Mental Health Assessment of Health Care Providers of Bangladesh during Covid-19 Pandemic: A Cross-Sectional Web-Based Study.

SARS COV 2 raised immense agony to everyone. As health care workers remains direct contact with patients, they are vulnerable to mental health disorder. The study was done to identify mental health status of health care providers of Bangladesh. It was a cross sectional web based survey among health care providers of bangladesh. A structured web based questionnaire was prepared both in Bangla and English version for better understanding. Total 2594 health care workers responded to online survey. Male were 1303(50.2%) and female were 1291(49.8%). Occupational demography shows 98.5% were physicians. Mean value of patient health questionnaire 9 (PHQ9) and generalized anxiety disorder (GAD) were 8.43±6.082 and 8.27±5.234 respectively among healthcare providers. To overcome anxiety and depression, 93.4% of HCW didn't receive any psychological material and 85.6% HCW didn't find physiological resources even through media. Despite many lacking, healthcare workers were resilient in our study.

Alleviation of cisplatin-induced hepatotoxicity and nephrotoxicity by L-carnitine.

Objectives: To assess the protective effect of L-carnitine in reducing cisplatin toxicity via estimating biochemical tests, histomorphometric, and immunohistochemistry (IHC) of ß-catenin and cyclin D. Materials and Methods: Fifteen adult male rabbits were used in this study and allocated into 3 groups; Group 1 (Control negative), rabbits of this group were not given any treatment. In group 2, the animals were injected with cisplatin single-dose/per week. Group 3 rabbits were treated with Cisplatin+L-carnitine orally by gavage tube for 29 days. At the end of the experiments, blood samples from all rabbits were taken from the earlobe, and then the biochemical test was done, the kidney and tissue sections were prepared for both H& E and IHC for both ß-catenin and cyclin D genes. Results: Treatment with L-carnitine reduced the injury effect of cisplatin via a decline in serum creatinine, urea, bilirubin, GPT, GOP, and ALP significantly (P<0.05). Also, administration of LC attenuates the histopathologic abnormality in the kidney (15.71% vs 85.18%) and liver (score 3 vs 15 ) induced by cisplatin. L-carnitine elevates the expression of ß-catenin and cyclin D in renal and hepatic parenchyma by diffuse, moderate-strong positivity vs cisplatin that showed local-weak staining. Conclusion: These findings imply that L-carnitine, by its pleiotropic actions in activating Wnt signaling, alleviates cisplatin-induced renal and hepatic destruction. It might be a method of preventing cisplatin-related nephrotoxicity and hepatotoxicity.

Clinical Presentation of Congenital Adrenal Hyperplasia in Children: Experience in a Tertiary Care Hospital of Bangladesh.

This cross sectional study was conducted in Paediatric Endocrine Outpatient Department of BIRDEM General Hospital, a tertiary care centre in Dhaka, Bangladesh among patients diagnosed with congenital adrenal hyperplasia (CAH) from January 2005 to December 2018. The study was aimed to find out the clinical and laboratory profile of all patients at presentation diagnosed with CAH during the study period. Data were obtained by reviewing the medical records of the patients. Total 102 children with CAH were diagnosed during the study period. Among them 68 were female and 34 were male (female to male ratio of 2:1). Median age was 3.5 month (range 0.2-158 month) and 5.5 month (range 1-108 month) in female and male respectively (p=0.42). Family history was available in 93 patients. Consanguinity was present in 16(17.2%), history of sib death in 12(12.9%), other family members were affected in 8(8.6%). Sixty patient (58.8%) had salt-wasting (SW), 39(38.2%) had simple virilizing (SV) and 3(2.9%) had non- classic form of CAH. Median age of presentation was 2 month (range 0.2-70 month) and 42 month (range 0.8-158 month) in SW and SV group respectively (p=0.001) and 119 month (range 108-152 month) in non- classic group. Common presentations were: genital ambiguity (64.7%), vomiting (46.5%), failure to thrive (41.6%), features of early puberty (precocious pseudopuberty) (24.5%), diarrhea (12.0%). Hyperpigmentation was noted in 49.0% of patients. Among the salt-wasting type in male failure to thrive (FTT) was the most common presentation (83.3%), followed by vomiting (75.0%). In female genital ambiguity was the commonest presentation (97.2%), followed by vomiting (77.1%). Among the simple virilizing type in male early puberty was the commonest presentation (100%) and genital ambiguity was the presenting feature in all the female (100.0%).

Prevalence of Sonologically Detected Non-alcoholic Fatty liver disease Among School Children of Sylhet City.

This cross sectional descriptive type of observational study was designed to see the prevalence of NAFLD among school children of Sylhet, Bangladesh from December 2019 to January 2020. School children of three private schools of Sylhet City were randomly selected. Socio-demographic data and anthropometric measurement of them were recorded. Then they underwent screening sonologically for NAFLD. Data analysis was done using SPSS version 20.0. P value <0.05 was taken as significant. Total 174 students, boys 99(56.9%) and girls 75(43.1%) were included. Age of them varied from nine years to 17 years (mean 13.408). In this series 82(47.1%), 52(29.9%) and 40(23.0%) were of normal weight, overweight and obese children respectively. In this study 29(16.7%) children had NAFLD and seven (4.0%) had biliary sludge in gall bladder. NAFLD was significantly higher among children with higher BMI (p=0.00). Female sex, children consuming fast food and playing video games were about two times more prone to develop NAFLD. Biliary sludge was found significantly higher among children consuming fast food (p=0.02). NAFLD among school going children is not uncommon. Higher BMI is a significant risk factor of developing NAFLD. Girls and children consuming fast food and playing video games also are more prone to develop NAFLD. Prevalence of sludge in gall bladder is low among school children.

A Clinical Study on Gallbladder Polyp.

There are different types of gallbladder polyps having variable morphology and nature. Symptoms are vague and mostly diagnosed incidentally on abdominal ultrasound. Risk factors are many. Indications of surgery are not clearly understood due to lack of clinical evidence. This study was carried out to analyze the management of gallbladder polyp in our settings and review of available evidence. Patients who underwent clip less mini laparoscopic cholecystectomy for gallbladder polyp were included. Clinical data including short history, age, gender, parity, lab tests, ultrasound report and macroscopy were recorded during surgery. Histopathology reports collected later. Total of 112 patients with mean age 42.6 years (range 22-82), 74 of whom were female. Most of them (46.42%) were asymptomatic. Single polyp was in 69, multiple in 32 and polyp with stone in 8 patients. Sessile polyp was 74.10%. Average polyp size was 7.6mm. Cholesterol polyp was in 82 patients. Four out of 5 malignant polyps were single and sessile. Abdominal ultrasound is the mainstay of diagnosis of gallbladder polyp but size, location, morphology, number and presence of associated gallstone has to be mentioned. Symptomatic gallbladder polyp benefited from surgery. Polyps more than 6mm, single and sessile morphology above the age 50 should undergo cholecystectomy.

Bovine neonate is deficient in innate immunity at birth.

With an objective to understand acquisition of innate immunity in bovine neonates, we analyzed perinatal expression of cytokine, adhesion molecule and complement component genes involved in innate and adaptive immune functions. Statistically robust transcriptomic analysis of 27 cytokines showed low IL1B, IL2 and IL7 but high IL23, TGFB1 and TGFB2 expression in bovine neonates post-birth. Unlike mice and humans, no TH2 polarizing cytokine expression occurs in bovine neonates. Further, TH17 and Treg differentiation in bovine neonates may differ from other species like mice and humans. Decreased IL7, IL23R, CXCR3 and increased TGFB1 and TGFB2 expression provides an immunosuppressive environment in the bovine neonate at birth. Transcriptomic analysis of 31 adhesion molecules showed rapid increase in ITGAL expression within a week post-birth in bovine neonates that permits acquisition of innate cytotoxic functions by granulocytes (antibody-mediated), cytotoxic T and NK cells. However, innate immune functions involving phagocytosis and platelet aggregation are deficient in bovine neonates at birth. Of twenty-seven, 18 complement component genes show no significant differential gene expression in neonates post-birth. But low expression of C1QA, C1QB, CQC, C1R and C2 compromises classical and lectin complement pathways mediated lytic function in bovine neonates. The complement-mediated cytotoxic functions, however, normalize between days 7 and 28 post-birth. To conclude, bovine neonate is immunosuppressed and deficient in innate immune competence at birth. Such differences with regard to global innate immune deficiency and lack of TH2 polarization in bovine neonates have profound implications for designing vaccines to prevent neonatal infections. To conclude, species-specific unique characteristics of developing innate and adaptive immune system need to be taken into consideration while designing new immunization strategies to prevent neonatal mortality from infections.

Preferential expression of IGHV and IGHD encoding antibodies with exceptionally long CDR3H and a rapid global shift in transcriptome characterizes development of bovine neonatal immunity.

With an objective to understand natural development of bovine neonatal immunity, we analyzed 18 RNA-seq libraries from peripheral blood lymphocytes of three neonatal calves pre- (day 0) and post-colostrum (7, 14 and 28) uptake as compared to their dams. A significant global shift in neonatal transcriptome occurs within first week post-birth, in contrast to dams, with an upregulation of 717 genes. Global pathway analysis of the transcriptome revealed 110 differentially expressed immune-related genes, such as, complement, MHCII, chemokine receptors, defensins and cytokines, at birth. The signaling molecules (LAX1, BLK) and transcription factors (GATA3, FOXP3) are expressed at high levels. High expression of GATA3 transcription factor at birth seems to skew the neonatal immune response towards TH2 type. The high levels of T-cell signaling molecules, CD3G and CD3D, at birth are important in neonatal T cell development. Unlike adults, IGKC expression is high in the neonates where IGKV12 is preferentially expressed at birth. But IGLC is predominant in both neonates and adult where IGLV3.4 is preferentially expressed in B cells at birth. Both IGHM and IGHD are expressed at birth and IGHM achieves adult levels by day 7. This is followed by IGHA and IGHG expression 14-28 days post-birth. Importantly, preferential expression of IGHV1S1(BF4E9) and longest IGHD2(DH2) genes that encode immunoglobulin with exceptionally long CDR3H at birth indicates their critical role, as B cell antigen receptor, in the B cell development via idiotype-anti-idiotype interactions. The transcriptome signatures described here permit assessment bovine neonatal immunocompetence. Bovine neonates acquire innate and IgM-mediated humoral immunocompetence within first week post-birth.

Exceptionally long CDR3H of bovine scFv antigenized with BoHV-1 B-epitope generates specific immune response against the targeted epitope.

We discovered that some bovine antibodies are amongst the largest known to exist due to the presence of an exceptionally long CDR3H (≥49 amino acids) with multiple cysteines that provide a unique knob and stalk structure to the antigen binding site. The large CDR3H size, unlike mouse and human, provides a suitable platform for antigenization with large configurational B-epitopes. Here we report the identification of a B-epitope on the gC envelope protein of bovine herpes virus type-1 (BoHV-1) recognized by a bovine IgG1 antibody. The identified 156 amino acid long gC fragment (gC156) was expressed as a recombinant protein. Subsequently, a functional scFv fragment with a 61 amino-acid long CDR3H (scFv1H12) was expressed such that gC156 was grafted into the CDR3H, replacing the "knob" region (gC156scFv1H12 or Ag-scFv). Importantly, the Ag-scFv could be recognized by a neutralizing antibody fragment (scFv3-18L), which suggests that the engraftment of gC156 into the CDR3H of 1H12 maintained the native conformation of the BoHV-1 B-epitope. A 3D model of gC156 was generated using fold-recognition approaches and this was grafted onto the CDR3H stalk of the 1H12 Fab crystal structure to predict the 3D structure of the Ag-scFv. The grafted antigen in Ag-scFv is predicted to have a compact conformation with the ability to protrude into the solvent. Upon immunization of bovine calves, the antigenized scFv (gC156scFv1H12) induced a higher antibody response as compared to free recombinant gC156. These observations suggest that antigenization of bovine scFv with an exceptionally long CDR3H provides a novel approach to developing the next generation of vaccines against infectious agents that require induction of protective humoral immunity.

Serum magnesium in hospital admitted diabetic patients.

Magnesium depletion has a negative impact on glucose homeostasis and insulin sensitivity in diabetic patients. Low plasma magnesium concentration is a highly specific indicator of poor magnesium status. This case control study was conducted in Mymensingh Medical College Hospital, Bangladesh, from May 2011 to April 2012. This study was done to assess the serum magnesium level in diabetic and non-diabetic patients and to find out the association between serum magnesium, blood glucose and duration of diabetes mellitus. Serum magnesium concentrations were determined in 50 diabetics and 50 age and sex matched non-diabetic patients or attendants for controls. Among the case and control group 32 were male and 18 were female. Mean age of control group was 54.42±2.33 and study group was 55.78±2.43 years. Duration of diabetes of case group in which 50% of patients have diabetes for up to 4 years, 18% of patients have diabetes for 5-9 years, 5% of patients have for 10-14 years and 2% of patients have diabetes for 15 years and above. Serum magnesium concentrations below the normal reference range for study group was 44% and control group was 6%. The mean serum magnesium level of study group was 0.70±0.01mmol/L and that of control group was 0.83±0.02mmol/L. So, the difference in serum magnesium level between two groups were statistically highly significant (p = 0.001). Correlation between FBS and SML of study group, which is negative (r = - 0.182). There is opposite relation among the characteristics as r is negative. And also correlation between 2HAFB and SML of study group, which is negative (r = - 0.06). There is opposite relation among the characteristics as r is negative.

Hypertriglyceridemia associated with eruptive xanthomas and lipemia retinalis in newly diagnosed diabetes mellitus.

Severe hypertriglyceridemia can cause the deposit of lipids in the dermis (eruptive xanthomas) and in the retina (lipemia retinalis). Lipemia retinalis and eruptive xanthoma are occasionally seen in patients with diabetes mellitus and are thought to be due to abnormalities in the serum lipid fractions. Here a 15 year's old girl presented with typical symptoms of diabetes with skin lesions over hands and feet. On examination skin lesions were non-tender yellow papules with creamy-colored centers on extensor surfaces of the arms, hands and feet. Ophthalmoscopic examination showed creamy white retinal vessels with a faded pinkish white retinal back ground both in the periphery and posterior pole of the retina. Laboratory findings showed a grossly lipemic serum with markedly elevated serum levels of triglycerides 8869mg/dl cholesterol 498mg/dl. Her fasting blood glucose was 20.8mmol/l, 2 hours after breakfast was 50.5mmol/l and HbA1c was 14.6%. Eruptive xanthomas and lipaemia retinalis can occur in primary and secondary hyperlipoproteinemias, therefore a careful personal and family history as well as laboratory investigations is recommended in order to detect an underlying cause.

Cataract in children and adolescents with fibrocalculous pancreatic diabetes.

Cataracts have been infrequently reported in Fibrocalculous pancreatic diabetes (FCPD) which is a form of diabetes secondary to chronic, non-alcoholic pancreatitis in tropical countries. A retrospective study was carried out to find out frequency of cataracts in children and adolescents with FCPD admitted in the Paediatric Unit at Bangladesh Institute of Research and Rehabilitation on Diabetes, Endocrine and Metabolic disorders (BIRDEM). A total of 83 patients with FCPD were admitted during the study period of 2000 to 2007. Among them twenty patients (24%) had bilateral cataracts who were enrolled in the study. Mean age at the onset of diabetes was 13.9±1.7 years. There was female preponderance. Ninety percent patients were girls. There was longer duration of symptoms in all patients. Mean duration of symptoms before diagnosis was 15.0±10.2 months. All patients had hyperglycaemia, mean HbA1c value was 18.4±5.1. Most of the patients had snowflake type cortical deposits and posterior subcapsular cataract.

Prevalence of impaired glucose tolerance among children and adolescents with obesity.

The study was undertaken to see the prevalence of impaired glucose tolerance among children and adolescents with obesity, attending the Paediatric Endocrine OPD, Bangladesh Institute of Research and Rehabilitation in Diabetes, Endocrine and Metabolic disorders (BIRDEM), Dhaka, Bangladesh. A cross sectional study from January 2006 to December 2008 was conducted among obese children and adolescents (6-18 years). Children with any other endocrine disorder, dysmorphism/syndrome were excluded. Obesity was defined as BMI ≥ 95th percentile for age and sex using CDC growth chart. Children underwent two hours oral glucose tolerance test with 1.75 gm/kg or 75 gm of glucose, anthropometric and blood pressure measurement. Fasting serum insulin and lipid profile were measured. Impaired glucose tolerance (IGT) was defined as fasting plasma glucose (FPG) <7 mmol/L and 2 hours post glucose load ≥ 7.8 mmol/L to <11.1 mmol/L. Diabetes mellitus (DM) was defined as FPG ≥ 7 mmol/L or 2 hours post glucose load ≥ 11.1 mmol/L. Homeostasis model assessment was used to estimate insulin resistance. A total of 161 children presented with obesity. Male to female ratio was 1.3:1. Mean age was 10.3 ± 2 .5 years. Mean BMI was 27.86 ± 4.1 kg/m². IGT was found in 16.9% of children and adolescents. In children aged 6-10 years IGT was detected in 16.1% and in adolescents aged 11-18 years IGT was detected in 20%. Diabetes mellitus was detected in 2.1% of subjects, all were adolescents. Serum fasting insulin and hip circumference were significantly higher among children and adolescents with IGT compared to that of normal glucose tolerance. The high rate of IGT among obese children and adolescents is of concern. Factors contributing towards obesity needs to be identified and strategies should be planned for prevention and management of this health problem.

Enhanced bovine herpesvirus type 1 neutralization by multimerized single-chain variable antibody fragments regardless of differential glycosylation.

Single-chain variable antibody fragments (scFvs) with a 2-amino-acid linker capable of multimerization as di-, tri-, or tetrabodies that neutralize bovine herpesvirus type 1 (BoHV-1) in vitro were constructed and expressed in Pichia pastoris. In contrast to the monomeric form, multimeric scFvs had a higher virus neutralization potency, as evidenced by a 2-fold increase in their ability to neutralize BoHV-1 due to avidity effects. Mass spectrum (quadrupole time of flight [Q-TOF]) analyses of multimeric scFv demonstrated extensive heterogeneity due to differential cleavage, variable glycosylation (1 to 9 mannose residues), and the incorporation of minor unidentified adducts. Regardless of the differential glycosylation patterns, the scFvs recognized non-gB or -gE target viral epitopes in the BoHV-1 envelope fraction in a Western blot and also neutralized BoHV-1 in infected Madin-Darby kidney (MDBK) cells in vitro. Indirect evidence for the noncovalent multimerization of scFv was the presence of a major peak of multimerized scFv without a His tag (due to differential cleavage) in the Q-TOF profile, unlike monomeric scFv, which copurified with normally His-tagged scFv and recognized the target antigen. Overall, differentially glycosylated recombinant scFvs against BoHV-1 with a short linker (2 amino acids) are capable of assembly into functional multimers that confer high avidity, resulting in increased virus neutralization in vitro compared to that of monovalent scFv with a long (18-amino-acid) flexible linker. Overall, recombinant multimerized scFv5-2L potentially provides a high-potency therapeutic and immunodiagnostic reagent against BoHV-1, which is suitable for passive immunization and topical application.

Epidermal growth factor receptor expression in mice skin upon ultraviolet B exposure - Seborrheic Keratosis as a coincidental and unique finding.

BACKGROUND: Ultraviolet B (UVB) is the most damaging component of sunlight. It rapidly activates the epidermal growth factor receptor (EGFR) and generates reactive oxygen species (ROS) in excessive quantities that quickly overwhelm tissue antioxidants. SETTING AND DESIGN: To demonstrate the effects of UVB radiation on EGFR expression in mice skin and to evaluate the role of antioxidants in the exposed group. MATERIALS AND METHODS: After obtaining the approval of the ethical committee, forty mice from BALB/c strain were used in this experiment and were allocated into 3 groups; 10 (control group); 15 (exposure group); and 15 (exposed and treated with antioxidants). Antioxidants were administered through subcutaneous injection. Skin biopsies from all groups were stained with EGFR antibodies. Total antioxidant status (TAS) was evaluated in all groups. STATISTICAL ANALYSIS: The data obtained were analyzed using ANOVA, Duncan's test, and Pearson's Correlation. RESULTS: The highest EGFR expression in exposure group was of score 3(+) (53%). The highest EGFR expression in treatment group was score 0 (40%). Apoptotic bodies and dermal mast cells increased in exposure group while decreased in treatment group. The mean values for TAS were measured for each group; control group = 1.2 mmol/l; exposure group = 0.87 mmol/l; treatment group =1.3 mmol/l. CONCLUSIONS: UVB led to Seborrheic Keratosis (SK) in mice through enhancement of EGFR expression. Antioxidants effectively reduced UVB-induced SK, reduced epidermal changes, apoptotic bodies, and decreased dermal mast cells. TAS measurement declined in exposure group, while it was within normal range in most treated cases.

A single point mutation in framework region 3 of heavy chain affects viral neutralization dynamics of single-chain Fv against bovine herpes virus type 1.

We constructed functional recombinant single chain Fv (scFv) against bovine herpes virus type 1 (BoHV-1), aetiological agent of respiratory and genital diseases in cattle for which available vaccines do not provide adequate protection. The scFv against BoHV-1 with 18 amino acid long flexible linker (scFv3-18L; monomeric form) recognized target antigen and, also, neutralized BoHV-1 in vitro. A comparison with recombinant scFv with 7 amino acid linker against BoHV-1 (scFv1-7L), capable of forming diabodies, indicated that a relatively higher concentration (two-fold) of monomer scFv3-18L is needed for virus neutralization as compared to scFv1-7L. A single point replacement mutation (Asp98 to Gly98) in the framework-3 (FR3) variable-region of scFv with 18 amino acid linker (scFv4m-18L), however, affected the viral neutralization in a dose-dependent manner where 2.7 fold higher mutant scFv concentration was required to achieve virus neutralization. Despite differences in dose-dependent viral neutralization of the mutant scFv-18L, it detected viral antigen in an immunofluorescent assay. The outlined experiments demonstrate that recombinant scFv against BoHV-1, whether expressed as scFv or diabody, provide an effective antibody based therapeutic and immunodiagnostic protein. Further, single point substitution mutation in the FR3 can affect viral neutralization dynamics without affecting qualitative viral antigen recognition.

Prevalence of obesity among affluent school children in Dhaka.

A cross sectional study was conducted in September 2006 in one of the private schools in Dhaka, Bangladesh to see the prevalence of obesity among affluent school children and adolescents. Informed consent was taken from school authority to take anthropometric measurement of all school children. Standing height was measured with a stadiometer and weight with a bathroom scale. Waist and hip circumference were measured with a measuring tape. The body mass index (BMI) was calculated using weight in kilogram/(height in meter)2 formula. Obesity was defined as BMI≥95th percentile for age and sex, over weight as BMI≥85th percentile for age and sex, normal weight as BMI between 5th and 84th percentile and underweight as BMI <5th percentile. Official centers for disease control (CDC) growth chart for boys and girls age 2-20 years was used. Children and adolescents were divided into group 1(3-5 years), Group 2(6-9years), group 3(10-13 years) and group 4(14-18 years). There were a total of 468 children and adolescents (male 266, female 202). In group 1 there were 110 children, in group 2 there were177 children, in group 3 and 4 there were 149 and 32 adolescents respectively. The prevalence of obesity was 17.9%, higher among males (19.9%), compared to females (15.3%). Obesity was highest (27.7%) in group 2, 14.5% in group 1, 10.7% in group 3 and 9.4% in group 4.

Organization and genomic complexity of bovine lambda-light chain gene locus.

Complete characterization and physical mapping of bovine lambda (lambda) light chain locus, spanning 412kbp, on chromosome 17, has revealed twenty-five V(lambda) genes, seventeen being functional, organized in three sub-clusters 23.7kbp 5' of the J(lambda)-C(lambda) units. Three V(lambda) sub-clusters are separated by two large introns of 126.8 and 138.3kbp. The predominantly expressed V(lambda)1 genes are present in the two 5' sub-clusters, while J(lambda)-proximal V(lambda) sub-cluster comprises rarely expressed V(lambda)2 and V(lambda)3 genes. The preferential expression of V(lambda)1 genes in the bovine immunoglobulin repertoire is influenced by the composition of recombination signal sequences (RSS). Of the J(lambda)-C(lambda) cluster, it is mainly J(lambda)3-C(lambda)3 unit that is expressed in reading frame 2, though J(lambda)2 and J(lambda)3 have identical RSS. The predominant expression of J(lambda)3-C(lambda)3 genes over J(lambda)2-C(lambda)2 is likely due to endogenous counter selection for J(lambda)2 encoded CDR3 and framework 4 regions. Differences in the genomic complexity of V(lambda) genes in Hereford and Holstein cattle are due to polymorphism at the lambda-light chain gene locus. Despite more potential germline encoded combinatorial diversity, restricted V(lambda)1-J(lambda)3-C(lambda)3 recombinations encode the most lambda-light chain repertoire in cattle.

Erratum to "Maternal, neonatal and child health interventions and services: moving from knowledge of what works to systems that deliver" [International Health 2 (2010) 87-98].

The Publisher regrets that an error occurred in the name of the 6th listed co-author for this paper. B. Matthias was listed in the original paper instead of M. Borchert; the correct listing can be seen above.

Construction of single-chain Fv with two possible CDR3H conformations but similar inter-molecular forces that neutralize bovine herpesvirus 1.

Bovine herpesvirus 1 (BoHV-1) causes respiratory and genital diseases in cattle for which available vaccines do not confer adequate protection. Since passive immunization with antibodies permits disease prevention, single-chain fragment variable (scFv), originating from a monoclonal bovine IgG1 antibody against BoHV-1, were constructed and expressed in Pichia pastoris in V(lambda)-V(H) orientation via a flexible seven-amino acid linker. Similar to the intact IgG, the purified recombinant scFv neutralized BoHV-1 in vitro and recognized viral antigens in BoHV-1 infected MDBK cells by immunofluorescence. Homology modeling of the Fv predicts two distinct conformations for CDR3H. Firstly, a long protruding CDR3H conformation where no disulfide linkage occurred between two "non-canonical" Cys residues resulted in a large binding cavity between V(lambda) and V(H). Secondly, a smaller potential antigen-binding cavity is predicted with a disulfide linkage between the two Cys residues of CDR3H creating a six-membered loop in the ascending polypeptide, which fitted into the space between V(lambda) and V(H). Despite such potential configurational diversity of the antigen-binding site, the electrostatic surface potentials that would interact with the BoHV-1 epitope are largely similar for both the topographies where salt-bridge type electrostatic interactions likely occur at the edges of the binding site. Given that IgG1 antibody against BoHV-1 is clonally selected, it is likely that disulfide-stabilized broader and flatter surface topography is specifically generated to accommodate the predicted carbohydrate neutralizing B-epitope on BoHV-1. The specificity and neutralizing capacity for BoHV-1 of the scFv should make this bovine antibody fragment a useful diagnostic and potential therapeutic candidate for an important viral pathogen in cattle.

Somatic hypermutations and isotype restricted exceptionally long CDR3H contribute to antibody diversification in cattle.

Antibody diversification in IgM and IgG antibodies was analyzed in an 18-month old bovine (Bos taurus) suffering from naturally occurring chronic and recurrent infections due to bovine leukocyte adhesion deficiency (BLAD). The BLAD, involving impaired leukocyte beta2 integrin expression on leukocytes, develops due to a single point mutation in conserved region of the CD18 gene resulting in substitution of aspartic acid128 with glycine (D128G). Twenty four VDJCmu and 25 VDJCgamma recombinations from randomly constructed cDNA libraries, originating from peripheral blood lymphocytes, were examined for the variable-region structural characteristics in IgM and IgG antibody isotypes. These analyses led to conclude that: (a) expression of exceptionally long CDR3H is isotype restricted to cattle IgM antibody; (b) VDJ recombinations encoding IgM with exceptionally long CDR3H undergo clonal selection and affinity maturation via somatic mutations similar to conventional antibodies; (c) somatic mutations contribute significantly to both IgM and IgG antibody diversification but significant differences exist in the patterns of 'hot spot' in the FR1, FR3 and CDR1H and, also, position-dependant amino acid diversity; and (d) transition nucleotide substitutions predominate over transversions in both VDJCmu and VDJCgamma recombinations consistent with the evolutionary conservation of somatic mutation machinery. Overall, these studies suggest that both somatic mutations and exceptional CDR3H size generation contribute to IgM and IgG antibody diversification in cattle during the development of immune response to naturally occurring chronic and multiple microbial infections.