Diagnostic accuracy of Dengue NS1 lateral flow immunoassay in comparison to reverse transcriptase polymerase chain reaction and enzyme linked Immunosorbent Assay.

The most widely used invitro diagnostic qualitative screening method for dengue virus infection is the lateral flow immunoassay technique. Testing of dengue non-structural antigen NS1 offers specificity in determining the active infection while testing of IgM and IgG helps in differentiating the primary and secondary dengue infections. The ELISA functions as the golden standard for dengue testing and PCR credits for the most accurate determination tool at the genetic level. The RT-PCR endorsed NS1 gene and in ELISA or LFIA NS1 antigen is used as the marker owing to the specificity and lesser chances of mutation effects. This study evaluated the performance of AG-Q Dengue NS1 LFIA kit in comparison with RT-PCR quantification cycle (Cq) Values and ELISA NS1 quantitation. The study also focused on differentiating the samples among dengue serotypes using the RealStar Dengue Type RT-PCR Kit 1.0. Dengue serotype 2 is the prominent viral strain in Kerala region succeeded by serotype 3 and 1 with a prevalence rate of 64 %, 20 % and 6 % respectively. Dengue serotype 4 was not reported during this study period. 10 % co-infection with DENV 1 & DENV 2 was also reported. The AG-Q Dengue NS1 kit stood as efficient in screening by providing positive results with samples having RT-PCR Cq values up to 43 and ELISA NS1 quantification minimum of 14 Panbio units.

A Study to Assess the Role of Date Syrup, Pineapple Juice, and Hematinic Syrup as Magnetic Resonance Oral Contrast Agents in Improving the Image Quality of Magnetic Resonance Cholangiopancreatography.

INTRODUCTION: Magnetic resonance cholangiopancreatography (MRCP) is an imaging technique that has advanced over the past few years. It still plays a crucial role in the study of numerous pancreaticobiliary diseases. This study aimed to evaluate the effects of hematinic syrup, date syrup, and pineapple juice on MRCP image quality. METHODOLOGY: This study involved a total of 90 participants, distributed evenly among three groups, with each group comprising 30 patients. Negative oral contrast solutions containing paramagnetic substances like Mn+2 and Fe+3, such as pineapple juice, date syrup, and hematinic syrup were imaged by 1.5 Tesla (T) magnetic resonance imaging (MRI) with T2-weighted (T2W) and MRCP sequences. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were computed. Ninety patients underwent MRCP 20-30 min after ingestion of 100 mL of date syrup, 30 ml of hematinic syrup diluted to 200 ml of water, and 200 mL of pineapple juice. MRCP images were taken to visualize various pancreaticobiliary structures (bile duct, stomach, and duodenum). RESULTS: The in vitro evaluation of the solutions showed that date syrup and hematinic syrup were hypointense in T2W sequences. The images obtained showed no significant difference in the CNR between the three solutions. However, the SNR was significantly higher for pineapple juice compared to date syrup and hematinic syrup in T2W and MRCP sequences. Images acquired post-administration of the oral contrast agents significantly improved the gastrointestinal tract signal suppression and increased visibility of the pancreaticobiliary structures (bile duct, stomach, and duodenum). No adverse events were observed among the participants. CONCLUSION: Pineapple juice was the best contrast agent. However, date syrup and hematinic syrup can also be used to improve the imaging quality.

Does surface enamel composition and characteristics vary with inter proximal enamel reduction?

OBJECTIVE: This study aimed to evaluate the chemical composition of the proximal enamel surface and the surface characteristics subjected to different extents of interproximal reduction (IPR) in a clinical setting. MATERIALS AND METHODS: Premolars of orthodontic patients which were designated for extraction were subjected to .2 mm, .3 mm, and .5 mm of IPR. After 1 month, the teeth were extracted and the teeth were subjected to scanning electron microscope (SEM) and energy-dispersive x-ray spectroscopy (EDX). RESULTS: The SEM images of the three experimental groups (taken at magnification of 500 × and 2000 ×) showed that the enamel surfaces were irregular and rough compared to the honey comb appearance of the unstripped group. Small areas of erosion of enamel surface were seen in Group I (0.2 mm) under 2000 × magnification compared to Group IV (control) which showed typical arrangement of enamel rods in alternating orientation. The enamel surfaces of stripped and unstripped enamel contained calcium, phosphorus, carbon, oxygen, and nitrogen. The differences were not statistically significant and neither were the calcium phosphorous stoichiometric ratios between the four groups. CONCLUSIONS: On analyzing the surface characteristics of enamel using SEM between the stripped and unstripped surfaces, there were irregularities and roughness seen in stripped surface whereas honey comb pattern was observed in unstripped enamel surfaces. The elements found in unstripped and stripped enamel surfaces were calcium, phosphorous, carbon, oxygen, and nitrogen. Although the calcium and phosphorus were high in the 0.5 mm IPR group, the difference between stripped and unstripped enamel surfaces was statistically not significant. CLINICAL RELEVANCE: There have been concerns that IPR can remove the superficial mineral-rich layer making the deeper layers more susceptible to carious attack. No study has evaluated the mineral content in different layers of enamel in response to IPR in vivo and this study found no significant difference between pristine enamel and enamel subjected to IPR. The results of this study strengthen the validity of the clinical protocol employed.

Antimicrobial activity of coral-associated beneficial bacteria against coral disease-causing microbial pathogens.

Microbial infection of immune-compromised corals influences disease severity, resulting in coral mortality. However, coral-associated beneficial bacteria are known to produce antimicrobial compounds that prevent the growth of potential pathogens and invading microbes. Hence, beneficial bacteria associated with coral Porites lutea were isolated and antimicrobial protein and bioactive secondary metabolites were extracted and tested for their antimicrobial activity against putative prokaryotic and eukaryotic coral pathogens. Bioactive secondary metabolites exhibited remarkable antagonism against various coral pathogens such as Serratia marcescens, Vibrio species, and Aspergillus sydowii. Besides, the metabolites of Cobetia marina, Cobetia amphilecti, Pseudoalteromonas neustonica, and Virgibacillus halodenitrificans manifested notable inhibition against the protozoan ciliates (Uronema marinum, Holosticha diademata, Cohnilembus verminus, and Euplotes vannus) and zooplankton that are known to be involved in the secondary pathogenesis in coral diseased lesion progression. Thus, the present study may benefit in understanding coral-associated beneficial bacteria for their antagonistic interactions with microbial pathogens, as well as their potential involvement in reducing coral disease severity.

Possible beneficial interactions of ciliated protozoans with coral health and resilience.

Microbial interactions contribute significantly to coral health in the marine environment. Most beneficial associations have been described with their bacterial communities, but knowledge of beneficial associations between protozoan ciliates and corals is still lacking. Ciliates are important bacterial predators and provide nutrition to higher trophic-level organisms. The mucus secreted by corals and the microenvironment of the coral surface layer attract ciliates based on their food preferences. The mixotrophic and heterotrophic ciliates play a major role in nutrient cycling by increasing nitrogen, phosphorus, and extractable sulfur, which can enhance the proliferation of coral beneficial microbe. Besides, bacterial predator ciliates reduce the pathogenic bacterial population that infects the coral and also act as bioindicators for assessing the toxicity of the reef ecosystem. Thus, these ciliates can be used as a beneficial partner in influencing coral health and resilience under various stress conditions. Herein, we explore the urgent need to understand the complex beneficial interactions of ciliates that may occur in the coral reef ecosystem.

Epilepsy in a mouse model of GNB1 encephalopathy arises from altered potassium (GIRK) channel signaling and is alleviated by a GIRK inhibitor.

De novo mutations in GNB1, encoding the Gß1 subunit of G proteins, cause a neurodevelopmental disorder with global developmental delay and epilepsy, GNB1 encephalopathy. Here, we show that mice carrying a pathogenic mutation, K78R, recapitulate aspects of the disorder, including developmental delay and generalized seizures. Cultured mutant cortical neurons also display aberrant bursting activity on multi-electrode arrays. Strikingly, the antiepileptic drug ethosuximide (ETX) restores normal neuronal network behavior in vitro and suppresses spike-and-wave discharges (SWD) in vivo. ETX is a known blocker of T-type voltage-gated Ca2+ channels and G protein-coupled potassium (GIRK) channels. Accordingly, we present evidence that K78R results in a gain-of-function (GoF) effect by increasing the activation of GIRK channels in cultured neurons and a heterologous model (Xenopus oocytes)-an effect we show can be potently inhibited by ETX. This work implicates a GoF mechanism for GIRK channels in epilepsy, identifies a new mechanism of action for ETX in preventing seizures, and establishes this mouse model as a pre-clinical tool for translational research with predicative value for GNB1 encephalopathy.

Soil polluted with nano ZnO reveals unstable bacterial communities and decoupling of taxonomic and functional diversities.

Due to relentless production and disposal of nano zinc oxide (nZnO), it has become critical to comprehend the serious risks large-scale accumulation of nZnO pose to bacterial communities in soil. The primary objective was to evaluate the changes in bacterial community structure and associated functional pathways through predictive metagenomic profiling and subsequent validation through Quantitative Realtime PCR in soil spiked with nZnO (0, 50, 200, 500 and 1000 mg Zn kg-1) and similar levels of bulk ZnO (bZnO). The results revealed that soil microbial biomass-C, -N, -P, soil respiration and enzyme activities decreased markedly at higher ZnO levels. The alpha diversity decreased with increasing ZnO level, with more impact under nZnO, while beta diversity analyses indicated a distinct dose- dependent separation of bacterial communities. The dominant taxa including Proteobacteria, Bacterioidetes, Acidobacteria and Planctomycetes significantly increased in abundance, while Firmicutes, Actinobacteria and Chloroflexi decreased in abundance with elevated nZnO and bZnO levels. Redundancy analysis indicated that changes in bacterial community structure instilled a greater dose- rather than size- specific response on key microbial parameters. Predicted key functions did not show a dose- specific response, and at 1000 mg Zn kg-1, methane metabolism as well as starch and sucrose metabolism were attenuated, while functions involving two component systems and bacterial secretion systems were enhanced under bZnO indicating better stress avoidance mechanism than under nZnO. Realtime PCR and microbial endpoint assays confirmed the metagenome derived taxonomic and functional data, respectively. Taxa and functions that varied substantially under stress were established as bioindicators to predict nZnO toxicity in soils. Taxon-function decoupling indicated that the soil bacterial communities deployed adaptive mechanisms under high ZnO, with lesser buffering capacity and resilience of communities under nZnO.

Impact of Covishield Vaccination in Terms of SARS CoV-2 Neutralizing Antibody Expression.

The vaccination efficacy can indirectly be assessed through the quantification of neutralizing antibodies. Very few data are available on Covishield efficacy in terms of neutralizing antibody expression upon vaccination. This study is focused on profiling of neutralizing antibody expression during and after the Covishield two shot vaccination and observing COVID-19 infection in vaccinated participants during the period. SARS CoV-2 neutralizing antibody concentrations in samples were estimated using electrochemiluminescence immunoassay kit for Lifotronics eCL8000. The sampling had been done sequentially at 45th, 85th day after 1st dose and 15th day after 2nd dose Covishield vaccination. Parallelly, in order to confirm the total SARS CoV-2 IgG response in COVID-19 infection, measured the IgG using SARS CoV-2 IgG lateral flow immunoassay test kit. The subjects previously infected with COVID-19 before 1st dose vaccination demonstrated high neutralizing antibody (> 10AU/ml). In COVID-19 uninfected subjects, there was a sudden incline in neutralizing antibody after the 2nd dose. Infection with SARS CoV-2 between 1st and 2nd dose of Covishield vaccination implicate that the level of neutralizing antibody in serum after 1st dose was not adequate to combat the virus and prevent infection. We observed COVID-19 infection in participants even after 2nd dose of vaccination. Interestingly, there was no protection against SARS CoV-2 even with a high neutralizing antibody expression of 188.5 AU/mL after the 2nd dose. Findings of Covishield efficacy in different cohort samples before and after 2 doses of Covishield vaccination provide impetus for improvement or development of next generation vaccines.

Metagenomics indicates abundance of biofilm related genes and horizontal transfer of multidrug resistant genes among bacterial communities in nano zinc oxide polluted soil.

The unsafe and reckless disposal of metal oxide nanoparticles like ZnO (nZnO) into the soil could seriously impact bacterial behavioural responses and functions. Under such stress, biofilm formation is considered to be a robust mechanism for bacterial survival in soil. We examined the response of bacterial metagenomes in soils exposed to varying levels of Zn (50, 200, 500 and 1000 mg kg-1) as nano Zn oxide (nZnO) in terms of biofilm genesis and regulation and their co-occurrences with multidrug resistance genes (MDRGs) and mobile genetic elements (MGEs). The size-specific effects of nZnO were verified using its bulk counterpart (bZnO). Both nZnO and bZnO facilitated profusion of biofilm related genes (BGs) especially at higher Zn levels (500 and 1000 mg kg-1 Zn), though maximum abundance was registered at a comparatively lower level under nZnO. In general, nZnO favoured an enhancement of genes involved in exopolysaccharide biosynthesis and attachment, while bZnO favoured genes related to capsule formation, chemotaxis and biofilm dispersion. Co-occurrence network analysis revealed significant positive correlations between abundances of BGs, MDRGs and MGEs, indicating an enhanced probability for horizontal gene transfer of MDRGs in nZnO polluted soils.

A selectivity filter mutation provides insights into gating regulation of a K+ channel.

G-protein coupled inwardly rectifying potassium (GIRK) channels are key players in inhibitory neurotransmission in heart and brain. We conducted molecular dynamics simulations to investigate the effect of a selectivity filter (SF) mutation, G154S, on GIRK2 structure and function. We observe mutation-induced loss of selectivity, changes in ion occupancy and altered filter geometry. Unexpectedly, we reveal aberrant SF dynamics in the mutant to be correlated with motions in the binding site of the channel activator Gßγ. This coupling is corroborated by electrophysiological experiments, revealing that GIRK2wt activation by Gßγ reduces the affinity of Ba2+ block. We further present a functional characterization of the human GIRK2G154S mutant validating our computational findings. This study identifies an allosteric connection between the SF and a crucial activator binding site. This allosteric gating mechanism may also apply to other potassium channels that are modulated by accessory proteins.

A revised mechanism of action of hyperaldosteronism-linked mutations in cytosolic domains of GIRK4 (KCNJ5).

G protein-gated, inwardly rectifying potassium channels (GIRK) mediate inhibitory transmission in brain and heart, and are present in the adrenal cortex. GIRK4 (KCNJ5) subunits are abundant in the heart and adrenal cortex. Multiple mutations of KCNJ5 cause primary aldosteronism (PA). Mutations in the pore region of GIRK4 cause loss of K+ selectivity, Na+ influx and depolarization of zona glomerulosa cells followed by hypersecretion of aldosterone. The concept of selectivity loss has been extended to mutations in cytosolic domains of GIRK4 channels, remote from the pore. We expressed aldosteronism-linked GIRK4R52H , GIRK4E246K and GIRK4G247R mutants in Xenopus oocytes. Whole-cell currents of heterotetrameric GIRK1/4R52H and GIRK1/4E246K channels were greatly reduced compared with GIRK1/4WT . Nevertheless, all heterotetrameric mutants retained full K+ selectivity and inward rectification. When expressed as homotetramers, only GIRK4WT , but none of the mutants, produced whole-cell currents. Confocal imaging, single-channel and Förster Resonance Energy Transfer (FRET) analyses showed: (1) reduction of membrane abundance of all mutated channels, especially as homotetramers, (2) impaired interaction with Gßγ subunits, and (3) reduced open probability of GIRK1/4R52H . VU0529331, a GIRK4 opener, activated homotetrameric GIRK4G247R channels, but not GIRK4R52H or GIRK4E246K . In the human adrenocortical carcinoma cell line (HAC15), VU0529331 and overexpression of heterotetrameric GIRK1/4WT , but not overexpression of GIRK1/4 mutants, reduced aldosterone secretion. Our results suggest that, contrary to pore mutants of GIRK4, non-pore mutants R52H and E246K mutants are loss-of-function rather than gain-of-function/selectivity-loss mutants. Hence, GIRK4 openers may be a potential course of treatment for patients with cytosolic N- and C-terminal mutations. KEY POINTS: Mutations in GIRK4 (KCNJ5) G protein-gated channels cause primary aldosteronism, a major cause of secondary hypertension. The primary mechanism is believed to be loss of K+ selectivity. R52H and E246K, aldosteronism-causing mutations in cytosolic N- and C- termini of GIRK4, were reported to cause loss of K+ selectivity. We show that R52H, E246K and G247R mutations render homotetrameric GIRK channels non-functional. In heterotetrameric context with GIRK1, these mutations impair membrane expression, interaction with Gßγ and open probability, but do not alter K+ selectivity or inward rectification. In the human aldosterone-secreting cell line, a GIRK4 opener and overexpression of heterotetrameric GIRK1/4WT , but not overexpression of GIRK1/4 mutants, reduced aldosterone secretion. Aldosteronism-causing mutations in the cytosolic domain of GIRK4 are loss-of-function mutations rather than gain-of-function, selectivity-loss mutations. Deciphering of exact biophysical mechanism that impairs the channel is crucial for setting the course of treatment.

Ciliated protozoan occurrence and association in the pathogenesis of coral disease.

Various microbial infections have significantly contributed to disease progression leading to the mortality of corals. However, the holobiont and the external surfaces of coral, including the secreted mucus, provide a varied microenvironment that attracts ciliates based on their feeding preferences. Besides, some ciliates (e.g., Philasterine scuticociliate) may enter through the injuries or lesions on corals or through their indirect interactions with other types of microbes that influence coral health. Thus, ciliates occurrence and association are described with 12 different diseases worldwide. White syndrome disease lesions have diverse ciliate associations, and higher ciliate diversity was observed with diseased genera Acropora. Also, it was described, about sixteen ciliate species ingest coral Symbiodiniaceae and histophagous ciliates for coral tissue loss as secondary invaders. However, the ciliates nature of association with the coral disease remains unclear for primary or opportunistic secondary pathogenicity. Herein, we explore the urgent need to understand the complex interactions of ciliates in coral health.

Encephalopathy-causing mutations in Gß1 (GNB1) alter regulation of neuronal GIRK channels.

Mutations in the GNB1 gene, encoding the Gß1 subunit of heterotrimeric G proteins, cause GNB1 Encephalopathy. Patients experience seizures, pointing to abnormal activity of ion channels or neurotransmitter receptors. We studied three Gß1 mutations (K78R, I80N and I80T) using computational and functional approaches. In heterologous expression models, these mutations did not alter the coupling between G protein-coupled receptors to Gi/o, or the Gßγ regulation of the neuronal voltage-gated Ca2+ channel CaV2.2. However, the mutations profoundly affected the Gßγ regulation of the G protein-gated inwardly rectifying potassium channels (GIRK, or Kir3). Changes were observed in Gß1 protein expression levels, Gßγ binding to cytosolic segments of GIRK subunits, and in Gßγ function, and included gain-of-function for K78R or loss-of-function for I80T/N, which were GIRK subunit-specific. Our findings offer new insights into subunit-dependent gating of GIRKs by Gßγ, and indicate diverse etiology of GNB1 Encephalopathy cases, bearing a potential for personalized treatment.

A rare coexistence of aspergillosis with actinomycosis.

Aspergillosis is a common systemic mycosis which affects immunocompromised and immunocompetent hosts. Aspergillus spp. is wide spread in the environment in most countries, which renders an invasive form of disease. The presence conidial heads are pathognomic to aspergillosis in diagnosis. Actinomycosis is a subacute-to-chronic infection that causes sinus fistula, tract or abscess due to the invasion surrounding the soft tissue. Cervicofacial infection accounts for 50%-60% of all actinomycosis cases. The mandible and nasopharynx are the sites of predilection, but maxillary infection is rare. Aspergillosis and Actinomycosis each of them was reported in case, but mixed infection of both organisms is rare, only one case has been reported. This paper discussed about a case report of coexistence of aspergillosis with actinomycosis in 38-year-old male.

The Neuropeptide Orexin-A Inhibits the GABAA Receptor by PKC and Ca2+/CaMKII-Dependent Phosphorylation of Its ß1 Subunit.

Orexin-A and orexin-B (Ox-A, Ox-B) are neuropeptides produced by a small number of neurons that originate in the hypothalamus and project widely in the brain. Only discovered in 1998, the orexins are already known to regulate several behaviours. Most prominently, they help to stabilise the waking state, a role with demonstrated significance in the clinical management of narcolepsy and insomnia. Orexins bind to G-protein-coupled receptors (predominantly postsynaptic) of two subtypes, OX1R and OX2R. The primary effect of Ox-OXR binding is a direct depolarising influence mediated by cell membrane cation channels, but a wide variety of secondary effects, both pre- and postsynaptic, are also emerging. Given that inhibitory GABAergic neurons also influence orexin-regulated behaviours, crosstalk between the two systems is expected, but at the cellular level, little is known and possible mechanisms remain unidentified. Here, we have used an expression system approach to examine the feasibility, and nature, of possible postsynaptic crosstalk between Ox-A and the GABAA receptor (GABAAR), the brain's main inhibitory neuroreceptor. When HEK293 cells transfected with OX1R and the α1, ß1, and γ2S subunits of GABAAR were exposed to Ox-A, GABA-induced currents were inhibited, in a calcium-dependent manner. This inhibition was associated with increased phosphorylation of the ß1 subunit of GABAAR, and the inhibition could itself be attenuated by (1) kinase inhibitors (of protein kinase C and CaM kinase II) and (2) the mutation, to alanine, of serine 409 of the ß1 subunit, a site previously identified in phosphorylation-dependent regulation in other pathways. These results are the first to directly support the feasibility of postsynaptic crosstalk between Ox-A and GABAAR, indicating a process in which Ox-A could promote phosphorylation of the ß1 subunit, reducing the GABA-induced, hyperpolarising current. In this model, Ox-A/GABAAR crosstalk would cause the depolarising influence of Ox-A to be boosted, a type of positive feedback that could, for example, facilitate the ability to abruptly awake.

Efficient visible upconversion luminescence in Er3+ and Er3+/Yb3+ co-doped Y2O3 phosphors obtained by solution combustion reaction.

Combustion derived Er(3+) -doped Y2O3 and Er(3+)/Yb(3+)co-doped Y2O3 powders have been characterized by X-ray diffraction, energy dispersive X-ray analysis, Fourier transform infrared spectroscopy and laser excited spectroscopy. Formation of Y2O3 phosphor was confirmed by X-ray diffraction and energy dispersive X-ray analysis. The vibrational properties of Y2O3 powder was studied by Fourier transform infrared spectroscopy. The luminescence spectra of Er(3+) -doped and Er(3+)/Yb(3+) co-doped Y2O3 powders were studied under 379nm excitation. The strong up-conversion luminescence for Er(3+) -doped and Er(3+)/Yb(3+) co-doped Y2O3 powders have been observed under 978nm laser excitation. The effect of Yb(3+) addition on optical and luminescence properties of Er(3+):Y2O3 powders were studied. The ratio of red to green intensity has been enhanced when Er(3+) -doped Y2O3 is co-doped with Yb(3+) ions. The effect of co-doping of Yb(3+) ions on the visible luminescence intensity of Er(3+) has been studied and the mechanism responsible for the variation in the green and red intensity is discussed.

Pseudoaneurysm of homograft placed in right ventricular outflow tract.

Pseudoaneurysm of the right ventricular outflow tract after homograft placement is an infrequent complication after intracardiac repair for tetralogy of Fallot. We report two cases of pseudoaneurysm of right ventricular outflow tract after homograft placement for surgical repair of tetralogy of Fallot with pulmonary atresia.