Emergence of a new genotype of clade 2.3.4.4b H5N1 highly pathogenic avian influenza A viruses in Bangladesh.

Influenza virological surveillance was conducted in Bangladesh from January to December 2021 in live poultry markets (LPMs) and in Tanguar Haor, a wetland region where domestic ducks have frequent contact with migratory birds. The predominant viruses circulating in LPMs were low pathogenic avian influenza (LPAI) H9N2 and clade 2.3.2.1a highly pathogenic avian influenza (HPAI) H5N1 viruses. Additional LPAIs were found in both LPM (H4N6) and Tanguar Haor wetlands (H7N7). Genetic analyses of these LPAIs strongly suggested long-distance movement of viruses along the Central Asian migratory bird flyway. We also detected a novel clade 2.3.4.4b H5N1 virus from ducks in free-range farms in Tanguar Haor that was similar to viruses first detected in October 2020 in The Netherlands but with a different PB2. Identification of clade 2.3.4.4b HPAI H5N1 viruses in Tanguar Haor provides continued support of the role of migratory birds in transboundary movement of influenza A viruses (IAV), including HPAI viruses. Domestic ducks in free range farm in wetland areas, like Tangua Haor, serve as a conduit for the introduction of LPAI and HPAI viruses into Bangladesh. Clade 2.3.4.4b viruses have dominated in many regions of the world since mid-2021, and it remains to be seen if these viruses will replace the endemic clade 2.3.2.1a H5N1 viruses in Bangladesh.

Aetiological Factors of Female Genitourinary Fistula.

The purpose of this observational cross-sectional study was to evaluate the histories of women with urinary incontinence caused by genitourinary fistulae in Bangladesh. This is an exploratory analysis to investigate possible contributing factors to the development of urogenital fistulae. Sixty four (64) women who were treated for urogenital fistulae at the National Fistula Center, Dhaka Medical College Hospital, Dhaka, Bangladesh were interviewed from July 2013 to December 2013. Their average age was 35.91 years; majorities were illiterate; had early marriage and came from poor family. Majority had height less than 150 cm, body weight 50 Kg or less and 67.19% of the women were multigravid. Obstetrical aetiology predominated in fistula causation and vesicovaginal fistula was the most frequent fistula type. Most of the women with fistulas did not receive antenatal care (88.46%); had developed a fistula after laboring for 48 hours or longer (59.61%) with vaginal delivery (59.61%) at home (57.69%) and within seven days of delivery (63.45%). Most women who experienced fistulae also had poor obstetric outcomes. Increasing access to emergency obstetric care is of paramount importance to prevent fistula formation.

Low Free T3 was Associated with Poor Glycemic Control in Type 2 Diabetes in a Hospital Based Study in Bangladesh.

Clinical significance of low free T3 (FT3) has not been well explained in patients with type 2 diabetes mellitus (T2DM); FT3 level may be associated with diabetes control and also with diabetic complications. This cross-sectional study was carried out among 153 (mean age 46.8±12 years; female 68%, mean duration of diabetes 5.5±0.53 years, 63.4% either overweight or obese) non-pregnant adults with T2DM who had no acute illness and were unaware about their thyroid function status from July 2016 to December 2016. Serum TSH, free T4 (FT4) and FT3 were measured in all patients by radioimmunoassay (RIA). Patients having subnormal FT3 level with normal TSH and FT4 levels were labeled as having low FT3 syndrome. The mean HbA1c of the study subjects was 8.3±1.7%, serum TSH 2.24±0.34µIU/mL, FT4 16.5±6.56fmol/mL and FT3 was 5.36±1.74fmol/mL. Among them, 9.15% were found to have low FT3 syndrome. There was no statistical difference of FT3 level between males and females (5.87±1.50 vs. 5.99±1.85fmol/mL, mean±SD; p=0.165) and among different BMI groups (p=0.179). Patients with uncontrolled diabetes had lower FT3 than those with controlled diabetes (5.91±1.83 vs. 6.15±1.21fmol/mL, mean±SD, p=0.024). The frequency of low FT3 syndrome was 11.1% in uncontrolled diabetic patients whereas none of the patients with controlled diabetes had low FT3. FT3 level showed positive correlations with the duration of diabetes (r=0.296, p=0.002) and FT4 level (r=0.490, p<0.001) only in female subjects. A fair number of clinically stable T2DM patients had low FT3 in our study. Patients with uncontrolled diabetes had lower FT3 than those with controlled diabetes.

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.

Van Wyk and Grumbach Syndrome: An Unusual Presentation of Hypothyroism.

An 18 years-old-girl presented one and half years back with the complaints of short stature, retarded growth, and menorrhagia with sudden severe lower abdominal pain; was diagnosed as bilateral ovarian cysts and underwent bilateral ovarian cystectomy. Later on she was incidentally diagnosed as a case of hypothyroidism when she had been experiencing slowly enlarging left lower abdominal mass with dull ache for the 5 month and then was transferred to the department of Endocrinology for further evaluation. Detailed work up revealed her short stature with obesity, delayed bone age and other features of hypothyroidism which was confirmed by thyroid function testing. She had enlarged left ovary with multiple follicles as shown in ultrasonography. Magnetic resonance imaging (MRI) showed sellar mass which was suspicious of macroadenoma. Levothyroxine replacement was started and she had a dramatic improvement of her problems with disappearance of the ovarian cysts and sellar mass.

Ramadan Perspective Epidemiology and Education in Diabetes (RAPEED) Study.

Ramadan fasting is one of the five pillars of Islam and is obligatory for all healthy adult and adolescent Muslims. A significant number of patients with diabetes observe fasting during Ramadan. The objectives of this Ramadan Perspective Epidemiology and Education in Diabetes (RAPEED) study were to find out the current knowledge, attitude, and practices about Ramadan fasting among people with diabetes in Bangladesh. This retrospective cross-sectional study recruited 648 subjects with diabetes mellitus attending the Endocrinology outpatient department of a tertiary level hospital of Bangladesh within two months of the end of Ramadan in 2016. Socio-demographic data, data related to diabetes treatment, complications and co-morbidities; changes in lifestyle and diabetes treatment during Ramadan and frequency of hypoglycemia were collected from all. The mean age of the study population was 50.32±12.1 years and the majority (98.6%) had type 2 diabetes and 63.9% were overweight or obese. The majority (89.35%) of the patients fasted in Ramadan and among them, more than half of the fasters received physicians' advice for Ramadan fasting (60.6%) and changed diabetes medication (69.90%) during Ramadan. Although the amount of total food consumption was unchanged in more than one half (60.6%) of the fasters, majority of them reduced sugar-sweet intake (75.5%), increased fluid drinking (75.8%) and decreased physical activity (75.8%) during the month. A large portion (37.48%) of them did not check blood glucose and more than half (54.06%) of fasters failed to visit their physicians during Ramadan. Among them 14.85% experienced mild to moderate episodes of hypoglycemia and none had severe hypoglycemia and the most (61.6%) episodes of hypoglycemia occurred in the late evening. Hypoglycemic episodes were more frequent among insulin users, patients who had received fasting advice from physicians and in those who had adjusted diabetes drugs before Ramadan fasting. Safe Ramadan fasting is a great challenge not only for the patients but also the consulting physicians. Current study reflects the necessity of both physicians' expertise and patients' awareness for safe fasting.

The replication of Bangladeshi H9N2 avian influenza viruses carrying genes from H7N3 in mammals.

H9N2 avian influenza viruses are continuously monitored by the World Health Organization because they are endemic; they continually reassort with H5N1, H7N9 and H10N8 viruses; and they periodically cause human infections. We characterized H9N2 influenza viruses carrying internal genes from highly pathogenic H7N3 viruses, which were isolated from chickens or quail from live-bird markets in Bangladesh between 2010 and 2013. All of the H9N2 viruses used in this study carried mammalian host-specific mutations. We studied their replication kinetics in normal human bronchoepithelial cells and swine tracheal and lung explants, which exhibit many features of the mammalian airway epithelium and serve as a mammalian host model. All H9N2 viruses replicated to moderate-to-high titers in the normal human bronchoepithelial cells and swine lung explants, but replication was limited in the swine tracheal explants. In Balb/c mice, the H9N2 viruses were nonlethal, replicated to moderately high titers and the infection was confined to the lungs. In the ferret model of human influenza infection and transmission, H9N2 viruses possessing the Q226L substitution in hemagglutinin replicated well without clinical signs and spread via direct contact but not by aerosol. None of the H9N2 viruses tested were resistant to the neuraminidase inhibitors. Our study shows that the Bangladeshi H9N2 viruses have the potential to infect humans and highlights the importance of monitoring and characterizing this influenza subtype to better understand the potential risk these viruses pose to humans.

Multiple introductions of highly pathogenic avian influenza H5N1 viruses into Bangladesh.

Highly pathogenic H5N1 and low pathogenic H9N2 influenza viruses are endemic to poultry markets in Bangladesh and have cocirculated since 2008. H9N2 influenza viruses circulated constantly in the poultry markets, whereas highly pathogenic H5N1 viruses occurred sporadically, with peaks of activity in cooler months. Thirty highly pathogenic H5N1 influenza viruses isolated from poultry were characterized by antigenic, molecular, and phylogenetic analyses. Highly pathogenic H5N1 influenza viruses from clades 2.2.2 and 2.3.2.1 were isolated from live bird markets only. Phylogenetic analysis of the 30 H5N1 isolates revealed multiple introductions of H5N1 influenza viruses in Bangladesh. There was no reassortment between the local H9N2 influenza viruses and H5N1 genotype, despite their prolonged cocirculation. However, we detected two reassortant H5N1 viruses, carrying the M gene from the Chinese H9N2 lineage, which briefly circulated in the Bangladesh poultry markets and then disappeared. On the other hand, interclade reassortment occurred within H5N1 lineages and played a role in the genesis of the currently dominant H5N1 viruses in Bangladesh. Few 'human-like' mutations in H5N1 may account for the limited number of human cases. Antigenically, clade 2.3.2.1 H5N1 viruses in Bangladesh have evolved since their introduction and are currently mainly homogenous, and show evidence of recent antigenic drift. Although reassortants containing H9N2 genes were detected in live poultry markets in Bangladesh, these reassortants failed to supplant the dominant H5N1 lineage.

Population dynamics of rhesus macaques and associated foamy virus in Bangladesh.

Foamy viruses are complex retroviruses that have been shown to be transmitted from nonhuman primates to humans. In Bangladesh, infection with simian foamy virus (SFV) is ubiquitous among rhesus macaques, which come into contact with humans in diverse locations and contexts throughout the country. We analyzed microsatellite DNA from 126 macaques at six sites in Bangladesh in order to characterize geographic patterns of macaque population structure. We also included in this study 38 macaques owned by nomadic people who train them to perform for audiences. PCR was used to analyze a portion of the proviral gag gene from all SFV-positive macaques, and multiple clones were sequenced. Phylogenetic analysis was used to infer long-term patterns of viral transmission. Analyses of SFV gag gene sequences indicated that macaque populations from different areas harbor genetically distinct strains of SFV, suggesting that geographic features such as forest cover play a role in determining the dispersal of macaques and SFV. We also found evidence suggesting that humans traveling the region with performing macaques likely play a role in the translocation of macaques and SFV. Our studies found that individual animals can harbor more than one strain of SFV and that presence of more than one SFV strain is more common among older animals. Some macaques are infected with SFV that appears to be recombinant. These findings paint a more detailed picture of how geographic and sociocultural factors influence the spectrum of simian-borne retroviruses.

Zoonotic simian foamy virus in Bangladesh reflects diverse patterns of transmission and co-infection.

Simian foamy viruses (SFVs) are ubiquitous in non-human primates (NHPs). As in all retroviruses, reverse transcription of SFV leads to recombination and mutation. Because more humans have been shown to be infected with SFV than with any other simian borne virus, SFV is a potentially powerful model for studying the virology and epidemiology of viruses at the human/NHP interface. In Asia, SFV is likely transmitted to humans through macaque bites and scratches that occur in the context of everyday life. We analyzed multiple proviral sequences from the SFV gag gene from both humans and macaques in order to characterize retroviral transmission at the human/NHP interface in Bangladesh. Here we report evidence that humans can be concurrently infected with multiple SFV strains, with some individuals infected by both an autochthonous SFV strain as well as a strain similar to SFV found in macaques from another geographic area. These data, combined with previous results, suggest that both human-facilitated movement of macaques leading to the introduction of non-resident strains of SFV and retroviral recombination in macaques contribute to SFV diversity among humans in Bangladesh.

CARF binds to three members (ARF, p53, and HDM2) of the p53 tumor-suppressor pathway.

The INK4a locus (chromosome 9p21) encodes two structurally distinct tumor-suppressor proteins, p16(INK4a) and the alternative reading frame protein, ARF (p19(ARF) in mouse and p14(ARF) in human). Each of these proteins has a major role in cell cycle control and senescence pathways. We originally identified a novel collaborator of ARF, CARF, from a two-hybrid interactive screen using p19(ARF) as bait and found that CARF interacts with ARF in the perinucleolar region and activates p53 function. In the absence of ARF, it interacts with p53 directly leading to ARF-independent enhancement of p53 function and in turn undergoes a negative feedback regulation. Very recently, we found that CARF interacts with HDM2 and undergoes degradation by an HDM2-dependent proteasome pathway. CARF may exert a vital control on p53-HDM2-p21(WAF1) pathway that is central to the cell cycle control, senescence, and DNA damage response of human cells.