Pain/Stress, Mitochondrial Dysfunction, and Neurodevelopment in Preterm Infants.

INTRODUCTION: Preterm infants experience tremendous early life pain/stress during their neonatal intensive care unit (NICU) hospitalization, which impacts their neurodevelopmental outcomes. Mitochondrial function/dysfunction may interface between perinatal stress events and neurodevelopment. Nevertheless, the specific proteins or pathways linking mitochondrial functions to pain-induced neurodevelopmental outcomes in infants remain unidentified. Our study aims to investigate the associations among pain/stress, proteins associated with mitochondrial function/dysfunction, and neurobehavioral responses in preterm infants. METHODS: We conducted a prospective cohort study, enrolling 33 preterm infants between September 2017 and July 2022 at two affiliated NICUs located in Hartford and Farmington, CT. NICU Network Neurobehavioral Scale (NNNS) datasets were evaluated to explore potential association with neurobehavioral outcomes. The daily pain/stress experienced by infant's during their NICU stay was documented. At 36-38 weeks post-menstrual age (PMA), neurobehavioral outcomes were evaluated using the NNNS and buccal swabs were collected for further analysis. Mass spectrometry-based proteomics was conducted on epithelial cells obtained from buccal swabs to evaluate protein expression level. Lasso statistical methods were conducted to study the association between protein abundance and infants' NNNS summary scores. Multiple linear regression and Gene Ontology (GO) enrichment analyses were performed to examine how clinical characteristics and neurodevelopmental outcomes may be associated with protein levels and underlying molecular pathways. RESULTS: During NICU hospitalization, preterm premature rupture of membrane (PPROM) was negatively associated with neurobehavioral outcomes. The protein functions including leptin receptor binding activity, glutathione disulfide oxidoreductase activity and response to oxidative stress, lipid metabolism, and phosphate and proton transmembrane transporter activity were negatively associated with neurobehavioral outcomes; in contrast, cytoskeletal regulation, epithelial barrier, and protection function were found to be associated with the optimal neurodevelopmental outcomes. In addition, mitochondrial function-associated proteins including SPRR2A, PAIP1, S100A3, MT-CO2, PiC, GLRX, PHB2, and BNIPL-2 demonstrated positive association with favorable neurodevelopmental outcomes, while proteins of ABLIM1, UNC45A, keratins, MUC1, and CYB5B showed positive association with adverse neurodevelopmental outcomes. CONCLUSION: Mitochondrial function-related proteins were observed to be associated with early life pain/stress and neurodevelopmental outcomes in infants. Large-scale studies with longitudinal datasets are warranted. Buccal proteins could be used to predict potential neurobehavioral outcomes.

Parental arsenic exposure and tissue-specific DNA methylation in Bangladeshi infants with spina bifida.

An emerging hypothesis linking arsenic toxicity involves altered epigenetic mechanisms, such as DNA methylation. In this study, we examined the relationship between parents' arsenic exposure and DNA methylation in tissues obtained from 28 infants with spina bifida from Bangladesh. We analyzed arsenic in parents' toenails using inductively coupled plasma mass spectrometry (ICP-MS). DNA methylation was measured in infants' dural tissue, buccal swabs, and whole blood using the Illumina Infinium MethylationEPIC BeadChip. We performed epigenome-wide association analyses (EWAS) and tested differentially methylated regions (DMRs). In EWAS, DNA methylation at cg24039697 in dural tissue was positively associated (ß = 0.59, p = 7.6 × 10-9) with father's toenail arsenic concentrations, adjusting for covariates. We did not identify any CpG sites related to father's arsenic exposure in the other tissues, or any CpG sites related to mother's arsenic exposure. Gene ontology analysis identified many biological pathways of interest, including the Wnt signaling pathways. We identified several DMRs across the tissues related to arsenic exposure that included probes mapping to genes that have previously been identified in studies of neural tube defects. This study emphasizes the potential impact of arsenic exposure in fathers, often understudied in epidemiological studies, on DNA methylation in a unique neurological tissue specific to spina bifida.

Antinociceptive and antioxidant potential of the crude ethanol extract of the leaves of Ageratum conyzoides grown in Bangladesh.

CONTEXT: Ageratum conyzoides Linn. (Asteraceae) is an annual herbaceous plant with a long history of traditional medicinal and agricultural uses; it is usually grown in the northeast part of Bangladesh. OBJECTIVE: The ethanol extract of the plant leaves was evaluated for preliminary phytochemical screening with its antinociceptive and antioxidant activities. MATERIALS AND METHODS: The preliminary phytochemical analysis was performed on the basis of standard procedures. The analgesic activity of the extract was investigated using the acetic acid-induced writhing method in mice. Five complementary tests such as DPPH free radical scavenging, nitric oxide (NO) scavenging, reducing power, Fe(++) ion chelating ability and total phenolic content were used for determining antioxidant activities. RESULTS: The results of preliminary phytochemical analysis showed the presence of alkaloids, reducing sugars, saponins, gums, steroids, tannins and flavonoids. The extract possessed a significant dose-dependent DPPH free radical scavenging activity with an IC50 value of 18.91 µg/ml compared to ascorbic acid (IC50: 2.937 µg/ml) and butylated hydroxyanisole (IC50: 5.10 µg/ml). The IC50 value of the extract for NO scavenging (41.81 µg/ml) was also found to be significant compared to the IC50 value of ascorbic acid (37.93 µg/ml). Moreover, the extract showed reducing power activity and Fe(++) ion chelating ability. The total phenolic amount was also calculated as quite high (378.37 mg/g of gallic acid equivalents) in the crude ethanol extract. DISCUSSION AND CONCLUSION: Therefore, the obtained results tend to suggest the antinociceptive and antioxidant activities of the ethanol extract of the plant leaves and justify its use in folkloric remedies.

Parental metal exposures as potential risk factors for spina bifida in Bangladesh.

BACKGROUND: Neural tube defects are a pressing public health concern despite advances in prevention from folic acid-based strategies. Numerous chemicals, in particular arsenic, have been associated with neural tube defects in animal models and could influence risk in humans. OBJECTIVES: We investigated the relationship between parental exposure to arsenic and 17 metals and risk of neural tube defects (myelomeningocele and meningocele) in a case control study in Bangladesh. METHODS: Exposure assessment included analysis of maternal and paternal toenail samples using inductively coupled plasma mass spectrometry (ICP-MS). A total of 278 participants (155 cases and 123 controls) with data collected from 2016 to 2020 were included in the analysis. RESULTS: In the paternal models, a one-unit increase in the natural logarithm of paternal toenail arsenic was associated with a 74% (odds ratio: 1.74, 95% confidence interval: 1.26-2.42) greater odds of having a child with spina bifida, after adjusting for relevant covariates. Additionally, paternal exposure to aluminum, cobalt, chromium, iron, selenium, and vanadium was associated with increased odds of having a child with spina bifida in the adjusted models. In the maternal models, a one-unit increase in the natural logarithm of maternal toenail selenium and zinc levels was related to a 382% greater (odds ratio: 4.82, 95% confidence interval: 1.32-17.60) and 89% lower (odds ratio: 0.11, 95% confidence interval: 0.03-0.42) odds of having a child with spina bifida in the adjusted models, respectively. Results did not suggest an interaction between parental toenail metals and maternal serum folate. DISCUSSION: Parental toenail levels of numerous metals were associated with increased risk of spina bifida in Bangladeshi infants. Paternal arsenic exposure was positively associated with neural tube defects in children and is of particular concern given the widespread arsenic poisoning of groundwater resources in Bangladesh and the lack of nutritional interventions aimed to mitigate paternal arsenic exposure. The findings add to the growing body of literature of the impact of metals, especially paternal environmental factors, on child health.

Topical Pilocarpine Formulation for Diagnosis of Cystic Fibrosis.

Cystic fibrosis is diagnosed in infants by estimating the levels of chloride ions present in the sweat induced by iontophoresis of pilocarpine solution. Elevated levels of chloride (≥60 mMol/L) in sweat are indicative of cystic fibrosis. However, the iontophoretic method of delivering pilocarpine is cumbersome and usually is associated with several side effects such as skin burn, skin rashes, erythema, and so forth. The objective of this study was therefore to develop a topical formulation that delivers adequate amount of pilocarpine. The drug delivery of formulation was compared with iontophoresis of aqueous solution of pilocarpine nitrate in vitro using porcine skin model. The pilocarpine levels in the skin exposed to topical pilocarpine solution under mild hyperthermia was 152.04 ± 52.23 µg/cm2 after 10 min of application, whereas it was 97.05 ± 27.93 µg/cm2 in the skin after 10 min of iontophoresis. The topical formulation was subjected to clinical evaluation to assess the efficacy of the product to induce sweat production. The average amount of the sweat secreted on application of topical formulation was found to be 77.28 ± 18.97 mg. Based on these results, it was found that the topical formulation was successful in delivering pilocarpine and to stimulate sweat secretion.

Assessment of physicians' knowledge to combat obesity in Bangladesh.

OBJECTIVE: Physicians need to play a proactive role to combat obesity and its associated comorbidities. The present survey was conducted to assess the awareness, knowledge, practice and attitude of the physicians in Bangladesh in the prevention and management of obesity. METHODS: Three hundred physicians were randomly selected from a medical university, a government medical college and a private medical college in Bangladesh to be included in this survey. All of them voluntarily participated in the survey upon the assurance of anonymity. All the selected physicians were provided with a questionnaire consisting of nine questions for assessing their awareness, knowledge, practice and attitude regarding obesity. RESULTS: Out of 300 participants, about 77% claimed that they know their own BMI and BMI cut-off points for overweight and obesity. But 38% physicians were unable to write the cut-off points correctly. Near about 50% physicians claimed that they know the BMI cut-off points for Asian population. However, only 7% were able to correctly write the BMI cut-off points for Asian population. About 47% physicians agreed that they do not calculate BMI or evaluate other measures of body fatness during clinical practice. However, 99% of the physicians considered that measuring BMI during consultation or clinical practice is important. CONCLUSIONS: It may be concluded that Bangladeshi physicians' have positive attitude for managing obesity but their practice is grossly inadequate. Most importantly, knowledge and awareness of the physicians about diagnosis of obesity is very poor.

Biomarkers of cardiovascular and metabolic diseases in otherwise healthy overweight subjects in Bangladesh.

Biomarkers of cardiovascular and metabolic diseases were assessed in Bangladeshi overweight subjects categorized by Asian BMI criteria. After screening 300 people, 90 apparently healthy subjects were enrolled and grouped into normal weight, overweight and obese. Compared to normal weight, the overweight and obese groups showed significant elevation of serum triglyceride, insulin, homocysteine, insulin resistance/sensitivity, and atherogenic index of plasma. Cardiometabolic biomarkers did not differ between overweight and obese groups. Some biomarkers showed correlation with BMI in overweight but not in obese group. It was concluded that cardiometabolic biomarkers are elevated in Bangladeshi overweight subjects categorized by Asian BMI criteria.

Vitamin D and Parathyroid Hormone Status in Female Garment Workers: A Case-Control Study in Bangladesh.

Despite the abundant sunlight, vitamin D deficiency is prevalent in South Asian countries including Bangladesh. Information on vitamin D level is insufficient in adults particularly in female garment workers in Bangladesh. This study was designed to evaluate the status of vitamin D, parathormone (PTH), calcium, and alkaline phosphatase (ALP) among the female garment workers in Bangladesh. Blood samples were collected from female garment workers (n = 40, case group) and general female workers (n = 40, control group) in Dhaka. Serum vitamin D, PTH, calcium, and ALP were measured by chemiluminescence microparticle immunoassay. The mean level of vitamin D was significantly (p < 0.001) lower in case (14.2 ± 2.6 ng/mL) than in the control (22.4 ± 2.4 ng/mL) group. No significant difference was found at mean of PTH and calcium between case (33.9 ± 17.2 pg/mL; 9.1 ± 0.6 mg/dL, resp.) and control (35.9 ± 16.3 pg/mL; 9.3 ± 0.6 mg/dL, resp.) group. The mean ALP in case (117.2 ± 14.4 U/L) group was significantly (p < 0.001) higher than the control group (80.5 ± 30.6 U/L). Overall, PTH level did not show significant correlation with vitamin D. However, calcium and ALP levels showed a significant positive (p < 0.05) and negative (p < 0.001) correlation with vitamin D, respectively. This study indicates a high prevalence of vitamin D deficiency in the female garment workers in Bangladesh.

Does the Interdependence between Oxidative Stress and Inflammation Explain the Antioxidant Paradox?

Oxidative stress has been implicated in many chronic diseases. However, antioxidant trials are so far largely unsuccessful as a preventive or curative measure. Chronic low-grade inflammatory process, on the other hand, plays a central role in the pathogenesis of a number of chronic diseases. Oxidative stress and inflammation are closely related pathophysiological processes, one of which can be easily induced by another. Thus, both processes are simultaneously found in many pathological conditions. Therefore, the failure of antioxidant trials might result from failure to select appropriate agents that specifically target both inflammation and oxidative stress or failure to use both antioxidants and anti-inflammatory agents simultaneously or use of nonselective agents that block some of the oxidative and/or inflammatory pathways but exaggerate the others. To examine whether the interdependence between oxidative stress and inflammation can explain the antioxidant paradox we discussed in the present review the basic aspects of oxidative stress and inflammation and their relationship and dependence.

Relationship of Soluble RAGE with Insulin Resistance and Beta Cell Function during Development of Type 2 Diabetes Mellitus.

This study examined whether circulating levels of soluble receptor for advanced glycation end products (sRAGE) alter in prediabetes and correlate with insulin resistance (IR) and beta cell function in prediabetes and newly diagnosed type 2 diabetes mellitus (T2DM). Subjects without previous history of diabetes were recruited and grouped as control, prediabetes, and newly diagnosed T2DM. The control subjects (n = 40) and people with prediabetes (n = 52) and diabetes (n = 66) were similar in terms of age, sex, BMI, systolic and diastolic BP, and fasting insulin level. HOMA-IR was found significantly higher in people with diabetes than control subjects (p < 0.001) and people with prediabetes (p = 0.005); and HOMA-%B was found significantly deteriorated in people with diabetes (p < 0.001) compared to control subjects and people with prediabetes. However, serum sRAGE levels did not show any significant alteration in people with prediabetes compared to control subjects. Moreover, univariate and multivariate analyses did not identify any significant correlation and statistical association of sRAGE with HOMA-IR and HOMA-%B in people with prediabetes and newly diagnosed T2DM. Our data suggest that serum sRAGE levels do not alter in people with prediabetes compared to control subjects and do not correlate or associate with IR and beta cell function during development of T2DM.

Performance of kala-azar surveillance in Gaffargaon subdistrict of Mymensingh, Bangladesh.

INTRODUCTION: Elimination of kala-azar is planned for South Asia requiring good surveillance along with other strategies. We assessed surveillance in Gaffargaon upazila (a subdistrict of 13 unions) of Mymensingh district, Bangladesh highly endemic for kala-azar. METHODS: In 4703 randomly sampled households, within nine randomly sampled villages, drawn from three randomly sampled unions, we actively searched for kala-azar cases that had occurred between January 2010 and December 2011. We then searched for medical records of these cases in the patient registers of Gaffargaon upazila health complex (UHC). We investigated factors associated with the medical recording by interviewing the cases and their families. We also did a general observation of UHC recording systems and interviewed health staff responsible for the monthly reports of kala-azar cases. RESULTS: Our active case finding detected 58 cases, but 29 were not recorded in the Gaffargaon UHC. Thus, only 50% (95% CI: 37%-63%) of kala-azar cases were reported via the government passive surveillance system. Interviews with health staff based in the study UHC revealed the heavy reporting burden for multiple diseases, variation in staff experience, high demands on the staff time and considerable complexity in the recording system. After adjusting for kala-azar treatment drug, recording was found more likely for those aged 18 years or more, males, receiving supply and administration of drug at the UHC, and more recent treatment. DISCUSSION: Fifty percent of kala-azar cases occurring in one highly endemic area of Bangladesh were recorded in registers that were the source for monthly reports to the national surveillance system. Recording was influenced by patient, treatment, staff and system factors. Our findings have policy implications for the national surveillance system. Future studies involving larger samples and including interviews with health authorities at more central level and surveillance experts at the national level will generate more precise and representative evidence on the performance of kala-azar surveillance in Bangladesh.

Phytochemical investigation and in vitro antinociceptive activity of Clerodendrum indicum leaves.

The crude ethanolic extracts of Clerodendrum indicum Linn. leaves were investigated for possible antinociceptive activity using acetic acid induced writhing model in mice. Phytochemical analysis was also carried out according to the standard procedures to identify the presence of different phytoconstituents in the ethanolic extract of the plant leaves. The study results showed 38.91 and 55.24% inhibition of writhings in the tested mice when ethanolic extract of Clerodendrum indicum Linn. leaves at doses of 250 and 500 mg kg(-1) body weight was given intraperitoneally, respectively. The study results were also compared with antinociceptive activity of the standard drug, Diclofenac sodium (68.37% inhibition) used at 25 mg kg(-1) body weight. At the above doses, the crude ethanolic extract of the plant showed significant antinociceptive activity in dose dependent fashion in acetic acid-induced writhing model in mice. The inhibition of writhings was calculated in respective to control group and it was found that p-values (<0.0001) obtained in all cases were extremely statistically significant. However, the phytochemical analysis showed the presence of alkaloid, steroid, saponin, tannin, reducing sugar and gum. The results suggest that crude ethanolic extracts of Clerodendrum indicum leaves possess significant antinociceptive properties justifying its folkloric use as analgesics and further research is necessary to isolate the principle phytochemical constituent(s) responsible for this activity.

Phytochemical screening, cytotoxicity and antibacterial activities of two Bangladeshi medicinal plants.

The objectives of the present study were to investigate phytochemical screening and to assay cytotoxicity and antibacterial activities of ethanolic extracts of leaves of two medicinal plants, Aglaonema hookerianum Schott (Family: Araceae) and Lannea grandis Engl. (Family: Anacardiaceae) available in Bangladesh. The brine shrimp lethality bioassay showed that the ethanolic extracts of Aglaonema hookerianum and Lannea grandis possessed cytotoxic activities with LC50 5.25 (microg mL(-1)) and 5.75 (microg mL(-1)) and LC90 10.47 (microg mL(-1)) and 9.55 (microg mL(-1)), respectively. Two extracts obtained from leaves were examined for their antibacterial activities against some gram positive bacteria such as Bacillus subtilis, Bacillus megaterium and Staphylococcus aureus, also gram negative strains of Pseudomonas aeruginosa, Escherichia coli, Shigella dysenteriae, Salmonella typhi, Salmonella paratyphi and Vibrio cholerae. Agar disc diffusion method was applied to observe the antibacterial efficacy of the extracts. Results indicated that both plant extracts (500 microg disc(-1)) displayed antibacterial activity against all of the tested microorganisms. These results were also compared with the zones of inhibition produced by commercially available standard antibiotic, Amoxicillin at concentration of 10 microg disc(-1). Observed antibacterial properties of the ethanolic extract of Aglaonema hookerianum Schott and Lannea grandis Engl. showed that both plants might be useful sources for the development of new potent antibacterial agents.