Radiation-Induced Hydrogel for Water Treatment.

Along with serving as drug delivery sensors and flexible devices, hydrogels are playing pioneering roles in water purification. Both chemical and radiation methods can produce hydrogels, with the latter method gaining preference for its pure adducts. The water treatment process entails the removal of heavy and toxic metals (above the threshold amount), dyes, and solid wastes from industrial effluents, seawater, and groundwater, as well as sterilization for microorganism destruction. This review analyzed the different types of hydrogels produced by applying various radiations for water treatment. Particularly, we examined the hydrogels created through the application of varying levels of gamma and electron beam radiation from the electron gun and Co-60 sources. Moreover, we discuss the optimized radiation doses, the compositions (monomers and polymers) of raw materials required for hydrogel preparation, and their performance in water purification. We present and predict the current state and future possibilities of radiation-induced hydrogels. We explain and compare the superiority of one radiation method over other radiation methods (UV-visible, X-ray, microwave, etc.) based on water treatment.

Anthropometric Study of Foot Breadth of 5-10 Years Aged Bangladeshi Children.

The present anthropometric study was designed to construct data of 5-10 years aged Bangladeshi children regarding foot breadth and an attempt has been made out to grow interest among the researchers for future study and also to compare the data with the data of the people of other races. This cross-sectional, descriptive and analytic type study was conducted among 5-10 years aged 109 Bangladeshi children (70 male and 39 female) at different areas of Mymensingh district (Fulpur, Muktagacha, Fulbaria, Trisal and Haluaghat), Bangladesh from January 2016 to December 2016. Non-random purposive sampling technique was taken for sample collection. Any kind of foot deformity due to either from congenital or physical injury was excluded to construct standard measurement. Foot breadth was measured using slide caliper. The children were asked to stand with weight distributed equally on both feet. The legs were perpendicular to the feet. The mean foot breadth of right side of 5, 6, 7, 8, 9 and 10 years aged male children were 7.08±.61 cm, 7.11±.30 cm, 7.61±.45 cm, 7.44±.43 cm, 8.12±.38 cm and 8.17±.44 cm respectively and those of female children were 6.55±.64 cm, 7.17±.27 cm, 7.04±.70 cm, 7.62±.54 cm, 7.48±.88 cm and 7.85±.72 cm respectively. The mean foot breadth of left side of 5, 6, 7, 8, 9 and 10 years aged male children were 7.04±.60 cm, 7.03±.30 cm, 7.52±.47 cm, 7.36±.45 cm, 8.03±.38 cm and 8.08±.43 cm respectively and those of female children were 6.46±.63 cm, 7.10±.31 cm, 6.95±.71 cm, 7.54±.52 cm, 7.37±.89 cm and 7.77±.71 cm respectively. Comparison of foot breadth between male and female children was done by Unpaired Students 't' test and the differences of means were statistically non-significant. Data were tabulated and statistically analyzed using Microsoft excel and SPSS software. The results of present study would be useful in anatomy, orthopedics, forensic science, plastic surgery, radiology, podiatry, archeology, anthropology and nutrition science.

Breast Cancer in Younger and Older Women: A Comparison of Clinicopathological Traits.

Breast cancer stands as the prevailing invasive cancer globally, bearing high mortality rates among women. Existing evidence indicates diminished survival rates in younger patients. Consequently, this study endeavors to assess and contrast the pathological features of breast cancer in women under 40 years of age with their older counterparts. Conducted as a cross-sectional analysis, this study encompasses 560 patients diagnosed with breast cancer, seeking treatment at Mymensingh Medical College Hospital (MMCH), Community Based Medical College Bangladesh (CBMCB) and several private hospitals in Mymensingh. The gathered data incorporates information such as age, residential area, occupation, tumor histopathology, TNM classification, staging and status of hormone receptor. The patients' mean age (standard deviation) was 49.7±11.9 years, with 20.5% below 40, most were from rural areas and were housewives. Ductal carcinoma prevailed as the most common histopathologic type (87.67%). However, younger patients exhibited a higher prevalence of lobular and other rare carcinomas compared to their older counterparts (p=0.04). Additionally, the younger group demonstrated larger tumor sizes (p=0.01), lymphatic node involvement (p=0.04) and advanced staging (p=0.004). Notably, younger age showed more negativity for estrogen and/or progesterone receptors. The results suggested that women under 40 years old exhibit more aggressive tumor characteristics and a more severe form of breast cancer compared to their older counterparts.

Primary Tuberculosis of Cervix Which Simulated Endocervical Polyp: A Case Study.

Tuberculosis has been described as the second great "Imitator" as it can imitate various other disease processes. The manifestations of genitourinary tuberculosis are protean in nature; still tuberculosis is a health concern in South-East Asia region. Tuberculosis of the cervix is rarely found and accounts for 5-10% among all types of genital tuberculosis. Despite meticulous history and clinical examination does not always lead to suspect this disease, the definitive diagnosis is based on the demonstration of the characteristic lesion on histopathology or on bacterial isolation. We are reporting a case of a 26-years-old woman who presented with secondary amenorrhea and a benign looking endocervical polyp. Diagnosis of cervical tuberculosis could be clinched after tissue biopsy which revealed caseous granuloma on histopathological examination along with other supportive laboratory investigation reports. Patient was subsequently started on antitubercular therapy (ATT) according to directly observed treatment schedule- category I, resulting in resumption of her menses after four months of starting of ATT. An awareness of the atypical clinical manifestations of tuberculosis is important, especially in regions where tuberculosis continues to be a major public health problem, such as Bangladesh. One should have high index of suspicion in order to diagnose tuberculosis of cervix in such cases, especially in high prevalence areas, so that patients can be managed appropriately with antitubercular therapy and complications can be prevented.

TRIB1 confers therapeutic resistance in GBM cells by activating the ERK and Akt pathways.

GBM (Glioblastoma) is the most lethal CNS (Central nervous system) tumor in adults, which inevitably develops resistance to standard treatments leading to recurrence and mortality. TRIB1 is a serine/threonine pseudokinase which functions as a scaffold platform that initiates degradation of its substrates like C/EBPα through the ubiquitin proteasome system and also activates MEK and Akt signaling. We found that increased TRIB1 gene expression associated with worse overall survival of GBM patients across multiple cohorts. Importantly, overexpression of TRIB1 decreased RT/TMZ (radiation therapy/temozolomide)-induced apoptosis in patient derived GBM cell lines in vitro. TRIB1 directly bound to MEK and Akt and increased ERK and Akt phosphorylation/activation. We also found that TRIB1 protein expression was maximal during G2/M transition of cell cycle in GBM cells. Furthermore, TRIB1 bound directly to HDAC1 and p53. Importantly, mice bearing TRIB1 overexpressing tumors had worse overall survival. Collectively, these data suggest that TRIB1 induces resistance of GBM cells to RT/TMZ treatments by activating the cell proliferation and survival pathways thus providing an opportunity for developing new targeted therapeutics.

Conversion of Laparoscopic to Open Cholecystectomy: Experience in A Tertiary Care Hospital.

Now-a-days Laparoscopic cholecystectomy is regarded as the gold standard treatment for benign gallbladder disease but in certain situations conversion to open cholecystectomy is extremely important for the safety of the patient. The objective of this study was to evaluate the reason for conversion of this operation to open surgery. This prospective study was carried out on 392 patients in a single unit of Department of Surgery, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh and in a private hospital from July 2013 to December 2018. Maximum (28.3%) patients were 31-40 years age group. Majority (75.3%) was female and 24.7% were male. It was observed that only 2.1% were converted due to dense adhesion (n=3), severe inflammation (n=2), difficult to define anatomy of Calot's triangle (n=2) and Mirizzi syndrome (n=1). Meticulous dissection and proper case selection can reduce the rate of conversion to open surgery.

Haploinsufficiency of NFKBIA reshapes the epigenome antipodal to the IDH mutation and imparts disease fate in diffuse gliomas.

Genetic alterations help predict the clinical behavior of diffuse gliomas, but some variability remains uncorrelated. Here, we demonstrate that haploinsufficient deletions of chromatin-bound tumor suppressor NFKB inhibitor alpha (NFKBIA) display distinct patterns of occurrence in relation to other genetic markers and are disproportionately present at recurrence. NFKBIA haploinsufficiency is associated with unfavorable patient outcomes, independent of genetic and clinicopathologic predictors. NFKBIA deletions reshape the DNA and histone methylome antipodal to the IDH mutation and induce a transcriptome landscape partly reminiscent of H3K27M mutant pediatric gliomas. In IDH mutant gliomas, NFKBIA deletions are common in tumors with a clinical course similar to that of IDH wild-type tumors. An externally validated nomogram model for estimating individual patient survival in IDH mutant gliomas confirms that NFKBIA deletions predict comparatively brief survival. Thus, NFKBIA haploinsufficiency aligns with distinct epigenome changes, portends a poor prognosis, and should be incorporated into models predicting the disease fate of diffuse gliomas.

Optimized Box-Behnken Design Combined Response Surface Methodology to Determine Calcium and Iron Contents Using Visible, Atomic Emission and Atomic Absorption Spectrophotometry in Vegetables and Wastewater Samples.

BACKGROUND: Calcium and iron are crucial essential minerals. Iron is mainly responsible for transporting oxygen in the body and the immune system. In comparison, calcium's primary function is in human bones and teeth. Due to that, it is vital to quantify the amount in vegetables. OBJECTIVE: Optimization and validation of three analytical procedures, visible, atomic emission spectrophotometry (AES), and atomic absorption spectrophotometry (AAS), were developed to determine calcium and iron in vegetables and wastewater samples using response surface methodology (RSM) via Box-Behnken design (BBD). The design helps to reduce experiment trials with selected variables to find a correlation between them and their respective dependent variables. METHODS: Method I was developed to quantify calcium in vegetables mixed with concentrated 3M HNO3 and heated to reflux as per the BBD. Then it was cooled, filtered, and completed with 3M HNO3 to be carried out utilizing AES and AAS. For method II, vegetables were mixed with nitric acid and sulfuric acid solution with an optimized 5M KSCN solution, which was computed using the AAS and visible spectrophotometry. RESULTS: First, percentage of water content was calculated for all vegetables, higher in malabar spinach and lower in peas. The calcium and iron contents were present within 0.59-2.68 mg and 35.8-211.5 mg, respectively, in 100 g of vegetables. The results showed a higher amount of iron was available in spinach and a lower amount in okra. In contrast, the highest calcium amount was present in broccoli and the lowest amount was in peas. The calcium and iron content were between 0.015-137.25 and 0.01-147.85 µg/mL in the wastewater samples. CONCLUSIONS: These methods can help to determine the amount of calcium and iron for the quality control samples in research and development, food, and the environmental industry. HIGHLIGHTS: Three validated analytical techniques quantify calcium and iron in vegetables and wastewater samples. The RSM-BBD optimized the method and determined its crucial factors.

MicroRNA-575 acts as a novel oncogene via targeting multiple signaling pathways in glioblastoma.

PURPOSE: Glioblastoma (GBM) patients currently face poor survival outcomes with an average survival period of <15 months, while only 3-5% of patients survive longer than 36 months. Although the mechanisms of tumorigenesis are still being elucidated, miRNAs are promising candidates to explore as novel and prognostic biomarkers in GBM. In this study, we identified the association between miR-575 expression and overall survival (OS) of primary GBM patients and undertook functional studies to discern the contribution of miR-575 to GBM tumorigenesis. METHODS: Total RNAs were isolated from 254 FFPE GBM tumor samples and miR expression was assayed (simultaneously) using NanoString Technologies. To determine the association between miR-575 and patients' prognosis, Kaplan-Meier, univariable and multivariable Cox regression analyses were performed. Cell proliferation, colony formation, migration assays were conducted to investigate the function of miR-575 in vitro and in vivo. In silico target gene network analysis was performed to identify the putative targets of miR-575 in GBM, which were further verified by luciferase reporter assay, as well as qPCR and immunoblotting. RESULTS: Our clinical data (n = 254) show that miR-575 is associated with worse GBM OS by univariable analysis (UVA, HR = 1.27, p-value<0.001) and multivariable (MVA, HR = 1.23, p = 0.007) analysis incorporating critical clinical variables. Functional studies indicated that overexpression of miR-575 significantly increased cell proliferation and migration of GBM cells in vitro, as well as tumor growth in vivo. Subsequent in silico target gene network and mechanistic studies identified CDKN1B/p27 and PTEN, as potential targets of miR-575 in GBM. MicroRNA-575 can also regulate the activity of AKT and ERK pathways in GBM. CONCLUSION: miR-575 has prognostic value in GBM, with higher expression associating with worse OS of patients, and contributes to GBM tumorigenesis by regulating multiple signaling pathways in GBM.

ADC Histogram Analysis of Pediatric Low-Grade Glioma Treated with Selumetinib: A Report from the Pediatric Brain Tumor Consortium.

BACKGROUND AND PURPOSE: Selumetinib is a promising MAP (mitogen-activated protein) kinase (MEK) 1/2 inhibitor treatment for pediatric low-grade gliomas. We hypothesized that MR imaging-derived ADC histogram metrics would be associated with survival and response to treatment with selumetinib. MATERIALS AND METHODS: Children with recurrent, refractory, or progressive pediatric low-grade gliomas who had World Health Organization grade I pilocytic astrocytoma with KIAA1549-BRAF fusion or the BRAF V600E mutation (stratum 1), neurofibromatosis type 1-associated pediatric low-grade gliomas (stratum 3), or sporadic non-neurofibromatosis type 1 optic pathway and hypothalamic glioma (OPHG) (stratum 4) were treated with selumetinib for up to 2 years. Quantitative ADC histogram metrics were analyzed for total and enhancing tumor volumes at baseline and during treatment. RESULTS: Each stratum comprised 25 patients. Stratum 1 responders showed lower values of SD of baseline ADC_total as well as a larger decrease with time on treatment in ADC_total mean, mode, and median compared with nonresponders. Stratum 3 responders showed a greater longitudinal decrease in ADC_total. In stratum 4, higher baseline ADC_total skewness and kurtosis were associated with shorter progression-free survival. When all 3 strata were combined, responders showed a greater decrease with time in ADC_total mode and median. Compared with sporadic OPHG, neurofibromatosis type 1-associated OPHG had lower values of ADC_total mean, mode, and median as well as ADC_enhancement mean and median and higher values of ADC_total skewness and kurtosis at baseline. The longitudinal decrease in ADC_total median during treatment was significantly greater in sporadic OPHG compared with neurofibromatosis type 1-associated OPHG. CONCLUSIONS: ADC histogram metrics are associated with progression-free survival and response to treatment with selumetinib in pediatric low-grade gliomas.

Polarization splitting with cubic crystals evaluated with synchrotron radiation.

X-ray polarization-splitting crystals separate incident x rays into two components with perpendicular polarization by Bragg reflections at 45° from paired sets of internal planes. Here, the polarization-splitting properties of a germanium crystal are verified using incompletely polarized synchrotron radiation. Cleaner data would have come from a beam with a higher degree of polarization, which is achievable with small changes in the experimental geometry.

Crosstalk of Cyclin-dependent kinase inhibitor 1A (CDKN1A) gene polymorphism with p53 and CCND1 polymorphism in breast cancer.

OBJECTIVE: Mutations and polymorphisms in genes of cell- cycle and apoptosis regulatory pathway influence the breast cancer risk. Analysis of single low penetrance mutant alleles may not reflect the precise risk association when analyzed alone. PATIENTS AND METHODS: A total of 115 DNA samples extracted from breast cancer patients and an equal number of age and sex-matched normal controls were used for polymorphic analysis. Genotyping for p21 rs1801270 and CCND1 rs603965 was done by PCR-RFLP method while AFLP method was used for p53 rs1042522 single nucleotide polymorphism detection. Statistical methods included simple mean±SD and correlation coefficient to analyze the risk of association of p21, p53 and CCND1 SNPs and breast cancer. RESULTS: Individuals harboring SNPs in p21, p53 and CCND1 genes namely rs1801270, rs1042522 and rs603965, respectively were rendered increasingly susceptible to developing breast cancer when compared with normal controls. CONCLUSIONS: Our report emphasizes the need of combinational analysis of low-penetrance mutant alleles to assess accurately their association with breast cancer risk. Future case-control studies analyzing gene-environment interactions across different populations may confirm reported risk associations of studied polymorphisms with developing breast cancer.

Chrysin inhibits propagation of HeLa cells by attenuating cell survival and inducing apoptotic pathways.

OBJECTIVE: Chrysin, one of the main active constituents of flavonoids, is known for demonstrating protective effects against various types of cancer including cervical cancer. The aim of this study was to determine apoptosis induction and antiproliferative action of chrysin on human cervical cancer cells. MATERIALS AND METHODS: In this study, attempts have been made to establish anticancer role of chrysin on HeLa cells. MTT, mitochondrial potential, DNA fragmentation, annexin V/propidium iodide assays, qPCR and protein profiling were performed. RESULTS: Chrysin treated HeLa cells showed time and dose dependent decrease in cell viability and demonstrated profound effects on nuclear morphology and DNA fragmentation. Chrysin treatment increased the expression of proapoptotic genes BAD, BAX, BID, BOK and APAF1, TNF, FASL, FAS, FADD and caspases (like caspase 3, caspase 7, caspase 8 and caspase 9), whereas it decreased the expression level of antiapoptotic genes MCL-1, NAIP, XIAP and Bcl-2 and cell cycle regulatory genes CCNB1, CCNB2, CCND1, CCND2, CCND3, CCNE2, CDK4 and CDK2 at transcript level. Furthermore, chrysin significantly upregulated pro-apoptotic proteins, like TRAILR2/DR5, TRAILR1/DR4, Fas/TNFRSF6/CD95, phosphoP53(S15), BAD, BAX, cleaved caspase 3, procaspase 3, HTRA2/Omi and SMAC/Diablo, while downregulated anti-apoptotic proteins like BCL-X, BCL2, XIAP and CIAPs that support chrysin mediated apoptosis in HeLa cells. Remarkably, chrysin downregulated the phosphorylated AKT pathway proteins, (p-473) AKT, (p-Ser 2448) mTOR, (p-Ser241) PDK1, (p-Ser112) BAD, and upregulated (p-Ser21) GSK3b, (p-Thr172) AMPKa, P27 (p-Thr198) and (p-Ser15) P53, which endorses chrysin mediated apoptosis. CONCLUSIONS: Chrysin significantly inhibited proliferation and induced apoptosis by modulation of various apoptotic genes and AKT/MAPK pathway genes.

Tumor Heterogeneity in Glioblastomas: From Light Microscopy to Molecular Pathology.

One of the main reasons for the aggressive behavior of glioblastoma (GBM) is its intrinsic intra-tumor heterogeneity, characterized by the presence of clonal and subclonal differentiated tumor cell populations, glioma stem cells, and components of the tumor microenvironment, which affect multiple hallmark cellular functions in cancer. "Tumor Heterogeneity" usually encompasses both inter-tumor heterogeneity (population-level differences); and intra-tumor heterogeneity (differences within individual tumors). Tumor heterogeneity may be assessed in a single time point (spatial heterogeneity) or along the clinical evolution of GBM (longitudinal heterogeneity). Molecular methods may detect clonal and subclonal alterations to describe tumor evolution, even when samples from multiple areas are collected in the same time point (spatial-temporal heterogeneity). In GBM, although the inter-tumor mutational landscape is relatively homogeneous, intra-tumor heterogeneity is a striking feature of this tumor. In this review, we will address briefly the inter-tumor heterogeneity of the CNS tumors that yielded the current glioma classification. Next, we will take a deeper dive in the intra-tumor heterogeneity of GBMs, which directly affects prognosis and response to treatment. Our approach aims to follow technological developments, allowing for characterization of intra-tumor heterogeneity, beginning with differences on histomorphology of GBM and ending with molecular alterations observed at single-cell level.

Role of endophytic Penicillium species and Pseudomonas monteilii in inducing the systemic resistance in okra against root rotting fungi and their effect on some physiochemical properties of okra fruit.

AIMS: The efficacy of three isolates of endophytic Penicillium species that have shown significant suppressive effect on root rotting fungi in our previous study were further evaluated in pots and field plot experiments for their effect on root diseases of okra, induction of systemic resistance and physiochemical properties of okra fruit. METHODS AND RESULTS: Aqueous suspensions of endophytic Penicillium and Pseudomonas monteilii were applied in pots and field plots using okra as test plant. Data on the fungal infection of roots, plant growth, plant resistance markers like polyphenol, salicylic acid and antioxidant status of plant were determined. These isolates significantly suppressed root diseases and induced systemic resistance via increasing level of resistance markers, polyphenol and salicylic acid besides improving antioxidant activity of Penicillium and P. monteilii-treated plants as compared to control plants. GC-MS analysis of n-hexane extract of mycelium of P. nigricans revealed the presence of 15 different volatile compounds. CONCLUSIONS: Endophytic Penicillium and P. monteilii have potential against root-infecting fungi of okra and can improve plant growth and yield. SIGNIFICANCE AND IMPACT OF THE STUDY: Endophytic Penicillium species and P. monteilii can suppress root rotting fungi by direct mechanism or induction of systemic resistance in plants.

A Novel miR-146a-POU3F2/SMARCA5 Pathway Regulates Stemness and Therapeutic Response in Glioblastoma.

Rapid tumor growth, widespread brain-invasion, and therapeutic resistance critically contribute to glioblastoma (GBM) recurrence and dismal patient outcomes. Although GBM stem cells (GSC) are shown to play key roles in these processes, the molecular pathways governing the GSC phenotype (GBM-stemness) remain poorly defined. Here, we show that epigenetic silencing of miR-146a significantly correlated with worse patient outcome and importantly, miR-146a level was significantly lower in recurrent tumors compared with primary ones. Further, miR-146a overexpression significantly inhibited the proliferation and invasion of GBM patient-derived primary cells and increased their response to temozolomide (TMZ), both in vitro and in vivo. Mechanistically, miR-146a directly silenced POU3F2 and SMARCA5, two transcription factors that mutually regulated each other, significantly compromising GBM-stemness and increasing TMZ response. Collectively, our data show that miR-146a-POU3F2/SMARCA5 pathway plays a critical role in suppressing GBM-stemness and increasing TMZ-response, suggesting that POU3F2 and SMARCA5 may serve as novel therapeutic targets in GBM. IMPLICATIONS: miR-146a predicts favorable prognosis and the miR-146a-POU3F2/SMARCA5 pathway is important for the suppression of stemness in GBM.

Phytochemicals induce apoptosis by modulation of nitric oxide signaling pathway in cervical cancer cells.

OBJECTIVE: Nitric Oxide (NO) is produced by NO synthases (NOS) and is a key signaling molecule that regulates tumorigenesis, both aiding and alleviating it. Elevated NO levels are cytotoxic to cancer cells, making NOS an important target for cancer treatment. In the present study, the modulatory effects of the phytochemicals, quercetin, sulforaphane, genistein, and epigallocatechin-3-gallate on NO pathway and apoptosis were shown in HeLa cervical cancer cells. MATERIALS AND METHODS: Fluorescent microscopy and flow cytometry were used to assess apoptosis. A Griess assay was used to quantitatively measure NO, quantitative PCR array was used to assess the expression levels of genes involved in the NO signaling pathway, and immunocytochemistry was used to determine NOS protein expression. The functional association among the modulated genes was evaluated using network biology analysis, gene set enrichment, and KEGG pathway analysis. RESULTS: Treatment with the phytochemicals elevated NO levels in HeLa cells and modulated various genes involved in nitric oxide biosynthesis, superoxide metabolism, and oxidative stress, including NOS1, NOS2, NOS3, ALOX12, and SOD2, with a concomitant increase in NOS2 and NOS3 protein expression levels; also, the phytochemicals were found to induce apoptosis. CONCLUSIONS: These results suggest that the phytochemical-induced cell death is partially attributed to the activation of the NO pathway and upregulation of pro-oxidant ROS generators. Further experimental studies are required to explore this mechanistic association of NO signaling pathway activation and induction of apoptosis in other types of cancer.

Densely Populated Water Droplets in Heavy-Oil Seeps.

Most of the microbial degradation in oil reservoirs is believed to take place at the oil-water transition zone (OWTZ). However, a recent study indicates that there is microbial life enclosed in microliter-sized water droplets dispersed in heavy oil of Pitch Lake in Trinidad and Tobago. This life in oil suggests that microbial degradation of oil also takes place in water pockets in the oil-bearing rock of an oil leg independent of the OWTZ. However, it is unknown whether microbial life in water droplets dispersed in oil is a generic property of oil reservoirs rather than an exotic exception. Hence, we took samples from three heavy-oil seeps, Pitch Lake (Trinidad and Tobago), the La Brea Tar Pits (California, USA), and an oil seep on the McKittrick oil field (California, USA). All three tested oil seeps contained dispersed water droplets. Larger droplets between 1 and 10 µl revealed high cell densities of up to 109 cells ml-1 Testing for ATP content and LIVE/DEAD staining showed that these populations consist of active and viable microbial cells with an average of 60% membrane-intact cells and ATP concentrations comparable to those of other subsurface ecosystems. Microbial community analyses based on 16S rRNA gene amplicon sequencing revealed the presence of known anaerobic oil-degrading microorganisms. Surprisingly, the community analyses showed similarities between all three oil seeps, revealing common OTUs, although the sampling sites were thousands of kilometers apart. Our results indicate that small water inclusions are densely populated microhabitats in heavy oil and possibly a generic trait of degraded-oil reservoirs.IMPORTANCE Our results confirmed that small water droplets in oil are densely populated microhabitats containing active microbial communities. Since these microhabitats occurred in three tested oil seeps which are located thousands of kilometers away from each other, such populated water droplets might be a generic trait of biodegraded oil reservoirs and might be involved in the overall oil degradation process. Microbial degradation might thus also take place in water pockets in the oil-bearing oil legs of the reservoir rock rather than only at the oil-water transition zone.