A comprehensive entomological survey was undertaken in Alipurduar District, West Bengal, from 2018 to 2020 and in 2022. This study was prompted by reported malaria cases and conducted across nine villages, seven Sub-Centres, and three Primary Health Centres (PHCs). Mosquitoes were hand-collected with aspirators and flashlights from human dwellings and cattle sheds during the daytime. Both morphological and molecular techniques were used for species identification. Additionally, mosquitoes were tested for Plasmodium parasites and human blood presence. Mosquito species such as An. barbirostris s.l., An. hyrcanus s.l., An. splendidus, and An. vagus were morphologically identified. For species like An. annularis s.l., An. minimus s.s., An. culicifacies s.l., and An. maculatus s.s., a combination of morphological and molecular techniques was essential. The mitochondrial cytochrome c oxidase gene subunit 1 (CO1) was sequenced for An. annularis s.l., An. maculatus s.s., An. culicifacies s.l., An. vagus, and some damaged samples, revealing the presence of An. pseudowillmori and An. fluviatilis. The major Anopheles species were An. annularis s.l., An. culicifacies s.l., and An. maculatus s.s., especially in Kumargram and Turturi PHCs. Plasmodium positivity was notably high in An. annularis s.l. and An. maculatus s.s. with significant human blood meal positivity across most species. Morphological, molecular, and phylogenetic analyses are crucial, especially for archived samples, to accurately identify the mosquito fauna of a region. Notably, this study confirms the first occurrence of An. pseudowillmori and An. sawadwongporni in West Bengal and implicates An. maculatus s.s., An. culicifacies s.l., and An. annularis s.l. as significant vectors in the Alipurduar region.
BACKGROUND: Pyruvate dehydrogenase complex deficiency (PDCD) is a mitochondrial neurometabolic disorder of energy deficit, with incidence of about 1 in 42,000 live births annually in the USA. The median and mean ages of diagnosis of PDCD are about 12 and 31 months, respectively. PDCD is a major cause of primary lactic acidosis with concomitant elevation in blood alanine (Ala) and proline (Pro) concentrations depending on phenotypic severity. Alanine/Leucine (Ala/Leu) ≥4.0 and Proline/Leucine (Pro/Leu) ≥3.0 combination cutoff from dried blood spot specimens was used as a biomarker for early identification of neonates/infants with PDCD. Further investigations were needed to evaluate the sensitivity (SN), specificity (SP), and clinical utility of such amino acid (AA) ratio combination cutoffs in discriminating PDCD from other inborn errors of metabolism (IEM) for early identification of such patients. METHODS: We reviewed medical records of patients seen at UPMC in the past 11 years with molecularly or enzymatically confirmed diagnosis. We collected plasma AA analysis data from samples prior to initiation of therapeutic interventions such as total parenteral nutrition and/or ketogenic diet. Conditions evaluated included organic acidemias, primary mitochondrial disorders (MtDs), fatty acid oxidation disorders (FAOD), other IEMs on current newborn screening panels, congenital cardiac great vessel anomalies, renal tubular acidosis, and non-IEMs. The utility of specific AA ratio combinations as biomarkers were evaluated using receiver operating characteristic curves, correlation analysis, principal component analysis, and cutoff SN, SP, and positive predictive value determined from 201 subjects with broad age range. RESULTS: Alanine/Lysine (Ala/Lys) and Ala/Leu as well as (Ala + Pro)/(Leu + Lys) and Ala/Leu ratio combinations effectively discriminated subjects with PDCD from those with other MtDs and IEMs on current newborn screening panels. Specific AA ratio combinations were significantly more sensitive in identifying PDCD than Ala alone or combinations of Ala and/or Pro in the evaluated cohort of subjects. Ala/Lys ≥3.0 and Ala/Leu ≥5.0 as well as (Ala + Pro)/(Leu + Lys) ≥2.5 and Ala/Leu ≥5.0 combination cutoffs identified patients with PDCD with 100% SN and ~85% SP. CONCLUSIONS: With the best predictor of survival and positive cognitive outcome in PDCD being age of diagnosis, PDCD patients would benefit from use of such highly SN and SP AA ratio combination cutoffs as biomarkers for early identification of at-risk newborns, infants, and children, for early intervention(s) with known and/or novel therapeutics for this disorder.
Metabolism, Inborn Errors , Pyruvate Dehydrogenase Complex Deficiency Disease , Child , Humans , Infant, Newborn , Infant , Child, Preschool , Amino Acids , Leucine , Pyruvate Dehydrogenase Complex Deficiency Disease/diagnosis , Metabolism, Inborn Errors/diagnosis , Alanine , Proline , Biomarkers
The current molecular classification divides breast cancer into four major subtypes, including luminal A, luminal B, HER2-positive, and basal-like, based on receptor gene expression profiling. Luminal A and luminal B are hormone receptor (HR, estrogen, and/or progesterone receptor)-positive and are the most common subtypes, accounting for around 50-60% and 15-20% of the total breast cancer cases, respectively. The drug treatment for HR-positive breast cancer includes endocrine therapy, HER2-targeted therapy (depending on the HER2 status), and chemotherapy (depending on the risk of recurrence). In this review, in addition to classification, we focused on discussing the important aspects of HR-positive breast cancer, including HR structure and signaling, genetics, including epigenetics and gene mutations, gene expression-based assays, the traditional and new drugs for treatment, and novel or new uses of technology in diagnosis and treatment. Particularly, we have summarized the commonly mutated genes and abnormally methylated genes in HR-positive breast cancer and compared four common gene expression-based assays that are used in breast cancer as prognostic and/or predictive tools in detail, including their clinical use, the factors being evaluated, patient demographics, and the scoring systems. All these topic discussions have not been fully described and summarized within other research or review articles.
Microfluidics (MF) is the science dealing with the behavior, precise control, and manipulation of fluids as well as particles on the scale of tens to hundreds of micrometers. It is also utilized for chemical and biological applications, usually called micro-total analysis systems (µTAS) or lab-on-a-chip (LOC). MF is a fascinating and capable technology with various superior benefits compared to conventional macro-scale platforms, such as the lesser requirement of sample and reagent volumes, higher sensitivity, low cost, portability, faster processing of samples and potential to be automated and highly integrated to reduce human errors. The concept of transformation of meso to nanoliters using MF technology has shown its potential in the healthcare system for early diagnosis, and personalized medicine. The integrated multifunctional system with parallelization provides a better and faster process control. Minimization of the consumption of fluid makes the technology safer in every aspect of the development process, analysis, and storage. The impressive improvement in patient care and monitoring has led to the commercial motivation of the pharmaceutical industry to develop new drugs and modify existing products with better efficacy and safety in a cost-effective manner using MF technologies. Hence, the present review briefs on the applications of MF technology in the key issues of the drug discovery process, overcoming the limitations of development of analytical procedures and prosperous pharmaceutical manufacturing for novel controlled and targeted release dosage forms to fabricate quality products.
A majority of pediatric outpatient practice involves managing familiar diseases that present in familiar ways. Occasionally, a familiar disease presents uniquely, which adds a diagnostic challenge and enhances the clinical experience of the clinician. We describe an 18-month-old male who presented to the clinic with a familiar disease but with unique additional findings. The patient had a one-day history of rash, subjective fever, and several episodes of non-bloody diarrhea. The rash included petechial lesions across his abdomen, groin, back, arms, and legs, as well as vesicular lesions in the mouth and on the palms and soles. A tentative diagnosis of hand, foot, and mouth disease (HFMD) was made. However, the presence of petechiae prompted further laboratory evaluation, including a complete blood count (CBC) and comprehensive metabolic panel (CMP). The CBC was unremarkable, but the CMP revealed an abnormally high serum alkaline phosphatase (ALP) level of 1,353 U/L (normal range: 53-128 U/L). The patient was subsequently diagnosed with an atypical presentation of HFMD associated with transient hyperphosphatasemia (TH). TH is characterized by a benign increase in serum alkaline phosphatase levels with an absence of liver or bone diseases. TH is usually clinically silent. Clinicians should consider the possibility of TH in pediatric patients who are found incidentally to have an elevated ALP, especially with a concomitant viral infection. An awareness and understanding of TH will prevent unnecessary additional testing and avoid undue parental anxiety.
