Molecular prevalence of Anaplasma marginale in ruminants and Rhipicephalus ticks in northern Pakistan.

Anaplasma marginale is the most prevalent tick-borne haemoparasite of cattle and causes huge economic losses to the dairy industry worldwide. This study aimed to determine the occurrence of A. marginale infection in blood and tick samples collected from livestock animals in the districts located in Khyber Pakhtunkhwa (KPK), Pakistan. A total of 184 blood and 370 tick samples were included in this study. It has never been reported that sheep, goats, and cattle in Tank, Ghulam Khan, Birmil and Miran Shah areas were infected with A. marginale. All samples of blood and ticks were collected through random sampling from March 2021 to January 2022 from cattle, sheep and goats and screened through PCR for anaplasmosis by using primer pairs of Anaplasma spp. Three hundred and seventy ticks were collected from infested hosts (120/184, 64.21%). Among the four morphologically identified tick species, the highest occurrence was recorded for Rhipicephalus sanguineus (n=138, 37.29%), followed by Rhipicephalus microplus (n=131, 35.4%), Rhipicephalus annulatus (n=40, 10.81%), Hyalomma anatolicum (n=31, 8.37%), and Hyalomma marginatum (n=30, 8.1%). The occurrence of female tick was highest (n=160, 43.24%), followed by nymphs (n=140, 37.38%) and males ticks (n=70, 18.9%). Among these ticks, A. marginale was detected in female ticks of R. microplus, and R. sanguineus. Molecular identification of A. marginale was confirmed in 120 out of 184 blood samples and 6 out of 74 tick samples. Overall, occurrence of A. marginale in blood and tick samples was found to be 65.21% and 8.1% respectively. Species-wise occurrence in blood samples of goats were 71.11% followed by sheep 68.31% and cattle 50%. Specie-wise occurrence of A. marginale in tick samples of cattle were 12.5% followed by goats 6.89%. The obtained sequence showed similarity with A. marginale reported from Kenya and USA. We report the first PCR based detection of A. marginale infection in blood samples and in R. sanguineus ticks of goats simultaneously.

Molecular prevalence of Anaplasma marginale in ruminants and Rhipicephalus ticks in northern Pakistan

@#Anaplasma marginale is the most prevalent tick-borne haemoparasite of cattle and causes huge economic losses to the dairy industry worldwide. This study aimed to determine the occurrence of A. marginale infection in blood and tick samples collected from livestock animals in the districts located in Khyber Pakhtunkhwa (KPK), Pakistan. A total of 184 blood and 370 tick samples were included in this study. It has never been reported that sheep, goats, and cattle in Tank, Ghulam Khan, Birmil and Miran Shah areas were infected with A. marginale. All samples of blood and ticks were collected through random sampling from March 2021 to January 2022 from cattle, sheep and goats and screened through PCR for anaplasmosis by using primer pairs of Anaplasma spp. Three hundred and seventy ticks were collected from infested hosts (120/184, 64.21%). Among the four morphologically identified tick species, the highest occurrence was recorded for Rhipicephalus sanguineus (n=138, 37.29%), followed by Rhipicephalus microplus (n=131, 35.4%), Rhipicephalus annulatus (n=40, 10.81%), Hyalomma anatolicum (n=31, 8.37%), and Hyalomma marginatum (n=30, 8.1%). The occurrence of female tick was highest (n=160, 43.24%), followed by nymphs (n=140, 37.38%) and males ticks (n=70, 18.9%). Among these ticks, A. marginale was detected in female ticks of R. microplus, and R. sanguineus. Molecular identification of A. marginale was confirmed in 120 out of 184 blood samples and 6 out of 74 tick samples. Overall, occurrence of A. marginale in blood and tick samples was found to be 65.21% and 8.1% respectively. Species-wise occurrence in blood samples of goats were 71.11% followed by sheep 68.31% and cattle 50%. Specie-wise occurrence of A. marginale in tick samples of cattle were 12.5% followed by goats 6.89%. The obtained sequence showed similarity with A. marginale reported from Kenya and USA. We report the first PCR based detection of A. marginale infection in blood samples and in R. sanguineus ticks of goats simultaneously.

Dental students' experience and perceived confidence level in different restorative procedures.

PURPOSE: Graduating confident students who are able to flourish and develop in their future careers is an importance outcome of the dental education. This study aims to evaluate self-perceived level of confidence of fifth-year dental students in different restorative-related tasks and the relationship between their clinical training and its corresponding confidence. METHODS: Fifth-year graduate dental students (n = 202) were asked to fill a questionnaire that rated their level of confidence in different clinical restorative dentistry tasks. Additionally, they were asked about the number of times they performed tasks prior to attempting the equivalent competency and in which sub-specialty they felt the most confident. RESULTS: Completed responses were received from 120 students giving a response rate of 59%. Students felt the most prepared and confident in direct restorations and simple endodontic treatment, and they seemed less confident in indirect restorations and complex endodontic treatment. Regarding the steps of indirect restorations, students expressed least confidence in crown preparation compared to impression making and try-in procedures (P < .05). Students rated their confidence highest in operative dentistry, followed by endodontics and lastly fixed prosthodontics. CONCLUSION: The results highlight the areas in which students exhibit the least confidence. Enhancement of student clinical skills and directed exposure is necessary to raise the level of perceived confidence which will reflect positively on their current and future professional performance.

Esthetic outcome of immediately placed and nonfunctionally loaded implants in the anterior maxilla utilizing a definitive abutment: A pilot clinical trial.

BACKGROUND: Immediate dental implants placement and loading utilizing definitive abutments might save time and cost when an esthetic final result is anticipated. The objective of this prospective clinical trial was to evaluate the esthetic outcome of immediate implantation and immediate nonfunctional loading utilizing definitive abutments, with and without bony substitutes filling the peri-implant gap. MATERIALS AND METHODS: In this clinical trial study a total of 11 implants were placed utilizing a flapless immediate post extraction approach in the maxilla (second premolar to second premolar). Atraumatic extraction was performed and implants were immediately placed. The gap was either left without grafting or filled with particulate bone material. Immediate nonfunctional loading was performed utilizing a definitive abutment. The pink esthetic scores (PESs) were assessed preoperatively, at 1- and 2-year follow-up periods. Dental casts were obtained at respective time intervals; scanned, registered, and closest point distances were measured. For all statistical tests, value of P = 0.05 was set as a statistical significance level. RESULTS: The mean of PES at baseline was 9.4 ± 1.69, at 1 year was 9.5 ± 2.07, at 2 years was 10.2 ± 2.75, for the graft group 10.3 ± 2.8, and for nongrafting group was 10.2 ± 2.59. There were no statistically significant differences in PESs at baseline when compared to 1- and 2-year intervals, and for grafting group versus nongrafting group (P = 0.24). Distances between the two time points for all cases were <1 mm in all reference planes. CONCLUSION: Immediate placement and nonfunctional loading utilizing a definitive abutment appear to result in a stable result as far as esthetic outcome and alveolar process sufficiency are concerned.

A Physically Based Constitutive Model and Continuous Dynamic Recrystallization Behavior Analysis of 2219 Aluminum Alloy during Hot Deformation Process.

The isothermal compression tests of the 2219 Al alloy were conducted at the temperature and the strain rate ranges of 623⁻773 K and 0.01⁻10 s-1, respectively, and the deformed microstructures were observed. The flow curves of the 2219 Al alloy obtained show that flow stress decreases with the increase in temperature and/or the decrease in strain rate. The physically based constitutive model is applied to describe the flow behavior during hot deformation. In this model, Young's modulus and lattice diffusion coefficient are temperature-dependent, and the creep exponent is regarded as a variable. The predicted values calculated by the constitutive model are in good agreement with the experimental results. In addition, it is confirmed that the main softening mechanism of the 2219 Al alloy during hot deformation is dynamic recovery and incomplete continuous dynamic recrystallization (CDRX) by the analysis of electron backscattered diffraction (EBSD) micrographs. Moreover, CDRX can readily occur under the condition of high temperatures, low strain rates, and large strains. Meanwhile, the recrystallization grain size will also be larger.

Bacterial count and predisposing factors of Clostridium perfringens (targeting CPA gene) infection along with antimicrobial sensitivity in diarrheic sheep in Pakistan.

Clostridium perfringens (C. perfringens) is a normal inhabitant in the gut of animals. It may proliferate rapidly in favorable conditions and produces lethal toxins. These toxins may cause lethal effects in the intestines and systemically it may cause enterotoxaemia. In disease conditions, the presence of C. perfringens CFU/g in fecal sample can be of diagnostic value. This study aims to determine the bacterial counts and predisposing factors of C. perfringens (targeting CPA gene) infection in addition to an in-vitro antimicrobial trial in entero-toxemic sheep in Pakistan. A total of 192 diarrheic sheep irrespective of age, gender and breed were selected and the CFU/g was determined from the fecal samples. The study showed that 34.9% of the samples had elevated level of bacterial count compared to the normal (104-107 CFU/g). Out of the total, 7.8% of the samples had subnormal bacterial count (CFU/g), while, 57.3% of the samples showed bacterial counts in the normal ranges. The confirmation of selectively isolated C. perfringens was done by amplification of 324bp CPA gene fragment using polymerase chain reaction (PCR). The in-vitro antimicrobial sensitivity trials showed that penicillin, ciprofloxacin and ceftriaxone are 100% efficacious against C. perfringens, while, bacitracin, ampicillin and amoxicillin were found to be least effective. The key determinants in this study which support the in-vivo growths of C. perfringens were; carbohydrate rich diet and overcrowding with the odds ratios (OR) of 5.44 and 2.26, respectively. This study concludes that C. perfringens is highly prevalent in sheep population of Pakistan. The incidence of enterotoxaemia can be minimized by controlling the factors which enhance its in-vivo growth. The diseased animal associated with elevated C. perfringens levels can be effectively cured using any one of the penicillin, ciprofloxacin and ceftriaxone.

Pressure-induced structural change in liquid GaIn eutectic alloy.

Synchrotron x-ray diffraction reveals a pressure induced crystallization at about 3.4 GPa and a polymorphic transition near 10.3 GPa when compressed a liquid GaIn eutectic alloy up to ~13 GPa at room temperature in a diamond anvil cell. Upon decompression, the high pressure crystalline phase remains almost unchanged until it transforms to the liquid state at around 2.3 GPa. The ab initio molecular dynamics calculations can reproduce the low pressure crystallization and give some hints on the understanding of the transition between the liquid and the crystalline phase on the atomic level. The calculated pair correlation function g(r) shows a non-uniform contraction reflected by the different compressibility between the short (1st shell) and the intermediate (2nd to 4th shells). It is concluded that the pressure-induced liquid-crystalline phase transformation likely arises from the changes in local atomic packing of the nearest neighbors as well as electronic structures at the transition pressure.

Trends in the lifetime risk of developing cancer in Great Britain: comparison of risk for those born from 1930 to 1960.

BACKGROUND: Typically, lifetime risk is calculated by the period method using current risks at different ages. Here, we estimate the probability of being diagnosed with cancer for individuals born in a given year, by estimating future risks as the cohort ages. METHODS: We estimated the lifetime risk of cancer in Britain separately for men and women born in each year from 1930 to 1960. We projected rates of all cancers (excluding non-melanoma skin cancer) and of all cancer deaths forwards using a flexible age-period-cohort model and backwards using age-specific extrapolation. The sensitivity of the estimated lifetime risk to the method of projection was explored. RESULTS: The lifetime risk of cancer increased from 38.5% for men born in 1930 to 53.5% for men born in 1960. For women it increased from 36.7 to 47.5%. Results are robust to different models for projections of cancer rates. CONCLUSIONS: The lifetime risk of cancer for people born since 1960 is >50%. Over half of people who are currently adults under the age of 65 years will be diagnosed with cancer at some point in their lifetime.

Predictors of paravalvular aortic regurgitation following self-expanding Medtronic CoreValve implantation: the role of annulus size, degree of calcification, and balloon size during pre-implantation valvuloplasty and implant depth.

OBJECTIVES: We sought to investigate the role of balloon size during pre-implantation valvuloplasty in predicting AR and optimal Medtronic CoreValve (MCS) implantation depth. BACKGROUND: Paravalvular aortic regurgitation (AR) is common following MCS implantation. A number of anatomical and procedural variables have been proposed as determinants of AR including degree of valve calcification, valve undersizing and implantation depth. METHODS: We conducted a multicenter retrospective analysis of 282 patients who had undergone MCS implantation with prior cardiac CT annular sizing between 2007 and 2011. Native valve minimum (Dmin), maximum (Dmax) and arithmetic mean (Dmean) annulus diameters as well as agatston calcium score were recorded. Nominal and achieved balloon size was also recorded. AR was assessed using contrast angiography at the end of each procedure. Implant depth was measured as the mean distance from the nadir of the non- and left coronary sinuses to the distal valve frame angiographically. RESULTS: 29 mm and 26 mm MCS were implanted in 60% and 39% of patients respectively. The majority of patients (N=165) developed AR <2 following MCS implantation. AR ≥3 was observed in 16% of the study population. High agatston calcium score and Dmean were found to be independent predictors of AR ≥3 in multivariate analysis (P<0.0001). Nominal balloon diameter and the number of balloon inflations did not influence AR. However a small achieved balloon diameter-to-Dmean ratio (≤0.85) showed modest correlation with AR ≥3 (P=0.04). This observation was made irrespective of the degree of valve calcification. A small MCS size-to-Dmean ratio is also associated with AR ≥3 (P=0.001). A mean implantation depth of ≥8+2mm was also associated with AR ≥3. Implantation depth of ≥12 mm was associated with small MCS diameter-to-Dmean ratio and increased 30-day mortality. CONCLUSION: CT measured aortic annulus diameter and agatston calcium score remain important predictors of significant AR. Other procedural predictors include valve undersizing and low implantation depth. A small achieved balloon diameter-to-Dmean ratio might also predict AR ≥3. Our findings confirm that a small achieved balloon size during pre-implantation valvuloplasty predicts moderate-severe AR in addition to previously documented factors.

PGD2 DP1 receptor stimulation following stroke ameliorates cerebral blood flow and outcomes.

Stroke is a leading cause of death and morbidity worldwide, yet effective treatments are lacking. The association of prostaglandin D2 and its DP1 receptor with vasculature and blood propelled us to examine whether the clinically tested DP1 receptor agonist BW245C had beneficial effects following stroke. To determine if BW245C affects basal cerebral blood flow (CBF), C57BL/6 WT and DP1(-/-) mice were given a single i.p. injection of vehicle or BW245C, and CBF was recorded for 2h. To test the effect of BW245C on stroke, WT and DP1(-/-) mice were subjected to middle cerebral artery occlusion followed by a single i.p. injection of vehicle or 0.02, 0.2, or 2.0-mg/kg BW245C immediately before reperfusion. Functional and anatomical outcomes were determined at 96h. We also determined the effect of BW245C on CBF in peri-infarct and core during occlusion and reperfusion. Furthermore, we tested the effect of BW245C on bleeding time and ex vivo coagulation. BW245C treatment increased the basal CBF significantly in WT but not in DP1(-/-) mice. The BW245C treatment also significantly improved functional outcome and lowered infarction volume. The multisite CBF monitoring by laser-Doppler flowmetry shows that BW245C significantly increased the CBF in peri-infarct, with a significant inverse correlation between infarction and CBF. The significantly higher infarction volume in DP1(-/-) mice remained unchanged with BW245C treatment. Moreover, BW245C preserves hemostasis in non-stroke conditions. Combined, these data suggest that the DP1 receptor is an endogenous target that can rescue the brain following stroke by regulating CBF and hemostasis.

Identification of MAGEA antigens as causal players in the development of tamoxifen-resistant breast cancer.

The antiestrogen tamoxifen is a well-tolerated, effective treatment for estrogen receptor-α-positive (ER+) breast cancer, but development of resistance eventually limits its use. Here we show that expression of MAGEA2, and related members of this cancer-testis antigen family, is upregulated in tamoxifen-resistant tumor cells. Expression of MAGEA2 in tumor lines grown in vitro or as xenografts led to continued proliferation in the presence of tamoxifen. At the molecular level, we demonstrate that MAGEA2 protein localizes to the nucleus and forms complexes with p53 and ERα, resulting in repression of the p53 pathway but increased ER-dependent signaling. In a series of ER+, tamoxifen-treated breast cancer patients, we show a highly significant (P=0.006) association between MAGEA (melanoma-associated antigen) expression and reduced overall survival, confirming the clinical significance of our observations.

Association between DNA methylation of HSPB1 and death in low Gleason score prostate cancer.

BACKGROUND: Heat shock protein 27 (Hsp-27) encoded by gene HSPB1 is a critical regulator of the behavioral phenotype of human prostate cancer (PCa) cells, enhanced expression being associated with highly aggressive disease and poor clinical outcome. In contrast, the protein is not expressed in PCas of low malignant potential. To gain insight into the mechanism regulating its expression, we tested the hypothesis that differential methylation of CpG islands within HSPB1 controls transcription and subsequent translation of the gene. METHODS: We studied prostate epithelial cell lines and tissue biopsies, including 59 BPH and 415 PCas, of which 367 were a cohort of men with up to 20 years of follow-up. Methylation across the gene (DNA methylation (DNAme)) was assayed by pyrosequencing. Hsp-27 expression was assessed by western blot and immunohistochemistry. RESULTS: In cancer tissues, methylation increased in a 3' direction (P < 0.0001) whereas in benign hyperplasia methylation was constantly below 5%, a cutoff giving a specificity of 100% and sensitivity of 50%. Although methylation of the promoter region was significantly discriminating between benign and malignant prostatic epithelia, it compared poorly with methylation of the first intron. The prognostic value of HSPB1 DNAme was confirmed by both univariate (hazard ratio 1.77 per 50% increment, P = 0.02) and multivariate models. Interaction between HSPB1 methylation and Gleason score revealed high DNAme to be a reliable prognostic marker of poor outcome in men with low Gleason score (P = 0.014). CONCLUSIONS: Our data indicate CpG methylation of the first HSPB1 intron to be an important biomarker that identifies aggressive PCas otherwise regarded as low risk by current clinical criteria but that, biologically, require immediate active management.

Hypertensive disorders in pregnancy and fetal death at different gestational lengths: a population study of 2 121 371 pregnancies.

OBJECTIVE: To compare the proportion of offspring that was stillborn in pregnancies with pre-eclampsia, gestational hypertension or chronic hypertension with those in normotensive pregnancies. DESIGN: Register-based observational study. SETTING: The Medical Birth Registry of Norway. POPULATION: All singleton births after 20 completed weeks of gestation in Norway from 1967 to 2006 (n = 2 121 371). METHODS: The proportion of stillborn offspring was estimated in normotensive pregnancies, and in pregnancies with pre-eclampsia, gestational and chronic hypertension at different gestational lengths. In addition, changes in the proportions of stillborn offspring by maternal hypertensive disorder from 1967-1986 to 1987-2006 were estimated. MAIN OUTCOME MEASURES: Fetal death. RESULTS: The prevalence of hypertensive disorders in pregnancy was 4.7%. In total, 17 933 fetal deaths occurred and 9.2% of these were in hypertensive pregnancies. In normotensive pregnancies, 0.8% (16 290/2 022 400) experienced fetal death. This was true for 1.9% (1170/62 261) of the pregnancies with pre-eclampsia, 1.2% (390/32 068) with gestational hypertension and 1.8% (83/4642) with chronic hypertension. There was a 44% overall reduction in fetal death rate from 1967-1986 to 1987-2006. The largest decline was in women with pre-eclampsia (80% reduction). In women with gestational hypertension and chronic hypertension, the overall reductions in fetal death rates were 49% and 57%, respectively, comparable with the 41% decline in normotensive pregnancies. CONCLUSIONS: In our nationwide study during 1967-2006, the risk of fetal death among women with hypertensive disorders in pregnancy has been greatly reduced, especially among pre-eclamptic women at term. The risk of fetal death among women with gestational or chronic hypertension has also decreased, but in a different manner.

Cancer incidence in the United Kingdom: projections to the year 2030.

BACKGROUND: Projections of cancer incidence are important for planning health services and to provide a baseline for assessing the impact of public health interventions. METHODS: Rates estimated from smooth function age-period-cohort modelling of cancer incidence data from Great Britain 1975 to 2007 are extrapolated to 2030 and applied to UK population projections. Prostate and breast cancer projections take into account the effect of screening. RESULTS: Overall rates of cancer are projected to be stable over the next 20 years, but this masks individual changes. In both sexes, age-standardised rates of cancers of the stomach, larynx, bladder and leukaemia are projected to fall by ≥1% per year, whereas cancers of the lip, mouth and pharynx (ICD-10 C00-C14) and melanoma are projected to increase by ≥1% per year. The growing and aging populations will have a substantial impact: numbers of cancers in men and women are projected to increase by 55% (from 149,169 to 231,026) and 35% (from 148,716 to 200,929), respectively, between 2007 and 2030. The model used yields similar results to those of Nordpred, but is more flexible. CONCLUSION: Without new initiatives for smoking and obesity reduction, the number of cancers in the United Kingdom will increase substantially reflecting the growing and aging populations.

Interleukin 13 receptors as biochemical markers in atopic patients.

BACKGROUND: Interleukin (IL) 13, a type 2 helper T cell (T(H)2), is an important regulator of inflammatory immune responses. It mediates its action through a receptor complex consisting of IL-13Ralpha1 and IL-4Ralpha. IL-13Ralpha2 binds IL-13 with high affinity and is thought to act primarily as a decoy receptor, sequestering IL-13 and thus inhibiting its action. Our aim was to clarify the role of these receptors in the diagnosis and follow-up of atopic patients. METHODS: We genotyped the 1398A>G polymorphism in the IL-13Ralpha1 gene using restriction fragment length polymorphism for causal genetic diversity and measured serum levels of IL-13Ralpha2 in 105 atopic patients suffering from atopic asthma, atopic dermatitis, and atopic rhinitis (35 each). We compared the results with those of 35 nonatopic control individuals. Total immunoglobulin (Ig) E and serum IL-13Ralpha2 were measured using enzyme-linked immunosorbent assay, and the eosinophil counts were recorded. RESULTS: A significant increase in serum IL-13Ralpha2 levels was recorded in the 3 atopic groups compared with the control group (P < .001), as well as a significant increase in total IgE levels and eosinophil counts. No significant association was found between 1398A>G and atopy other than a suggestive association between this polymorphism and raised total serum IgE levels in all 3 atopic groups (P < .001). CONCLUSIONS: These findings indicate that IL-13Ralpha2 plays an important role in atopy and that increased levels in different groups highlight its regulatory role in the development of atopic symptoms. The 1398A>G polymorphism might be involved in the production of IgE.

Interleukin 10 receptor alpha subunit (IL-10RA) gene polymorphism and IL-10 serum levels in Egyptian atopic patients.

OBJECTIVES: To assess the value of serum interleukin (IL) 10 levels as an immunological marker in atopy and to determine the role of an IL-10RA gene single nucleotide polymorphism (SNP) (serine 138-to-glycine exchange [S138G]) in the pathogenesis of atopic diseases. METHODS: Seventy-five patients with atopic disorders were compared with 25 age-matched healthy volunteers. Serum total immunoglobulin (Ig) E and IL-10 levels were measured by enzyme-linked immunosorbent assay and the IL-10RA gene S138G variant was screened by multiplex allele-specific polymerase chain reaction. RESULTS: There was a significant association between G allele frequencies of the S138G variant (62%, 60% and 68% for atopic asthma, atopic dermatitis, and allergic rhinitis, respectively) in atopic patients compared to in controls. There were significant differences in mean IgE levels but not mean serum IL-10 levels between the allelic variants in atopy groups. CONCLUSION: The IL-10RA gene SNP S138G may contribute to susceptibility to atopic diseases but serum IL-10 level is not a sensitive indicator in atopy.

Chemical composition of the stem bark and leaves of Ficus pandurata Hance.

A new compound, 3-O-alpha-L-arabinopyranosyl-4-hydroxybenzoic acid (13), in addition to 16 newly reported compounds: alpha-amyrin acetate (1), beta-amyrone (2), 3beta-acetoxy-20-taraxasten-22-one (3), alpha-amyrin (4), ceryl alcohol (5), stigmasterol (6), beta-sitosterol (7), 2alpha,3alpha-dihydroxy-lup-20(29)-en-28-oate (8), ursolic acid (9), beta-sitosterol-3-O-glucosoide (10), protocatechuic acid (11), betulinic acid (12), quercetin (14), quercetin-3-O-beta-D-glucoside (15), kampferol-3-O-beta-neohesperidoside (16) and rutin (17) were isolated from the stem bark and leaves of Ficus pandurata (Hance) cultivated in Egypt. Identification of these compounds has been established by physical, chemical and spectral data (UV, IR, MS, (1)H- and (13)C-NMR), as well as comparison with authentic samples.

Interferon gamma gene polymorphism as a biochemical marker in Egyptian atopic patients.

OBJECTIVES: The aim of this study was to clarify the role of interferon (IFN) gamma in the diagnosis and follow-up of atopic patients. We genotyped the IFN-gamma polymorphism at position +874 to examine the relationship between serum levels of IFN-gamma and disease severity and the role of IFN-gamma as a biochemical and immunologic marker. METHODS: The study population comprised 75 patients suffering from atopic asthma, atopic dermatitis, and allergic rhinitis (25 each), and 25 control participants. Total immunoglobulin (Ig) E and serum IFN-gamma were measured by enzyme-linked immunosorbent assay, the IFN-gamma polymorphism at position +874 was determined by amplification refractory mutation system-polymerase chain reaction, and eosinophil counts were recorded. RESULTS: There was a significant association between genotype and the frequency of the A allele of the +874T/A polymorphism in atopic patients when compared with controls (P < .001). In all 3 groups, there was a significant increase in total IgE levels and eosinophil counts, and a decrease in serum IFN-gamma levels towards the presence of homozygous AA compared with homozygous TT. CONCLUSIONS: The IFN-gamma gene polymorphism at position +874 contributes to susceptibility to atopic diseases by decreasing the amount of IFN-gamma. Identification of variants of IFN-gamma gene signalling and its role in the development of atopic diseases provides a focus for the development of novel diagnostic and therapeutic strategies for these diseases.

Heme oxygenase-1 protects brain from acute excitotoxicity.

Heme oxygenase is a rate-limiting enzyme that degrades heme, a pro-oxidant, into carbon monoxide, iron, and bilirubin. Heme oxygenase has two active isoforms: heme oxygenase-1 and heme oxygenase-2. Heme oxygenase-1 can be induced by various insults. Several investigators have postulated that it has cytoprotective activities, although its role in the nervous system is not fully understood, especially considering that normally heme oxygenase-2 accounts for the vast majority of heme oxygenase activity in the brain. Here, the basal effect of heme oxygenase-1 was investigated in acute glutamatergic excitotoxicity to test the hypothesis that N-methyl-D-aspartate-induced acute toxicity in brain is attenuated by heme oxygenase-1. N-methyl-D-aspartate was unilaterally injected into the striatum of wildtype and heme oxygenase-1 knockout mice. After 48 h, brains were harvested, sectioned, and stained with Cresyl Violet to measure the lesion size. Lesion volume was significantly (P<0.05) greater in brains of heme oxygenase-1 knockout mice (15.2+/-3.1 mm(3); n=10) than in those of wildtype mice (6.2+/-1.5 mm(3); n=11). In addition, Western blot analysis indicated no detectable differences between wildtype and heme oxygenase-1 knockout mouse brains in the levels of the glutamate or N-methyl-D-aspartate receptors studied. To test whether heme oxygenase-1 could specifically protect neurons, mouse primary neuronal cell cultures of wildtype and heme oxygenase-1 knockout mice were treated with or without N-methyl-D-aspartate. Cell viability of the heme oxygenase-1 knockout neurons was significantly less than that of wildtype neurons at each of the N-methyl-D-aspartate concentrations tested (12.8+/-1.3%, 16.0+/-1.4%, and 18.4+/-1.8% at 30, 100, and 300 microM N-methyl-D-aspartate, respectively). These results indicate that heme oxygenase-1 provides neuroprotection against acute excitotoxicity and suggest that potential intervention that can increase heme oxygenase-1 activity within the brain should be considered as a therapeutic target in acute and potentially chronic neurological disorders.