Analysis of Polymorphisms in Genes (AGT,MTHFR, GPIIIa,and GSTP1) Associated with Hypertension,Thrombophilia and Oxidative Stress in Mestizo and Amerindian Populations of México

Several polymorphisms related to hypertension, thrombophilia, and oxidative stress has been associated with the development of cardiovascular disease. We analyzed the frequency of M235T angiotensinogen (AGT), A222V 5,10 methylenete-trahydrofolate reductase (MTHFR), L33P glycoprotein IIIa (GPIIIa), and I105V glutathione S-transferase P1 (GSTP1) polymorphisms in 285 individuals belonging to Mexican-Mestizo and five Amerindian population from México, by real time PCR allelic discrimination. Allele and genotype frequencies were compared using χ² tests.All populations followed the Hardy Weinberg equilibrium for assay markers with the exception of the Triki, whose were in Hardy Weinberg dysequilibrium for the glutathione S-transferase P1 polymorphism.Interestingly, according to all the analyzed single nucleotide polymorphisms (SNPs), the Triki population was the most differentiated and homogeneous group of the six populations analyzed. A comparison of our data with those previously published for some Caucasian, Asian and Black populations showed quite significant differences. These differences were remarkable with all the Mexican populations having a lower frequency of the 105V allele of the glutathione S-transferase P1 and reduced occurrence of the 222A allele of the 5,10 methylenetetrahydrofolate reductase. Our results show the genetic diversity among different Mexican populations and with other racial groups.     

Efficacy of deltamethrin (Butox® 7.5 pour on) against nymphs and adults of ticks (Ixodes ricinus, Rhipicephalus sanguineus) in treated hair of cattle and sheep

Ticks are known to be able to transmit a broad spectrum of agents of diseases in cattle or sheep. Therefore, measurements are needed to keep ticks away from the body of any ruminant belonging to the agricultural life stock. The present study dealt with investigations to measure the efficacy of the insecticide deltamethrin (Butox? 7.5 pour on) against specimens of two important species (Ixodes ricinus and Rhipicephalus sanguineus). Four sheep and four young cattle were treated lege arte along the vertebral column with 10 ml Butox? (deltamethrin) per sheep or 30 ml Butox? per cattle. Day 7, 14, 21, and 28 after the treatment, hair was shaved off from the head, ears, the back, belly, and the feet being collected in separate, suitable plastic bags, and transported to the institute, where these hair were brought into close contact with either adult and/or nymph stages of I. ricinus and R. sanguineus. As results, strong, acaricidal effects were seen, which varied according to the parasite species, the origin of the hair (e.g., head, leg, etc.) and according to the period after the treatment. In sheep, the acaricidal effect was noted for the whole period of 28 days along the whole body with respect to adults and nymphs of I. ricinus, while the acaricidal effects of deltamethrin were reduced for R. sanguineus stages beginning at day 21 after treatment. In cattle, the full acaricidal effect was seen for 21 days in I. ricinus stages and for 14 days in R. sanguineus, while the acaricidal efficacy became reduced after these periods of full action—beginning at the hair taken from the legs. Only R. sanguineus adults did not show any reaction on day 28 after treatment. Besides these acaricidal effects, repellent effects were also noted. Full repellency for both species was seen during the first 14 days in sheep and cattle against Ixodes and Rhipicephalus, while the repellency was later reduced, especially in contact with hair from the legs. As conclusion, deltamethrin, besides its very good effects against biting insects, brings acaricidal as well as repellent effects against ticks, thus protecting the sheep and cattle from transmission of agents of diseases.     

Long-term changes in body composition and prevalence of overweight and obesity in girls (aged 3–18 years) from Kraków (Poland) from 1983, 2000 and 2010

Background: Body mass disorders are an increasing problem, especially in industrialized countries.

Aim: Determination of time- and age-related differences in the prevalence of overweight, obesity and in body composition in girls from 1983, 2000 and 2010.

Subject and methods: In 2010 an anthropological study was conducted on 1970 girls aged 3–18 years living in Kraków (Poland). Data on selected skin-folds, BMI, muscle mass and bone mass were compared to two studies on analogous populations carried out in 1983 and 2000.

Results: Compared to 1983, the share of overweight girls in 2010 had decreased (from 12.4% to 11.2%), while the obesity rate had increased slightly from 2.3% to 3.2%. Girls from 2010 had lower overall body fat content than their peers studied in 1983. However, in 2010 suprailiac adiposity prevalence was higher, while triceps, subscapular and abdominal adiposity rates were lower. They also had higher muscle mass and lower bone mass.

Conclusion: The prevalence of overweight and obesity has not changed significantly over the last 30 years. However, significant changes have occurred in body composition. Girls measured in 2010 had lower total adipose tissue, although they also showed a tendency towards increased central adiposity.     

Biodegradation, biocompatibility, and drug delivery in poly(ω-pentadecalactone-co-p-dioxanone) copolyesters

Poly(ω-pentadecalactone-co-p-dioxanone) [poly(PDL-co-DO)] copolyesters are copolymers of an isodimorphic system, which remain semicrystalline over the whole range of compositions. Here, we evaluated enzymatically synthesized poly(PDL-co-DO) copolymers as new materials for biomedical applications. In vivo experiments using mice, showed that the copolyesters are well tolerated, with tissue responses that are comparable to poly(p-dioxanone). In addition, the copolymers were found to degrade hydrolytically at controlled rates over a period of several months under physiological conditions. The poly(PDL-co-DO) copolymers with up to 69 mol% DO units were successfully transformed to free-standing nanoparticles that are capable of encapsulating an anticancer drug, doxorubicin, or a polynucleotide, siRNA. Drug- or siRNA-loaded nanoparticles exhibited controlled and continuous release of agent over many weeks. In addition, siLUC-encapsulated poly(PDL-co-DO) nanoparticles were active in inhibiting luciferase gene expression in LUC-RKO cells. Because of substantial differences in structure and hydrophobicity between PDL and DO units, poly(PDL-co-DO) biodegradation rate and physical properties can be tuned over a wide range depending on the copolymer composition. Our results demonstrate that the semicrystalline and biodegradable poly(PDL-co-DO) copolyesters are promising biomaterials to serve as drug carriers, as well as potential raw materials for constructing bioabsorbable sutures and other medical devices.     

Analysis of TNFα promoter SNPs and the risk of cervical cancer in urban populations of Posadas (Misiones, Argentina)

Background

Human papillomavirus (HPV) plays a central role in cervical cancer development. However, only a small fraction of infected women develop the disease. Additional risk factors, including SNPs in immune system and cytokine genes, are likely to be important determinants.

Objective

We investigated the potential role of cytokine TNF-α promoter SNPs (TNFα-375A, TNFα-307A, TNFα-243A, and TNFα-237A) in the development of high-grade cervical lesions and cancer in urban women from Posadas (Misiones, Argentina).

Study design

Fifty-six cases (CINIII and invasive carcinoma) and 113 age-matched controls were included in the study. HPV genotype detection was conducted by PCR. TNFα SNP genotyping was conducted through PCR amplification and direct sequencing of genomic DNA.

Results

We observed differences in the allelic distribution of TNFα-307A and TNFα-375A SNPs among cases and controls (p < 0.05). The TNFα-307A variant was associated with cervical cancer at an OR 2.4 (CI 95% 1.1-5.4), while the TNFα-375A SNP was identified in 8.8% of the controls and none of the cases. Moreover, the TNFα-375A always occurred in association with the TNFα-237A SNP, indicating linkage disequilibrium between them.

Conclusion

Our study suggests that the presence of the high producer allele TNFα-307A is associated with an increased risk for the development of cervical cancer in the Posadas population. We also speculate that the “protective effect” of the TNFα-375A/-237A haplotype, which was restricted to controls, may be related to HLA genes linked on chromosome 6. These findings contribute to our understanding of immune gene variation in an Argentinean population, and its role in disease susceptibility.     

Comparative study on the effects of three insecticides (fipronil, imidacloprid, selamectin) on developmental stages of the cat flea (Ctenocephalides felis Bouché 1835): a light and electron microscopic analysis of in vivo and in vitro experiments

The effects of three insecticides (fipronil, imidacloprid and selamectin) on developmental stages of cat fleas (Ctenocephalides felis) were studied in vivo, in vitro and by means of light and electron microscopy. The results were documented by video. Adult fleas were attached to the skin of dogs that had been treated 7 days before with one of the three compounds. Furthermore, adult fleas were exposed exclusively to the hair and skin debris of such treated dogs or were placed on filter papers that had been impregnated with one of these three compounds or with the blood of treated dogs. Larval fleas were exposed to hair of treated dogs, to debris obtained by combing treated dogs, to dried blood samples of treated dogs or were placed onto filter papers impregnated with one of the three compounds. In these experiments with adult and larval fleas, it was noted that none of the three insecticides had a repellent effect on adult or larval fleas. Imidacloprid was the only compound that acted exclusively by body contact, and was apparently taken up by adult and larval fleas via the thin, non-sclerotized intersegmental membranes of the flea's body, shown when flea stages were exposed to hairs taken from dogs treated with one of the compounds or placed onto drug-impregnated filter papers. Imidacloprid killed larvae and adult fleas within 1 h, while it took at least 24 h until all adult fleas had died on fipronil- or selamectin-treated dogs, thus allowing longer feeding periods, increasing the risk of transmission of flea-derived diseases. Flea larvae covered with debris from dogs topically treated 7 days before with fipronil, imidacloprid or selamectin died, like the untreated control, within 16–28 h after exposure. This was, however, probably mainly due to a drying effect. Adult and larval fleas exposed to filter papers impregnated with the blood of treated dogs survived longer than 7 days, as did the untreated controls. All three drugs apparently acted on nerves and muscles and thus stopped motility. Received: 7 October 2000 / Accepted: 18 October 2000     

Record of Evlachovaea sp. (Hyphomycetes) on Triatoma sordida in the state of Goiás, Brazil, and its activity against Triatoma infestans (Reduviidae, Triatominae)

A fungal isolate was detected on a dead Triatoma sordida (St?l) collected in a peridomestic area in central Brazil. The fungus belongs to Evlachovaea Borisov and Tarasov, a new genus that was recently described in Russia. The isolate seems to be a third species and the second new and undescribed species from Brazil. The fungus was shown to be active against Triatoma infestans (Klug) third-instar nymphs at a humidity close to saturation. However, activity was reduced at a lower humidity (75%). Values of LC50 varied between 1.1 x 10(5) and 1.5 x 10(4) conidia/cm2 treated surface, 15 and 20 d after fungal application and incubation at humidity close to saturation. This new fungus may have a potential for biological control of peridomestic Chagas' disease vectors during the rainy season.     

Knowledge, attitudes and practices of populations towards mosquitoes in urban and rural area (Côte d'Ivoire--West Africa)

Three surveys related to the knowledge, attitude and practices of populations towards mosquitoes were conducted in C?te d'Ivoire. The first survey was undertaken in June 1994 in the city of Bouaké and the other two respectively in January 1996 and in June 1999 in the village of Kafiné. The surveys have revealed that whatever the environment, nuisance due to mosquitoes constitutes a great concern for the population of the villages. The majority of people interviewed in urban (94%), peri-urban (68%) zones and in Kafiné (94%) associates mosquito with malaria against 18% only in Kabolo. Furthermore, these surveys have allowed the inventory of methods of controlling mosquito currently used by the communities in both urban and rural areas. Generally mosquito coils as well as insect sprays are currently used because of their cheapness and their availability on the local market. in Bouaké, insect sprays (97% to 99%) are used as much as mosquito coils (87% to 98%). However, in Kafiné and Kabolo, insect sprays are only used by respectively 76% and 69% and the mosquito coils by respectively 68% and 71%. In urban zones, the insect sprays are more common than mosquito coils. On the contrary mosquito coils are currently used in villages due to their low price (40 to 50 FCFA) and their constant availability on the local market. Furthermore, local plants characterized by a repulsive essence are known by 48% and 75% of people interviewed respectively in the urban and peri-urban area of the city of Bouak6, by 74 % of the population of Kafiné and 73% of people living in Kabolo. In the last village 57% of people who were interviewed do not use any means of orotection due to financial reason; only 14% use mosquito coils and 20% use local plants with a repulsive essence. Mosquito net is known by the majority of people interviewed (82 - 98%) as an excellent means of orotection against mosquito bites. However only 27%, 13%, 26% and 5% of people respectively living in the urban and peri-urban areas of Bouaké and in the villages of Kafiné and Kabolo own their mosquito net. Its cost is the main reason for not using it as mentionned by 25% of people interviewed in the urban neighborhood, by 48% in the peri-urban areas, respectively 46% and 89% in the villages of Kafiné and Kabolo. Although its effectiveness has been proved by the communities, mosquito net still remains less common. Its price is relatively high (3500 - 5000 FCFA). It is not constantly available on the local market. Other socio-cultural factors such as local beliefs and taboos hinder the use of mosquito nets by the communities.     

Frequency of HLA-A, -B and -DRB1 specificities and haplotypic associations in the population of Castilla y León (northwest-central Spain)

The frequencies of human leukocyte antigen (HLA) class I and class II specificities and haplotypic associations were determined in 1940 unrelated donors from Castilla y León and compared with other Iberian, Mediterranean and European populations. Specificities were determined using polymerase chain reaction reverse sequence-specific oligonucleotide or polymerase chain reaction sequence-specific primer techniques. In the analysis, 19, 29 and 13 specificities were found for HLA-A, -B and -DRB1, respectively, with HLA-A*02 (26%), -A*01 (11%), -B*44 (16%), -B*35 (10%), -DRB1*07 (16%) and -DRB1*13 (14%) showing the highest frequencies. In addition, 10 common HLA-A-B-DRB1 haplotypic associations were observed, A*01-B*08-DRB1*03 (3%) and A*29-B*44-DRB1*07 (3%) being the most frequent ones. These findings indicate that the population of Castilla y León is genetically equidistant from the Portuguese and other Spanish populations and shares a common origin with other Iberian populations, in which European, Mediterranean and North African genetic components are present; this is in agreement with the historical and genetic background of the population. These data contribute to a better understanding of the genetic structure of the Iberian Peninsula and provide a healthy control population from our region that should be useful for the study of disease associations.     

Evaluation of Quinolones for Use in Detection of Determinants of Acquired Quinolone Resistance,Including the New Transmissible Resistance Mechanisms qnrA,qnrB, qnrS,and aac(6′)Ib-cr,in Escherichia coli and Salmonella enterica and Determinations of Wild-Type Distributions

Fluoroquinolone resistance in members of the Enterobacteriaceae family is mostly due to mutations in the quinolone resistance-determining regions of the topoisomerase genes. However, transferable genes encoding quinolone resistance have recently been described. The current methods for susceptibility testing are not adapted to the detection of new resistance determinants, which confer low levels of resistance. The aim of this study was to compare the ability of the screening of the different quinolones by disk diffusion assays and MIC determinations to detect fluoroquinolone resistance. Sixty-nine Escherichia coli strains and 62 Salmonella strains, including strains fully susceptible to quinolones, nalidixic acid-resistant strains, strains with resistance to fluoroquinolones (resistant to nalidixic acid), and strains showing low-level resistance to fluoroquinolones conferred by transferable quinolone resistance genes, including qnrA, qnrB, qnrS, and aac(6′)Ib-cr, were selected. Disk diffusion assays and MIC determinations by the agar dilution method were performed, according to CLSI standards, with nalidixic acid, flumequine, oxolinic acid, ciprofloxacin, enrofloxacin, marbofloxacin, norfloxacin, ofloxacin, and levofloxacin. The MIC of levofloxacin was determined by an Etest. The results showed a trimodal distribution of the MICs for both E. coli and Salmonella. The MIC distributions for the isolates varied with the compounds tested. Screening for nalidixic acid resistance by MIC testing or disk diffusion assay was not efficient for the detection of some of the isolates carrying qnr and aac(6′)Ib-cr. Transferable resistance genes would best be detected by testing for the MIC of ciprofloxacin or norfloxacin, as testing for the MICs of the other compounds would fail to detect isolates carrying aac(6′)Ib-cr because the enzyme produced is able to reduce the activities of these two compounds only due to their chemical structures. In conclusion, screening with nalidixic acid is efficient for the detection of mutants, but it is not so efficient for the detection of qnr and aac(6′)Ib-cr. Detection would be maximized by screening with either ciprofloxacin or norfloxacin by both MIC determination and disk diffusion assays. Furthermore, a low concentration of ciprofloxacin (1 μg) in the disks seemed to increase the sensitivity of the disk diffusion assay.Fluoroquinolone resistance in members of the Enterobacteriaceae family has until recently been attributed to mutations in the gyrase and topoisomerase genes quinolone resistance-determining regions (QRDRs). Stepwise increases in resistance have been described for both Escherichia coli and Salmonella enterica species as a result of the accumulation of topoisomerase mutations (14). Efflux pump mechanisms were also described to act in resistance alone or in combination with decreased levels of expression of outer membrane porins (20, 21, 27). However, in the late 1990s, Martinez-Martinez et al. found a new plasmid-mediated mechanism, qnrA1, that is able to increase the MICs of quinolones through a target protection mechanism (18). Other related qnr genes (qnrA, qnrB, qnrC, and qnrS) and numerous variants have been described (13, 15, 16). Furthermore, a new resistance mechanism, aac(6′)Ib-cr, based on the enzymatic modification of some fluoroquinolones was described (23). However, the reduction in the susceptibility to fluoroquinolones might not be observed when other quinolones, such as enrofloxacin, pefloxacin, gemifloxacin, and levofloxacin, are tested (22, 23).The actual screening methods include nalidixic acid as the first screening drug, since the sensitivity of the MIC test is higher for the detection of first-step mutants. However, the use of nalidixic acid might not be appropriate for the detection of these recently described resistance determinants that do not affect the MIC of nalidixic acid in the same way that mutations in the QRDRs of topoisomerases do. Furthermore, the breakpoints for quinolones for the detection of reduced susceptibility in surveillance programs have been under discussion to improve the detection of resistance determinants (5, 7).The methods for the detection of recently described transferable resistance are not well established, and if susceptibility testing is performed according to the CLSI guidelines and interpretations (8, 9, 10), the isolates carrying these resistant mechanisms would be classified as susceptible since they mediate a reduction in susceptibility which is below the actual clinical breakpoints, although their clinical importance is still unclear.Given their transferability and the possibility that they cause increases in resistance that might affect the clinical response to treatment, the detection of quinolone resistance should routinely be performed. Therefore, laboratories will have to adapt the detection methods that they use. The detection of quinolone resistance might be facilitated through the selection of the drugs that are the most adequate for use in susceptibility tests and by the recognition of the expected phenotypes, isolates with which can then be further studied for their genetic backgrounds.As part of this study, we measured the MICs and inhibition zone diameters for a panel of quinolones used in human and veterinary medicine to observe the distribution of quinolone-susceptible isolates and isolates harboring different mechanisms of resistance. This information could be useful for the optimization of quinolone resistance detection and the establishment of cutoff values and the interpretative criteria to be used for the detection of quinolone resistance for clinical and surveillance purposes.     

Roles of RseB, σE,and DegP in Virulence and Phase Variation of Colony Morphotype of Vibrio vulnificus

Vibrio vulnificus is an estuarine bacterium capable of causing serious and often fatal wound infections and primary septicemia. We used alkaline phosphatase insertion mutagenesis to identify genes necessary for the virulence of this pathogen. One mutant had an in-frame fusion of ′phoA to the gene encoding RseB, a periplasmic negative regulator of the alternative sigma factor σ^E. σ^E controls an extensive regulon involved in responding to cell envelope stresses. Colonies of the rseB mutant were less opaque than wild-type colonies and underwent phase variation between translucent and opaque morphologies. rseB mutants were attenuated for virulence in subcutaneously inoculated iron-dextran-treated mice. To obtain insight into the role of rseB and the extracytoplasmic stress response in V. vulnificus, mutants with defined mutations in rseB and two important members of the extracytoplasmic stress regulon, rpoE and degP, were constructed for analysis of virulence, colony morphology, and stress-associated phenotypes. Deletion of rseB caused reversible phase variation in the colony morphotype that was associated with extracellular polysaccharides. Translucent and transparent morphotype strains were attenuated for virulence. rpoE and degP deletion mutants were sensitive to membrane-perturbing agents and heat but were not significantly attenuated for V. vulnificus virulence in mice. These results reveal complex relationships between regulation of the extracytoplasmic stress response, exopolysaccharides, and the virulence of V. vulnificus.Vibrio vulnificus is a gram-negative estuarine bacterium responsible for severe opportunistic infections (for a review, see reference 17). Ingestion of raw contaminated seafood can lead to septicemia in susceptible patients, while contact with contaminated seawater or seafood can cause wound infection, which may progress to necrotizing fasciitis and sepsis. Mortality rates for sepsis and wound infection can be as high as 75% and 50%, respectively. Predisposing conditions for septicemia include liver disease, cirrhosis, hemochromatosis, diabetes, and immune compromise, while wound infection can occur in otherwise healthy persons.Several virulence factors have been identified for V. vulnificus, most notably the antiphagocytic capsule (55, 65), RtxA toxin (26, 28, 32), and iron acquisition systems (31, 64). For a review, see reference 17. However, much is yet to be discovered, particularly the mechanisms of extreme tissue damage and rapid growth in host tissues (17). To examine these traits, we focused on the factors that are localized to the bacterial cell surface or are secreted into the extracellular space, considering that most virulence factors are exported to interact with the host. Alkaline phosphatase (phoA) mutagenesis is a useful tool for identifying genes encoding exported products (33), based on the principle that alkaline phosphatase is active only when it is exported beyond the bacterial cytoplasm. Randomly generated phoA gene fusions, most often generated via TnphoA (33), must be in genes encoding exported proteins to have enzyme activity detected by plating on the chromogenic substrate 5-bromo-4-chloro-3-indolylphosphate (BCIP). In our studies, TnphoA did not work effectively in V. vulnificus for unknown reasons. We therefore created a mini-Tn5 transposon-based ′phoA delivery system, miniTn5phoA (8), that works well in V. vulnificus.Using this method, we identified a phoA mutant that had an in-frame fusion of ′phoA to the gene encoding RseB, a periplasmic negative regulator of sigma E (σ^E) activity. The rseB mutant exhibited several interesting phenotypes, including phase variation between translucent and opaque colony morphologies and attenuated virulence. σ^E is an alternative RNA polymerase sigma factor that controls an extensive regulon involved in responding to cell envelope stresses (48). This response, termed the extracytoplasmic stress response (ESR), is essential for maintaining the envelope integrity of gram-negative bacteria under certain stress conditions (for a review, see reference 48). Because rseB is involved in the ESR, we determined the role of the σ^E-mediated ESR in V. vulnificus. We also investigated the possible reasons for the translucent morphology of RseB variants by comparing these variants with an acapsular translucent mutant of V. vulnificus. This study uncovered a possible role for RseB in phase variation of extracellular polysaccharide (EPS) expression and is the first study to investigate the role of the ESR in the virulence of V. vulnificus.     

Value of (1-3)-β-d-glucan, Candida mannan and Candida DNA detection in the diagnosis of candidaemia

This study determined the value of (1,3)-β-d-glucan (BDG), Candida mannan (MN) and Candida species-specific DNA as surrogates for diagnosis of candidaemia. Thirty-nine patients yielding Candida species in blood cultures were investigated for presence of BDG, MN and Candida species-specific DNA in serum samples. The Candida spp. bloodstream isolates included C. albicans (n = 16), C. tropicalis (n = 10), C. parapsilosis (n = 7), C. glabrata (n = 3) and C. dubliniensis (n = 3). Positivity of the three markers was as follows: Candida DNA for corresponding Candida species, 100%; BDG, 87%; MN, 59%. Despite varying sensitivities of these biomarkers, they provided a useful adjunct to the diagnosis of candidaemia.     

Effects of a neem seed extract (MiteStop®) on mallophages (featherlings) of chicken: in vivo and in vitro studies

Mallophages of birds (featherlings) are mostly very tiny and can even as adults better be recognized by their movements than by their elongate body shape when using just the naked eye. Since some species (e.g., the “shaft louse” Menopon gallinae, the elongate feather louse Lipeurus caponis, or Columbicola sp.) may pierce the pulp of feathers or the skin by their biting or scratching mandibles and thus lick the excreted blood, they may be extremely dangerous especially to young birds, even if they only feed by nibbling along the feather surface and/or eat epidermal debris. The present paper reports on the successful treatment of different races of fowls being severely infested with both above cited species. This in vivo treatment was done either by a short dipping of the whole fowl into the 1:33 dilution (with tap water) of a neem seed extract (MiteStop®) or by spraying them with the freshly diluted product. It was seen that the dead mallophages dropped down from the feathers as soon as they were dry again. As a precaution, a second treatment was done by some owners 1 week after the first one in order to eliminate all stages, which eventually might have hatched from untouched nits during the time interval between the two treatments. When controlling the treated fowls 4 weeks after the treatment, in no case (treated once or twice), living motile stages were diagnosed indicating the high efficacy of this nontoxic neem seed extract. When treating in vitro cutoff feathers contaminated with L. caponis, it was seen under the stereomicroscope, that the mallophages tried to run away from the 1:33 water-diluted active compound indicating that there is also a repellent effect. Treated L. caponis stopped leg movements within 3 min and died on their feathers within 1–20 min. Then, the last slight trembling movements of their legs and convulsions of their intestine stopped finally.     

Expression of inhibin/activin subunits alpha (-α), BetaA (-βA), and BetaB (-βB) in placental tissue of normal, preeclamptic, and HELLP pregnancies

During human pregnancy the placenta produces a variety of proteins for the establishment of the fetoplacental unit, including inhibins and activins. Inhibins are dimeric glycoproteins, composed of an α-subunit and one of two possible β-subunits (βA or βB). Aims of the present study were (a) the determination of the frequency and tissue distribution patterns of the inhibin/activin subunits in human placental tissue of normal pregnancies and pregnancies complicated with preeclampsia and HELLP syndrome (hemolysis, elevated liver enzymes, low platelets) and (b) the assessment of a combined expression of inhibin-α- and both β-subunits (βA- and βB-subunits) using double immunofluorescence technique. A significant lower expression of the inhibin-α subunit in preeclamptic and HELLP placental tissue compared to normal pregnancies was observed, while the inhibin-α immunostaining was significantly upregulated in syncytotrophoblast. Additionally, we demonstrated a significant down-regulation of inhibin-βB subunit in extravillous trophoblast cells between normal and preeclamptic compared to HELLP placental tissue, while inhibin-βA-subunit was significantly higher in preeclamptic syncytotrophoblast cells. A colocalization of inhibin-α and the beta-subunits could be demonstrated, suggesting a production and secretion of intact inhibin A and inhibin B. Therefore, inhibin A and activin A might be useful markers in preeclampsia. Valuable parameters in HELLP syndrome could be inhibin A, rather than inhibin B, and activin B. Furthermore, the lower βB-subunit production in extravillous trophoblast cells demonstrates that this subunit might have an important role in the pathogenesis of HELLP syndrome. Additionally, the higher production of the βA-subunit in syncytotrophoblast cells suggest a higher production of activin A rather than inhibin A in preeclampsia that might be utilized as a marker of placental function.     

Co–circulation of different genotype,sub-genotype and serotypes of hepatitis B virus (HBV) and its epidemiology in Assam: A north-eastern state of India

PurposeHepatitis B virus (HBV) infection is a global health problem. HBV has different genotypes and subgenotypes with geographical distinctiveness. Aims: To study the molecular epidemiology and distribution pattern of HBV in Assam; a distinct state of India that may have different genotypic divergence. Settings and design: Patients attending a tertiary care hospital susceptive of Hepatitis B were included, irrespective of age and sex in different agro-climatic zones of Assam.MethodsSamples positive for Hepatitis B surface antigen test and COBAS®TaqMan® HBV tests were further confirmed by PCR and sequencing followed by phylogenetic analysis.Statistical analysis used: Chi-square test was used to determine whether there was a significant difference among the results in this study.Results: An increase trend of HBV positive cases has been observed in the state. The incidence in female was lower than that of male and age group 26–35 years was most vulnerable. Genotype D, subgenotype D2 and serotype ayw3 were predominant genotype, subgenotype and serotype. The prevalence of subgenotype C3 was a new finding. Phylogenetic analysis showed that the genotypes of HBV prevailing in the state have close relationship with neighboring countries of India which may be due to increased cross border migration of populationConclusionsThis comprehensive study of HBV in Assam described the distribution of HBV genotypes and subgenotypes and serotypes in different agro-climatic zones of Assam. These findings will help to formulate the roadmap for prevention and control of the disease as well as targeted therapy of HBV in this State.