Trends of Infectious Disease Markers Among Healthy Blood Donors In Northern India: A Ten Year Study.

OBJECTIVE: Transfusion transmitted infection (TTI) is a major hazard for blood transfusion. The present retrospective chart review was undertaken to study the demographic profile and TTI trends among blood donors to see impact of interventions on blood safety. METHODS: Data of donors and TTI screening results from 2010 to 2019 were analyzed. Degree of significance was determined by Chi square test. RESULTS: Out of 1,68,570 donors, 33,227 (19.7%) were voluntary and 1,35,343 (80.3%) were replacement with 2.8% females and 54% belonging to the age group 18-29 years. Voluntary donation increased by only 3% in ten years and total reactivity rate was 1.6%. The reactive rate for all infections was 0.8% in volunteer donors and 1.95% in replacement donors (p-value < 0.001). The prevalence of HBsAg, HCV, HIV and syphilis showed a significant decline from 2010 to 2014. Of the donors who were reactive for HBV, 8.7% were missed by ELISA but detected by NAT. Donor reactivity for malaria remained the same in this period. CONCLUSION: Newer strategies and effort to increase voluntary donation helping the general public adopt a healthy lifestyle is urgently needed in India. Higher prevalence of TTI among replacement donors is substantiated by this study. Role of counseling of donors cannot be overemphasized. Utility of malaria screening for blood donors needs to be reexamined by evaluating evidences from other blood banks. A rational policy approach, based on a careful assessment of epidemiological data, cost effectiveness analysis, and opinion of stakeholders is necessary for universal adoption of NAT.

-174G/C polymorphism in the interleukin-6 promoter is differently associated with prostate cancer incidence depending on race.

Interleukin-6 (IL-6), a pro-inflammatory cytokine, is involved in prostate cancer progression, including androgen independence. Serum IL-6 levels also correlate with prostate tumor burden, prostate-specific antigen levels and metastasis. Since circulating cytokine levels vary considerably inter-individually, such variation could be linked to genetic factors, including genetic polymorphism. The -174G>C/rs1800795 polymorphism in the IL-6 promoter is functionally relevant in terms of transcriptional regulation and disease association. We investigated a possible association of the -174G/C polymorphism with prostate cancer. Since significant racial disparities exist in prostate cancer incidence, we also investigated this association between the -174G/C polymorphism and prostate cancer in Caucasians and African-Americans, separately. Direct sequencing of the PCR amplicon from genomic DNA was used for genotyping rs1800795 in all subjects [age-matched controls (N = 140) and prostate cancer patients (N = 164)]. Sample size and power was calculated using the PGA software. We found the GG genotype to be associated with increased risk of prostate cancer in Caucasian subjects, whereas the CC genotype was associated with increased risk in the African-American sample set. Such a dimorphic genotypic association with cancer and race is unique and suggests a complex gene-gene and gene-environment interaction.

Indigenous chicken production system in different agro-ecology of Indian Himalayan Region: implication on food and economic security.

The indigenous chicken production system (ICPS) has several use values and ecosystem services. In the last few years, ICPS has been recognized for its possible contribution to household food security, income generation, wildlife protection, and bettering the women's lives. This study aimed to collect, for the first time, comprehensive information about ICPS in three different agro-ecologies (tropical, sub-tropical, and sub-temperate) of the Indian Himalayan Region (IHR) and its role in food and economic security of traditional communities. In this study region, ICPS is semi-extensive, providing homegrown feed and temporary night shelter. In sub-temperate agro-ecology, females owned non-significant (p = 0.170) more indigenous chicken flocks than males. Households in sub-temperate agro-ecologies had significantly (p ≤ 0.001) larger flock sizes and tropical livestock units (chicken-TLU). However, the livestock diversity index (LDI) was significantly higher (p ≤ 0.001) in tropical and subtropical agro-ecology. The households in the sub-temperate region highly (p ≤ 0.001) valued indigenous chicken because of its survivability and adaptability. In absolute numbers significant (p ≤ 0.001) higher numbers of adult birds died in past 1 year in sub-temperate agro-ecology. The mortality rate of adult birds in sub-temperate agro-ecology was 9%, and it was 14 and 15% in tropical and sub-tropical agro-ecologies, respectively. In sub-temperate agro-ecology, larger flock size translated into significantly higher (p ≤ 0.001) egg production and subsequently a significant (p ≤ 0.001) higher egg consumption per household per month. In sub-temperate agro-ecology, households' dietary diversity score was significantly (p ≤ 0.001) higher. Similarly, the average annual income from ICPS was significantly higher (p ≤ 0.001) in sub-temperate agro-ecology and accounted for 18% of household income. ICPS' marketing chain was relatively short in the sub-temperate region. In all agro-ecologies, indigenous chicken and egg demand was significantly higher (p ≤ 0.001) in the winter. ICPS litter is used as farmyard manure, enhancing ecological resilience. In all agro-ecologies, the three most frequently cited obstacles to extending the indigenous chicken production system are illnesses, predators, and a lack of chicks availability. ICPS contributes to food and nutritional security, economic stability, and ecological resilience in this hilly and fragile ecosystem. Even though the system is self-sustaining, management and health interventions can increase production and productivity.

Linseed oil in boar's diet during high temperature humidity index (THI) period improves sperm quality characteristics, antioxidant status and fatty acid composition of sperm under hot humid sub-tropical climate.

Environmental heat stress in sub-tropical climates negatively impacts boar semen production and its quality. The present study aimed to examine the heat stress alleviating effects of dietary linseed oil on semen quality and antioxidant status of boar, in the summer and winter seasons in sub-tropical climate. Six Hampshire crossbreed boars were fed with 90 mL linseed oil (treatment) whereas six boars of the same breed were fed 90 mL vegetable oil (control) for sixteen weeks during both season. Sperm quality was assessed for motility, viability, abnormality, acrosomal integrity, and Hypo-osmotic swelling test (HOST). Sperm velocity attributes were assessed by computer-assisted semen analysis (CASA). Antioxidants (glutathione peroxidase; GPx, catalase; CAT, total antioxidant capacity; TAC and nitric oxide; NO) and lipid peroxidation (malondialdehyde; MDA) were measured in seminal plasma and serum. Gas chromatography-mass spectrometry (GC-MS) was used for the estimation of fatty acid composition of seminal plasma and spermatozoa. Feeding linseed oil to the boars significantly (p < 0.05) improved sperm quality at the fresh stage and after 72 h of liquid storage in both season. There was a significant (p < 0.01) effect of treatment and season on semen quality parameters. Significant boar (p < 0.05) effect was recorded on reaction time, semen volume, sperm abnormality, acrosomal integrity and HOST reactive sperm. There was a significant (p < 0.01) effect of treatment and season on the velocity attributes viz. VAP, VSL, VCL, ALH, BCF and STR%. Linseed oil supplementation significantly (p < 0.01) enhanced antioxidant and lowered MDA levels in serum as well as seminal plasma. The concentration of alpha-linolenic (ALA), arachidonic and docosahexaenoic (DHA) fatty acids were significantly (p < 0.01) increased in seminal plasma and sperm after linseed oil supplementation. In conclusion, linseed oil supplementation to boar during high THI months improved the semen quality parameters viz. semen volume, sperm concentration, and progressive motile sperm, along with enhanced antioxidant capacity.

Applied anatomy and clinical significance of the maxillofacial and mandibular regions of the barking deer (Muntiacus muntjak) and sambar deer (Rusa unicolor).

BACKGROUND: There is no previously reported information on the applied anatomy and clinical significance of the maxillofacial and mandibular regions of the barking deer and sambar deer. MATERIALS AND METHODS: Therefore, the present study was designed to provide some important clinical landmarks related to tracking of the infraorbital, mental and mandibular nerves with its clinical implications in regional anaesthesia in both the species. RESULTS: In the present study, the distance between the most lateral bulging of the facial tuberosity to the infraorbital foramen and from the latter to the root of the alveolar tooth directly ventral to it was found to be 2.65 ± 0.01 cm and 0.90 ± ± 0.02 cm in males; 2.75 ± 0.01 cm, 1.11 ± 0.01 cm in females of barking deer and 4.57 ± 0.01 cm and 1.83 ± 0.02 cm in males; 4.52 ± 0.02 cm and 1.76 ± 0.02 cm in females of sambar deer. The infraorbital foramen was small, elliptical and was located at the level of first superior premolar teeth in barking deer and sambar deer. The facial tuberosity was located above the third superior premolar teeth in the barking deer but was located at the level of the first superior molar teeth in sambar deer. The distance between the lateral alveolar root of the third inferior incisor tooth to the mental foramen was 2.84 ± 0.01 cm in males, 2.78 ± 0.01 cm in females of barking deer and 3.04 ± 0.02 cm in males, 2.96 ± 0.01 cm in females of sambar deer which is an important landmark for achieving the location of the mental foramen nerve for the regional nerve block in both the species. The mandible of both the species showed oval-shaped mental foramen with unossified mandibular symphysis. CONCLUSIONS: The present study revealed that most of the parameters showed a statistically significant difference between the sexes in barking deer and sambar deer; however, from the practical point of view, these differences were meager. The results were discussed with regard to their clinical applications in various regional anaesthesia performed in maxillofacial and mandibular regions of both the species.

Small intestinal mucosal cells in piglets fed with probiotic and zinc: a qualitative and quantitative microanatomical study.

BACKGROUND: Probiotics and zinc are commonly used and beneficial in pig production. This work aimed to assess the effects of probiotic and zinc on the mucosal cells of the small intestine in respect to digestive capacity and immunity in pre- and post-weaned piglets. MATERIALS AND METHODS: Eighteen Large White Yorkshire piglets were divided equally into control and treatment groups. The piglets were maintained in standard management conditions and were weaned at 28 days of age. The treatment group of piglets fed a mixture of probiotics orally at 1.25 × 109 CFU/day and zinc at 2000 ppm/day from birth to 10 days of age. At three different age-groups viz. day 20 (pre-weaning) and, day 30 and day 60 (post-weaning), the animals were sacrificed. For histomorphology, the tissue samples were processed and stained with Mayer's haematoxylin and eosin for routine study, combined periodic acid-Schiff-Alcian blue for mucopolysaccharides and Masson-Hamperl argentaffin technique for argentaffin cells. The stained slides were observed under the microscope. The samples were processed as per the standard procedure for scanning and transmission electron microscopy. The statistical analysis of the data using the appropriate statistical tests was also conducted. RESULTS: The mucosal epithelium of villi and crypts were lined by enterocytes, goblet cells, argentaffin cells, microfold (M-cell) cells, tuft cells and intraepithelial lymphocytes. The multipotent stem cells were located at the crypt base. The length of the enterocyte microvilli was significantly longer (p < 0.05) in the treatment group of piglets. The number of different types of goblet cells and argentaffin cells was more in treated piglets irrespective of segments of intestine and age. The intraepithelial lymphocytes were located in apical, nuclear and basal positions in the lining epithelium of both villus tip and base with their significant increase in the treatment group of piglets. The transmission electron microscopy revealed the frequent occurrence of tuft cells in the lining mucosa of the small intestine in treated piglets. CONCLUSIONS: Dietary supplementation of probiotic and zinc induced the number of different mucosal cells of villi and crypts in the small intestine that might suggest the greater absorptive capacity of nutrients and effective immunity in critical pre and post-weaned piglets.

Androgens regulate the immune/inflammatory response and cell survival pathways in rat ventral prostate epithelial cells.

A major hurdle in understanding the role of androgens is the heterogeneity of androgen receptor (AR) expression in the prostate. Because the majority of prostate cancer arises from the AR-positive secretory luminal epithelial cells, identifying the androgen-mediated pathways in the prostate epithelium is of great significance to understanding their role in prostate pathogenesis. To meet this objective, the current study was designed to identify immediate-early genes expressed in response to the synthetic androgen R1881 in cultured rat ventral prostate epithelial cells. Rat ventral prostate epithelial cells, purified from 20-d-old rats, were cultured, and the presence of AR and the response to androgen were established. The cells were then treated with R1881 for 2 and 12 h to capture immediate-early genes in an Affymetrix-based gene chip platform. A total of 66 nonredundant genes were identified that were responsive to R1881. The functional androgen response elements were identified in the proximal promoter to determine possible molecular mechanism. Cluster analysis identified five distinct signatures of R1881-induced genes. Pathway analysis suggested that R1881 primarily influences cell proliferation/differentiation and inflammatory/immune response pathways. Androgens appear to regulate cell renewal by regulating differentiation, cell proliferation, and apoptosis. Two mutually exclusive inflammatory response pathways were observed. The interferon pathway was up-regulated, and the ILs were down-regulated. The data identified novel androgen-regulated genes (e.g. Id1, Id3, IL-6, IGF-binding protein-2 and -3, and JunB). The loss of androgen regulation of these genes can have important consequences for cellular transformation and transition to androgen-independent growth and survival.

Vertebral scale system to measure heart size in thoracic radiographs of Indian Spitz, Labrador retriever and Mongrel dogs.

AIM: To establish reference values of vertebral heart score (VHS) in Indian Spitz, Labrador retriever, and Mongrel dogs; to assess applicability of VHS in these three dog breeds; to determine if breed, recumbency side, gender, body weight, and thoracic depth (TD) to thoracic width (TW) ratio has an influence on the VHS measurement in these dog breeds. MATERIALS AND METHODS: A total of 60, client owned, clinically healthy Indian Spitz (n=20, mean age = 4.25±2.15 years, body weight = 11.87±2.7 kg), Labrador retriever (n=20, mean age = 4.75±1.91 years, body weight = 27.31±5.43 kg), and Mongrel dogs (n=20, mean age = 4.25±1.52 years, body weight = 16.25±3.99 kg), having no radiological and clinical signs of cardiovascular or pulmonary disease were included in the study. All dogs were restrained manually and left lateral (LL) and right lateral (RL) radiographic views were obtained. The size of heart in lateral radiographs was calculated using VHS method. Besides, the TD, TW and TD: TW were calculated to determine the type of thoracic conformation in the dog breeds. In addition to this, the effect of breed, side of recumbency, gender, body weight, and TD to TW ratio on the calculation of VHS was determined. RESULTS: VHS was calculated in all the animals of the breeds. VHS in Spitz and Labrador retriever was significantly (p<0.0001, p<0.0001, respectively) >9.7±0.5 v. RL and LL VHS in Mongrel dog was significantly (p<0.037) >9.7±0.5 v. Significant (p<0.05) differences in the VHS were observed among Spitz, Labrador retriever and Mongrel dogs, being higher for Labrador retriever followed by Spitz and Mongrel dogs. VHS in RL recumbency was significantly (p<0.001) greater than VHS in LL recumbency in all three breeds. LL and RL VHS correlated significantly with each other in Spitz (r=0.58; p=0.02), Labrador retriever (r=0.87; p<0.0001), and Mongrel dogs (r=0.93; p<0.0001). Significant (p<0.05) differences in the TD and TW were observed among Spitz, Labrador retriever, and Mongrel dogs. Non-significant effect of gender, body weight, and TD to TW ratio on the VHS measurement was observed in each dog breed. CONCLUSION: Breed-specific VHS reference ranges should be used for the objective measurement of heart size in dogs. Furthermore, the radiographic view should also be taken into consideration to avoid any erroneous interpretation of cardiac enlargement in dogs.

Control of Trachoma from Achham District, Nepal: A Cross-Sectional Study from the Nepal National Trachoma Program.

BACKGROUND: The WHO seeks to control trachoma as a public health problem in endemic areas. Achham District in western Nepal was found to have TF (trachoma follicular) above 20% in a 2006 government survey, triggering 3 annual mass drug administrations finishing in 2010. Here we assess the level of control that has been achieved using surveillance for clinical disease, ocular chlamydia trachomatis infection, and serology for antibodies against chlamydia trachomatis protein antigens. METHODS: We conducted a cross-sectional survey of children aged 1-9 years in communities in Achham District in early 2014 including clinical examination validated with photographs, conjunctival samples for Chlamydia trachomatis (Amplicor PCR), and serological testing for antibodies against chlamydia trachomatis protein antigens pgp3 and CT694 using the Luminex platform. FINDINGS: In 24 randomly selected communities, the prevalence of trachoma (TF and/or TI) in 1-9 year olds was 3/1124 (0.3%, 95% CI 0.1 to 0.8%), and the prevalence of ocular chlamydia trachomatis infection was 0/1124 (0%, 95% CI 0 to 0.3%). In 18 communities selected because they had the highest prevalence of trachoma in a previous survey, the prevalence of TF and/or TI was 7/716 (1.0%, 95% CI 0.4 to 2.0%) and the prevalence of ocular chlamydia trachomatis infection was 0/716 (0%, 95% CI 0 to 0.5%). In 3 communities selected for serological testing, the prevalence of trachoma was 0/68 (0%, 95% CI 0 to 5.3%), the prevalence of ocular chlamydia trachomatis infection was 0/68 (0%, 95% CI 0 to 0.5%), the prevalence of antibodies against chlamydia trachomatis protein antigen pgp3 was 1/68 (1.5%, 95% CI 0.04% to 7.9%), and the prevalence of antibodies against chlamydia trachomatis protein antigen CT694 was 0/68 (0%, 95% CI 0 to 5.3%). CONCLUSION/SIGNIFICANCE: This previously highly endemic district in Nepal has little evidence of recent clinical disease, chlamydia trachomatis infection, or serological evidence of trachoma, suggesting that epidemiological control has been achieved.

Suppression of c-Fos gene transcription with malignant transformation of human bronchial epithelial cells.

The Activator Protein-1 (AP-1) complex is a dimeric transcription factor composed of fos and jun proteins that regulates cellular growth and differentiation. We previously demonstrated a reduction in basal AP-1 transcriptional activity associated with the malignant transformation of human bronchial epithelial (HBE) cells that was, in part, a consequence of decreased c-fos expression. In this study, we investigated the mechanisms underlying the reduction in c-fos expression associated with the malignant transformation of HBE cells. c-Fos gene transcription was lower in tumorigenic HBE cells than in normal HBE cells, and the reduction in transcription involved c-fos gene promoter elements from -327 to +40. DNaseI footprinting and band shift analyses of motifs within this c-fos promoter region, including a cyclic AMP response element (CRE), serum response element (SRE), sis-inducible element (SIE), and a YY1 site, revealed that binding to these motifs was greater in tumorigenic HBE cells than in normal HBE cells. Site-directed mutagenesis of the CRE partially relieved the repression of c-fos promoter activity in tumorigenic HBE cells. Further, the activity of the Jun N-terminal Kinase (JNK)-dependent pathway, which was a positive regulator of the c-fos promoter, was greater in normal HBE cells than in tumorigenic HBE cells. These findings demonstrate a transcriptionally-mediated suppression of c-fos gene expression associated with the malignant transformation of HBE cells. The decreased activity of the c-fos promoter in tumorigenic 1170I cells appeared to involve suppression through a CRE site and reduced activation by JNK-dependent pathways.

Topical ocular antibiotics induce bacterial resistance at extraocular sites.

AIM: To compare the prevalence of antibiotic resistance found in nasopharyngeal Streptococcus pneumoniae between villages treated with topical tetracycline or systemic azithromycin as part of a trachoma control programme. METHODS: All children aged 1-10 years were offered either single dose oral azithromycin treatment (20 mg/kg) or a course of topical 1% tetracycline ointment, depending on the area. Treatment was given annually for 3 years. Six months after the third annual treatment in each village, children were surveyed for nasopharyngeal carriage of S pneumoniae and resistance was determined using broth dilution MIC technique. Children in two additional villages, which had not yet been treated, were also surveyed. RESULTS: Nasopharyngeal carriage of S pneumoniae was similar in the tetracycline treated, azithromycin treated, and untreated areas (p=0.57). However, resistance to tetracycline and azithromycin was distributed differently between the three areas (p=0.004). The village treated with topical tetracycline had a higher prevalence of tetracycline resistance than the other villages (p=0.010), while the oral azithromycin treated village had a higher prevalence of macrolide resistance than the other villages (p=0.014). CONCLUSIONS: Annual mass treatment with oral azithromycin may alter the prevalence of drug resistant S pneumoniae in a community. Surprisingly, topical tetracycline may also increase nasopharyngeal pneumococcal resistance. Topical antibiotics may have an effect on extraocular bacterial resistance.

Basic helix-loop-helix proteins can act at the E-box within the serum response element of the c-fos promoter to influence hormone-induced promoter activation in Sertoli cells.

The Sertoli cell is a terminally differentiated testicular cell in the adult required to maintain the process of spermatogenesis. Previously basic helix-loop-helix (bHLH) factors and c-fos have been shown to influence Sertoli cell-differentiated functions. The induction of Sertoli cell differentiation appears to involve the serum response element (SRE) of the c-fos promoter to activate c-fos and intermediate bHLH factor(s) that regulate down-stream Sertoli cell-differentiated genes (e.g. transferrin expression). The SRE of the c-fos promoter is influenced through the serum response factor (SRF). Interestingly, an E-box nucleotide sequence is present within the SRE. bHLH proteins act through E-box elements, and the current study investigates the possibility that bHLH proteins may directly influence the SRE of the c-fos promoter. The activation of the c-fos promoter in Sertoli cells was found to be inhibited with the overexpression of the inhibitory HLH protein Id. Analysis of major response elements within the c-fos promoter demonstrated that the expression of Id specifically inhibited the activation of SRE in Sertoli cells and no other elements tested. Mutations in the E-box of the SRE also inhibited the activation of SRE, suggesting the direct role of bHLH proteins in regulating SRE activity in Sertoli cells. In contrast, the activation of SRE containing a mutated E-box was comparable to wild-type SRE in control stromal cells. Analysis of SRE oligonucleotide gel mobility shift assays with nuclear extracts from Sertoli cells demonstrated the presence of both the SRF and the ubiquitously expressed bHLH protein E12/E47. In contrast, no E12/E47 was detected in the SRE oligonucleotide gel shift using control stromal cell nuclear extracts. Observations suggest the binding of E12/E47 to SRE may be a cell-specific event. The SRF and bHLH proteins appear to bind to the SRE and activate the c-fos promoter in Sertoli cells. Observations provide evidence that a bHLH protein can interact with the SRE of the c-fos promoter to influence hormone-induced promoter activation. Cross-talk between these nuclear transcription factors appears to be instrumental in the control of Sertoli cell-differentiated functions.

Virulence genes detection of Salmonella serovars isolated from pork and slaughterhouse environment in Ahmedabad, Gujarat.

AIM: The aim was to detect virulence gene associated with the Salmonella serovars isolated from pork and Slaughterhouse environment. MATERIALS AND METHODS: Salmonella isolates (n=37) used in this study were isolated from 270 pork and slaughter house environmental samples collected from the Ahmedabad Municipal Corporation Slaughter House, Ahmedabad, Gujarat, India. Salmonella serovars were isolated and identified as per BAM USFDA method and serotyped at National Salmonella and Escherichia Centre, Central Research Institute, Kasauli (Himachal Pradesh, India). Polymerase chain reaction technique was used for detection of five genes, namely invA, spvR, spvC, fimA and stn among different serovars of Salmonella. RESULTS: Out of a total of 270 samples, 37 (13.70%) Salmonella were isolated with two serovars, namely Enteritidis and Typhimurium. All Salmonella serovars produced 284 bp invA gene, 84 bp fimA and 260 bp amplicon for enterotoxin (stn) gene whereas 30 isolates possessed 310 bp spvR gene, but no isolate possessed spvC gene. CONCLUSION: Presence of invA, fimA and stn gene in all isolates shows that they are the specific targets for Salmonella identification and are capable of producing gastroenteric illness to humans, whereas 20 Typhimurium serovars and 10 Enteritidis serovars can able to produce systemic infection.

Detection of Salmonella spp. from chevon, mutton and its environment in retail meat shops in Anand city (Gujarat), India.

AIM: The aim of this study was (i) To attempt isolation and identification of Salmonella species from samples. (ii) Serotyping of Salmonella isolates. (iii) Detection of virulence factor associated genes by polymerase chain reaction (PCR). MATERIALS AND METHODS: A total of 284 samples comprised of chevon and mutton (112 samples each) as well as 60 samples (20 each of retail meat shops environment samples viz. Butchers' hands, knives and log swabs) were collected from the retail meat shops in and around Anand City under aseptic precautions. Rappaport-vassiliadis soy bean meal broth and tetrathionate broth was used for the enrichment of all the samples and inoculation was done on brilliant green agar and xylose lysine deoxycholate agar. This was followed by the confirmation of isolates using biochemical tests. For the serotyping, isolates were sent to the National Salmonella and Escherichia Centre, Central Research Institute, Kasauli, Himachal Pradesh. Detection of virulence genes was performed by PCR technique using previously reported primer. RESULT: Of 284 meats and retail meat shops environment samples, 13 (4.58%) samples were found positive for Salmonella. It was interesting to know that incidence of Salmonella was more in mutton (6.25%) than chevon (3.57%). In case of meat shop environmental samples 1 (5.00%) sample observed positive for Salmonella separately among the butchers' hands and knives swabs (Each of 20 samples) examined. Out of 13, eleven isolates detected as Salmonella Typhimurium, whereas only two isolates were detected as Salmonella Enteritidis. All Salmonella isolates possess invA and stn genes, whereas nine isolates had a presence of spvR gene while only five of the isolates revealed the presence of spvC gene as shown by in vitro detection of virulence genes by PCR. CONCLUSION: Therefore, might be suggested that the good hygiene practices and effective control measures should be taken to encourage clean meat production with prolonged shelf-life.

Serum metabolic and minerals profile in norgestomet primed postpartum anestrous surti buffaloes.

AIM: The study was undertaken to find out the serum metabolic and minerals profile in postpartum anestrous surti buffaloes treated with norgestomet ear implants alone and in combination with pregnant mare serum gonadotropin (PMSG). MATERIALS AND METHODS: The study was conducted on 18 postpartum anestrous Surti buffaloes divided into three groups of six animals each at random to conduct the experiment. The buffaloes in Group-I and Group-II were implanted with Crestar ear implant for 9 days together with 2 ml injection of Crestar solution given i/m on the day of the implant insertion. In Group-II, additionally 500 IU PMSG was given i/m on the day of implant removal, whereas the buffaloes in Group-III served as anestrous control group and received 5 ml Normal Saline i/m on day 0 and 9 as a placebo treatment. RESULTS: The overall serum total protein values did not differ significantly (p > 0.05) between time (days) intervals in any of the groups. The mean serum total cholesterol levels at 10(th) day and on the day of estrus were found significantly lower (p < 0.05) in the control group as compared to treatment Groups I and II. However, there was no significant difference (p > 0.05) at 10(th) day and on the day of estrus between treatment groups (T1 and T2). The overall mean serum cobalt, zinc, iron, and manganese values did not differ significantly (p > 0.05) between different time intervals among any of the groups, except copper which was significantly lower (p < 0.05) at 10(th) day in control group as compared to treatment groups. CONCLUSION: Microelements cannot be synthesized in the body. Hence, it is concluded that the mineral mixture should be supplied daily in the animals ration to suffice the requirement of the trace elements. The mean serum metabolic and micro-minerals profiles in treatment and control groups revealed that overall mean serum total protein, cholesterol, copper, and zinc levels were apparently higher in treatment groups whereas, mean serum cobalt, iron, and manganese concentration had no consistent trend between treatment and control groups of Surti buffaloes.

Mechanism of action of the novel plasma membrane Ca(2+)-pump inhibitor caloxin.

Caloxin 2A1 is a novel inhibitor of the plasma membrane (PM) Ca(2+)-pump [Am. J. Physiol. Cell Physiol. 280 (2001) C1027]. The PM Ca(2+)-pump is a Ca(2+)-Mg(2+)-ATPase that expels Ca(2+) from cells to help them maintain low concentrations of cytosolic Ca(2+). Caloxin 2A1 inhibits Ca(2+)-Mg(2+)-ATPase in human erythrocyte leaky ghosts. Here we report that this inhibition is non-competitive with respect to the substrates Ca(2+) and ATP and the activator calmodulin. This was anticipated since the high affinity binding site for Ca(2+) and sites for ATP and calmodulin are intracellular whereas caloxin 2A1 is a peptide selected for binding to the second extracellular domain of the pump. Caloxin 2A1 also inhibited the Ca(2+)-dependent formation of the acid stable 140 kDa acylphosphate intermediate from 32P-gamma-ATP. However, it did not inhibit the formation of the acylphosphate intermediate in the reverse direction-from 32P-orthophosphate. Consistent with results on mutagenesis of transmembrane residues in the pump protein, we suggest that caloxin 2A1 inhibits conformational changes required during the reaction cycle of the pump.

E-box and cyclic adenosine monophosphate response elements are both required for follicle-stimulating hormone-induced transferrin promoter activation in Sertoli cells.

Sertoli cells are the epithelial cells responsible for the onset of pubertal development and maintenance of spermatogenesis in the adult. Transferrin is one of the major secretory products expressed by differentiated Sertoli cells. Investigation of the transcriptional control of transferrin gene expression provides insight into the regulation of Sertoli cell differentiation. Analysis of the mouse transferrin (mTf) promoter reveals the presence of a number of conserved response elements that have previously been shown to regulate cell specific expression of the human transferrin (hTf) promoter. One of these elements is the human PRII region, which is a cAMP response element (CRE)-like element that is more than 80% conserved in the mTf promoter. The activation of the hTf promoter by FSH and cAMP in rat Sertoli cells has been shown to be mediated in part through the CRE-like PRII region and binding of the CRE binding protein (CREB). The present study investigates the role of PRII in the activation of mTf promoter by FSH and cAMP in rat Sertoli cells. Mutations in the PRII of the mTf promoter reduced FSH activation by only 50% and cAMP activation by more than 90%. In contrast, the mutant PRII mTf promoter construct was fully activated by a partially purified testicular paracrine activity PModS(S300). Gel shift experiments demonstrated that proteins that can bind a consensus CRE oligonucleotide also bind the PRII region of the mTf promoter. An immunoblot confirmed that CREB binds the PRII and promotes the gel shift observed. The hypothesis developed was that another cis-acting element in addition to the CRE-like PRII is also involved in FSH actions. A conserved response element in both the mTf and hTf promoters is the basic helix-loop-helix (bHLH) responsive E-box sequence. Both FSH and PModS (S300) activity were found to promote a mTf E-box gel shift that contained the E2A gene product the bHLH protein E47. Interestingly, mutations in the E-box of the mTf promoter completely abolished the PModS(S300) activation and partially (52%) inhibited the activation by FSH. In contrast, the mutant E-box mTf promoter construct was fully activated by cAMP. Finally a double mutation of both the PRII and the E-box completely abolished FSH activation of the mTf promoter. These results suggest that optimal activation of the mouse transferrin promoter by FSH requires both CREB binding to the CRE-like PRII region and bHLH binding to the E-box. Information is provided that indicates a number of Sertoli cell promoters contain a close association of E-box and CRE-like elements. Observations are discussed in regards to the potential interactions of the CRE and E-box response elements in mediating FSH actions in Sertoli cells.

Role of basic-helix-loop-helix transcription factors in Sertoli cell differentiation: identification of an E-box response element in the transferrin promoter.

Sertoli cells are critical for testicular function and maintenance of the spermatogenic process. The induction of Sertoli cell differentiation in the embryo promotes testicular development and male sex determination. The progression of Sertoli cell differentiation during puberty promotes the onset of spermatogenesis. The maintenance of optimal Sertoli cell differentiation in the adult is required for spermatogenesis to proceed. The current study was designed to investigate the transcriptional regulation of Sertoli cell differentiation through the analysis of a previously identified marker of differentiation, transferrin gene expression. Sertoli cells produce transferrin to transport iron to developing spermatogenic cells sequestered within the blood-testis barrier. The transferrin promoter was characterized and found to contain two critical response elements, designated Sertoli element 1 (SE1) and Sertoli element 2 (SE2). Through sequence analysis, SE2 was found to contain an E-box response element, which has been shown to respond to basic-helix-loop-helix (bHLH) transcription factors. The bHLH proteins are a class of transcription factors associated with the induction and progression of cell differentiation. bHLH proteins dimerize through the conserved helix-loop-helix region and bind DNA through the basic region. Nuclear extracts from Sertoli cells were found to cause an E-box gel shift when the cells were stimulated to differentiate in culture, but not under basal conditions. The SE2 gel shift of Sertoli nuclear extracts was competed with excess unlabeled SE2 or E-box DNA fragments. Several Sertoli nuclear proteins associate with the SE2 gel shifts, including 70-, 42-, and 25-kDa proteins. Therefore, the critical SE2 element in the transferrin promoter is an E-box element capable of binding bHLH transcription factors. The ubiquitously expressed E12 bHLH protein dimerizes with numerous cell-specific bHLH factors. A Western blot analysis demonstrated that E12 was present in Sertoli cell nuclear extracts and associated with the SE2 gel shift. A ligand blot of Sertoli cell nuclear extracts with radiolabeled E12 had apparent bHLH proteins when the cells were stimulated to differentiate. The E-box sequence in the SE2 fragment of the transferrin promoter was CATCTG and was similar in gel shifts to the consensus E-box elements (CANNTG) previously characterized. A bHLH inhibitory factor (Id) competed and inhibited formation of the Sertoli cell nuclear extract E-box gel shift. To extend this observation, Id protein was overexpressed in cultured Sertoli cells. A transferrin promoter chloramphenicol acetyltransferase construct was used to monitor Sertoli cell function. The presence of Id suppressed the activation of the promoter induced by Sertoli differentiation factors. Therefore, the inhibition of Sertoli bHLH factors by Id suppressed Sertoli cell differentiated function, as measured by transferrin expression. An E-box-chloramphenicol acetyltransferase construct was also found to be active in Sertoli cells when cells were induced to differentiate. Screening the computerized nucleotide data bases demonstrated that putative E-box response elements are present in the promoters of a large number of Sertoli cell differentiated genes. In summary, a critical E-box response element has been identified in the transferrin promoter that can be activated by bHLH factors (e.g. E12) present in Sertoli cells. Inhibition of Sertoli bHLH factors by Id suppresses Sertoli cell differentiated function (i.e. transferrin expression), suggesting that bHLH transcription factors may be important in regulating Sertoli cell differentiated functions.

Role of winged helix transcription factor (WIN) in the regulation of Sertoli cell differentiated functions: WIN acts as an early event gene for follicle-stimulating hormone.

Members of the winged helix transcription factor family are known to regulate epithelial cell differentiation by regulating cell-specific gene expression. rWIN is a newly discovered member of the winged helix family shown to be present in the adult rat testis. In the testis the human homolog of rWIN, HFH-11, was localized to the germ cells (i.e. spermatocytes and spermatids) undergoing spermatogenesis. In the present study we show that rWIN is also expressed in testicular Sertoli cells. Sertoli cells are the epithelial component of the seminiferous tubule and provide both the cytoarchitectural support and the microenvironment for developing germ cells. The presence of rWIN in Sertoli cells was confirmed by Northern blot and RT-PCR analysis. The rWIN transcript size in the Sertoli cells was different from the germ cell transcript that is probably due to alternative splicing or modifications of the 3'-untranslated region. At least two spliced variants of rWIN were observed in the Sertoli cells corresponding to the deletion of an exon in the DNA-binding region. Long term stimulation of cultured Sertoli cells with the gonadotropin FSH down-regulated rWIN expression. In contrast, short-term stimulation (2 h) transiently up-regulated rWIN expression. The FSH-induced transient stimulation of rWIN precedes expression of the transferrin gene that is a marker of Sertoli cell differentiation. FSH-induced transferrin promoter activity was inhibited when cultured Sertoli cells were treated with an antisense oligonucleotide to rWIN. Interestingly, the constitutive overexpression of the DNA-binding domain of rWIN also down-regulated transferrin promoter activity. Analysis of the transferrin promoter with various deletion mutations suggested that rWIN acts at an upstream gene of the transferrin promoter. The results indicate that a transient up-regulation of rWIN in part mediates the ability of FSH to activate the transferrin promoter, which can be inhibited with a rWIN antisense oligonucleotide or constitutive expression of the rWIN DNA-binding domain. The current study demonstrates that rWIN acts as an early event gene for FSH actions on Sertoli cells and that rWIN appears to have a role in the regulation of Sertoli cell differentiated functions.