Leick, C M; Behrends, J M; Solaiman, S G; Broadway, P R; Min, B R; Mikel, W B; Williams, J B; Schilling, M W
Intact male Boer and Kiko goats (n=48) were harvested after 0, 4, 8, or 12 weeks on a 16% crude protein concentrate diet. Boneless goat carcass left sides were ground and formed into patties to evaluate cook loss, texture profile analysis, and descriptive sensory characteristics. Increasing feeding duration increased percent fat and decreased moisture in raw ground meat (P<0.05). Boer ground meat had more fat and less moisture than Kiko meat (P<0.05). Breed and feeding duration did not affect cook loss (P>0.05). Increased feeding duration increased aroma intensity and goaty, bloody, musty, and liver/organy aromas; salty, bitter, umami, grassy, goaty, fat, liver/organy, metallic, earthy, and chemical flavors; and juiciness and oiliness, while decreasing chewiness and crumbliness (P<0.05). Boer and Kiko patties had similar sensory properties after 0 and 4weeks on feed, but breeds were more distinguishable after 8 or 12 weeks on feed. PMID:22417728
Background Metabolic engineering projects often require integration of multiple genes in order to control the desired phenotype. However, this often requires iterative rounds of engineering because many current insertion approaches are limited by the size of the DNA that can be transferred onto the chromosome. Consequently, construction of highly engineered strains is very time-consuming. A lack of well-characterised insertion loci is also problematic. Results A series of knock-in/knock-out (KIKO) vectors was constructed for integration of large DNA sequences onto the E. coli chromosome at well-defined loci. The KIKO plasmids target three nonessential genes/operons as insertion sites: arsB (an arsenite transporter); lacZ (β-galactosidase); and rbsA-rbsR (a ribose metabolism operon). Two homologous ‘arms’ target each insertion locus; insertion is mediated by λ Red recombinase through these arms. Between the arms is a multiple cloning site for the introduction of exogenous sequences and an antibiotic resistance marker (either chloramphenicol or kanamycin) for selection of positive recombinants. The resistance marker can subsequently be removed by flippase-mediated recombination. The insertion cassette is flanked by hairpin loops to isolate it from the effects of external transcription at the integration locus. To characterize each target locus, a xylanase reporter gene (xynA) was integrated onto the chromosomes of E. coli strains W and K-12 using the KIKO vectors. Expression levels varied between loci, with the arsB locus consistently showing the highest level of expression. To demonstrate the simultaneous use of all three loci in one strain, xynA, green fluorescent protein (gfp) and a sucrose catabolic operon (cscAKB) were introduced into lacZ, arsB and rbsAR respectively, and shown to be functional. Conclusions The KIKO plasmids are a useful tool for efficient integration of large DNA fragments (including multiple genes and pathways) into E. coli. Chromosomal
Hewitt, Sylvia C; Li, Leping; Grimm, Sara A; Winuthayanon, Wipawee; Hamilton, Katherine J; Pockette, Brianna; Rubel, Cory A; Pedersen, Lars C; Fargo, David; Lanz, Rainer B; DeMayo, Francesco J; Schütz, Günther; Korach, Kenneth S
Estrogen receptor α (ERα) interacts with DNA directly or indirectly via other transcription factors, referred to as "tethering." Evidence for tethering is based on in vitro studies and a widely used "KIKO" mouse model containing mutations that prevent direct estrogen response element DNA- binding. KIKO mice are infertile, due in part to the inability of estradiol (E2) to induce uterine epithelial proliferation. To elucidate the molecular events that prevent KIKO uterine growth, regulation of the pro-proliferative E2 target gene Klf4 and of Klf15, a progesterone (P4) target gene that opposes the pro-proliferative activity of KLF4, was evaluated. Klf4 induction was impaired in KIKO uteri; however, Klf15 was induced by E2 rather than by P4. Whole uterine chromatin immunoprecipitation-sequencing revealed enrichment of KIKO ERα binding to hormone response elements (HREs) motifs. KIKO binding to HRE motifs was verified using reporter gene and DNA-binding assays. Because the KIKO ERα has HRE DNA-binding activity, we evaluated the "EAAE" ERα, which has more severe DNA-binding domain mutations, and demonstrated a lack of estrogen response element or HRE reporter gene induction or DNA-binding. The EAAE mouse has an ERα null-like phenotype, with impaired uterine growth and transcriptional activity. Our findings demonstrate that the KIKO mouse model, which has been used by numerous investigators, cannot be used to establish biological functions for ERα tethering, because KIKO ERα effectively stimulates transcription using HRE motifs. The EAAE-ERα DNA-binding domain mutant mouse demonstrates that ERα DNA-binding is crucial for biological and transcriptional processes in reproductive tissues and that ERα tethering may not contribute to estrogen responsiveness in vivo. PMID:24713037
Li, Leping; Grimm, Sara A.; Winuthayanon, Wipawee; Hamilton, Katherine J.; Pockette, Brianna; Rubel, Cory A.; Pedersen, Lars C.; Fargo, David; Lanz, Rainer B.; DeMayo, Francesco J.; Schütz, Günther; Korach, Kenneth S.
Estrogen receptor α (ERα) interacts with DNA directly or indirectly via other transcription factors, referred to as “tethering.” Evidence for tethering is based on in vitro studies and a widely used “KIKO” mouse model containing mutations that prevent direct estrogen response element DNA- binding. KIKO mice are infertile, due in part to the inability of estradiol (E2) to induce uterine epithelial proliferation. To elucidate the molecular events that prevent KIKO uterine growth, regulation of the pro-proliferative E2 target gene Klf4 and of Klf15, a progesterone (P4) target gene that opposes the pro-proliferative activity of KLF4, was evaluated. Klf4 induction was impaired in KIKO uteri; however, Klf15 was induced by E2 rather than by P4. Whole uterine chromatin immunoprecipitation-sequencing revealed enrichment of KIKO ERα binding to hormone response elements (HREs) motifs. KIKO binding to HRE motifs was verified using reporter gene and DNA-binding assays. Because the KIKO ERα has HRE DNA-binding activity, we evaluated the “EAAE” ERα, which has more severe DNA-binding domain mutations, and demonstrated a lack of estrogen response element or HRE reporter gene induction or DNA-binding. The EAAE mouse has an ERα null–like phenotype, with impaired uterine growth and transcriptional activity. Our findings demonstrate that the KIKO mouse model, which has been used by numerous investigators, cannot be used to establish biological functions for ERα tethering, because KIKO ERα effectively stimulates transcription using HRE motifs. The EAAE-ERα DNA-binding domain mutant mouse demonstrates that ERα DNA-binding is crucial for biological and transcriptional processes in reproductive tissues and that ERα tethering may not contribute to estrogen responsiveness in vivo. PMID:24713037
Hewitt, Sylvia C.; Li, Yin; Li, Leping; Korach, Kenneth S.
Estrogen enables uterine proliferation, which depends on synthesis of the IGF1 growth factor. This proliferation and IGF1 synthesis requires the estrogen receptor (ER), which binds directly to target DNA sequences (estrogen-responsive elements or EREs), or interacts with other transcription factors, such as AP1, to impact transcription. We observe neither uterine growth nor an increase in Igf1 transcript in a mouse with a DNA-binding mutated ERα (KIKO), indicating that both Igf1 regulation and uterine proliferation require the DNA binding function of the ER. We identified several potential EREs in the Igf1 gene, and chromatin immunoprecipitation analysis revealed ERα binding to these EREs in wild type but not KIKO chromatin. STAT5 is also reported to regulate Igf1; uterine Stat5a transcript is increased by estradiol (E2), but not in KIKO or αERKO uteri, indicating ERα- and ERE-dependent regulation. ERα binds to a potential Stat5a ERE. We hypothesize that E2 increases Stat5a transcript through ERE binding; that ERα, either alone or together with STAT5, then acts to increase Igf1 transcription; and that the resulting lack of IGF1 impairs KIKO uterine growth. Treatment with exogenous IGF1, alone or in combination with E2, induces proliferation in wild type but not KIKO uteri, indicating that IGF1 replacement does not rescue the KIKO proliferative response. Together, these observations suggest in contrast to previous in vitro studies of IGF-1 regulation involving AP1 motifs that direct ERα-DNA interaction is required to increase Igf1 transcription. Additionally, full ERα function is needed to mediate other cellular signals of the growth factor for uterine growth. PMID:19920132
The meat goat industry is growing rapidly in the U.S., particularly on small farms. Weight gain and carcass parameters were determined for traditional lambs (Suffolk, SX), hair sheep lambs (Katahdin, KA), and Boer x Kiko meat goats (GX) finished on a mixed pasture of orchardgrass (Dactylis glomerat...
...The following applicants filed AM or FM proposals to change the community of license: 1TV.COM, INC., Station KIKO, Facility ID 72477, BP-20100824ABA, From MIAMI, AZ, To SUPERIOR, AZ; AIRWAVES FOR JESUS, INC., Station NONE, Facility ID 176879, BMPED-20100831AAQ, From CRAIGSVILLE, WV, To WEBSTER SPRINGS, WV; ENTRAVISION HOLDINGS, LLC, Station KVVA-FM, Facility ID 1331, BPH-20100817ABA, From......
Spataro, B.; Alesini, D.; Chimenti, V.; Dolgashev, V.; Haase, A.; Tantawi, S.G.; Higashi, Y.; Marrelli, C.; Mostacci, A.; Parodi, R.; Yeremian, A.D.; /SLAC
Two 11.424 GHz single cell standing wave accelerating structures have been fabricated for high gradient RF breakdown studies. Both are brazed structures: one made from copper and the other from sintered molybdenum bulk. The tests results are presented and compared to those of similar devices constructed at SLAC (Stanford Linear Accelerator Center) and KEK (Ko Enerugi Kasokuki Kenkyu Kiko). The technological issues to build both sections are discussed.
...The following applicants filed AM or FM proposals to change the community of license: 1TV.COM, INC., Station KIKO, Facility ID 72477, BP-20100824ABA, From MIAMI, AZ, To APACHE JUNCTION, AZ; LA NUEVA CADENA RADIO LUZ, INC., Station KLIT, Facility ID 86722, BPED- 20110502AET, From DEL RIO, TX, To EAGLE PASS, TX; LOUT, JAMES M, Station NEW, Facility ID 170971, BMPH-20100301ABS, From PINELAND, TX,......
Min, Byeng R.; Solaiman, Sandra; Shange, Raymon
Eighteen Kiko-cross meat goats (n = 6) were used to collect gastrointestinal (GI) bacteria and methanogenic archaea for diversity measures when fed condensed tannin-containing pine bark (PB). Three dietary treatments were tested: control diet (0% PB and 30% wheat straw (WS); 0.17% condensed tannins (CT) dry matter (DM)); 15% PB and 15% WS (1.6% CT DM), and 30% PB and 0% WS (3.2% CT DM). A 16S rDNA bacterial tag-encoded FLX amplicon pyrosequencing technique was used to characterize and elucidate changes in GI bacteria and methanogenic archaea diversity among the diets. Proteobacteria was the most dominant phylum in goats with mean relative abundance values ranging from 39.7 (30% PB) to 46.5% (control) and 47.1% (15% PB). Other phyla individually accounted for fewer than 25% of the relative abundance observed. Predominant methanogens were Methanobrevibacter (75, 72, and 49%), Methanosphaera (3.3, 2.3, and 3.4%), and Methanobacteriaceae (1.2, 0.6, and 0.7%) population in control, 15, and 30% PB, respectively. Among methanogens, Methanobrevibacter was linearly decreased (P = 0.05) with increasing PB supplementation. These results indicate that feeding PB selectively altered bacteria and methanogenic archaeal populations in the GI tract of goats. PMID:24669219
Natochin, Iu V; Berger, V Ia
Marine molluscs exhibit relative constancy in intracellular potassium at the back ground of significant changes in intracellular sodium during acclimation to differen salinities. These changes, which were observed in cells of the adductor muscle and hepa topancreas, result mainly from active extrusion of sodium (possibly, of chloride as well) from the cell at low salinities and accumulation of these ions within the cell during the increase in salinity. Changes in intracellular concentration of sodium and chloride ions serve presumably as one of the main mechanisms of volume stabilization of cells, which together with the amino acid regulation alleviates the hydration of cells at different salinities. The capacity of cells to keep their potassium accounts for the maintenance of Ki/Ko ratio during changes in cellular volume induced by osmotic effects. These data are discussed in relation to two alternative hypotheses of the decreased and close to the present salinity of ocean at the initial stages of formation of the ionic composition of cells. PMID:473987
Farfan, L. M.; Cosio, M. A.
General characteristics of tropical weather systems were documented on a real-time basis. The geographical area of interest is the Baja California peninsula, located in northwestern Mexico. This study covers the warm season of 2007, from May through October, and includes observations derived from radar and satellite imagery as well as reports from a network of rain gauges. A set of graphical products were generated and they were available to the public through the internet. The analysis system has been in operation since the summer of 2005 and it is focused to document the development of tropical cyclones in eastern Pacific Ocean. During the season of 2007, this basin had a total of 11 tropical storms and four of them were within 800 km from the west coast of Mexico (Dalila, Ivo, Juliette and Kiko). Only one system made landfall in the area of interest: Hurricane Henriette which moved across Baja California, the Gulf of California and a portion of the state of Sonora. This presentation provides an overview of the graphical products along with lessons learned from the season studied, collaborations with local emergency managers and plans for the upcoming season of 2008.
Terrill, T H; Dykes, G S; Shaik, S A; Miller, J E; Kouakou, B; Kannan, G; Burke, J M; Mosjidis, J A
Gastrointestinal nematodes (GIN) parasitism is the greatest threat to economic sheep and goat production in the southern USA, and there is widespread prevalence of GIN resistance to broad-spectrum anthelmintics in this region. A natural alternative for controlling GIN in small ruminants is feeding hay of sericea lespedeza [SL, Lespedeza cuneata (Dum.-Cours., G. Don)], a perennial warm-season legume high in condensed tannins. To determine the level of SL needed to reduce GIN infection, a confinement study was completed with 32 Spanish/Boer/Kiko cross yearling bucks offered one of four diets with 75% hay and 25% concentrate (n=8, 2 pens/treatment, 4 goats/pen). The hay portion of each diet consisted of a combination of ground SL (0%, 25%, 50%, and 75% of the diet) and bermudagrass [BG, Cynodon dactylon (L.) Pers.; 75%, 50%, 25%, and 0% of the diet]. The bucks were allowed to acquire a natural GIN infection on pasture prior to moving to the pens. After a 3-week adjustment period in the pens, the goats were stratified by fecal egg count (FEC) and packed cell volume (PCV), randomly assigned to treatments and pens, and then fed the treatment diets for six weeks. During the experimental period, fecal and blood samples were collected from individual animals weekly to determine FEC and PCV, respectively. Adult worms from abomasum and small intestines were collected for counting and identification of species at slaughter. Goats fed SL hay at 25%, 50%, and 75% of the diet had 45.3% (P=0.2048), 66.3% (P=0.0134), and 74.5% (P=0.0077) lower FEC than control animals (75% BG hay) after 21 days. The 50% and 75% SL goats had 84.6% (P=0.0625) and 91.9% (P=0.0340) lower FEC than controls by day 42. The 75% SL-fed goats tended to have higher (P=0.0624) PCV and had fewer (P=0.035) abomasal worms than control animals, while PCV and adult worm numbers of the 50% and 25% SL goats were not different from controls. The optimum level of SL hay in the diet for reducing worm numbers of small
Moore, D A; Terrill, T H; Kouakou, B; Shaik, S A; Mosjidis, J A; Miller, J E; Vanguru, M; Kannan, G; Burke, J M
Goat production is increasing in the United States due to high ethnic demand, but infection with gastrointestinal nematode (GIN) parasites is a major constraint to the industry. Increasing GIN resistance to chemical anthelmintics worldwide has led to the development of alternative control strategies, including use of forages containing condensed tannins (CT). An experiment was designed using infected and dewormed male kids (Kiko x Spanish, 6 mo old, 18.9 +/- 3.25 kg) fed diets containing 25% concentrate and either 75% sericea lespedeza [SL; Lespedeza cuneata (Dum-Cours.) G. Don], a high CT forage (87 to 181 g of CT/kg), or 75% bermudagrass [BG; Cynodon dactylon (L.) Pers.] hay (n = 10/treatment). The kids were weighed every 14 d, and fecal and blood samples were taken weekly for fecal egg counts and packed cell volume determination, respectively. Fecal cultures were processed every 14 d to determine CT effect on larval development. At slaughter, adult GIN were collected from the abomasum and small intestines for counting and speciation. Blood samples were also analyzed for plasma urea-N, and ruminal VFA and pH were determined. The infected SL-fed kids had consistently lower (P < 0.05) fecal egg counts than the infected BG goats throughout the trial and greater (P < 0.05) packed cell volume beginning by d 77. Average daily gain was greater (P < 0.001) in kids fed SL- than BG-based diets, regardless of infection status (104.3 +/- 5.0 and 75.5 +/- 4.8 g/d, respectively). Total VFA and acetate concentrations were greater (P < 0.001) in the BG- than in SL-fed goats, whereas propionate levels were unaffected by diet. Acetate:propionate ratio (P = 0.01) and plasma urea-N (P = 0.03) levels were greater in BG-fed goats, whereas rumen pH was greater (P < 0.001) in the SL-fed goats. Feeding SL hay can reduce GIN infection levels and increase performance of goats compared with BG hay. PMID:18469053
Kommuru, D S; Barker, T; Desai, S; Burke, J M; Ramsay, A; Mueller-Harvey, I; Miller, J E; Mosjidis, J A; Kamisetti, N; Terrill, T H
Infection with Eimeria spp. (coccidia) can be devastating in goats, particularly for young, recently-weaned kids, resulting in diarrhea, dehydration, and even death. Feeding dried sericea lespedeza [SL; Lespedeza cuneata (Dum.-Cours.) G. Don.] to young goats has been reported to reduce the effects of internal parasites, including gastrointestinal nematodes (GIN) but there have been no reports of the effects of feeding this forage on Eimeria spp. in goats. Two confinement feeding experiments were completed on recently-weaned intact bucks (24 Kiko-cross, Exp. 1; 20 Spanish, Exp. 2) to determine effects of SL pellets on an established infection of GIN and coccidia. The bucks were assigned to 1 of 2 (Exp. 1) or 3 (Exp. 2) treatment groups based upon the number of Eimeria spp. oocysts per gram (OPG) of feces. In Exp. 1, the kids were fed 1 of 2 pelleted rations ad libitum; 90% SL leaf meal+10% of a liquid molasses/lignin binder mix and a commercial pellet with 12% crude protein (CP) and 24% acid detergent fiber (n=12/treatment group, 2 animals/pen). For Exp. 2, treatment groups were fed (1) 90% SL leaf meal pellets from leaves stored 3 years (n=7), (2) 90% SL pellets from leaf meal stored less than 6 months, (n=7), and the commercial pellets (n=6) ad libitum. For both trials, fecal and blood samples were taken from individual animals every 7 days for 28 days to determine OPG and GIN eggs per gram (EPG) and packed cell volume (PCV), respectively. In Exp. 2, feces were scored for consistency (1=solid pellets, 5=slurry) as an indicator of coccidiosis. In Exp. 1, EPG (P<0.001) and OPG (P<0.01) were reduced by 78.7% and 96.9%, respectively, 7 days after initiation of feeding in goats on the SL pellet diet compared with animals fed the control pellets. The OPG and EPG remained lower in treatment than control animals until the end of the trial. In Exp. 2, goats fed new and old SL leaf meal pellets had 66.2% and 79.2% lower (P<0.05) EPG and 92.2% and 91.2% lower (P<0.05) OPG