Role of combination of mifepristone and misoprostol versus misoprostol alone in the management of late intrauterine fetal death

Background: Intrauterine fetal death (IUFD) occurs in 1% of pregnancies and has devastating consequences. Previous methods for inducing labor in IUFD involved oxytocin and prostaglandins. The combination of mifepristone and misoprostol is commonly used for early first-trimester termination. This study aimed to compare the effectiveness of mifepristone and misoprostol combination versus misoprostol alone for labor induction in intrauterine fetal death. Methods: A randomized controlled clinical trial was conducted at Sir Salimullah Medical College, Mitford Hospital, Dhaka, from January 2017 to June 2017. Sixty-four pregnant women with intrauterine fetal death after 28 weeks of gestation were included. Participants were randomly assigned to either group-I (mifepristone and misoprostol) or group-II (misoprostol alone). Statistical analyses were performed using statistical package for the social sciences (SPSS) version 20.0 for Windows. Results: The mean age was 27.7±5.6 years in group I and 27.5±4.3 years in group II. Majority of patients in group I were housewives (87.5%), while in group II, it was 78.1%. Most patients in group I (56.3%) came from lower-income families, compared to 65.6% in group II. The gestational age did not significantly differ between the groups. The induction to delivery interval was significantly shorter in group I (8.6±2.0 hours) compared to group II (15.1±3.5 hours). The dose administration pattern of misoprostol differed significantly between the groups. Conclusions: Both methods are equally safe and effective for managing intrauterine fetal death. However, the combination of mifepristone and misoprostol showed greater efficacy in terms of reducing the induction to delivery interval and requiring a lower dose of misoprostol

Molecular detection of a potentially toxic diatom species

A few diatom species produce toxins that affect human and animal health. Among these, members of the Pseudo-nitzschia genus were the first diatoms unambiguously identified as producer of domoic acid, a neurotoxin affecting molluscan shell-fish, birds, marine mammals, and humans. Evidence exists indicating the involvement of another diatom genus, Amphora, as a potential producer of domoic acid. We present a strategy for the detection of the diatom species Amphora coffeaeformis based on the development of species-specific oligonucleotide probes and their application in microarray hybridization experiments. This approach is based on the use of two marker genes highly conserved in all diatoms, but endowed with sufficient genetic divergence to discriminate diatoms at the species level. A region of approximately 450 bp of these previously unexplored marker genes, coding for elongation factor 1-a (eEF1-a) and silicic acid transporter (SIT), was used to design oligonucleotide probes that were tested for specificity in combination with the corresponding fluorescently labeled DNA targets. The results presented in this work suggest a possible use of this DNA chip technology for the selective detection of A. coffeaeformis in environmental settings where the presence of this potential toxin producer may represent a threat to human and animal health. In addition, the same basic approach can be adapted to a wider range of diatoms for the simultaneous detection of microorganisms used as biomarkers of different water quality levels

Risk factors of mortality in hospitalized children with severe acute malnutrition

Background: Severe acute malnutrition (SAM) is a major cause of morbidity and mortality in children around the world. It is critical to identify the factors that contribute to mortality to reduce SAM related mortality. This study aimed to analyze the risk factors of mortality in hospitalized children with SAM. Methods: This case-control study was conducted in the SAM unit, department of pediatrics, institute of child and mother health, Matuail-1362, Dhaka, from January 2021 to December 2021. Data analysis was conducted using SPSS version 22. Univariate analysis was done to determine factors affecting mortality, and multivariate logistic regression was used to determine significant independent risk factors. Results: Mean age of the study subject was 6.38±3.45 months and 10.90±10.00 months in the case and control groups respectively. So, death was more common in younger children. The percentage of death was more (61.5% vs 54.6%) in males. Mortality was more common in family income <10,000 Tk/ month, 53.8% in the case group and 21.9% in the control group. The mean age of the mother was 19.23±0.60 years and 21.78±4.78 years in the death and survived group. Among risk factors of mortality, dermatosis (46.2% vs 4.9%), oral ulcer (46.2% vs 5.5%), hypoglycemia (46.2% vs 3.8%), severe anemia (38.5% vs 2.2%), septicemia (76.9% vs 29.5%) in case and control group respectively. These risk factors were significantly higher in the death group compared to the surviving group. After doing multivariate logistic regression analysis it was observed that hypoglycemia (OR=9.17 with 95% CI 1.44 to 58.29) and severe anemia (OR=13.42 with 95% CI 1.42 to 126.13) were the strongest predictors of mortality among the hospitalized children with SAM. Conclusions: Hypoglycemia and severe anemia were the main contributing factors of mortal among the children with SAM in the hospital

Identification of freshwater diatoms in environmental samples to assess water quality using oligonucleotide probes\ud and microarrays

Monitoring the quality of water is of paramount importance for public health. According to the Water Framework Directive 2000/60/EC, “water is not a commercial product but a heritage that must be protected, defended and treated as such”. In Europe, about 40% of the drinking water is derived from surface waters and pathogenic organisms occurring in lakes and rivers represent a particularly serious health-hazard. In fact, approximately 100 million\ud Europeans lack safe and reliable water supplies and microbial pathogens in drinking water are causing significant morbidity and mortality with >170.000 estimated cases of water-related disease in Europe (UN News Centre, Protocol on Water and Health). The potential threat of\ud water contamination and the spread of waterborne diseases are becoming more serious than ever before. This is the result of a number of factors, including the increase in population, globalisation and the movement of more people across borders and between continents, and the effects of global warming. Furthermore, both abundance and distribution of all microbes are expected to change as a result of global climate change that will cause, for instance, increased or decreased rainfall, temperature and radiation.\ud As the nature of the diatom communities reflects environmental conditions and since they react rapidly and sensitively to water quality changes, these microorganisms have become widely used over the last 50 years as biomarkers for water quality assessment. Moreover, diatom doubling time is one of the quickest among various biological indicators and rapidly reveals a change in water quality. Diatoms are excellent bioindicators [...]. Traditional methods for diatom classification are based on microscopic frustule morphology but are laborious, time-consuming and not suitable for routine monitoring programs. Molecular biological tools have now greatly enhanced our ability to investigate biodiversity by identifying species and estimating gene flow and distribution of species.\ud Therefore, there is a strong incentive to explore and develop alternative methods that are faster, less expensive, reliable and efficient for identification of diatoms in complex environmental samples.\ud Microarrays have been the subject of several EU projects, like EU PICODIV and MICROPAD. Microarrays were developed for the detection of algae and protozoa, and results\ud from chip hybridization were favourably compared to other measurements of diversity, (i.e., direct cell counts and clone libraries). In these two projects, the microarrays were in early stages of development and proof of principle was the major outcome, because it was discovered that probes previously made for fluorescence in situ hybridisation (FISH) could not be directly transferred to a microarray chip format. With a few exceptions, nearly every probe had to be modified for successful use in the microarray chip format. Problems with transferring FISH probes to a microarray chip format led workers in the EU project MIDICHIP to modify their probes and microarrays, especially those designed for detection of cyanobacteria. The FP6 project FISH AND CHIPS made use of prototype findings to develop a microarray chip for phytoplankton at the class level and field data were analysed over three years with rRNA as the preferred target molecule [...] In the EU project AQUACHIP, pathogenic bacteria were the target of interest. This project developed a chip for five bacteria but they were not widely tested with environmental samples. In addition, the detection system developed for this chip was based on a microtiter plate system with detection under a fluorescent microscope. This, however, is not a standard protocol that can be used in a commercial microarray chip reader.\ud The results obtained in the earlier European Union (EU) projects were promising and contributed to the success of an application for an EU-funded grant. μAQUA is the acronym\ud for the EU research project "Universal microarrays for the evaluation of fresh-water quality based on detection of pathogens and their toxins" funded by the 7th Framework Programme of the European Commission and coordinated by Dr. Claudio Gualerzi and Dr. Roberto Spurio of the University of Camerino (Italy). μAQUA aims to design and develop a universal microarray chip for the high-throughput detection in freshwater of known and emerging pathogens (bacteria, viruses, protozoa and cyanobacteria) that cause human diseases and to assess the water quality by monitoring the presence of select bioindicators (diatoms). Existing\ud techniques are laborious and time-consuming, requiring labor-intensive cultivation and microscopic examination of potential pathogens from water samples. The innovative\ud molecular biological techniques being investigated by the μAQUA, by contrast, should enable the rapid - and more reliable - detection of pathogens and diatoms in large volumes of water.\ud The design of probes specifically targeting rDNA or rRNA from higher rank down to species level is made possible by the steadily growing number of rDNA-sequences available\ud (e.g. in the Ribosomal Database Project) [...]. The rDNAspecific probes are used to analyse phytoplankton communities with detection by flow cytometry, epifluorescence microscopy [...] or other methods that take advantage of the specificity ensured by nucleic acid hybridisation. However, a major drawback of these methods is that they are very time-consuming vis-à-vis the complexity of a microbial sample insofar as they can only be used to identify one or few organisms at a time [...]. By contrast, the DNA-microarray chip technology, which allows the rapid and simultaneous analysis of up to 200,000 probes without a cultivation step [...], has enormous potential as a method of choice for the analysis of complex environmental samples

Improved Canker Processing and Viability Droplet Digital PCR Allow Detection of <i>Erwinia amylovora</i> Viable Nonculturable Cells in Apple Bark

The bacterium Erwinia amylovora causes fire blight and continues to threaten global commercial apple and pear production. Conventional microbiology techniques cannot accurately determine the presence of live pathogen cells in fire blight cankers. Several factors may prevent E. amylovora from growing on solid culture media, including competing microbiota and the release of bacterial-growth-inhibitory compounds by plant material during sample processing. We previously developed a canker processing methodology and a chip-based viability digital PCR (v-dPCR) assay using propidium monoazide (PMA) to bypass these obstacles. However, sample analysis was still time-consuming and physically demanding. In this work, we improved the previous protocol using an automatic tissue homogenizer and transferred the chip-based v-dPCR to the BioRad QX200 droplet dPCR (ddPCR) platform. The improved sample processing method allowed the simultaneous, fast, and effortless processing of up to six samples. Moreover, the transferred v-ddPCR protocol was compatible with the same PMA treatment and showed a similar dynamic range, from 7.2 × 102 to 7.6 × 107 cells mL−1, as the previous v-dPCR. Finally, the improved protocol allowed, for the first time, the detection of E. amylovora viable but nonculturable (VBNC) cells in cankers and bark tissues surrounding cankers. Our v-ddPCR assay will enable new ways to evaluate resistant pome fruit tree germplasm, further dissect the E. amylovora life cycle, and elucidate E. amylovora physiology, epidemiology, and new options for canker management

Molecular tools for the selective detection of nine diatom species biomarkers of various water quality levels

Our understanding of the composition of diatom communities and their response to environmental changes is currently limited by laborious taxonomic identification procedures. Advances in molecular technologies are expected to contribute more efficient, robust and sensitive tools for the detection of these ecologically relevant microorganisms. There is a need to explore and test phylogenetic markers as an alternative to the use of rRNA genes, whose limited sequence divergence does not allow the accurate discrimination of diatoms at the species level. In this work, nine diatom species belonging to eight genera, isolated from epylithic environmental samples collected in central Italy, were chosen to implement a panel of diatoms covering the full range of ecological status of freshwaters. The procedure described in this work relies on the PCR amplification of specific regions in two conserved diatom genes, elongation factor 1-a (eEF1-a) and silicic acid transporter (SIT), as a first step to narrow down the complexity of the targets, followed by microarray hybridization experiments. Oligonucleotide probes with the potential to discriminate closely related species were designed taking into account the genetic polymorphisms found in target genes. These probes were tested, refined and validated on a small-scale prototype DNA chip. Overall, we obtained 17 highly specific probes targeting eEF1-a and SIT, along with 19 probes having lower discriminatory power recognizing at the same time two or three species. This basic array was validated in a laboratory setting and is ready for tests with crude environmental samples eventually to be scaled-up to include a larger panel of diatoms. Its possible use for the simultaneous detection of diatoms selected from the classes of water quality identified by the European Water Framework Directive is discussed