A middle aged lady with recurrent low trauma fracture due to parathyroid adenoma.

A 48 year old lady was referred to BIRDEM Hospital, Dhaka, Bangladesh by her local physician for evaluation of hypercalcaemia and increased serum parathyroid hormone (PTH) in the background history of low trauma fracture. Ultrasound of neck and parathyroid scintigraphy with 99mTc-MIBI revealed a parathyroid adenoma. Parathyroidectomy was done. Histopathology report showed features consistent with parathyroid adenoma. Primary hyperparathyroidism should be kept in mind in all patients presenting with history of bone problems ranging from simple bone pain to spontaneous or low trauma fracture associated with hypercalcemia. By the help of appropriate localization technique it can be localized and cured by parathyriodectomy.

Effects of Indole-3-Butyric Acid (IBA) and rooting media on rooting and survival of air layered wax apple (Syzygium samarangense) CV Jambu Madu.

The wax apple or jambu madu, is a non-climacteric tropical fruit from Myrtaceae family and widely cultivated in South East Asia. The limited availability of good quality seedlings of wax apple is the main problem to development of flourish it's market share in the current fruit industry. Therefore, in order to produce good quality planting materials, a study aimed at optimizing propagation and adventitious rooting technique and survivability of wax apple air layer was conducted. In this study, four different levels of Indole-3-Butyric Acid (IBA) concentration (0, 1000, 1500 and 2000 mg L-1) and three rooting media (sphagnum moss, vermicompost and garden soil) were applied after removal of bark (phloem) on the shoot to determine the effect on rooting and survivability of the wax apple air layer under field conditions. The results showed that the wax apple shoots treated with 2000 mg L-1 IBA produced the significantly higher number of roots, increased length of root, diameter of branch, length of branch, number of leaf and leaf area of air layers. In addition, the highest chlorophyll content and stomatal aperture were recorded in 2000 mg L-1 IBA treatment compared to other treatments including control. Vermicompost medium was better than garden soil and sphagnum moss in respect of rooting and survivability of air layers. The results showed that the combination of 2000 mg L-1 IBA and vermicompost as rooting media give the best combination to root initiation, root number, root length and survival rate (100%) of wax apple air layers. From this study, it can be concluded that 2000 mg L-1 IBA and vermicompost treatment enhance the root initiation, early establishment and survivability of wax apple air layered under field conditions.

Efficacy and Safety of Empagliflozin in Patients with Type 2 Diabetes Mellitus Fasting During Ramadan: A Real-World Study from Bangladesh.

Aim: In Bangladesh, there is a large population of Muslims with type 2 diabetes mellitus (T2DM) who fast during Ramadan. Changes in the pattern of meal and fluid intake during this long-fasting hours may increase the risk of hypoglycaemia, hyperglycaemia, and dehydration. Our key point of focus was to evaluate the efficacy and safety of Empagliflozin, a sodium-glucose co transporter 2 inhibitor (SGLT2i), in patients with T2DM while fasting during Ramadan. Methods: This was a 24-weeks, multi-centre, open-label, two-arm parallel-group study. In this prospective type of observational study, we enrolled patients taking Empagliflozin and Metformin with or without a DPP-4 inhibitor in one group (n = 274) and a parallel group (n = 219) who were treated with Metformin with or without a DPP-4 inhibitor. The primary endpoint of this study was HbA1c reduction, weight loss and the number of reported or symptomatic hypoglycemic events. In secondary endpoints, we evaluated the changes from baseline in blood pressure, estimated glomerular filtration rate (eGFR), serum creatinine, and serum electrolyte, the proportion of volume depletion (≥1 event) and incidence of other adverse events (AEs) of interest potentially related to SGLT2 inhibitor. Results: During Ramadan, HbA1c reduction was significant in Empagliflozin arm (-0.49% vs -0.12%); [p < 0.001]. From before to the end of the study, significant weight reduction was seen in the Empagliflozin arm (-1.4 kg vs -0.09 kg); [p < 0.001]. We observed no significant increase in the incidence of hypoglycemia (0.7% vs 0.4%, p = 0.267) and volume depletion (2.6% vs 1.8%; p = 0.55) in both arm. All these milder forms events did not require any hospital admission. There was no report of serious adverse events or any discontinuation, or reduction of prescribed doses of empagliflozin during Ramadan. Conclusion: Empagliflozin is efficacious and safe for treating adults with T2DM during Ramadan.

Bangladesh Endocrine Society (BES) Position Statement for Management of Diabetes and Other Endocrine Diseases in Patients with COVID-19.

BACKGROUND: The year 2020 witnessed a largely unprecedented pandemic of coronavirus disease (COVID-19), caused by SARS COV-2. Many people with COVID-19 have comorbidities, including diabetes, hypertension and cardiovascular diseases, which are significantly associated with worse outcomes. Moreover, COVID-19 itself is allied with deteriorating hyperglycemia. Therefore, Bangladesh Endocrine Society has formulated some practical recommendations for management of diabetes and other endocrine diseases in patients with COVID-19 for use in both primary and specialist care settings. OBJECTIVE: The objective of the article is to develop a guideline to protect the vulnerable group with utmost preference - the elderly and those with comorbid conditions. Therefore, to ensure the adequate protective measures and timely treatment for COVID-19 patients with diabetes, other endocrine diseases or any other comorbidities. CONSIDERING AND MONITORING ISSUES: The risk of a fatal outcome from COVID-19 may be up to 50% higher in patients with diabetes than in non-diabetics.Patients with diabetes and COVID had CFR 7.3-9.2%, compared with 0.9-1.4% in patients without comorbidities.Diabetic ketoacidosis may be one of the causes of mortality in COVID-19.There is wide fluctuation of blood glucose in these patients, probably due to irregular diet, reduced exercise, increased glucocorticoids secretion, and use of glucocorticoids. HbA1c should be <7.0% for the majority of the patients, this target may be relaxed in appropriate clinical settings.More emphasis should be given on day-to-day blood glucose levels. Hypoglycemia (<3.9 mmol/l) must be avoided.Frequent monitoring of blood glucose is needed in critically ill patients. CONCLUSION: The fight against COVID-19 has been proven to be a challenging one. Therefore, all healthcare personnel should make the best use of updated knowledge and skills to ensure adequate protective measures and timely treatment for COVID-19 patients with diabetes, other endocrine diseases or any other comorbidities.

Role of virion-associated glycosylphosphatidylinositol-linked proteins CD55 and CD59 in complement resistance of cell line-derived and primary isolates of HIV-1.

This study investigates whether cell-derived glycosylphosphatidylinositol-linked complement control proteins CD55 and CD59 can be incorporated into HIV-1 virions and contribute to complement resistance. Virus was prepared by transfection of cell lines with pNL4-3, and primary isolates of HIV-1 were derived from patients' PBMCs. Virus was tested for sensitivity to complement-mediated virolysis in the presence of anti-gp160 antibody. Viral preparations from JY33 cells, which lack CD55 and CD59, were highly sensitive to complement. HIV-1 preparations from H9 and U937 cells, which express low levels of CD55 and CD59, had intermediate to high sensitivity while other cell line-derived viruses and primary isolates of HIV-1 were resistant to complement-mediated virolysis. Although the primary isolates were not lysed, they activated complement as measured by binding to a complement receptor positive cell line. While the primary isolates were resistant to lysis in the presence of HIV-specific antibody, antibody to CD59 induced lysis. Likewise, antibody to CD55 and CD59 induced lysis of cell line-derived virus. Western blot analysis of purified virus showed bands corresponding to CD55 and CD59. Phosphatidylinositol-specific phospholipase C treatment of either cell line-derived or primary isolates of HIV-1 increased sensitivity to complement while incubation of sensitive virus with purified CD55 and CD59 increased resistance to complement. These results show that CD55 and CD59 are incorporated into HIV-1 particles and function to protect virions from complement-mediated destruction, and they are the first report of host cell proteins functioning in protection of HIV-1 from immune effector mechanisms.

Macaca fascicularis are highly susceptible to an RT-SHIV following intravaginal inoculation: a new model for microbicide evaluation.

BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is a major target for antiretroviral strategy to block or curtail HIV infection. A suitable RT-SHIV/macaque model is urgently needed for the evaluation of HIV/AIDS therapies and microbicides specifically targeting HIV-1 RT. METHODS: Fifteen cynomolgus macaques (Macaca fascicularis) were divided into three groups (n = 5) and intravaginally inoculated with 4800, 1200, or 300 TCID(50) of RT-SHIVtc. Systemic infections of RT-SHIVtc exposed macaques were determined by both virological and immunologic parameters during 24 weeks post-challenge. RESULTS: Within 2 weeks post-inoculation, 13 of 15 macaques became infected as confirmed by virus isolation, plasma viral RNA, proviral DNA, declined CD4(+)T cell counts in peripheral blood and seroconversion. CONCLUSIONS: Results serve to validate the infectivity and pathogenicity of RT-SHIVtc following vaginal exposure in M. fascicularis. This RT-SHIVtc/macaque model could be suitable for the pre-clinical evaluation of non-nucleoside RT inhibitor-based anti-HIV microbicides.

Frequency and Predictors of Erectile Dysfunction in Bangladeshi Men with Type 2 Diabetes Mellitus: Experience from a Tertiary Center.

Erectile dysfunction (ED) is a common complication of diabetes mellitus. But it is frequently under diagnosed and may result in poor quality of life. Previous studies have shown a high frequency of ED in diabetic men. The aim of this cross-sectional study was to determine the frequency of ED and explore its risk factors in type 2 diabetic (T2DM) men in Bangladesh. During August 2013 to July 2014, 508 diabetic men aged 30-69 years were interviewed at the outpatient and inpatient departments of Bangladesh Institute of Research and Rehabilitation in Diabetes, Endocrine and Metabolic Disorder (BIRDEM), Dhaka, Bangladesh. Recent biochemical data (within last 6 months) were collected from the patient's diabetes guide book and hospital records. Erectile function (EF) was assessed using the validated Bengali version of the International Index of Erectile Function-15 (IIEF-15) questionnaire. Out of 508 type 2 diabetic men, ED was found in 306(60.2%) patients. The frequency of ED was increased with age from 35.5% in men aged 28-39 years to 100% in those aged 60 years and above (p<0.001). Increasing age was also associated with an increase in the severity of ED (5.5% severe ED in 28-39 years age group vs. 77.4% in 60-69 years group, p=0.000). Duration of diabetes was also associated with the increase in both frequency and severity of ED (20.2% ED and 2.4% severe ED in diabetes duration 0-5 years vs. 100% ED and 100% severe ED in diabetes duration >20 years, p=0.000). The frequency of ED in patients with good and poor glycemic control was 3.5% and 71.6% respectively (p=0.000); frequency of severe ED was also higher in uncontrolled diabetic males (0% vs. 28.4% in controlled vs. uncontrolled DM, p=0.000). The characteristics found to be significantly associated with erectile dysfunction were: patient's age, housebound bedridden status, sedentary work, diabetes duration, HbA1c level, microvascular complications, IHD, and diuretic drugs use. Moderate physical activity was found to be inversely related to the frequency of erectile dysfunction. The frequency of ED is very high among Bangladeshi T2DM male and the frequency and severity of ED may be reduced by improving glycemic status.

Transmigrated and Impacted Upper 2nd Premolar Associated with Dentigerous Cyst.

A male patient about 45 years old came to my private clinic "The Dental Aid", Dhanmondi, Dhaka, Bangladesh last year (February 2016) with the complaint of pain at right palatal premolar region. On clinical examination we found the absence of upper left 2nd premolar from the series of dentition but on the right side premolars are present. The oral mucosa was intact and normal in color. There was no caries, dental pocket or any other pathology in the teeth of both jaws. On palpation we found soft area on the right palatal side at the premolar area. We took intraoral peri-apical radiograph and found impacted left 2nd premolar whose crown was encircled by radiolucent area at the palatal side. Here, we present a rare case of transmigrated and impacted left 2nd maxillary premolar associated with dentigerous cyst. The cyst was successfully treated under general anaesthesia by enucleation along with extraction of the involved maxillary transposed and impacted left second premolar.

Prevalence and Predictors of Depression among Patients with Type 2 Diabetes: A Multicenter Cross-sectional Study from Bangladesh.

Depression is a common comorbidity of type 2 diabetes mellitus (T2DM) which adversely affects diabetes management and outcome. Identifying and treating comorbid depression may improve diabetes care. This cross-sectional study was conducted in several tertiary hospitals throughout Bangladesh from July 2017 to April 2018. Nine hundred (900) adult patients with T2DM aging ≥25 years having diabetes for at least 6 months and equal numbers of non-diabetic otherwise healthy controls were recruited from the outpatient departments of these centers. Depression was assessed in all consenting patients and controls by administering the Bangla (local language) version of the PRIME-MD Patient Health Questionnaire (PHQ-9); participants obtaining a score of 5 or more were labeled to have depression. Depression was present in 60.3% of T2DM patients and in 29.4% of controls. Statistically significant difference was found in age, marital status, occupation, body mass index (BMI), waist circumference, systolic blood pressure (BP), diastolic BP and PHQ-9 score between diabetic and non-diabetic subjects (<0.001). T2DM subjects had 4.71-fold higher odds of depression in comparison to the controls (95% CI: 3.76-5.90; p<0.001). Age ≥50 years, unmarried status, years of schooling ≤10 years, underweight, abdominal obesity, and hypertension appeared to be the significant predictors of depression in the study subjects. In T2DM subjects, diabetes in the family members, the presence of other comorbidities, diabetic complications, diabetes duration >5 years, insulin use, using insulin syringe for injection, albuminuria and CKD were the important predictors of depression. Our study found higher prevalence and risk of depression in T2DM patients than their non-diabetic counterparts. T2DM patients should be screened for depression in order to achieve and maintain the treatment goals.

Vitamin D Status in Newly Diagnosed Type 2 Diabetes Patients Attending in a Tertiary Hospital of Bangladesh.

Vitamin D deficiency is common in patients with type 2 diabetes mellitus (T2DM). Vitamin D status may have a causal role in the onset of T2DM and may influence glycaemic control in these patients. However, data on vitamin D status among Bangladeshi T2DM patients are scarce. The present cross-sectional study was done among newly diagnosed 50 T2DM patients attending outpatient department of Comilla Diabetic Hospital, Bangladesh to address this lacuna. Serum 25(OH)D was measured in all and classified as normal (≥30ng/mL), insufficient (>20 to 29.9ng/mL), and deficient (≤20ng/mL). Mean serum 25(OH)D level was 27.91±2.58ng/mL (mean±SEM). Among the study subjects, 30% were D-deficient, 36% were D-insufficient and 34% had normal 25(OH)D. There was no statistical difference of 25(OH)D level between the younger (age <40 years) and older (≥40 years) patients (28.31±4.3 vs. 27.44±2.6ng/mL, mean±SEM; p=0.869); males and females (26.79±2.1 vs. 31.09±8.2ng/mL, mean±SEM; p=0.470); among smokers, non-smokers and ex-smokers (26.86±4.31, 27.10±2.49 and 42.62±1.71ng/mL respectively, mean±SEM; p=0.363); among normal weight, overweight and obese (30.61±6.16, 35.61±9.52 and 24.27±1.71ng/mL respectively, mean±SEM; p=0.191); and among normotensive, borderline hypertensive and hypertensive (25.29±2.46, 32.57±5.32 and 20.84±3.66ng/mL respectively, mean±SEM; p=0.277) patients. 25(OH)D level showed significant negative correlation with body mass index (r= -0.391, p=0.017) and positive correlation (r=0.334, p=0.044) with fasting plasma glucose in male subjects. Age, sex, smoking status, BMI, systolic BP, diastolic BP, family history of DM and smoking status were not found to influence vitamin D level independently. Our study found high frequency of subnormal vitamin D in newly diagnosed T2DM patients. Screening for vitamin D status may be beneficial in T2DM patients.

Thyroid Dysfunction and Autoimmunity in First Trimester of Pregnancy, Single Center Experience in Bangladesh.

Study on thyroid function and thyroid autoantibody status in pregnancy considering the trimester specific range, is scarce in Bangladesh. This cross sectional study done in Bangabandhu Sheikh Mujib Medical University, Bangladesh from August 2012 to June 2013 encompassed 186 pregnant women of first trimester for study of thyroid function (TSH, FT4) and anti-thyroid antibodies (anti-TPO, anti-TG). Age of the subjects was 25.4±4.9 years (mean±SD), median gestational age was 9 weeks. Applying the trimester-specific normal reference range set by American Thyroid Association (ATA), 48(25.8%) of the women were found to have thyroid dysfunction; 40(21.5%) subclinical hypothyroid (SCH), 1(0.5%) overt hypothyroid (OH) and 7(3.8%) hyperthyroid; 40(21.5%) women had goiter. If non-pregnant adult normal rage is used, 22 of SCH women as per ATA criteria will be labeled as normal and 19 normal women as per ATA cut off will be labeled as hyperthyroid. There was statistically significant disparity for functional status defined by these two references cut off value (p<0.001). 29(15.6%) women had thyroid autoimmunity and the autoantibody positivity was more frequent in women with thyroid dysfunction than euthyroid women (22.92% vs. 13.04%, p<0.001). Even though universal screening for thyroid dysfunction is not yet a recommendation, it should be considered in our population.

Effects of Indole-3-Butyric Acid (IBA) and rooting media on rooting and survival of air layered wax apple (Syzygium samarangense) CV Jambu Madu / Efeitos do Ácido Indol-3-Butírico (AIB) e meio de enraizamento no enraizamento e sobrevivência de maçã de cera em camada de ar (Syzygium samarangense) CV Jambu Madu

The wax apple or jambu madu, is a non-climacteric tropical fruit from Myrtaceae family and widely cultivated in South East Asia. The limited availability of good quality seedlings of wax apple is the main problem to development of flourish it's market share in the current fruit industry. Therefore, in order to produce good quality planting materials, a study aimed at optimizing propagation and adventitious rooting technique and survivability of wax apple air layer was conducted. In this study, four different levels of Indole-3-Butyric Acid (IBA) concentration (0, 1000, 1500 and 2000 mg L-1) and three rooting media (sphagnum moss, vermicompost and garden soil) were applied after removal of bark (phloem) on the shoot to determine the effect on rooting and survivability of the wax apple air layer under field conditions. The results showed that the wax apple shoots treated with 2000 mg L-1 IBA produced the significantly higher number of roots, increased length of root, diameter of branch, length of branch, number of leaf and leaf area of air layers. In addition, the highest chlorophyll content and stomatal aperture were recorded in 2000 mg L-1 IBA treatment compared to other treatments including control. Vermicompost medium was better than garden soil and sphagnum moss in respect of rooting and survivability of air layers. The results showed that the combination of 2000 mg L-1 IBA and vermicompost as rooting media give the best combination to root initiation, root number, root length and survival rate (100%) of wax apple air layers. From this study, it can be concluded that 2000 mg L-1 IBA and vermicompost treatment enhance the root initiation, early establishment and survivability of wax apple air layered under field conditions.

A maçã de cera, ou jambu madu, é uma fruta tropical não climatérica da família Myrtaceae e amplamente cultivada no Sudeste Asiático. A disponibilidade limitada de mudas de macieira de boa qualidade é o principal problema para o desenvolvimento de sua participação de mercado na fruticultura atual. Portanto, com o objetivo de produzir materiais de plantio de boa qualidade, foi realizado um estudo visando otimizar a técnica de propagação e enraizamento adventício e a sobrevivência da camada aérea da cera de macieira. Neste estudo, quatro diferentes níveis de concentração de ácido indol-3-butírico (AIB) (0, 1000, 1500 e 2000 mg L-1) e três meios de enraizamento (musgo esfagno, vermicomposto e solo de jardim) foram aplicados após a remoção da casca (floema) na parte aérea para determinar o efeito no enraizamento e capacidade de sobrevivência da camada de ar da macieira em condições de campo. Os resultados mostraram que os brotos de macieira tratados com 2000 mg L-1 de AIB produziram significativamente maior número de raízes, maior comprimento de raiz, diâmetro de galho, comprimento de galho, número de folhas e área foliar das camadas aéreas. Além disso, o maior teor de clorofila e abertura estomática foram registrados no tratamento 2000 mg L-1 IBA em comparação com outros tratamentos, incluindo o controle. O meio de vermicomposto foi melhor do que o solo de jardim e o musgo esfagno em relação ao enraizamento e capacidade de sobrevivência das camadas aéreas. Os resultados mostraram que a combinação de 2000 mg L-1 de AIB e vermicomposto como meio de enraizamento proporciona a melhor combinação para iniciação radicular, número de raízes, comprimento radicular e taxa de sobrevivência (100%) das camadas aéreas de macieira. A partir deste estudo, pode-se concluir que 2.000 mg L-1 de AIB e tratamento com vermicomposto melhoram a iniciação radicular, o estabelecimento precoce e a capacidade de sobrevivência de macieiras em camadas de ar em condições de campo.

HTLV-I activates complement leading to increased binding to complement receptor-positive cells.

This investigation was performed to determine whether HTLV-I can activate complement, since previous studies show that complement activation by some viruses, including HIV-1, can enhance binding to, and infection of complement receptor-positive (CR+) cells. Complement treatment increased binding of HTLV-I to CR+ HPB-ALL cells by approximately 5-fold. In contrast, increased binding was not observed with H9 cells, which lack CR. Heat inactivation or EDTA treatment of complement blocked this increased binding while EGTA treatment only partially blocked binding. Anti-CR2 antibody significantly blocked binding of complement-treated HTLV-I to HPB-ALL cells. Since previous studies showed that HIV-1 could activate complement, activation of complement by this virus was compared with HTLV-I. It was observed that binding of HTLV-I to HPB-ALL cells was enhanced by highly dilute complement (> or = 1:810) while HIV-1 required much higher concentrations of complement (> or = 1:30), indicating that HTLV-I is a much stronger complement activator. Treatment with complement transiently increased the ability of HTLV-I to infect CR+ cell lines as judged by provirus formation (4- to 8-fold increase) and p24 production (5- to 10-fold increase). In contrast, complement treatment did not increase infection of CR- cells. In conclusion this study shows that HTLV-I activates complement leading to increased binding to, and transiently increased infection of, CR+ cells. This complement-mediated increased binding of HTLV-I may dramatically affect viral trafficking and immunological reactivity of virus in vivo.

Host cell components affect the sensitivity of HIV type 1 to complement-mediated virolysis.

An infection-competent, full-length HIV-1 clone (pNL4-3) was expressed in seven human cell lines and in peripheral blood mononuclear cells in order to assess the contribution of host cell components toward interaction of free virus with the complement system. HIV-1 expressed in the H9 cell line, which is frequently used for in vitro infection, was relatively susceptible to complement-mediated virolysis in the presence of both HIV antibody-positive patient serum and an anti-V3 monoclonal antibody. Expression of complement receptors 1, 2, and 3, complement control proteins membrane inhibitor of reactive lysis (MIRL, CD59) and decay-accelerating factor (DAF, CD55), and HLA-DR was assessed on host cells. There was an inverse relationship between the sensitivity of virus to complement and the amount of expression of MIRL and DAF on cells. HIV derived from the JY cell line and the mutant JY33 cell line, which is deficient in expression of phosphatidylinositol (PI)-linked proteins including MIRL and DAF, were also evaluated for complement-mediated virolysis. Virus expressed in the mutant cell line was more sensitive to antibody-independent as well as antibody-dependent complement-mediated virolysis than virus expressed in the wild-type cells. Direct demonstration of the presence of MIRL and DAF on the viral surface was obtained by showing that anti-MIRL or anti-DAF antibody induced complement-mediated virolysis. These experiments show that the host cell type can substantially influence the susceptibility of HIV to complement-mediated virolysis and suggest that PI-linked complement control proteins play an important role in this resistance.

Regulation of HIV-1 transcription.

Human immunodeficiency virus type-1 (HIV-1) is a highly pathogenic lentivirus that requires transcription of its provirus genome for completion of the viral life cycle and the production of progeny virions. Since the first genetic analysis of HIV-1 in 1985, much has been learned about the transcriptional regulation of the HIV-1 genome in infected cells. It has been demonstrated that HIV-1 transcription depends on a varied and complex interaction of host cell transcription factors with the viral long terminal repeat (LTR) promoter. The regulatory elements within the LTR interact with constitutive and inducible transcription factors to direct the assembly of a stable transcription complex that stimulates multiple rounds of transcription by RNA polymerase II (RNAPII). However, the majority of these transcripts terminate prematurely in the absence of the virally encoded trans-activator protein Tat, which stimulates HIV-1 transcription elongation by interacting with a stem-loop RNA element (TAR) formed at the extreme 5' end of all viral transcripts. The Tat-TAR interaction recruits a cellular kinase into the initiation-elongation complex that alters the elongation properties of RNAPII during its transit through TAR. This review summarizes our current knowledge and understanding of the regulation of HIV-1 transcription in infected cells and highlights the important contributions human lentivirus gene regulation has made to our general understanding of the transcription process.

Isolation and characterization of a hemorrhagin from the venom of Ophiophagus hannah (king cobra).

The major hemorrhagin (termed hannahtoxin) of the venom of Ophiophagus hannah (king cobra) was purified to electrophoretic homogeneity by DEAE-Sephacel ion exchange chromatography, Sephadex G-200 gel filtration followed by a second DEAE-Sephacel chromatography. Proteolytic activity was associated with the hemorrhagic activity throughout the purification procedures. Hannahtoxin constituted approximately 2% of the crude venom. It had an isoelectric point of 5.3, a carbohydrate content of 12%, a mol. wt of 66,000 as determined by SDS-polyacrylamide gel electrophoresis and 63,000 as determined by gel filtration. It contains 1 mole of Zn per mole of protein. The minimum hemorrhage doses for hannahtoxin are 0.7 microgram and 75 micrograms, respectively, in rabbits and in mice. Hannahtoxin was not lethal to mice at a dose of 2 mg/kg (i.v.) but killed rabbits at doses above 0.18 mg/kg (i.v.). It liberated protein from rabbit glomerular basement membrane but not rat glomerular basement membrane. Treatment of the hemorrhagin with EDTA and 1,10-phenanthroline eliminated both the proteolytic and hemorrhagic activities completely.

Preparation of antibodies to king cobra (Ophiophagus hannah) venom hemorrhagin and investigation of their cross-reactivity.

Hannahtoxin, the major hemorrhagin purified from king cobra (Ophiophagus hannah) venom, elicits hemorrhages in rabbits but not in mice. Two antisera against hannahtoxin were prepared: one raised against purified hannahtoxin, while the other was raised against glutaraldehyde cross-linked and detoxified hannahtoxin. The antisera were refined by pepsin digestion and ammonium sulfate precipitation. They are of approximately equal potency in their ability to neutralize the hemorrhagic activity of king cobra venom in rabbits. The antisera did not form a precipitin line with venom of snakes of the Viperidae family nor neutralize hemorrhages elicited in mice by any of these venoms. However, when the hemorrhagic activity was assayed in rabbits, both antisera were able to abolish the hemorrhages elicited by all of the venoms tested. These results suggest that hannahtoxin displays few epitopes in common with hemorrhagins of viperid venoms, except those involved in the neutralization of hemorrhagic activity in rabbits. The epitopes of viperid venom hemorrhagins involved in the neutralization reaction in rabbits are different from those in mice.

Development of an immunocytochemical procedure to detect adenoviral antigens in chicken tissues.

An immunocytochemical technique utilizing an avidin-biotin peroxidase complex was developed to detect viral antigens in various tissues following oral administration of a locally isolated serotype 8 avian adenovirus (AAV) in specific pathogen-free (SPF) chickens. A strong color reaction was obtained with tissues from infected birds that contained a minimal amount of AAV antigens as determined by an indirect enzyme-linked immunosorbent assay. No reaction was detected in sections of tissues obtained from SPF chickens, and the reactivity with infected tissues could be removed by prior absorption of the primary antibody with purified AAV. A group-specific antigen common to the 12 serotypes of AAV was demonstrated by this technique. Because of the high sensitivity and broad-spectrum reactivity, this technique could be useful for studying the pathogenesis and laboratory diagnosis of inclusion body hepatitis caused by several serotypes of AAV.

A primary epidemic of inclusion body hepatitis in broilers.

Inclusion body hepatitis (IBH) was diagnosed in 15 broiler flocks supplied by one breeder in the South Island of New Zealand. The affected flocks suffered mortality up to 30%. Malaise and slightly increased mortality were noticed by growers from about day 12 post-hatch; mortality peaked in the fourth week, and, in most flocks, declined to normally accepted levels from day 33 on. Gross signs seen at necropsy usually included bone-marrow aplasia, atrophy of the bursa of Fabricius and the thymus, and swollen hemorrhagic livers with focal necrosis. Jaundice was seen in many surviving birds. In some flocks, there was also proventricular hemorrhage, mild tracheitis, and airsacculitis. Downgrading and condemnation rates were increased in all flocks. Eosinophilic intranuclear inclusion bodies were seen in hepatocytes of some affected birds. An adenovirus was isolated from a number of cases investigated. The disease in broilers was preceded by production drops associated with feed refusal and increased mortality in the breeder stock.

Development of an enzyme-linked immunosorbent assay to detect and quantify adenovirus in chicken tissues.

An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect and quantify avian adenovirus (AAV) in various chicken tissues, including blood. A positive ELISA absorbance value was obtained with suspensions of infected liver tissue that contained less than 100 mean tissue-culture infective doses per gram. A positive correlation was observed between the absorbance values and titer of infectious virus in infected liver tissue. A group-specific antigen common to the 12 serotypes of AAV tested was demonstrated by this ELISA. Because of the high sensitivity and broad-spectrum reactivity, this ELISA could be useful for the study of AAV pathogenesis, for laboratory diagnosis of inclusion body hepatitis irrespective of the serotype of AAV involved, and for screening commercial and specific-pathogen-free flocks for the presence of AAV.