isolates from two hospitals. ISEcp1 carrying blaCTX-M-group 9 was the predominant transposable elements of the ESBL genes. Among the thirteen fosfomycin resistant isolates, functionless transporters were identified in 9 isolates. Three isolates contained novel amino acid substitutions (Asn67Ile, Phe151Ser and Trp164Ser, Val146Ala and His159Tyr, respectively) in MurA (the target of fosfomycin). Four isolates had fosfomycin modified enzyme (fosA3) in their plasmids. The fosA3 gene was harboured in an IncN-type plasmid (101 kbp) in the three pig isolates and an IncB/O-type plasmid (113 kbp) in the human isolate. In conclusion, we identified that 6% and 23% of the ESBL-producing E. coli from human and pigs were resistant to fosfomycin, respectively

  18. Growth Factor Content in Human Sera Affects the Isolation of Mesangiogenic Progenitor Cells (MPCs) from Human Bone Marrow

    PubMed Central

    Montali, Marina; Barachini, Serena; Panvini, Francesca M.; Carnicelli, Vittoria; Fulceri, Franca; Petrini, Iacopo; Pacini, Simone

    2016-01-01

    Mesangiogenic Progenitor Cells (MPCs) are human bone marrow-derived multipotent cells, isolated in vitro under selective culture conditions and shown to retain both mesengenic and angiogenic potential. MPCs also co-isolated with multipotent stromal cells (MSCs) when bone marrow primary cultures were set up for clinical applications, using human serum (HS) in place of fetal bovine serum (FBS). MPC culture purity (over 95%) is strictly dependent on HS supplementation with significant batch-to-batch variability. In the present paper we screened different sources of commercially available pooled human AB type serum (PhABS) for their ability to promote MPC production under selective culture conditions. As the majority of “contaminating” cells in MPC cultures were represented by MSC-like cells, we hypothesized a role by differentiating agents present in the sera. Therefore, we tested a number of growth factors (hGF) and found that higher concentrations of FGF-2, EGF, PDGF-AB, and VEGF-A as well as lower concentration of IGF-1 give sub-optimal MPC recovery. Gene expression analysis of hGF receptors was also carried out both in MSCs and MPCs, suggesting that FGF-2, EGF, and PDGF-AB could act promoting MSC proliferation, while VEGF-A contribute to MSC-like cell contamination, triggering MPC differentiation. Here we demonstrated that managing hGF contents, together with applying specific receptors inhibitors (Erlotinib-HCl and Nintedanib), could significantly mitigate the batch-to-batch variability related to serum supplementation. These data represent a fundamental milestone in view of manufacturing MPC-based medicinal products. PMID:27800477

  19. Isolation of a new herpes virus from human CD4 sup + T cells

    SciTech Connect

    Frenkel, N.; Schirmer, E.C.; Wyatt, L.S.; Katsafanas, G.; Roffman, E.; Danovich, R.M. ); June, C.H. )

    1990-01-01

    A new human herpes virus has been isolated from CD4{sup +} T cells purified from peripheral blood mononuclear cells of a healthy individual (RK), following incubation of the cells under conditions promoting T-cell activation. The virus could not be recovered from nonactivated cells. Cultures of lymphocytes infected with the RK virus exhibited a cytopathic effect, and electron microscopic analyses revealed a characteristic herpes virus structure. RK virus DNA did not hybridize with large probes derived from herpes simplex virus, Epstein-Barr virus, varicella-zoster virus, and human cytomegalovirus. The genetic relatedness of the RK virus to the recently identified T-lymphotropic human herpes virus 6 (HHV-6) was investigated by restriction enzyme analyses using 21 different enzymes and by blot hydridization analyses using 11 probes derived from two strains of HHV-6 (Z29 and U1102). Whereas the two HHV-6 strains exhibited only limited restriction enzyme polymorphism, cleavage of the RK virus DNA yielded distinct patterns. Of the 11 HHV-6 DNA probes tested, only 6 cross-hybridized with DNA fragments derived from the RK virus. Taken together, the maximal homology amounted to 31 kilobases of the 75 kilobases tested. The authors conclude that the RK virus is distinct from previously characterized human herpesviruses. The authors propose to designate it as the prototype of a new herpes virus, the seventh human herpes virus identified to date.

  20. Human intestinal cells modulate conjugational transfer of multidrug resistance plasmids between clinical Escherichia coli isolates.

    PubMed

    Machado, Ana Manuel Dantas; Sommer, Morten O A

    2014-01-01

    Bacterial conjugation in the human gut microbiota is believed to play a major role in the dissemination of antibiotic resistance genes and virulence plasmids. However, the modulation of bacterial conjugation by the human host remains poorly understood and there is a need for controlled systems to study this process. We established an in vitro co-culture system to study the interaction between human intestinal cells and bacteria. We show that the conjugation efficiency of a plasmid encoding an extended spectrum beta-lactamase is reduced when clinical isolates of Escherichia coli are co-cultured with human intestinal cells. We show that filtered media from co-cultures contain a factor that reduces conjugation efficiency. Protease treatment of the filtered media eliminates this inhibition of conjugation. This data suggests that a peptide or protein based factor is secreted on the apical side of the intestinal cells exposed to bacteria leading to a two-fold reduction in conjugation efficiency. These results show that human gut epithelial cells can modulate bacterial conjugation and may have relevance to gene exchange in the gut.

  1. Comparative Genomic and Phylogenetic Analysis of the First Usutu Virus Isolate from a Human Patient Presenting with Neurological Symptoms

    PubMed Central

    Gaibani, Paolo; Cavrini, Francesca; Gould, Ernest A.; Rossini, Giada; Pierro, Anna; Landini, Maria Paola; Sambri, Vittorio

    2013-01-01

    Usutu virus (USUV) is a mosquito-borne flavivirus, belonging to the Japanese encephalitis antigenic complex, that circulates among mosquitoes and birds. We describe and analyze the complete genome sequence of the first USUV strain isolated from an immunocompromised patient with neuroinvasive disease. This USUV isolate showed an overall nucleotide identity of 99% and 96%, respectively, with the genomes of isolates from Europe and Africa. Comparison of the human USUV complete polyprotein sequence with bird-derived strains, showed two unique amino acid substitutions. In particular, one substitution (S595G) was situated in the DIII domain of the viral Envelope protein that is recognized by flavivirus neutralizing antibodies. An additional amino acid substitution (D3425E) was identified in the RNA-dependent RNA polymerase (RdRp) domain of the NS5 protein. This substitution is remarkable since E3425 is highly conserved among the other USUV isolates that were not associated with human infection. However, a similar substitution was observed in Japanese encephalitis and in West Nile viruses isolated from humans. Phylogenetic analysis of the human USUV strain revealed a close relationship with an Italian strain isolated in 2009. Analysis of synonymous nucleotide substitutions (SNSs) among the different USUV genomes showed a specific evolutionary divergence among different countries. In addition, 15 SNSs were identified as unique in the human isolate. We also identified four specific nucleotide substitutions in the 5′ and 3′ untranslated regions (UTRs) in the human isolate that were not present in the other USUV sequences. Our analyses provide the basis for further experimental studies aimed at defining the effective role of these mutations in the USUV genome, their potential role in the development of viral variants pathogenic for humans and their evolution and dispersal out of Africa. PMID:23741387

  2. [Metastatic penile lesions secondary to transitional cell carcinoma of the bladder: a rare cause of "malignant priapism"].

    PubMed

    Casoli, Eugenio; Di Fiore, Francesco; Longobardi, Santina; Intilla, Oliviero; Pone, Domenico

    2002-03-01

    Metastases to the penis from transitional cell carcinoma of the bladder are rare. In the literature about 300 cases of secondary penile malignancies were described; 35% out of these cases were from primary neoplasms of the bladder. The Authors describe a case of priapism secondary to penile metastases from a transitional cell carcinoma of the bladder.

  3. Simultaneous penile prosthesis and male sling/artificial urinary sphincter

    PubMed Central

    Lee, Dominic; Romero, Claudio; Alba, Frances; Westney, O Lenaine; Wang, Run

    2013-01-01

    Erectile dysfunction (ED) and stress urinary incontinence (SUI) from urethral sphincteric deficiency is not an uncommon problem. The commonest etiology is intervention for localized prostate cancer and/or radical cystoprostatectomy for muscle invasive bladder cancer. Despite advances in surgical technology with robotic assisted laparoscopic prostatectomy and nerve sparing techniques, the rates of ED and SUI remain relatively unchanged. They both impact greatly on quality of life domains and have been associated with poor performance outcomes. Both the artificial urinary sphincter and penile prosthesis are gold standard treatments with proven efficacy, satisfaction and durability for end-stage SUI and ED respectively. Simultaneous prosthesis implantation for concurrent conditions has been well described, mostly in small retrospective series. The uptake of combination surgery has been slow due in part to technical demands of the surgery and to an extent, a heightened anxiety over potential complications. This paper aims to discuss the technical aspect of concurrent surgery for both disease entity and the current published outcomes of the various surgical techniques with this approach. PMID:23202702

  4. Clinical predictive factors of sildenafil response: a penile hemodynamic study.

    PubMed

    Elhanbly, S M; Elkholy, A A-M; Alghobary, M; Abou Al-Ghar, M

    2015-03-01

    Phosphodiestrase-5 inhibitors are an important line of treatment for erectile dysfunction (ED). To detect the clinical and hemodynamic predictors of sildenafil response, we conducted this study on 124 Egyptian men with ED. All patients were evaluated by thorough history and clinical assessment with measurement of the abridged international index of erectile function-5 (IIEF-5) score. All patients were then subjected to intracavernosal injection (ICI) of trimix and pharmaco-penile duplex ultrasonography (PPDU). Patients were then classified into sildenafil responders and non-responders after six consecutive doses of 100 mg sildenafil. On doing the binary logistic stepwise regression analysis, only ED duration, IIEF-5 score, and response to ICI were the significant independent predictors of sildenafil response. These three parameters together correctly predicted the sildenafil response by 81.5% (p value <0.001). With the receiver operator characteristic curve analysis, the cut-off value of ED duration was 2.5 years and it was 14 for the IIEF-5 score. These findings indicate that ED duration, the IIEF-5 score and response to ICI are more significant predictors of sildenafil response than the more expensive and time-consuming PPDU testing. PMID:25644869

  5. Butea superba (Roxb.) improves penile erection in diabetic rats.

    PubMed

    Tocharus, C; Sooksaen, P; Shimbhu, D; Tocharus, J

    2012-05-01

    The objective of the present study was to investigate the effect of ethanolic extract of Butea superba (Roxb.) on erectile dysfunction in diabetic rats by the measurement of intracavernous pressure (ICP) and on cavernosal smooth muscle relaxation. Male Sprague-Dawley rats were induced to become diabetic by a single intravenous injection of Streptozotocin (55 mg kg(-1) body weight). The ethanolic extract at the concentration of 1, 10 and 100 mg kg(-1) BW was administered orally once a day to diabetic rats in each group for 4 weeks. Diabetic rats showed a significant decrease in both ICP and the relaxation of the cavernosal smooth muscle compared with the normal rats. The extract of B. superba significantly increased the ICP with the effective dose of 10 mg kg(-1) BW (61.00 ± 11.11 mmHg versus 39.61 ± 11.01 mmHg in the diabetic control group). Moreover, the B. superba-treated group also showed enhanced relaxation of the cavernosal smooth muscle with EC(50) of 1.17 mg ml(-1). These results suggest that the extract of B. superba enhanced penile erection in diabetic rats by increasing the ICP. This might be explained by the increased blood flow as a result of the relaxation of the cavernous smooth muscle.

  6. Multiple-Locus Variable-Number Tandem-Repeat Analysis Genotyping of Human Brucella Isolates from Turkey▿†

    PubMed Central

    Kılıç, Selçuk; Ivanov, Ivan N.; Durmaz, Rıza; Bayraktar, Mehmet Refik; Ayaşlıoğlu, Ergin; Uyanık, M. Hamidullah; Alışkan, Hikmet; Yaşar, Ekrem; Bayramoğlu, Gülçin; Arslantürk, Ahmet; Vergnaud, Gilles; Kantardjiev, Todor V.

    2011-01-01

    A multiple-locus variable-number tandem-repeat analysis (MLVA) was applied to investigate the epidemiological relationship and genetic diversity among 162 human Brucella isolates collected from all geographic regions of Turkey in an 8-year period (2001 to 2008). The isolates were genotyped by using an MLVA assay developed in Orsay, France (MLVA-16Orsay) including eight minisatellite (panel 1) and eight microsatellite (panel 2, subdivided into 2A and 2B) markers. Panels 1 and 2A distinguish 14 genotypes; two of these represented 85% of the strains. Panel 2B displayed a very high discriminatory power. Three loci from panel 2B had diversity index values higher than 0.74. MLVA-16Orsay yielded 105 genotypes; 73 were represented by a unique isolate, and 32 included two to eight isolates. The isolates from different patients within the same outbreak or from the same patient before first-line therapy and after relapse showed identical genotypes. A number of MLVA genotypes appeared to be partially restricted to some geographic areas and displayed no annual variation, possibly reflecting persistence of genotypes in certain areas for a time span of at least a decade. This study, representing the first molecular typing results of human Brucella isolates from Turkey, indicated that Turkish human Brucella melitensis isolates were most closely related to the neighboring countries' isolates included in the East Mediterranean group. PMID:21795514

  7. Multiple-locus variable-number tandem-repeat analysis genotyping of human Brucella isolates from Turkey.

    PubMed

    Kiliç, Selçuk; Ivanov, Ivan N; Durmaz, Riza; Bayraktar, Mehmet Refik; Ayaslioglu, Ergin; Uyanik, M Hamidullah; Aliskan, Hikmet; Yasar, Ekrem; Bayramoglu, Gülçin; Arslantürk, Ahmet; Vergnaud, Gilles; Kantardjiev, Todor V

    2011-09-01

    A multiple-locus variable-number tandem-repeat analysis (MLVA) was applied to investigate the epidemiological relationship and genetic diversity among 162 human Brucella isolates collected from all geographic regions of Turkey in an 8-year period (2001 to 2008). The isolates were genotyped by using an MLVA assay developed in Orsay, France (MLVA-16(Orsay)) including eight minisatellite (panel 1) and eight microsatellite (panel 2, subdivided into 2A and 2B) markers. Panels 1 and 2A distinguish 14 genotypes; two of these represented 85% of the strains. Panel 2B displayed a very high discriminatory power. Three loci from panel 2B had diversity index values higher than 0.74. MLVA-16(Orsay) yielded 105 genotypes; 73 were represented by a unique isolate, and 32 included two to eight isolates. The isolates from different patients within the same outbreak or from the same patient before first-line therapy and after relapse showed identical genotypes. A number of MLVA genotypes appeared to be partially restricted to some geographic areas and displayed no annual variation, possibly reflecting persistence of genotypes in certain areas for a time span of at least a decade. This study, representing the first molecular typing results of human Brucella isolates from Turkey, indicated that Turkish human Brucella melitensis isolates were most closely related to the neighboring countries' isolates included in the East Mediterranean group.

  8. Cryopreservation of isolated human hepatocytes for transplantation: State of the art.

    PubMed

    Terry, Claire; Dhawan, Anil; Mitry, Ragai R; Hughes, Robin D

    2006-10-01

    Hepatocytes isolated from unused donor livers are being used for transplantation in patients with acute liver failure and liver-based metabolic defects. As large numbers of hepatocytes can be prepared from a single liver and hepatocytes need to be available for emergency and repeated treatment of patients it is essential to be able to cryopreserve and store cells with good thawed cell function. This review considers the current status of cryopreservation of human hepatocytes discussing the different stages involved in the process. These include pre-treatment of cells, freezing solution, cryoprotectants and freezing and thawing protocols. There are detrimental effects of cryopreservation on hepatocyte structure and metabolic function, including cell attachment, which is important to the engraftment of transplanted cells in the liver. Cryopreserved human hepatocytes have been successfully used in clinical transplantation, with evidence of replacement of missing function. Further optimisation of hepatocyte cryopreservation protocols is important for their use in hepatocyte transplantation.

  9. Genetic diversity of Dirofilaria spp. isolated from subcutaneous and ocular lesions of human patients in Ukraine.

    PubMed

    Rossi, Alice; Peix, Álvaro; Pavlikovskaya, Tamara; Sagach, Olga; Nikolaenko, Svetlana; Chizh, Nina; Kartashev, Vladimir; Simón, Fernando; Siles-Lucas, Mar

    2015-02-01

    This short communication describes the phylogenetic analysis of 48 Dirofilaria worms isolated from human patients in Ukraine. 102 cases were both of subcutaneous (47; 46.1%) and ocular (54; 52.9%) locations. Worms from 44 patients (15 subcutaneous and 29 ocular) were subjected to DNA extraction and amplification of a specific fragment of the 12S rRNA subunit, and sequences were used for phylogenetic analysis. Results showed that 13.8% of the ocular cases analyzed at molecular level were caused by Dirofilaria immitis. Very few cases of ocular human dirofilariosis due to D. immitis have been described in the literature to date, majority of them attributed to Dirofilaria repens. Our results show that ocular dirofilariosis cannot be excluded in areas of low endemicity for D. repens were D. immitis is also present. PMID:25447827

  10. Cryopreservation of isolated human hepatocytes for transplantation: State of the art.

    PubMed

    Terry, Claire; Dhawan, Anil; Mitry, Ragai R; Hughes, Robin D

    2006-10-01

    Hepatocytes isolated from unused donor livers are being used for transplantation in patients with acute liver failure and liver-based metabolic defects. As large numbers of hepatocytes can be prepared from a single liver and hepatocytes need to be available for emergency and repeated treatment of patients it is essential to be able to cryopreserve and store cells with good thawed cell function. This review considers the current status of cryopreservation of human hepatocytes discussing the different stages involved in the process. These include pre-treatment of cells, freezing solution, cryoprotectants and freezing and thawing protocols. There are detrimental effects of cryopreservation on hepatocyte structure and metabolic function, including cell attachment, which is important to the engraftment of transplanted cells in the liver. Cryopreserved human hepatocytes have been successfully used in clinical transplantation, with evidence of replacement of missing function. Further optimisation of hepatocyte cryopreservation protocols is important for their use in hepatocyte transplantation. PMID:16793034

  11. Pancreatic Ductal Perfusion at Organ Procurement Enhances Islet Yield in Human Islet Isolation

    PubMed Central

    Shimoda, Masayuki; Kanak, Mazhar A.; Shahbazov, Rauf; Kunnathodi, Faisal; Lawrence, Michael C.; Naziruddin, Bashoo; Levy, Marlon F.

    2015-01-01

    Objective Pancreas preservation is a major factor influencing the results of islet cell transplantation. This study evaluated the effects of two different solutions for pancreatic ductal perfusion (PDP) at organ procurement. Methods Eighteen human pancreases were assigned to three groups: non-PDP (control), PDP with ET-Kyoto solution, and PDP with cold storage/purification stock solution. Pancreatic islets were isolated according to the modified Ricordi method. Results No significant differences in donor characteristics, including cold ischemia time, were observed between the three groups. All islet isolations in the PDP groups had >400,000 IEQ in total islet yield post-purification, a significant increase when compared with the control (P = 0.04 and <0.01). The islet quality assessments—including an in vivo diabetic nude mice assay and the response of high-mobility group box protein 1 to cytokine stimulation—also showed no significant differences. The proportion of TUNEL-positive cells showing apoptosis in islets in the PDP groups was significantly lower than in the control group (P < 0.05). Conclusion Both ET-Kyoto solution and cold storage/purification stock solution are suitable for PDP and consistently resulted in isolation success. Further studies with a larger number of pancreas donors should be done to compare the effects of the PDP solutions. PMID:25058879

  12. Taxonomic status and ecologic function of methanogenic bacteria isolated from the oral cavity of humans

    SciTech Connect

    Kemp, C.W.

    1985-01-01

    The detection of methane gas in samples of dental plaque and media inoculated with dental plaque was attributed to the presence of methane-producing bacteria in the plaque microbiota. The results of a taxonomic analysis of the 12 methanogenic isolates obtained from human dental plaque, (ABK1-ABK12), placed the organisms in the genus Methanobrevibacter. A DNA-DNA hybridization survey established three distinct genetic groups of oral methanogens based on percent homology values. The groups exhibited less than 32% homology between themselves and less than 17% homology with the three known members of the genus methanobrevibacter. The ecological role of the oral methanogens was established using mixed cultures of selected methanogenic isolates (ABK1, ABK4, ABK6, or ABK7) with oral heterotrophic bacteria. Binary cultures of either Streptococcus mutans, Streptococcus sanguis, Veillonella rodentium, Lactobacillus casei, or Peptostreptococcus anaerobius together with either methanogenic isolates ABK6 or ABK7 were grown to determine the effect of the methanogens on the distribution of carbon end products produced by the heterotrophs. Binary cultures of S. mutans and ABK7 exhibited a 27% decrease in lactic acid formation when compared to pure culture of S. mutans. The decrease in lactic acid production was attributed to the removal of formate by the methanogen, (ABK7), which caused an alteration in the distribution of carbon end products by S. mutans.

  13. Isolation and mapping of the human eukaryotic translation initiation factor 5 to chromosome 14

    SciTech Connect

    Romano, D.M.; Wasco, W.; Murell, J.

    1994-09-01

    Eukaryotic translation initiation factor 5 (eIF-5) is essential for the initiation of protein synthesis. eIF-5 catalyzes the hydrolysis of GTP on the 40S ribosomal initiation complex. Subsequent to GTP hydrolysis and the release of eIF-2-GDP, the 60S ribosomal subunit is joined to the 40S subunit to form an 80S initiation complex which can engage in peptide transfer. In an effort to isolate the major early-onset familial Alzheimer`s disease (FAD) gene on chromosome 14, we have isolated expressed sequences from this autosome in the form of exons `trapped` from chromosome 14-specific cosmids (library provided by L. Deaven, Los Alamos, NM). One cosmid yielded multiple exons displaying strong DNA and amino acid homology (>90%) with the rat eIF-5 gene. These exons were used to isolate full-length cDNAs from a human brain library. The eIF-5 message is approximately 3.6 kB in size and is ubiquitously expressed. The predicted amino acid sequence reveals multiple phosphorylation sites which may be involved in regulation of activity of eIF-5 and regions with homology to the GTPase superfamily, consistent with eIF-5`s role in GTP hydrolysis. Further studies are underway to determine whether the eIF-5 gene resides within the FAD minimal candidate region on chromosome 14q24.3.

  14. Virulence of Yersinia spp. strains isolated from non-human sources.

    PubMed

    Apfel, M I; Noleto, A L

    1991-01-01

    1. Strains of Yersinia enterocolitica, Y. intermedia, Y. frederiksenii and Y. kristensenii from non-human sources were examined for virulence factors. Four of these strains were positive for autoagglutination, three were calcium dependent at 37 degrees C, three produced lipase, five had the ability to bind Congo red, six had pyrazinamidase activity, and two had a 42 MDa plasmid. 2. One strain (Y. enterocolitica 0:5) was lethal for mice and had the ability to invade guinea pig eyes and HeLa cells. After inoculation of mice by the intravenous route, this strain was isolated from the cecum. The spleen, liver, kidneys and lymph nodes presented necrosis. After intragastric inoculation, the strain was isolated from all of the organs and tissues examined. 3. Three of the remaining strains invaded HeLa cells but none caused guinea pig conjunctivitis. 4. After intragastric inoculation, all the strains were isolated from the cecum but disappeared between days 3 and 6. After intravenous infection, three strains produced necrosis of the spleen and were more invasive, eliciting infection in various organs. The remaining strains caused hypertrophy and hyperplasia of Peyer's patches and/or lymph nodes. 5. These results indicate that Y. enterocolitica 0:5 can be considered as virulent as typical European Y. enterocolitica strains. The remaining strains perhaps induce a weak immune response independent of virulence factors.

  15. Identification of Clinical Staphylococcal Isolates from Humans by Internal Transcribed Spacer PCR

    PubMed Central

    Couto, Isabel; Pereira, Sandro; Miragaia, Maria; Sanches, Ilda Santos; de Lencastre, Hermínia

    2001-01-01

    The emergence of coagulase-negative staphylococci not only as human pathogens but also as reservoirs of antibiotic resistance determinants requires the deployment and development of methods for their rapid and reliable identification. Internal transcribed spacer-PCR (ITS-PCR) was used to identify a collection of 617 clinical staphylococcal isolates. The amplicons were resolved in high-resolution agarose gels and visually compared with the patterns obtained for the control strains of 29 staphylococcal species. Of the 617 isolates studied, 592 (95.95%) were identified by ITS-PCR and included 11 species: 302 isolates of Staphylococcus epidermidis, 157 of S. haemolyticus, 79 of S. aureus, 21 of S. hominis, 14 of S. saprophyticus, 8 of S. warneri, 6 of S. simulans, 2 of S. lugdunensis, and 1 each of S. caprae, S. carnosus, and S. cohnii. All species analyzed had unique ITS-PCR patterns, although some were very similar, namely, the group S. saprophyticus, S. cohnii, S. gallinarum, S. xylosus, S. lentus, S. equorum, and S. chromogenes, the pair S. schleiferi and S. vitulus, and the pair S. piscifermentans and S. carnosus. Four species, S. aureus, S. caprae, S. haemolyticus, and S. lugdunensis, showed polymorphisms on their ITS-PCR patterns. ITS-PCR proved to be a valuable alternative for the identification of staphylococci, offering, within the same response time and at lower cost, higher reliability than the currently available commercial systems. PMID:11526135

  16. Schistosoma mansoni Sambon, 1907: morphometric differences between adult worms from sympatric rodent and human isolates.

    PubMed

    Neves, R H; Pereira, M J; de Oliveira, R M; Gomes, D C; Machado-Silva, J R

    1998-01-01

    A computer software for image analysis (IMAGE PRO PLUS, MEDIA CYBERNETICS) was utilized in male and females adult worms, aiming the morphological characterization of Schistosoma mansoni samples isolated from a slyvatic rodent, Nectomys squamipes, and humans in Sumidouro, Rio de Janeiro, Brazil and recovered from Mus musculus C3H/He. The following characters for males's testicular lobes were analyzed: number, area, density, larger and smaller diameter, longer and shorter axis and perimeter and extension; for females: area, longer and shorter axis, larger and smaller diameter and perimeter of the eggs and spine; oral and ventral suckers area and distance between them in both sex were determined. By the analysis of variance (one way ANOVA) significant differences (p < 0.05) were observed in all studied characters, except for the density of testicular lobes. Significant differences (p < 0.05) were detected for all characters in the female worms. Data ratify that sympatric isolates present phenotypic differences and the adult female characters are useful for the proper identification of S. mansoni isolates.

  17. Characterisation of Aeromonas spp. isolated from frozen fish intended for human consumption in Mexico.

    PubMed

    Castro-Escarpulli, G; Figueras, M J; Aguilera-Arreola, G; Soler, L; Fernández-Rendón, E; Aparicio, G O; Guarro, J; Chacón, M R

    2003-07-15

    A total of 82 strains of presumptive Aeromonas spp. were identified biochemically and genetically (16S rDNA-RFLP). The strains were isolated from 250 samples of frozen fish (Tilapia, Oreochromis niloticus niloticus) purchased in local markets in Mexico City. In the present study, we detected the presence of several genes encoding for putative virulence factors and phenotypic activities that may play an important role in bacterial infection. In addition, we studied the antimicrobial patterns of those strains. Molecular identification demonstrated that the prevalent species in frozen fish were Aeromonas salmonicida (67.5%) and Aeromonas bestiarum (20.9%), accounting for 88.3% of the isolates, while the other strains belonged to the species Aeromonas veronii (5.2%), Aeromonas encheleia (3.9%) and Aeromonas hydrophila (2.6%). Detection by polymerase chain reaction (PCR) of genes encoding putative virulence factors common in Aeromonas, such as aerolysin/hemolysin, lipases including the glycerophospholipid-cholesterol acyltransferase (GCAT), serine protease and DNases, revealed that they were all common in these strains. Our results showed that first generation quinolones and second and third generation cephalosporins were the drugs with the best antimicrobial effect against Aeromonas spp. In Mexico, there have been few studies on Aeromonas and its putative virulence factors. The present work therefore highlights an important incidence of Aeromonas spp., with virulence potential and antimicrobial resistance, isolated from frozen fish intended for human consumption in Mexico City. PMID:12781953

  18. Molecular characterization of rabies virus isolates from Mexico: implications for transmission dynamics and human risk.

    PubMed

    De Mattos, C C; De Mattos, C A; Loza-Rubio, E; Aguilar-Setién, A; Orciari, L A; Smith, J S

    1999-10-01

    Twenty-eight samples from humans and domestic and wild animals collected in Mexico between 1990 and 1995 were characterized by using anti-nucleoprotein monoclonal antibodies and limited sequence analysis of the nucleoprotein gene. The variants of rabies viruses identified in these samples were compared with other isolates from Mexico and the rest of the Americas to establish epidemiologic links between cases and outbreaks and to increase the understanding of rabies epidemiology in the Western Hemisphere. Antigenic and genetic diversity was found in all samples from dogs and dog-related cases, suggesting a long-term endemic situation with multiple, independent cycles of virus transmission. Two isolates from bobcats were antigenically and genetically homologous to the rabies variant circulating in the Arizona gray fox population, indicating a wider distribution of this variant than previously reported. Rabies isolates from skunks were unrelated to any variant analyzed in this study and represent a previously unrecognized cycle of rabies transmission in skunks in Baja California Sur. Two antigenic and genetic variants co-circulating in southern and eastern Mexico were found in viruses obtained from cases epidemiologically related to vampire bats. These results serve