Chicken viperin inhibits Newcastle disease virus infection in vitro: A possible interaction with the viral matrix protein.

Viperin is an interferon-inducible protein that helps in protecting mammals against various virus infections. Viperin is a highly conserved member of the interferon-stimulated genes (ISG) family in many species. Viperin has been shown to play a pivotal role in the innate immunity of chicken; however, its role has not been explored in its antiviral potential. Newcastle disease virus (NDV) is the causative agent of an infectious disease in poultry. In the present study, we have shown the anti-NDV effect of chicken viperin (cViperin). The impact of cViperin upon NDV infection was investigated in chicken embryo fibroblast. The modeling of the cViperin protein was done using I-TASSER and ZDOCK is used to predict the possible interaction with the matrix protein of NDV. The interaction was further confirmed by co-immunoprecipitation assay using recombinant matrix protein of NDV with the recombinant cViperin. The recombinant NDV expressing cViperin showed reduced replication of the virus upon its growth kinetics. Our results suggest downregulation of NDV replication in the presence of cViperin. The study will be critical to elaborate our understanding of the chicken innate immune system which could help develop antiviral strategies against NDV infection.

Understanding the maturity of coconut water through 1H NMR profiling and MPAES analyses.

This study investigated the relationship between coconut maturity stages and the sugar, amino acid, and mineral profiles of coconut water (CW). Metabolite profiles were analysed using 1H NMR, covering glucose (G), fructose (F), sucrose (S), reducing sugars (RS), total sugars (TS), amino acids, and organic acids. Mineral composition was measured using Microwave Plasma Atomic Emission Spectroscopy (MPAES). The results revealed distinct metabolite and mineral profiles across different maturity stages. Immature CW had high G/F and RS/TS ratios but low S/G ratios. Conversely, mature CW showed decreased G/F and RS/TS ratios but an increase in S/G. Mineral analysis revealed potassium as the predominant mineral in CW, peaking in the youngest stage and declining with maturity. Sodium, magnesium, and calcium showed a similar pattern, with higher concentrations in early than in later stages. The study identifies the age of 9-10 months as optimal stages for selecting tender coconut water.

Rapid screening for the detection of HLA-B57 and HLA-B58 in prevention of drug hypersensitivity.

HLA-B57 and HLA-B58 are major histocompatibility class (MHC)-I allotypes that are potentially predictive of important clinical immune phenotypes. HLA-B*5701 is strongly associated with hypersensitivity to the HIV drug abacavir, liver toxicity from the antibiotic flucloxacillin and is a marker for slow progression of HIV AIDS. HLA-B*5801 is associated with hypersensitivity to allopurinol used to treat hyperuricaemia and recurrent gout. Here we describe a monoclonal antibody (mAb) specific for HLA-B57 and HLA-B58 that provides an inexpensive and sensitive screen for these MHC-I allotypes. The usefulness of HLA-B57 screening for prediction of abacavir hypersensitivity was shown in three independent laboratories, including confirmation of the mAb sensitivity and specificity in a cohort of patients enrolled in the PREDICT-1 trial. Our data show that patients who test negative by mAb screening comprise 90%-95% of all individuals in most human populations and require no further human leukocyte antigen (HLA) typing. Patients who test positive by mAb screening should proceed to high-resolution typing to ascertain the presence of HLA-B*5701 or HLA-B*5801. Hence, mAb screening provides a low-cost alternative to high-resolution typing of all patients and lends itself to point-of-care diagnostics and rapid ascertainment of low-risk patients who can begin immediate therapy with abacavir, flucloxacillin or allopurinol.

Technetium-99m-HMPAO SPECT cerebral blood flow study in children with craniosynostosis.

UNLABELLED: Premature closure of cranial sutures (primary craniosynostosis) in children leads to characteristic skull deformities and prevents the constricted brain from growing normally. Although the cause remains unknown, several etiological factors have been cited. Recently, hypovascularity has been reported as a possible cause of craniosynostosis. METHODS: In a prospective study regional cerebral blood flow studies were carried out with 99mTc-HMPAO SPECT in seven children with craniosynostoses. Five preoperative and six postoperative studies were conducted and the results correlated with radiological and surgical findings. RESULTS: Preoperative studies revealed regional hypovascularity in the underlying cerebral hemisphere, corresponding to the fused sutures. Postoperative studies revealed disappearance of these perfusion defects in most cases, indicating normalization of perfusion following surgical decompression. CONCLUSION: This study establishes the presence of cerebral hypovascularity in craniosynostoses and suggests that early surgery and release of craniostenosis is essential to achieve optimum perfusion and brain development.

Monocyte derived IL 10 and PGE2 are associated with the absence of Th 1 cells and in vitro T cell suppression in lepromatous leprosy.

Our previous studies had shown that the clinicopathological spectrum in leprosy was associated with discrete T cell subsets in circulation, with tuberculoid patients having antigen-induced Th 1, whereas lepromatous leprosy patients with antigen-specific T cell anergy possessed Th 2 cells. The present study shows that infected monocytes from lepromatous but not tuberculoid leprosy patients released soluble factors (MoF(s)) containing IL-10 and PGE2 which inhibited M. leprae induced in vitro lymphoproliferation of previously sensitised healthy or tuberculoid leprosy subjects. A strong negative correlation was observed between adherent cell derived IL-10 and IL-2 at the level of both the product and cytokine mRNA. Moreover, anti-IL-10 antibodies and indomethacin partially reversed the suppressor effects of MoF(s). Taken together these studies indicate that infected monocytes contribute to the development of T cell anergy by releasing factors that affect regulatory cytokines and T cell subset differentiation in lepromatous leprosy.

Rio Mamore virus: genetic characterization of a newly recognized hantavirus of the pygmy rice rat, Oligoryzomys microtis, from Bolivia.

Human hantavirus disease occurs throughout much of South America. The rodent hosts and the specific etiologic agent(s) are largely unknown, but many reported cases occurred within the habitation ranges of oryzomine rodents (rice rats). We have identified a genetically novel hantavirus (Rio Mamore virus [RM]) of the pygmy rice rat Oligoryzomys microtis in Bolivia. The complete sequence of the small (S) genome and the partial sequence of the medium (M) genome are described. This virus is closely related to the newly identified human pathogen Andes virus from Patagonia. To facilitate improved diagnosis of hantavirus infections in South America, we have expressed the complete nucleocapsid protein of RM in Escherichia coli and affinity-purified it for use in an ELISA and Western blot assays for antibodies to RM.

Cocirculation of multiple hantaviruses in Texas, with characterization of the small (S) genome of a previously undescribed virus of cotton rats (Sigmodon hispidus).

An environmental and laboratory investigation was conducted after a fatal childhood case of hantavirus pulmonary syndrome occurred in Deaf Smith County, Texas in May 1995. A trapping campaign was conducted to identify possible rodent carriers. Six species of murid and heteromyid rodents were collected, and at least one hantavirus-seropositive specimen was found in each of the five murid species. Tissues from a selection of 11 seropositive specimens were examined by the polymerase chain reaction (PCR) and sequencing of viral genetic material. The predominant hantavirus was El Moro Canyon virus (ELMCV), which occurred in three of three harvest mice (Reithrodontomys megalotis) and in three of four deer mice (Peromyscus maniculatus) examined. Sin Nombre virus (SNV) was found in one deer mouse and one white-footed mouse (P. leucopus). A seropositive house mouse (Mus musculus) was negative by PCR. Two cotton rats (Sigmodon hispidus) were infected by a virus of novel genotype (Muleshoe virus [MULEV]) that bears closet resemblance to Bayou hantavirus. The sequence of the complete small genomic segment was determined for one MULEV, and high-level expression of its nucleocapsid protein was induced in Escherichia coli. Serologic studies indicated that the most likely etiologic agent in the human infection was SNV.

High prevalence of hantavirus infection in Indian communities of the Paraguayan and Argentinean Gran Chaco.

Serologic evidence of past infection with a Sin Nombre-like hantavirus(es) was demonstrated in 78 (40.4%) of 193 Indians living in western Paraguay and in 38 (17.1%) of 222 Indians inhabiting the Salta province of northern Argentina. In both populations seroprevalence increased with age, with the most striking increase occurring at 18 years of age in the Paraguayan population and at 35 years of age in the Salta population. The peak prevalences in both populations (66.6% and 44.0%, respectively) were seen in Indians > 53 years old. Although no sex difference was observed in the Paraguayan Indians, in the Salta population seroprevalence was greater in males than in females. Familiar clustering of the infection was observed. The data indicate that the Indian populations of the Gran Chaco are frequently exposed to and survive infection with a Sin Nombre-like virus(es). Possible explanations of this novel epidemiology are discussed.

Dysregulation of IL-4 expression in lepromatous leprosy patients with and without erythema nodosum leprosum.

In order to increase our understanding of the immunological basis of erythema nodosum leprosum (ENL), we studied Th-like cytokine profiles in 130 leprosy patients, employing both the conventional and a novel, real-time, fluorogenic reverse transcriptase-based PCR (RT-PCR). The concomitant expression of both Th-like cytokines, interferon-gamma and IL-4, and the regulatory cytokines, IL-10 and IL-12, was studied in the peripheral blood cells of leprosy patients with and without ENL. In the conventional RT-PCR, varied cytokine profiles were observed in individual patients of all clinical types. Fifty-three percent of lepromatous patients without ENL and 59% of tuberculoid leprosy patients showed co-expression of IFN gamma and IL-4, indicating a non-polarized Th 0 pattern. Of the 36 patients with ENL, 58% demonstrated a polarized Th 1 pattern, with only 30% expressing both cytokines. Semiquantitative RT-PCR indicated a lower expression of IL-4 compared to that of IFN gamma in the lepromatous patients without ENL; the difference was even greater among those with ENL. The sensitive, real-time PCR confirmed the down-regulation of IL-4 and IL-10, with absence of IL-4 in half of the patients, resulting in skewing of the cytokine response toward a Th 1-like profile.

Will a vaccine to nasopharyngeal carcinoma retain orphan status?

Epstein-Barr virus (EBV) infection is associated with infectious mononucleosis (IM) and with a number of human malignancies including nasopharyngeal carcinoma (NPC), Hodgkin's disease (HD) and immunoblastic lymphoma (IL). Their potential for immunotherapeutic treatment by cytotoxic T cells (CTL) is dependent on the degree of EBV antigen expression, with the best prospect revolving around IM where a vaccine is under development and IL of transplant patients where adoptive transfer of in vitro reactivated CTL has already been demonstrated to be effective. The opportunities for effective immunotherapy in the treatment of NPC is reduced since the available targets are limited to relatively non-immunogenic proteins. Perhaps more importantly, the development of immunotherapeutics is not considered a realistic commercial proposition. The best chance of developing an effective vaccine is to exploit the similarities in phenotype between HD and NPC since a vaccine to the former is likely to have more commercial appeal.

Management of hydrocephalus.

The present study is an analysis of 747 patients with hydrocephalus, treated and followed up in the Hydrocephalus Clinic run by the department of Paediatric Surgery at the All India Institute of Medical Sciences, New Delhi. The distribution of patients was: congenital-46%, post-meningomyelocoele excision-28%, post-meningitic-21% and others-5% (including post haemorrhagic and post encephalocoele excision hydrocephalus. The average age was 7 months in the shunted group and 10 months in the medical group with overall male to female ratio of 2.3:1. The data were analysed to study the effect of treatment on ventriculomegaly and mental development with special reference to the type of treatment (shunt versus medical) and age at starting treatment. The probability of shunt failure was also studied. A comparison of ventricular size in US/CAT scans between the time of starting treatment and last follow-up revealed improvement in ventriculomegaly in 60% of the shunted patients but only 30% of the medically treated patients. A significant difference was particularly noted in patients with severe hydrocephalus, 72% and 22%, respectively. Comparison of the mean Mental Performance Quotient (MPQ) scores in the shunted & medically treated patients also revealed significantly better MPQ scores in the shunted group (p = < 0.001). Probability of shunt survival, as depicted by the Kaplan-Meier survival curve, revealed that there is a high rate of shunt failure in the first 12 months, followed by a dramatic slowing down. Our observations support the contention that CSF shunt surgery offers better outcome than medical management even when ventriculomegaly is severe at the time of presentation.

Decidualized pseudopregnant rat uterus shows marked reduction in Ang II and Ang-(1-7) levels.

UNLABELLED: Previous studies showed that angiotensin (Ang) II and Ang-(1-7) concentrations were reduced in the implantation site at day 7 of pregnancy in Sprague-Dawley rats as compared to the site immediately adjacent to it, which does not have the embryo attached, clearly showing the importance of the blastocyst in the regulation of renin-angiotensin system (RAS). OBJECTIVE: The objective of this study was to evaluate the regulation of the RAS in the decidualized uterus in the pseudopregnant rat, a model without the presence of a conceptus. METHODS: Ovariectomized, adult female rats were sensitized for the decidual cell reaction with steroid treatments; decidualization was induced by oil-injection of the right horn; the left horn served as a control. The uterine content of Ang I, Ang II, and Ang-(1-7) was examined in the decidualized and non-decidualized uteri. RESULTS: Both Ang-(1-7) and Ang II and ACE and ACE2 mRNA were significantly reduced in the decidualized horn as compared to the non-decidualized horn. Immunocytochemical characterization of Ang II, Ang-(1-7), ACE and ACE2 demonstrated that Ang-(1-7), Ang II, and ACE2 polarize to the anti-mesometrial pole with decidualization. CONCLUSION: The decidualization process elicits marked reduction in uterine Ang II and Ang-(1-7) content as compared to the non-decidualized horn. The differential immunocytochemical expression of Ang II and Ang-(1-7) with ACE2, but not ACE in the anti-mesometrial pole of the decidualized horn may favor the formation and action of Ang-(1-7) in the anti-mesometrial pole, an area which plays a role in triggering the decidualization process.

Markers and risk factors for HCV, HBV and HIV in a network of injecting drug users in Melbourne, Australia.

BACKGROUND AND AIMS: Current injecting drug users (IDU) in major street drug markets within greater Melbourne were recruited to a longitudinal study on blood borne viruses. Here we investigated risk factors for hepatitis C virus (HCV), hepatitis B virus (HBV) and HIV infection in these IDU at the time of their recruitment. METHODS: Three hundred and eighty-two IDU completed detailed questionnaires on their drug use and risk behaviours, and provided blood samples for serology testing. These data were analysed using univariate and multivariate techniques. RESULTS: The overall prevalence of exposure to HCV, HBV and HIV was estimated at 70%, 34% and <1%, respectively. Independent predictors of HCV exposure were history of imprisonment (RR 1.34, 95% CI 1.19-1.52), use of someone else's needle or syringe (RR 1.23, 95% CI 1.07-1.42), >7.6years length of time injecting (RR 1.21, 95% CI 1.07-1.37), and originating from Vietnam (RR 1.12, 95% CI 1.07-1.18). Independent predictors of HBV exposure were HCV exposure (RR 2.15, 95% CI 1.35-3.43), >7.6years length of time injecting (RR 1.57, 95% CI 1.17-2.13) and originating from outside Australia (RR 1.60, 95% CI 1.22-2.10). Neither prison- nor community-applied tattoos predicted HCV or HBV exposure. Up to 31% of IDU who injected for 1year or less were HCV antibody positive, as were 53% of those who injected for 2years or less. CONCLUSIONS: Ongoing engagement with young IDU, through the provision of harm reduction education and resources, is critical if we are to address blood borne viral infections and other health and social harms associated with injecting drug use.

Allelic variations in CYP2D6 gene and susceptibility to cervical cancer.

Epidemiological studies have identified a number of risk factors that contribute to the development of cervical cancer precursors and cervical cancer. These include infection with certain oncogenic types of human papillomaviruses (HPVs) and other socio-economic factors. Tobacco smoking is an independent risk-factor for cervical neoplasia. It has been found that polymorphism at loci that encode carcinogen-metabolizing enzyme such as cytochrome P450 2D6 (CYP2D6) catalyzing the detoxification of carcinogens may determine susceptibility to cervical cancer. Therefore, it is likely that an understanding of these allelic differences is important for determining an individual's risk of cancer and susceptibility to potentially toxic agents. The aim of the present study was to elucidate the role of CYP2D6 polymorphism and susceptibility to squamous cell carcinoma of the uterine cervix in Indian population. Therefore, the genotype frequencies at this locus in females suffering with low-grade CIN, high-grade CIN and squamous cell carcinoma were compared. The control group consisted of 77 females with normal cervical cytology and the cases comprised of 61 mild/moderate dysplasia, 48 severe dysplasia and 45 cases of squamous cell carcinoma of uterine cervix. The individuals were divided into poor metabolizers (PM) and extensive metabolizers (EM) on the basis of their ability to metabolize certain drugs and carcinogens. Comparison of the frequency distribution for the combination of CYP2D6 EM genotype and smoking between mild/moderate and severe dysplasia was statistically significant (p=0.047) suggesting that women with cervical intraepithelial neoplasia I/II (CIN I/ CIN II) and CYP2D6 EM genotype who smoke appears to have more chances for the lesions to progress to CIN III. Whereas, frequency distribution for the same combination between severe dysplasia and squamous cell carcinoma failed to attain any statistical significance suggesting that CIN III with CYP2D6 EM genotype has less chance to progress to cervical cancer. Increased frequency of CYP2D6 EM and tobacco smoking show strong association with CIN III, indicating that not all lesions with the histopathological high grade CIN are premalignant. Conversely some squamous cell carcinomas may not be preceded by CIN.

Myelin P0 glycoprotein and a synthetic peptide containing the palmitoylation site are both autoacylated.

P0, the major protein of the PNS myelin, is palmitoylated at the cytoplasmic Cys153. To gain insights into the mechanism of P0 acylation, the in vitro palmitoylation of both P0 and a synthetic Cys153-containing octapeptide was studied. Incubation of PNS myelin membranes or isolated P0 with [3H]palmitoyl-CoA resulted in specific labeling of this protein, suggesting that the reaction is nonenzymatic. Incorporation of the labeled fatty acid into P0 was not affected by boiling the isolated P0 for 15 min before incubation or by adding sciatic nerve homogenate to the reaction mixture, which confirms the nonenzymatic nature of the reaction. After chemical deacylation, P0 was palmitoylated at a higher rate, suggesting that the original site was reacylated. Furthermore, tryptic digestion and peptide mapping showed that the same sites are acylated in vitro as in nerve slices indicating that the reaction has physiological significance. On incubation with [14C]palmitoyl-CoA, the synthetic peptide encompassing the natural P0 acylation site (I150RYCWLRR157) was also spontaneously acylated at the cysteine residue. Thus, the integrity of the protein is not required for the nonenzymatic transacylation reaction. At pH 7.4 and 37 degrees C, peptide palmitoylation followed a second-order reaction (k2 = 246 +/- 6 M-1 min-1) and is likely a bimolecular nucleophilic substitution with the peptide thiolate attacking the highly reactive thioester bond in palmitoyl-CoA. The activation energy calculated from the Arrhenius plot is approximately 2 kcal/mol and much lower than that of enzyme-catalyzed transacylations. Finally, two other P0 peptides (V121PTRYG126 and K109TSQVTL115) as well as various unrelated thiol-containing compounds, including cysteine, glutathione, pressinoic acid (CYFQNC), and crustacean cardioactive peptide (PFCNAFTGC), were not autoacylated. These results indicate that the IRYCWLRR peptide represents a particular structural motif and/or has some chemical features that allow the reaction to occur spontaneously.

Morphological changes in the testis and epididymis of camels (Camelus dromedarius). Part I.

The normal anatomy and morphological changes in the testis and epididymis, due to age and seasonal variations, were studied in camels (Camelus dromedarius) between 4 and 20 years of age. The testes had a dark gray colour and were situated in the perineum. The weight ranged from 32 to 225 g. The length and maximum cross-diameter were 6--13 and 3--6.5 cm, respectively. The maximum weight of the left testis in 12-year-old camels during the winter months (December to March) was 225 g, and the minimum weight in the camels of the same age group during the summer months (April to July) was 181 g. The seasonal influence on the weight of the testis was very profound in the camels of 9--14 years. The weight of the epidiymis ranged from 10 to 40 g; their weight increased in all age groups during the winter months.