Microbial sensitivity of the common pathogens for UTIs are declining in diabetic patients compared to non-diabetic patients in Bangladesh: An institution-based retrospective study.

Background: Urinary tract infection (UTI) is one of the most recurrent infections in the community and healthcare settings. Although many studies related with microbial sensitivity (MS) of uropathogens (UPs) to antibiotics have been done in Bangladesh, no conclusive study has compared antibiotic sensitivity (AS) to UPs in diabetic and non-diabetic patients. The aim of the study is to find out whether there is a difference in AS in common UPs between diabetic and non-diabetic UTI patients. Methods: A retrospective review was conducted on 833 patients. The data was collected from different diagnostic centers located within Dhaka city in Bangladesh, and the data was analyzed using convenient statistical tools. Results: We have studied a total of 833 UTI patients. Out of 833 patients, 664 were diabetic and 169 were non-diabetic patients respectively. Among the studied population, females were found to be more inclined to have UTIs as compared to males. E. coli was found to be the leading UPs in our study. Patients within the age of 20-34 were more vulnerable to UTI in both groups. Imipenem and meropenem showed 100% sensitivity against E. coli, Staphylococcus and Klebsiella in non-diabetic patients, while both antibiotics showed lower sensitivity to the same organisms in diabetic patients. Antibiotics like nitrofurantoin (p ≤ 0.0002), ceftazidime (p ≤ 0.0124) and ceftriaxone (p ≤ 0.0168) showed less sensitivity to E. coli in diabetic UTI patients as compared to non-diabetic UTI patients. Overall sensitivity patterns elucidated that all the studied antibiotics, except ciprofloxacin and levofloxacin, showed lower sensitivity against UPs in diabetic while compared to non-diabetic UTI patients (p= <0.05 to 0.0001). Conclusion: We found significant difference in microbial sensitivity in patients with diabetes compared to non-diabetic UTI patients. Diabetes changes the pathophysiological state of the uropathogens leading to the declining sensitivity of the antibiotics in diabetic patients with UTIs.

Effects of cholesterol on the anionic magnetite nanoparticle-induced deformation and poration of giant lipid vesicles.

We have investigated the effects of cholesterol on the deformation and poration of giant unilamellar vesicles (GUVs) induced by anionic magnetite nanoparticles (NPs). Negatively charged lipid, neutral lipid, and cholesterol were used to prepare the charged GUVs (surface charge density of membranes - 0.16 C m-2), while only neutral lipid and cholesterol were used to prepare the neutral GUVs. Cholesterol content varied from 0 to 40 mole% for preparing the biologically relevant membranes. The degree of deformation has been characterized by compactness, the value of which remains at 1.0 for spherical GUVs. The value of compactness increases with time for both membranes, but this increase depends on cholesterol content. The average compactness decreases with cholesterol content, and at 60 min, the values are 1.280 ± 0.002 and 1.131 ± 0.010 for 0 and 40 mole% cholesterol containing charged GUVs. The average compactness is relatively lower for neutral GUVs for the corresponding cholesterol. Membrane poration has been investigated by the leakage of calcein, which indicates a two-state transition model. The fraction of deformation is higher for charged GUVs than for neutral ones, while the fraction of poration shows the opposite result. Both the fractions decrease with cholesterol content.

Influence of sugar concentration on the vesicle compactness, deformation and membrane poration induced by anionic nanoparticles.

Sugar plays a vital role in the structural and functional characteristics of cells. Hence, the interaction of NPs with cell membranes in the presence of sugar concentrations is important for medicinal and pharmacological innovations. This study integrated three tools: giant unilamellar vesicles (GUVs), anionic magnetite nanoparticles (NPs), and sugar concentrations, to understand a simplified mechanism for interactions between the vesicle membranes and NPs under various sugar concentrations. We focused on changing the sugar concentration in aqueous solution; more precisely, sucrose inside the GUVs and glucose outside with equal osmolarity. 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (sodium salt) (DOPG) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were used to prepare the charged membranes of 40mole%DOPG/60mole%DOPC-GUVs, whereas only DOPC was used to prepare the neutral membranes. Phase contrast fluorescence microscopy shows that the adherence of 18 nm magnetite NPs with anionic charge depends on the sugar concentration. The alterations of GUVs induced by the NPs are characterized in terms of i) vesicle compactness, ii) deformation, and iii) membrane poration. The presence of sugar provides additional structural stability to the GUVs and reduces the effects of the NPs with respect to these parameters; more precisely, the higher the sugar concentration, the smaller the alteration induced by the NPs. The differences in NPs effects are explained by the change in the type of interaction between sugar molecules and lipid membranes, namely enthalpy and entropy-driven interaction, respectively. In addition, such alterations are influenced by the surface charge density of the lipid bilayer. The surface pressure of membranes due to the adsorption of NPs is responsible for inducing the poration in membranes. The differences in deformation and poration in charged and neutral GUVs under various sugar concentrations are discussed based on the structure of the head of lipid molecules.

Effects of hydrocarbon chain on the vesicle size distribution, kinetics of average size, bending modulus, and elastic modulus of lipid membranes.

The effects of the hydrocarbon chain of lipids on the size distribution of giant unilamellar vesicles (GUVs), kinetics of average size, bending modulus, and elastic modulus of membranes have been investigated. 1,2-dioleoyl-sn-glycero-3-phosphocholine (18:1 (Δ9-Cis) PC (DOPC)), 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine (16:1 (Δ9-Cis) PC), and 1,2-ditridecanoyl-sn-glycerol-3-phosphocholine (13:0 PC (DTPC)) lipids were considered. The number of hydrocarbons in a chain of the corresponding lipid was 18, 16, and 13. GUVs were prepared using the natural swelling method under incubation times of 20, 40, 60, 90, 120, and 180 min. The size distribution of vesicles was fitted using the lognormal distribution. The average sizes of DOPC, 16:1 (Δ9-Cis) PC, and DTPC-GUVs increased with the incubation time until 120 min, and then remained steady at 16.7 ± 0.2, 15.2 ± 0.4 and 12.0 ± 0.3 µm for the corresponding lipids. The average size at equilibrium state increased with the number of hydrocarbons. The incubation time-dependent average size was fitted with an exponential growth equation, and then the kinetic constants of 0.028 ± 0.004, 0.036 ± 0.007, and 0.083 ± 0.009 min-1 for DOPC, 16:1 (Δ9-Cis) PC, and DTPC-GUVs, respectively, were obtained. The equilibrium size distribution was fitted by the theoretical equation, and the bending modulus for DOPC, 16:1 (Δ9-Cis) PC, and DTPC membranes were 19.5 ± 0.2, 18.5 ± 0.1 and 14.3 ± 0.1 kBT, respectively. The bending modulus increased with the number of hydrocarbons. The elastic modulus of these membranes was 261 mN/m with a 4% fluctuation. The correlation between the average size and the square root of the bending modulus was supported by theoretical analysis.

A new purification technique to obtain specific size distribution of giant lipid vesicles using dual filtration.

A new purification technique is developed for obtaining distribution of giant unilamellar vesicles (GUVs) within a specific range of sizes using dual filtration. The GUVs were prepared using well known natural swelling method. For filtration, different combinations of polycarbonate membranes were implemented in filter holders. In our experiment, the combinations of membranes were selected with corresponding pore sizes-(i) 12 and 10 µm, (ii) 12 and 8 µm, and (iii) 10 and 8 µm. By these filtration arrangements, obtained GUVs size distribution were in the ranges of 6-26 µm, 5-38 µm and 5-30 µm, respectively. In comparison, the size distribution range was much higher for single filtration technique, for example, 6-59 µm GUVs found for a membrane with 12 µm pores. Using this technique, the water-soluble fluorescent probe, calcein, can be removed from the suspension of GUVs successfully. The size distributions were analyzed with lognormal distribution. The skewness became smaller (narrow size distribution) when a dual filtration was used instead of single filtration. The mode of the size distribution obtained in dual filtration was also smaller to that of single filtration. By continuing this process of purification for a second time, the GUVs size distribution became even narrower. After using an extra filtration with dual filtration, two different size distributions of GUVs were obtained at a time. This experimental observation suggests that different size specific distributions of GUVs can be obtained easily, even if GUVs are prepared by different other methods.

Millions of Bangladeshi Children Missed Their Scheduled Vaccination Amidst COVID-19 Pandemic.

The Government of Bangladesh imposed a movement control order as a mass quarantine strategy to control the outbreak of coronavirus disease 2019 (COVID-19). Adherence to the home quarantine may put children at risk by missing routine vaccination. In this study, we investigated the impact of COVID-19 on child routine immunization in a rural area of Bangladesh and consider the broader implications. Data for this study comes from the Chakaria Health and Demographic Surveillance System (HDSS) of icddr,b with a population of 90,000 people residing in 16,000 households in 49 villages in a rural, coastal area of Southeast Bangladesh. We used an explanatory sequential mixed methods design which involved two phases between March 1, 2020, and May 31, 2020: first, we observed 258 outreach sessions of 86 EPI centers. We calculated the number of Expanded Program on Immunization (EPI) outreach sessions suspended and the number of children who missed their routine vaccination due to the COVID-19. We extrapolated the number of Bangladeshi children who missed their routine vaccination using Chakaria HDSS observations. Secondly, we conducted in-depth interviews to explain the quantitative results. The EPI outreach session (EOS) declined to 74.42% (95% CI 63.97-83.22), 10.45% (95% CI 5.00-18.94), and 3.45% (95% CI 1.00-9.75) from 2019 levels in March, April, and May 2020, respectively. By extrapolation, in Bangladesh, between March and May 2020, 3.2 million children missed their scheduled vaccination compared to 2019. Results from in-depth interviews showed that the unwillingness of villagers to hold EOS and the absenteeism of the vaccinators due to social distancing recommendations and lack of personal safety measures were the main reasons for the discontinuation of the EOS. Resuming EPI outreach sessions and introducing a special catch-up program is essential to prevent future outbreaks and deaths due to vaccine-preventable diseases in Bangladesh and the countries where children missed their routine vaccination due to COVID-19. This health system failure should be considered a factor in all future pandemic preparedness plans.

The Effect of Gestational Diabetes Mellitus on the Chorionic Villi of Human Placenta Among Saudi Arabian Mothers: A Quantitative and Comparative Study.

Background The placenta is a temporary pivotal organ - the gate between the mother and the fetus. It has multiple functions such as nutrient uptake, elimination of waste products, gas exchange, and production of some vital hormones. However, the pregnancy state is a diabetogenic condition caused by insulin resistance, resulting from physiological variations. Gestational diabetes mellitus (GDM) can have an impact on both the mother and the fetus by causing numerous complications. In our research study, we aim to study and compare the quantitative effect of GDM at the microscopical level within the chorionic villi of the placenta of both mothers diagnosed with GDM and healthy mothers as well as the clinical correlation. Methods After applying the inclusion and exclusion criteria, we collected 84 placental samples from February 2017 until May 2017, which were composed of 42 GDM samples and 42 healthy samples. All of these samples have been studied under a light microscope for measuring different parameters. Results We found that some of the measured parameters among diabetic villi were lower than those of healthy villi with a p-value < .05 being significant. These include the surface area of the blood vessels (P = .008), the perimeters of the blood vessels (P = .002), the placental barrier thickness/perimeters of the villous blood vessels ratio (P ≤ .001), the placental barrier thickness/surface area of the blood vessels ratio (P ≤ .001), the number of Hofbauer cells/surface area of the villous ratio (P ≤ .001), the number of the blood vessels/surface area of the villous ratio (P = .001), the surface area of the blood vessels/surface area of the villous ratio (P = .004), and the perimeters of the blood vessels/surface area of the villous ratio (P ≤ .001). These parameters have significant effects on fetal development as well as the mother's status. Conclusions GDM is associated with multiple changes in the placenta. Moreover, these changes can impact the fetoplacental circulation and cause multiple complications for the mother and the fetus. Therefore, identifying pregnant women with GDM and controlling hyperglycemia will improve the outcomes of the pregnancy.

The Body-wide Transcriptome Landscape of Disease Models.

Virtually all diseases affect multiple organs. However, our knowledge of the body-wide effects remains limited. Here, we report the body-wide transcriptome landscape across 13-23 organs of mouse models of myocardial infarction, diabetes, kidney diseases, cancer, and pre-mature aging. Using such datasets, we find (1) differential gene expression in diverse organs across all models; (2) skin as a disease-sensor organ represented by disease-specific activities of putative gene-expression network; (3) a bone-skin cross talk mediated by a bone-derived hormone, FGF23, in response to dysregulated phosphate homeostasis, a known risk-factor for kidney diseases; (4) candidates for the signature activities of many more putative inter-organ cross talk for diseases; and (5) a cross-species map illustrating organ-to-organ and model-to-disease relationships between human and mouse. These findings demonstrate the usefulness and the potential of such body-wide datasets encompassing mouse models of diverse disease types as a resource in biological and medical sciences. Furthermore, the findings described herein could be exploited for designing disease diagnosis and treatment.

Protein-based open sandwich immuno-PCR for sensitive detection of small biomarkers.

Open sandwich (OS) immunoassay utilizes antigen-dependent stabilization of an antibody variable region to quantify various antigens, enabling noncompetitive detection of small molecules with a broad working range. For further improvement of its sensitivity, OS Immuno-PCR was attempted with recombinant fusion proteins. The maltose binding protein-fused heavy chain variable region (MBP-V(H)) of an antibody that recognizes the C-terminal fragment of human osteocalcin (bone Gla protein, BGP), a biomarker for bone-related diseases, was immobilized onto microplate wells, and the antigen together with streptavidin (SA)-fused light chain variable region of the same antibody (SA-V(L)) was added and incubated. The amount of immobilized SA-V(L) was quantified by tethered biotinylated DNA, which was used to estimate the amount of antigen by realtime PCR. When BGP C-terminal peptide was detected, the limit of detection was 100 fg/mL, which was superior than that of our previously reported phage-based OS Immuno-PCR. The developed OS Immuno-PCR system will be useful for the detection of small molecule biomarkers for disease prevention.

Profile: the Chakaria Health and Demographic Surveillance System.

Chakaria Health and Demographic Surveillance System (CHDSS), located on the south-eastern coast of the Bay of Bengal, was established in 1999 and is one of the field sites of International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDRB). The surveillance covers 118 315 residents living in 19 847 households. Data on socio-demographic and health indicators including birth, death, migration, marriage, maternal health, education and employment are recorded through quarterly household visits. The primary objective of CHDSS is to monitor the changes in socio-demographic indicators, inequalities in health and impact of public health interventions. A demographic change was accompanied by a shift from traditional to modern society during the past decade, but inequality in health still persists. The findings from the surveillance are shared regularly among the local and global communities. Data are also available upon request to ICDDRB and INDEPTH for use by researchers and policy makers.

Detection of small molecule diagnostic markers with phage-based open-sandwich immuno-PCR.

Open sandwich immunoassay (OS-IA) utilizes antigen-dependent stabilization of antibody variable region to quantify various antigens, enabling noncompetitive detection of small molecules with a broad working range. To further improve its detection sensitivity, here we employed phage-based immuno-PCR approach, wherein OS-IA and quantitative PCR methodologies were combined with the use of immobilized V(L) fusion protein and filamentous phages displaying V(H) fragment, whose DNA was extracted for PCR amplification. This approach significantly enhanced the assay sensitivity for small molecule antigens osteocalcin (BGP) peptide and 17beta-estradiol.