A study of [2 + 2] cycloaddition-retroelectrocyclization in water: observation of substrate-driven transient-nanoreactor-induced new reactivity.

Organic solvents limit [2 + 2] cycloaddition-retroelectrocyclization (CA-RE) in biological fields. We examined the formation of 1,1,4,4-tetracyanobuta-1,3-dienes (TCBDs) through CA-RE reactions and their unusual reactivity to produce N-heterocyclic compounds when the nature of the surfactant and the concentrations were varied in the aqueous phase. An environment in which transient self-assemblies (vesicles) were induced by the substrate and surfactant molecules initiated new reactivity through H2O addition on the TCBD, generating the enol form of the intermediate, which results in the formation of the 6,6-dicyano-heteropentafulvene (amidofulvene) compound, while lamellar sheets at higher concentrations favored TCBD generation. Interestingly, the amidofulvene underwent a clean transformation to 6-membered heterocycles that resemble cardiotonic drugs (milrinone, amrinone) via keto-enol tautomerism mediated by a polar aprotic solvent, opening up a new avenue for drug discovery. Unlike organic-solvent-mediated CA-RE reactions, the present nanoreactor-mediated approach enabled the selective production of TCBDs as well as new heterocycles using H2O as a green solvent. In addition to the widely explored organic electronics/materials, we believe that this study will help to overcome the long-standing limitation of CA-RE reaction applicability in biological fields.

Shifts in Bacterial Community Composition and Functional Traits at Different Time Periods Post-deglaciation of Gangotri Glacier, Himalaya.

Climate change causes an unprecedented increase in glacial retreats. The melting ice exposes land for colonization and diversification of bacterial communities leading to soil development, changes in plant community composition, and ecosystem functioning. Although a few studies have focused on macro-level deglaciation impacts, little is known about such effects on the bacterial community succession. Here, we provide meta-barcoding-based insight into the ecological attributes of bacterial community across different retreating periods of the Gangotri glacier, western Himalaya. We selected three sites along a terminal moraine representing recent (~ 20 yrs), intermediate (~ 100 yrs), and late (~ 300 yrs) deglaciation periods. Results showed that the genus Mycobacterium belonging to phylum Actinobacteria dominated recently deglaciated land. Relative abundance of these pioneer bacterial taxa decreased by 20-50% in the later stages with the emergence of new and rising of the less abundant members of the phyla Proteobacteria, Firmicutes, Planctomycetes, Acidobacteria, Verrucomicrobia, Candidatus TM6, and Chloroflexi. The community in the recent stage was less rich and harbored competitive interactions, while the later stages experienced a surge in bacterial diversity with cooperative interactions. The shift in α-diversity and composition was strongly influenced by soil organic carbon, carbon to nitrogen ratio, and soil moisture content. The functional analyses revealed a progression from a metabolism focused to a functionally progressive community required for bacterial co-existence and succession in plant communities. Overall, the findings indicate that the bacterial communities inhabit, diversify, and develop specialized functions post-deglaciation leading to nutrient inputs to soil and vegetation development, which may provide feedback to climate change.

Bilateral Microvascular Submandibular Gland Transfer with Implantation of Wharton's Duct in Superior Conjunctival Fornix: A Vision-Saving Procedure for Severe Dry Eye Disease.

Dry eye can initially cause mild symptoms of irritation and may rapidly progress to corneal scarring and blindness. Tear substitutes can only help for mild cases. With the advancement in microsurgical techniques, an option of transferring vascularized salivary glands has shown positive results. We present a case of a 5-year-old boy with congenital alacrimia with ocular surface damage. Vascularized autologous submandibular gland transfer was considered as a viable option for this patient. We performed the gland transfer in two separate stages for the two eyes (1 year 5 months apart). The patient was evaluated for up to 2 years for the right eye and for 7 months for the left eye. Dry eye workup showed drastic improvement (right > left). Biochemical analysis showed gradual transition to resemble that of natural tears. This procedure can result in significant symptomatic improvement and can be a promising treatment option for cases of severe dry eye.

Clickable, selective, and cell-permeable activity-based probe of human cathepsin B - Minimalistic approach for enhanced selectivity.

Human cathepsin B is a cysteine-dependent protease whose roles in both normal and diseased cellular states remain yet to be fully delineated. This is primarily due to overlapping substrate specificities and lack of unambiguously annotated physiological functions. In this work, a selective, cell-permeable, clickable and tagless small molecule cathepsin B probe, KDA-1, is developed and kinetically characterized. KDA-1 selectively targets active site Cys25 residue of cathepsin B for labeling and can detect active cellular cathepsin B in proteomes derived from live human MDA-MB-231 breast cancer cells and HEK293 cells. It is anticipated that KDA-1 probe will find suitable applications in functional proteomics involving human cathepsin B enzyme.

Idiopathic CD4+ T lymphocytopenia: A case report.

Idiopathic CD4+ T lymphocytopenia (ICL) is a very rare immunodeficiency syndrome with an unexplained depletion of CD4+ T lymphocytes and no evidence of Human Immunodeficiency Virus (HIV) infection. Here we report a 29-year-old male patient who had severe ulcerative colitis with low level CD4+ count of 254 cells/mm3, and had no evidence of HIV or Human T cell Lymphotrophic virus type I or II infections. He had recurrent Candidiasis infection and his CD4 count was just 53 cells/mm3 after 3 months. The cause for the decline of CD4 T lymphocytes was unknown.

Quantification of functional crosslinker reaction kinetics via super-resolution microscopy of swollen microgels.

Super resolution microscopy (SRM) brings the advantages of optical microscopy to the imaging of nanostructured soft matter, and in colloidal microgels, promises to quantify variations of crosslink densities at unprecedented length scales. However, the distribution of all crosslinks does not coincide with that of dye-tagged crosslinks, and density quantification in SRM is not guaranteed due to over/under-counting dye molecules. Here we demonstrate that SRM images of microgels encode reaction rate constants of functional cross linkers, which hold the key to correlating these distributions. Combined with evolution of microgel particle radii, the functional cross linker distributions predict consumption versus time with high fidelity. Using a Bayesian regression approach, we extract reaction rate constants for homo and cross propagation of the functional crosslinker, which should be widely useful for predicting spatial variations in crosslink density of gels.

Revascularization in Immature and Mature Teeth with Necrotic Pulp: A Clinical Study.

AIM: The aim of this in vivo study was to evaluate the revascularization procedure both in immature and mature teeth with necrotic pulp and open apices, disinfected with triple antibiotic paste followed by inducing blood clot in the root canal. MATERIALS AND METHODS: Fifteen patients were selected who presented with immature and mature permanent teeth with pulpal necrosis and open apices. In the first visit, the root canal was accessed with LA and rubber dam isolation; then the canal was disinfected using triple antibiotic paste containing ciprofloxacin, metronidazole, and clindamycin in the ratio of 1:1:1 and closed with IRM. In the second visit, after administering local anesthesia and isolating with a rubber dam, the triple antibiotic paste was washed out by saline irrigation, and apical papilla beyond the confines of the root canal was stimulated with sterile H file to produce a blood clot. Finally, the access was closed using a double seal with mineral trioxide aggregate (MTA) placed apical to cementoenamel junction and resin bonded cement over the MTA. Radiographic examination and pulp sensibility test was done during the follow-up period of 2,4,6,8 and 10 months. RESULT: After 10 months follow-up, 10 out of 13 patients showed root development and apical closure. The eight patients out of 13 showed root development, apical closure and lateral thickening of radicular dentin and 2 out of 13 patients showed a positive response to electric sensibility test. CONCLUSION: Within the limitation of this study, it can be concluded that there is evidence of root development, increase in lateral wall thickness, apical closure and positive response to pulp sensibility test in both mature and immature teeth with necrotic pulp. CLINICAL SIGNIFICANCE: The conventional approach for management of teeth with necrotic pulp and the open apex is altered with the possibility of tissue regeneration within the pulp space and continued root development through revascularization procedures. It also re-establishes the vitality in a previously nonvital and necrosed tooth.

International cancer seminars: a focus on esophageal squamous cell carcinoma.

The International Agency for Research on Cancer (IARC) and the US National Cancer Institute (NCI) have initiated a series of cancer-focused seminars [Scelo G, Hofmann JN, Banks RE et al. International cancer seminars: a focus on kidney cancer. Ann Oncol 2016; 27(8): 1382-1385]. In this, the second seminar, IARC and NCI convened a workshop in order to examine the state of the current science on esophageal squamous cell carcinoma etiology, genetics, early detection, treatment, and palliation, was reviewed to identify the most critical open research questions. The results of these discussions were summarized by formulating a series of 'difficult questions', which should inform and prioritize future research efforts.

Quantitative X-ray microradiography for high-throughput phenotyping of osteoarthritis in mice.

OBJECTIVE: To investigate and validate digital X-ray microradiography as a novel, high-throughput and cost-effective screening approach to identify abnormal joint phenotypes in mice. METHOD: Digital X-ray microradiography was used to quantify the subchondral bone mineral content (BMC) in the medial tibial plateau. Accuracy and reproducibility of the method were determined in 22 samples from C57BL/6(B6Brd;B6Dnk;B6N-Tyr(c-Brd)) wild-type mice. The method was then validated in wild-type mice that had undergone surgical destabilisation of medial meniscus (DMM) and in a genetically modified mouse strain with an established increase in trabecular bone mass. RESULTS: The measurement of subchondral BMC by digital X-ray microradiography had a coefficient of variation of 3.6%. Digital X-ray microradiography was able to demonstrate significantly increased subchondral BMC in the medial tibial plateau of male mice 4 and 8 weeks after DMM surgery and in female mice 8 weeks after surgery. Furthermore, digital X-ray microradiography also detected the increase in subchondral BMC in a genetically modified mouse strain with high trabecular bone mass. CONCLUSION: Quantitation of subchondral BMC by digital X-ray microradiography is a rapid, sensitive and cost-effective method to identify abnormal joint phenotypes in mice of both genders at several ages.

Incidence of tardive dyskinesia: a comparison of long-acting injectable and oral paliperidone clinical trial databases.

BACKGROUND: To assess the tardive dyskinesia (TD) rate in studies of once-monthly long-acting injectable (LAI) paliperidone palmitate (PP) and once-daily oral paliperidone extended release (Pali ER). METHODS: Completed schizophrenia and bipolar studies for PP and Pali ER (≥ 6 month duration with retrievable patient-level data) were included in this post hoc analysis. Schooler-Kane research criteria were applied using Abnormal Involuntary Movement Scale (AIMS) scores to categorise probable (qualifying AIMS scores persisting for ≥ 3 months) and persistent TD (score persisting ≥ 6 months). Spontaneously reported TD adverse events (AEs) were also summarised. Impact of exposure duration on dyskinesia (defined as AIMS total score ≥ 3) was assessed by summarising the monthly dyskinesia rate. RESULTS: In the schizophrenia studies, TD rates for PP (four studies, N = 1689) vs. Pali ER (five studies, N = 2054), were: spontaneously reported AE, 0.18% (PP) vs. 0.10% (Pali ER); probable TD, 0.12% (PP) vs. 0.19% (Pali ER) and persistent TD, 0.12% (PP) vs. 0.05% (Pali ER). In the only bipolar study identified [Pali ER (N = 614)], TD rate was zero (spontaneously reported AE reporting, probable and persistent TD assessments). Dyskinesia rate was higher within the first month of treatment with both PP (13.1%) and Pali ER (11.7%) and steadily decreased over time (months 6-7: PP: 5.4%; Pali ER: 6.4%). Mean exposure: PP, 279.6 days; Pali ER, 187.2 days. CONCLUSIONS: Risk of TD with paliperidone was low (< 0.2%), regardless of the formulation (oral or LAI), in this clinical trial dataset. Longer cumulative exposure does not appear to increase the risk of dyskinesias.

Constructing a database of alien plants in the Himalaya to test patterns structuring diversity.

Differences in the number of alien plant species in different locations may reflect climatic and other controls that similarly affect native species and/or propagule pressure accompanied with delayed spread from the point of introduction. We set out to examine these alternatives for Himalayan plants, in a phylogenetic framework. We build a database of alien plant distributions for the Himalaya. Focusing on the well-documented regions of Jammu & Kashmir (west) and Bhutan (east) we compare alien and native species for (1) richness patterns, (2) degree of phylogenetic clustering, (3) the extent to which species-poor regions are subsets of species-rich regions and (4) continental and climatic affinities/source. We document 1470 alien species (at least 600 naturalised), which comprise ~14% of the vascular plants known from the Himalaya. Alien plant species with tropical affinities decline in richness with elevation and species at high elevations form a subset of those at lower elevations, supporting location of introduction as an important driver of alien plant richness patterns. Separately, elevations which are especially rich in native plant species are also rich in alien plant species, suggesting an important role for climate (high productivity) in determining both native and alien richness. We find no support for the proposition that variance in human disturbance or numbers of native species correlate with alien distributions. Results imply an ongoing expansion of alien species from low elevation sources, some of which are highly invasive.

Harmony in nature: understanding the cultural and ecological aspects of plant use in Ladakh.

BACKGROUND: Traditional knowledge (TK) in Ladakh encapsulates a repository of experimental wisdom cultivated over millennia. Despite this cultural wealth, dwindling interest among the younger generations in the region's age-old practices underscores the urgency to document TK. The current study investigates the diverse usage of plants in Surru, Wakha and Lower Indus valleys of Western Ladakh exploring the influence of socioeconomic and ecological factors. METHODS: A stratified random sample approach was adopted to select 540 respondents for gathering information of useful plants through interviews and questionnaires. Participant observation, questionnaires, open-ended and semi-structured interviews were conducted for data collection. Free listing was done to create an extensive list of plants and their uses. Ethnobotanical metrics such as relative frequency of citation (RFC), relative importance index (RI), cultural value (CV) index and cultural importance (CI) index were computed to assess species applicability. Additionally, one-way analysis of variance (ANOVA) was utilized to discern significant differences in knowledge levels based on valleys, gender, education and religion using TK as a response variable. RESULTS: Altogether, we recorded 246 plant species under various ethnobotanical uses from Western Ladakh. These include medicinal (126), fodder (124), wild ornamentals (86), food (81), fuel wood (54), dye (20), religious (31) and others (34). Novel plant reports include Berberis brandisiana Ahrendt and Dactylorhiza kafiriana Renz. The dominant plant family is Asteraceae with 35 species. Suru valley exhibits the highest number of cited plants followed by Wakha-chu and Lower Indus valleys (192, 168 and 152 species, respectively). CONCLUSION: Disparities in plant use understanding are evident among different groups, prompting further investigation through intercultural comparisons. Plants such as Arnebia euchroma, Juniperus semiglobosa, and Artemisia species emerge with cultural importance. Gender, valley affiliation, religious background and the remoteness of a village all influence local plant knowledge. These variations are linked to socioeconomic disparities among communities.

Long-term survival following medical management of Aspergillus endocarditis with dissemination as a consequence of steroid therapy in severe COVID-19 pneumonia.

A male in his 40's with no known comorbidities developed severe COVID-19 pneumonia and received a four-week course of methylprednisolone. The patient subsequently developed disseminated Aspergillus endocarditis, manifesting as multiple organ involvement including the heart, eyes, and brain. Despite the poor prognosis generally associated with fungal endocarditis, the patient survived following aggressive medical management with a combination of liposomal amphotericin b and voriconazole therapy and is now doing well for over two years and is off antifungal therapy for a year.

Aminoacylase efficiently hydrolyses fatty acid amino acid conjugates of Helicoverpa armigera potentially to increase the pool of glutamine.

One of the most prevalent bioactive molecules present in the oral secretion (OS) of lepidopteran insects is fatty acid amino acid conjugates (FACs). Insect dietary components have influence on the synthesis and retaining the pool of FACs in the OS. We noted differential and diet-specific accumulation of FACs in the OS of Helicoverpa armigera by using Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry. Interestingly, we identified FACs hydrolyzing enzyme aminoacylase (HaACY) in the OS of H. armigera through proteomic analysis. Next, we have cloned, expressed, and purified active recombinant HaACY in the bacterial system. Recombinant HaACY hydrolyzes all the six identified FACs in the OS of H. armigera larvae fed on host and non-host plants and releases respective fatty acid and glutamine. In these six FACs, fatty acid moieties vary while amino acid glutamine was common. Glutamine obtained upon hydrolysis of FACs by HaACY might serve as an amino acid pool for insect growth and development. To understand the substrate choices of HaACY, we chemically synthesized, purified, and characterized all the six FACs. Interestingly, rHaACY also shows hydrolysis of synthetic FACs into respective fatty acid and glutamine. Our results underline the importance of diet on accumulation of FACs and role of aminoacylase(s) in regulating the level of FACs and glutamine.

Ocular surface changes in patients who have undergone head and neck radiation therapy.

PURPOSE: Dry eye syndrome (DES) is a familiar sequelae of radiation therapy (RT) for head and neck cancers (HNC). Ocular surface changes such as DES occur due to injury to the conjunctival epithelium, goblet cells, corneal surface, lacrimal glands, and meibomian glands. This study aimed at the evaluation and early detection of changes in ocular surface parameters in patients receiving RT for extraocular HNC. METHODS: Forty-two eyes of 21 patients undergoing HNC RT were evaluated. Radiation technique and dose of radiation to the lens and eye were recorded. Subjects were evaluated for meibomian gland changes by meiboscore grading, ocular surface disease index (OSDI) questionnaire, Schirmer's test, tear film break-up time (TBUT), and slit-lamp examination before RT, immediately post RT, and 6 weeks post RT. A comparison of the ipsilateral eye on the irradiated side to the contralateral eye was done. RESULTS: A significant reduction in TBUT was seen immediately post RT and 6 weeks post RT ( P < 0.001 and 0.008, respectively), with an increase in meiboscore at both visits ( P < 0.001). An OSDI score of >13 was seen in 23.80% of patients post RT, with a significant difference from baseline ( P < 0.001). On comparing ipsilateral and contralateral eye groups, a significant difference from baseline was seen in TBUT ( P < 0.001 and 0.033, respectively) and meiboscore ( P < 0.001 for both eyes). A significant change of >1 second in TBUT and >1.7 in meiboscore was seen with a mean dose of around 8 Gy to the lens. CONCLUSION: All patients undergoing HNC RT should be followed up for ocular surface and meibomian gland changes. The contralateral eye should also be evaluated. Patients receiving lower doses to the ocular structures should also be kept under follow-up.

Knowledge and awareness of clinical trials among trial participants in India: A multicentric questionnaire-based cross-sectional study.

PURPOSE: The knowledge and attitude of participants toward clinical trials (CTs) are a key determinant in successful recruitment and retention. This study aimed to evaluate knowledge and awareness-attitude among the recruited CT participants about CTs. METHODS: This was a cross-sectional face-to-face survey comprising CT participants involved in the past or currently a part of a CT during this study within the Indian Ophthalmology Clinical Trial Network (IOCTN). A previously validated questionnaire was used, and data regarding demographics, knowledge, and awareness-attitude about CTs were collected. The total awareness-attitude score was used to determine the knowledge and awareness of the participants. RESULTS: A total of 121 subjects had participated in the study who were part of ongoing trials, of which only five participants had prior experience of CTs. The majority (90%) had knowledge about CTs, whereas only 7% had confirmed signing consent forms. The total awareness-attitude score significantly varied across locations (27% for the southern zone, 53% for the central zone, and 52% for the western zone), and this was negatively associated with education. The most negative attitude (38.6%) was observed regarding adequate information sharing among participants, whereas non-disruption of their routine family life (60.7%) and financial stability (67.6%) were the top most positive attitudes revealed during their participation. The participants' pre- and post-participation attitude toward CTs revealed a positive attitude. CONCLUSION: The factors influencing a participant's knowledge and attitudes toward CTs were revealed in the study. Furthermore, the need for awareness regarding benefits of CTs to the public, participant's rights, and their voluntary power were the major highlights.

Brolucizumab-associated intraocular inflammation in Indian patients by VRSI study group.

CONTEXT: Concerns about brolucizumab's (Pagenax®) association with intraocular inflammation (IOI) limit its use despite its cost-effectiveness and efficacy. This multicentric study analyzes IOI incidence across 21 tertiary eyecare centers in India since its introduction in October 2020. PURPOSE: To determine the real-world incidence rate of IOI in Indian patients secondary to intravitreal brolucizumab across 21 tertiary eye care centers in India. SETTINGS AND DESIGN: Retrospective multicentric, survey-based study. METHODS: Data including number of patients treated, clinical indications, side effects encountered, and IOI case details was collected via Google Forms in 21 Indian tertiary eye care centers since October 2020. Mean, median, frequency, and standard deviation were calculated for statistical analysis. RESULTS: All centers used pro re nata protocol for brolucizumab injections with a minimum injection interval of 8 weeks. The incidence of IOI was 0.79% (21 events out of 2655 eyes). Treatment indications included idiopathic polypoidal choroidal vasculopathy, neovascular age-related macular degeneration, diabetic macular edema, and off-label uses. IOI was experienced after the first injection (57%) in majority of cases with a median onset of 14 days (range: 1-65 days). IOI was mild in 28.5%, moderate in 33%, and severe in 38% of cases. Eighteen out of 21 IOI eyes recovered preinjection best corrected visual acuity or better. CONCLUSIONS: Our study found a lower IOI incidence (0.79%) with brolucizumab (Pagenax) in Indian patients compared to previously reported literature. IOI events were mostly mild to moderate, and post-treatment, most patients improved or maintained BCVA. Larger prospective multicentric studies with PRN dosing protocol are needed to confirm these findings.

Homology-based annotation yields 1,042 new candidate genes in the Drosophila melanogaster genome.

The approach to annotating a genome critically affects the number and accuracy of genes identified in the genome sequence. Genome annotation based on stringent gene identification is prone to underestimate the complement of genes encoded in a genome. In contrast, over-prediction of putative genes followed by exhaustive computational sequence, motif and structural homology search will find rarely expressed, possibly unique, new genes at the risk of including non-functional genes. We developed a two-stage approach that combines the merits of stringent genome annotation with the benefits of over-prediction. First we identify plausible genes regardless of matches with EST, cDNA or protein sequences from the organism (stage 1). In the second stage, proteins predicted from the plausible genes are compared at the protein level with EST, cDNA and protein sequences, and protein structures from other organisms (stage 2). Remote but biologically meaningful protein sequence or structure homologies provide supporting evidence for genuine genes. The method, applied to the Drosophila melanogaster genome, validated 1,042 novel candidate genes after filtering 19,410 plausible genes, of which 12,124 matched the original 13,601 annotated genes. This annotation strategy is applicable to genomes of all organisms, including human.

Evaluation of fracture resistance of reattached vertical fragments bonded with fiber-reinforced composites: an in vitro study.

AIM: Aim of this in vitro study was to evaluate the resistance to fracture of vertically fractured and reattached fragments bonded with fiber-reinforced composites. MATERIALS AND METHODS: Root canals of 45 teeth were prepared, and the teeth were intentionally fractured into two separate fragments. Control groups (n = 15 each) consisted unfractured teeth with instrumented and obturated. Fractured teeth were divided into three groups (n = 15) and were attached using (1) dual-cure resin cement (RelyX U100), (2) dual-cure resin cement and polyethylene fiber (Ribbond), (3) dual-cure resin cement and glass fibers (stick-net). Force was applied at a speed of 0.5 mm/min to the root until fracture. RESULTS AND STATISTICAL ANALYSIS: Group 1 (RelyX U100 group) demonstrated lowest fracture resistance. Group 4 (control group) showed highest fracture resistance followed by group 2 (Ribbond group) and group 3 (Stick-Net groups). Statistically no significant difference was there between groups 2, 3 and 4. CONCLUSION: Vertically fractured teeth can be treated by filling the root canal space with dual-cure adhesive resin cement or by adding polyethylene fiber or glass fiber to increase the fracture resistance of the reattached tooth fragments, an alternative to extraction.