Lasiodiplodia theobromae is a fungus of the family Botryosphaeriaceae that causes Botryosphaeria dieback of grapevine worldwide. Abiotic stress may change host vitality and impact susceptibility to the pathogen and/or change the pathogen’s life cycle. However, the interaction between both stress drivers is poorly understood for woody plants. We addressed the hypothesis that distinct morpho-physiological and biochemical responses are induced in grapevine (Vitis vinifera)–L. theobromae interactions depending on when water deficits are imposed. Grapevines were submitted to water deficit either before or after fungus inoculation. Water deficit led to the reduction of the net photosynthetic rate, stomatal conductance, and transpiration rate, and increased the abscisic acid concentration regardless of fungal inoculation. L. theobromae inoculation before water deficit reduced plant survival by 50% and resulted in the accumulation of jasmonic acid and reductions in malondialdehyde levels. Conversely, grapevines inoculated after water deficit showed an increase in proline and malondialdehyde content and all plants survived. Overall, grapevines responded differently to the primary stress encountered, with consequences in their physiological responses. This study reinforces the importance of exploring the complex water deficit timing × disease interaction and the underlying physiological responses involved in grapevine performance.Diseases and climate change are major factors limiting grape productivity and fruit marketability.  Vitis vinifera L. is a very important commercial fruit crop worldwide, covering a global area of approximately 7.5 million hectares. Besides grapevine berries being mainly used for wine production, grapevine is also exploited for the production of fresh table grapes, dried fruit (raisins), juice, tannins, and antioxidants . B. B40]. BOne of the most important responses of grapevines to WD is the overproduction of total free amino acids and proline ,43. The w/w) peat:perlite and acclimated for three months (April to July 2018) under greenhouse conditions: natural light, 60/65% relative humidity (day/night), and approximately 25 °C day/15 °C night. Plants were watered once a week until rooting and the development of the first leaves. Afterwards, all grapevines were watered up to 70% field capacity (FC) and fertigated twice a week with 5 mL/L of N:P:K nutrient solution .Dormant cuttings of grapevine rootstocks (1103-P) and Touriga Nacional scion  were rooted in 2 L plastic pots filled with 3:2 . Grapevine plants were randomly allocated to five different treatments  were maintained gravimetrically every day to keep the soil water content at 70% FC by manually watering. To evaluate the effects of water deficit timing on host–fungi interactions, watering was withheld until the soil moisture content reached 15% FC, and then maintained gravimetrically daily for 14 days before inoculation (WD-F) or after inoculation F-WS; .L. theobromae Bt105 isolate used in this study was isolated from V. vinifera cv. Castelão in Portugal  and B = [(Abs 440+TBA − Abs 600+TBA) × 0.0571].Lipid peroxidation was estimated by measuring the amount of malondialdehyde (MDA) in the leaves following the protocol described by Hodges et al. , using 5−1. The gradient used was: (0–2 min) 90:10 (A:B), (2–6 min) 10:90 (A:B), and (6–7 min) 90:10 (A:B). Hormones were quantified with a Quattro LC–triple quadrupole mass spectrometer  connected online to the output of the column through an orthogonal Z-spray electrospray ion source. The analytes were quantified after external calibration against the standards.Salicylic acid (SA), jasmonic acid (JA), and abscisic acid (ABA) were extracted from five leaves per treatment based on Durgbanshi et al. . After fp ≤ 0.05), statistical differences among all treatments were evaluated by one-way analysis of variance  followed by post-hoc multiple comparisons using the Tukey test. Different lower cases indicate significant differences among the treatments  at p ≤ 0.05.Data are presented as mean ± standard error (SE) of five independent biological replicates. Statistical procedures were performed using SigmaPlot . After testing for ANOVA assumptions (homogeneity of variances with the Levene’s mean test, and normality with the Kolmogorov–Smirnov test, There have been very few studies experimentally addressing the effects of the timing of drought on physiological responses of woody plants to pathogen infection ,50. WithL. theobromae inoculation, contrasting with the priming effect verified when WD was imposed prior to fungal inoculation. Overall, grapevines respond differently to the primary stress encountered, with consequences at different physiological and—mainly—biochemical responses, which were not found in internal stem lesions, but were clear in the grapevines’ survival rate.Our results illustrate different specific responses in the interactions between biotic and abiotic stress in grapevines that are dependent on the order of stress imposition. The percentage of survival decreased only when WD was imposed after


Lespedeza buergeri is presented in this report. It is 149,065 bp in length and divided into four distinct regions: a small single copy (SSC) region of 18,934 bp, a large single copy (LSC) region of 82,476 bp, and a pair of inverted repeat regions of 23,826 bp. The annotation of the L. buergeri complete chloroplast genome predicted a total of 123 genes . Phylogenetic analysis with the reported chloroplast genomes revealed that L. buergeri is nested in the genus Lespedeza of Fabaceae family. Furthermore, L. buergeri exhibited a close relationship with Lespedeza bicolor and Lespedeza cuneata. This results in this study might contribute to further investigating the evolutionary relationship of family Fabaceae.The complete chloroplast genome sequence of  Lespedeza buergeri Miq.1876 belongs to the genus Lespedeza, and this genus contains about 60 species within the family Fabaceae , and the specimen was deposited in the Center of Herbarium, Anhui University of Traditional Chinese Medicine, Hefei, China, under accession number 20211019 . Genomic DNA was extracted by DNAsecure Plant Kit . A total of 200 μg of genomic DNA was randomly fragmented to an average size of 300–350 bp, and the obtained DNA was constructed into paired-end (PE) libraries of an average of 500 bp. DNA libraries with different indices were multiplexed and loaded on an Illumina HiSeq instrument according to manufacturer’s instructions . Sequencing was carried out using a 2 × 150 paired-end configuration, image analysis and base calling were conducted by the HiSeq control software (HCS) + OLB + GAPipeline-1.6 (Illumina) on the HiSeq instrument. The reads were quality controlled and then assembled using velvet (version 1.2.10), gaps filled with SSPACE (version 3.0) and GapFiller (version 1–10)  was utilized as a reference genome for predicting genes by Prodigal (version 3.02)  region of 82,476 bp, a small single copy (SSC) region of 18,934 bp, and a pair of inverted repeat regions of 23,826 bp. Overall, the GC content of this Cp genome was 35.82%. The Cp genome of L. buergeri encoded a total of 123 genes, which belong to three categories, including 77 protein coding genes, 8 ribosomal RNA (rRNA) genes and 38 transfer RNA (tRNA) genes. The assembled complete Cp genome sequences of L. buergeri had been submitted to NCBI.The Cp genome of L. buergeri and other related species (Katoh and Standley L. buergeri closely grouped with L. bicolor and L. cuneata in genus Lespedeza. This complete Cp genome can contribute to future population genomic studies, DNA barcoding, and conservation genetics.Furthermore, the Cp genome sequences of 21 species were aligned by MAFFT v7 to understand the phylogenetic relationship between


Ocimum species (Lamiaceae) has been reported, however, its anti-filarial effect has not been explored to date.The anthelminthic effect of Ocimum sanctum L. (OS) against lymphatic filarial parasites.This study evaluates the effect of Setaria cervi. Equal size and number (n = 10) of adult female S. cervi worms were incubated in 125, 250 or 375 μg/mL EOS extract for 6 h at 37 °C. The OS bioactive components were identified by UPLC-ESI-MS/MS and subjected to docking and molecular dynamics (MD) simulation against filarial antioxidant proteins.The ethanol extract of OS (EOS) leaves was tested for anti-filarial activity against S. cervi after 6 h of incubation. The motility was found to be reduced by 53.7% in 375 µg/mL and 43.8% in 250 µg/mL EOS after 6 h of treatment. The UPLC-ESI-MS/MS analysis of ethanol extract of O. sanctum revealed the presence of 13 bioactive compounds. The docking analysis showed eight OS bioactive compounds to have high binding affinity  towards antioxidant proteins of filarial parasites. Additionally, MD simulation studies showed significant impact of (RMSD ≤ 10 Å) chlorogenic acid, luteolin and ursolic acid on filarial antioxidant enzymes/proteins. To our knowledge, this is the first report of the anti-filarial activity of Ocimum sanctum.The EOS significantly inhibited the motility of adult female The effect of EOS and OS bioactive components on human filarial parasites can be further evaluated for the development of new anti-filarial formulations. Wuchereria bancrofti, Brugia malayi, and Brugia timori. Presently 893 million people in 49 countries are living at the risk of LF  is a major health concern of tropical and sub-tropical countries. The disease is caused by three nematode worms: Adult filarial worms are long lived and easily survive for a period of 8–10 years inside their human hosts. Filarial worms have efficient antioxidant systems which protect them from reactive oxygen species generated by host macrophages, neutrophils, and eosinophils. The antioxidant enzymes superoxide dismutase (SOD), glutathione S-transferase (GST), glutathione reductase (GR), and thioredoxin reductase (TrxR) are established helminth drug targets and studies confirm that inhibiting one or more of these activities proves fatal for the parasites (Gupta and Rathaur Ocimum (Lamiaceae) encompasses most popular medicinal herbs and culinary ingredients. Ocimum basilicum L., Ocimum gratissimum L., and Ocimum sanctum L. are extensively studied for their therapeutic properties. The ethanol extract of Ocimum basilicum (OB) is hypocholesterolemic, hypolipidemic, and can modulate the activity of macrophage surface scavenger receptors  is widely consumed across the world for its medicinal and nutritional value and its alcohol extract can counteract cyclophosphamide induced nephrotoxicity  is an herbaceous plant, rich in medicinal properties. Ocimum sanctum is also known as the elixir of life in Ayurveda because of its curative effects. In India OS is considered a holy plant and is grown in almost every household. Ocimum species are found in India, Malaysia, Saudi Arabia and is widely distributed in tropical and sub-tropical countries. The plant is a potent antioxidant and has immunomodulatory, antimicrobial, anticancer, anti-HIV properties , Varanasi, UP, India. The botanical identification was made by the taxonomist Prof. Shashi Pandey at the Botany Department, Institute of Science, Banaras Hindu University. Leaves were washed, shade-dried, ground to a fine powder in mortar and pestle, and stored at 4 °C for extract preparation. A 10% ethanol extract was prepared from OS leaf powder in a Soxhlet apparatus  and B-phase, methanol and water (95:5) with 0.1% formic acid. Mass Lynx 4.1 software was used for data acquisition and processing. RIKEN tandem mass spectral database (ReSpect) for phytochemicals software was used for detailed analysis of UPLC-ESI-MS/MS data , coupled to a Quadrupole-Time of Flight mass spectrometer , equipped with electrospray ionization (ESI). Chromatographic separation of OS extract was performed by ACQUITY UPLC BEH CSetaria cervi is a widely used WHO recommended model for lymphatic filariasis  supplemented with streptomycin, penicillin, glutamine, and 0.5% glucose (KRB maintenance medium). Worms were washed with KRB and incubated in KRB maintenance medium at 37 °C in a water bath for 1 h before subsequent treatments (Singh and Rathaur n = 10) of adult female S. cervi worms were incubated in supplemented KRB having 125, 250, or 375 μg/mL EOS leaf extract for 6 h at 37 °C in a CO2 incubator at 95% humidity. Worms incubated in supplemented KRB medium with 0.4% DMSO served as vehicle control.Equal size and number   and thioredoxin (Trx)  were retrieved from Protein Data Bank . 3D structure of Brugia malayi glutathione reductase (GR) (PM0077742) was retrieved from Protein Model Database (srv00.recas.ba.infn:it/PMDB). The three-dimensional structure of any filarial superoxide dismutase (SOD) was not available in these databases, hence the template sequence of SOD (Accession no. CTP82144.1) was selected based on maximum score and query coverage for protein structure modelling. LOMETS, a meta server approach to template-based protein structure prediction was used to model the structure of B. malayi SOD. Validation of protein models was done by Rampage server (mordred.bioc.cam.ac.uk), and PROCHECK servers . Lipinski filter was used to predict drug like behaviour of OS bioactive compounds   . The statistical significance between control and EOS treated worms was calculated with Student’s t-test. *p < 0.05, **p < 0.01 and ***p < 0.001 was considered statistically significant.All S. cervi were incubated for 6 h in KRB maintenance medium at 37 °C under 5% CO2 and 95% humidity. After completion of incubation, the parasites were transferred to a fresh medium for assessing the recovery. At EOS concentrations of 250 µg/mL and above adult female S. cervi became non-motile after 5 h of incubation  and 53.7% in 375 µg/mL  EOS-treated worms. The increase in ROS generation upon EOS treatment varied in a concentration-dependent manner.The production of reactive oxygen species by adult female arasites . The ROSS. cervi worms were treated with different concentrations of EOS and DNA fragmentation was confirmed by extraction of total genomic DNA content of treated and control parasites. The genomic DNA analysis showed dose dependent nucleosomal DNA degradation after 6 h of treatment with EOS. The maximum DNA laddering was observed at 375 µg/mL and least at 125 µg/mL concentration of EOS  software. The ethanol fraction contained 13 metabolites, i.e., apigenin, caffeic acid, chlorogenic acid, eugenol, ferulic acid, kaempferol, luteolin, quercetin, rosmarinic acid, rutin, sinapic acid, ursolic acid, and vitexin, which were identified in the positive ion mode  values were below grade III except for vitexin and caffeic acid, thus confirming that the above mentioned OS compounds can be considered for further studies  representing the compactness of the complexed filarial antioxidant proteins with OS bioactive compounds during MD simulation is given in Molecular dynamic simulations of only 8 OS bioactive compounds, apigenin, chlorogenic acid, kaempferol, luteolin, quercetin, rosmarinic acid, ursolic acid, and vitexin was performed. These compounds were chosen for MD run based on higher binding energy and GSC scores in molecular docking. The MD simulations of individual filarial GST, Trx, GR and SOD with water were utilized as a control for respective targets. The MD run was carried out for 50 nanoseconds (ns) in an isothermal-isobaric (NPT) ensemble (310 K and 1 bar). The RMSF graphs of GST, Trx, GR and SOD are represented in LF infection results in severe pathologies like lymphedema, hydrocele, lymphangitis, elephantiasis, and tropical pulmonary eosinophilia. The WHO recommends IDA program  for LF elimination and control. These drugs are only effective against microfilariae and have negligible effect on adult worms. The IDA treatment is often associated with various side effects like transient fever, headache, dizziness, malaise, myalgia, fatigue, and gastrointestinal problems  plant has been shown to have anthelminthic effects against gastrointestinal nematodes of sheep and ruminants  is widely known since ancient times for its medicinal properties. OS is a diverse medicinal plant having antioxidant, anti-inflammatory, immunomodulatory, antidiabetic, and antimicrobial properties . We observed that treatment of S. cervi with EOS resulted in DNA fragmentation and increase in ROS levels both of which are hallmarks for apoptosis and programmed cell death.In the study, treatment of female Ocimum sanctum is rich in phytochemicals and contains many bioactive and nutraceuticals molecules that differ from strain to strain and even differ from place to place. The variation in bioactive compounds of Ocimum is common to different growing locations, environmental conditions, and geographical origins. Although most researchers reported ursolic acid, eugenol and rosmarinic acid as major constituents of OS leaf extract , thioredoxin (Trx), glutathione reductase (GR), and superoxide dismutase (SOD) which are key antioxidant enzymes/proteins involved in the defense system of filarial worms. Molecular docking is a faster option for the discovery of novel drugs and is an inexpensive alternative to tedious in vitro drug screening procedures. The maximum interactions of Ocimum bioactive compounds with filarial antioxidant proteins were present within the active site of GST, Trx, GR, and SOD  has preventive and curative properties against many diseases affecting the liver and brain, and also in cancers and metabolic syndromes etc. (Seo et al. Ocimum species for the treatment of parasitic diseases is being explored extensively due to higher efficiency and fewer side effects as compared to allopathic drugs. In this report, the three most promising OS bioactive compounds having anti-filarial activity were identified as chlorogenic acid, luteolin, and ursolic acid. The overall findings from our study provide an understanding of the effect of OS and EOS on filarial parasites. The antioxidant properties and multi-inhibitory potential of EOS and OS bioactive compounds against filarial proteins/enzymes indicates that further studies may lead to development of better anti-filarial treatment strategies.Lately, the use of


Osteoarthritis (OA) is a common type of arthritis characterized by degeneration of the articular cartilage and joint dysfunction. Various pharmacological and non-pharmacological techniques have been used to manage these diseases. Due to the diverse therapeutic properties of marine collagen, it has received considerable attention in its pharmacological application. Thus, the purpose of this study was to compare the efficacy of jellyfish collagen, collagen peptide, other sources of marine collagen, and glycine in treating OA. In the OA rat model, an anterior cruciate ligament transection combined with medial meniscectomy surgery (ACLT + MMx) was used to induce osteoarthritis in rats. Two weeks before surgery, male Sprague–Dawley rats were fed a chow-fat diet. After 6 weeks of treatment with collagen, collagen peptide, and glycine, the results show that they could inhibit the production of proinflammatory cytokines and their derivatives, such as COX-2, MMP-13, and CTX-II levels; therefore, it can attenuate cartilage degradation. Moreover, collagen peptides can promote the synthesis of collagen type II in cartilage. These results demonstrate that collagen and glycine have been shown to have protective properties against OA cartilage degradation. In contrast, collagen peptides have been shown to show cartilage regeneration but less protective properties. Jellyfish collagen peptide at a dose of 5 mg/kg b. w. has the most significant potential for treating OA because it protects and regenerates cartilage in the knee.  Osteoarthritis (OA), the most common form of arthritis, is generally characterized by slowly progressive degeneration of articular cartilage, particularly in weight-bearing joints . The comMost current treatment modes aim to alleviate pain and reduce inflammation, thereby improving function and reducing disability with fewer adverse effects . A more in vitro, suggesting that it may be used to treat osteoarthritis  and medial meniscectomy (MMx)–induced osteoarthritis in Sprague–Dawley rats.This research aims to investigate the effect of Rhopilema esculentum was obtained from Fuzhou Jellyfish Investment Co., Ltd. Seventy male Sprague–Dawley rats (10-weeks old) were purchased from BioLASCO Co. Ltd. . Glucosamine sulfate (GS) was purchased from Viatril-S (United Kingdom). Glycine was brought from Yeongca Co., Ltd. Tilapia peptide (IXOS HDL-50SP) was obtained from Toong Yeuan Enterprise Co., Ltd. Matrix™ Neo was a kind gift from Kanematsu Chemical Corp, Collactive™ was obtained from Green Strong International Co., Ltd. The standard laboratory chow-fed diet (Laboratory Rodent Diet 5001) was purchased from PMI Nutrition International, Inc. . Matrix metalloproteinase-13 (MMP-13), prostaglandin E2, C-Terminal Cross-Linked Telopeptide of Type II Collagen, and Tumor Nuclear Factor Alpha ELISA kits were purchased from Finetest biotechnology Inc. . Zoletil 50 was purchased from Virbac . Lofalin injections (cefazolin sodium) were purchased from Gentle Pharm Co. Ltd. . Formaldehyde solution was purchased from Avantor Performance Materials Inc. .Rhopilema esculentum collagen extraction was performed according to previously used methods . methods , with moRhopilema esculentum were extracted using previously described methods . After discarding the jellyfish, the collagen peptide is freeze-dried to obtain the final jellyfish collagen peptide (JCP).Collagen peptides from  methods  with mod−1 and a resolution of 1 cm−1.Proximate analysis was carried out following the standard of AOAC  for moisad libitum. The rats were acclimatized for 2 weeks and were housed one rat per cage in a room maintained at 25 ± 2°C under a 12 h day/night cycle throughout experimentation. The Institutional Animal Care and Use Committee of the National Taiwan Ocean University reviewed and approved all protocols. The rats were fed for 6 weeks accordingly. They were subdivided into ten groups (n = 7 rats/group): the control group (surgery with an opened knee-joint capsule without ACLT + MMx) and nine groups receiving surgery with ACLT + MMx.Ten-week-old male Sprague–Dawley rats were purchased from the National Laboratory Animal Center . All rats were fed a standard chow-fed diet (Laboratory Rodent Diet 5001) and water 2) in an empty chamber at week 9 of the experiment (treatment with samples or GS for 6 weeks). The rats were starved for 12 h before surgery and euthanization, and on the day of euthanization, the body weight was measured using a weighing scale. Whole blood and knee joints were collected and stored for further analysis. The whole blood of rats was collected from the abdominal aorta of the rats on the day of euthanization using a heparinized syringe. The plasma or serum was separated from whole blood by centrifugation  at 3,000 rpm and 4°C for 15 min. The supernatant was gently collected by using a micropipette and stored at −20°C for future analysis  as a positive control . The dose of JCP was evaluated from some previous studies related to oral administration of jellyfish collagen and jellyfish collagen peptide in rats . The dosanalysis .According to previous methods, the ACLT + MMx surgery to induce OA was performed . BrieflyThe serum tumor necrosis factor-alpha, C-telopeptide fragments of type II collagen Matrix metalloproteinase-13, and cyclooxygenase-2 were measured following Finetest’s protocols. According to p < 0.05 was considered significantly different.All data were expressed as the mean ± standard deviation (SD). The experimental data were analyzed by Statistical Package software such as Statistical Product and Services Solution  and GraphPad Prism 5 for statistical analysis of variance (ANOVA). Duncan’s multiple range test was carried out to analyze the significant difference; Proximate analysis from jellyfish bell shows that ash content was observed at about 72.88% dry weight, followed by protein with 13.93%, carbohydrate with 12.47%, and lipid with 0.71%, with a water content of 74.59% . The exp50 value is a widely used parameter for determining the antioxidant activity of test samples. It is calculated as the antioxidant concentration required for reducing the initial DPPH concentration by 50% . As shown in Rhopilema esculentum collagen peptide (5 mg/kg b. w.) was observed compared to the control group. There is no significant difference in the weight of the kidney, spleen, abdominal fat, and epididymis fat (p < 0.05).As illustrated in p < 0.05). However, TNF-α levels significantly decrease after treatment of jellyfish collagen, collagen peptide, and glycine compared to the OA group (p < 0.05) and show no significant difference from the control group (p < 0.05). In addition, COX-2 and NO levels were also significantly decreased in treated groups (p < 0.05). . .p < 0.05). Moreover, CTX-II levels also significantly decreased in the collagen and glycine group (p < 0.05)   comparedThe articular cartilage in the control group was well-stained with Safranin O (which stains PG red) with no apparent loss of staining intensity, indicating that the matrix’s PG content was normal. The osteoarthritis group’s articular cartilage significantly reduced Safranin O staining intensity in the non-calcified region and a slight reduction in the calcified region, indicating a significant decrease in the matrix’s PG content.Rhopilema collagen, it showed a better histological appearance with a smoother surface. However, a notched surface was still observed, recovery of chondrocytes, and less degenerative change in subchondral bone, but it shows an unclear tide mark  guidelines. A rank order is made as follows JCP5 > GS > TC > JC > glycine > matrix > JCP1 > collactive > OA, as shown in Collagen is the most abundant protein in mammals. It is mainly found as part of the extracellular matrix in connective tissues such as cartilage, tendons, ligaments, bones, skin, etc. . CollageRhopilema esculentum collagen , leak from the liver cells into the bloodstream. GGT can be utilized in multiple diagnostic and prognostic algorithms or scores for liver disease. It is one of the most powerful predictors of the development of existence of liver disease . No signA complex network of factors regulates the anabolic/catabolic balance in articular cartilage, and both sides are impacted during the progression of degeneration. Numerous proinflammatory cytokines, including tumor necrosis factor-α, contribute to the maintenance of the catabolic phenotype during the progression of osteoarthritis . TNF-α lNitric oxide (NO) is an inflammatory catabolic mediator that is produced by osteoarthritic chondrocytes . NO has Undenatured type II collagen provided symptom relief, an action attributed to oral tolerance and modulating inflammatory pathways . Anti-inCollagen peptides can inhibit the activation of JNK and ERK pathways . In the MMP-13 is by far the most investigated metalloproteinase in cartilage pathology . Since MWe have used Safranin-O staining in OA cartilage to confirm that collagen, collagen peptide, and glycine can recover cartilage metabolism’s homeostasis and suppress OA development. The articular cartilage in the control group was well-stained with Safranin O (which stains PG red) with no apparent loss of staining intensity, indicating that the matrix’s PG content was normal. The osteoarthritis group’s articular cartilage significantly reduced Safranin O staining intensity in the non-calcified region and a slight reduction in the calcified region, indicating a significant decrease in the matrix’s PG content. In addition, the articular cartilage’s surface was fibrillated.The treated group shows a recovery of cartilage and less fibrillation in every group, especially in the Rhopilema collagen group, glycine group, glucosamine group, tilapia collagen peptide, and Rhopilema collagen 5x dose group. It showed that cartilage of the treated group had better conservation than that of the untreated group . When itDecreased level of secreted MMP13  observedIn addition, both Gly-Pro-Hyp and Pro-Hyp can operate as inducers of ECM production in cells . DiffereThe morphological changes and proteoglycan loss in stained knee joints were then measured, and OA cartilage pathologies were graded using the Osteoarthritis Research Society International (OARSI) guidelines. A rank order is made as JCP5 > GS > TC > JC > glycine > matrix > JCP1 > collactive > OA, and the same trend as OARSI scoring the incapacitance test between rats have the same trend.Ganoderma lucidum fermentation. As a result of their ability to inhibit the production of proinflammatory cytokines such as COX-2, NO, MMP-13, and CTX-II while also increasing cartilage thickness, collagen and collagen peptides can be used as an osteoarthritis treatment. Jellyfish collagen peptide at a dose of 5 mg/kg b. w., compared to other collagen peptides, has the greatest potential for treating osteoarthritis (OA) in the knee because of its protective and cartilage-regenerating properties.The collagen from jellyfish was successfully extracted using papain-assisted acid extraction, and the collagen peptide from jellyfish was successfully produced using


Research Methods. Eggs studied were exotic chicken, local chicken, turkey, quail, and guinea fowl eggs. The eggs were shelled and their contents (boiled and raw) lyophilized. The standard AOAC method (Kjeldahl) was used to determine protein content, while the amino acid composition was measured using an AA analyzer. The total AA scores were calculated based on the whole hen's egg AA profiles. Statistical significance was accepted at p < 0.05.  Food proteins provide amino acids (AAs) and serve as building blocks of all vital organs, muscles, hormones, and biological fluids such as blood. Eggs are known as a good source of protein. This study compared the protein quality of bird eggs (raw and boiled), using their AA scores since some individuals consume raw eggs for various reasons.  The Guinea fowl egg had the highest total amino acid score (TAAS) of 0.92. The other scores ranged as follows: 0.82 (quail) >0.81 (turkey) >0.75 (exotic chicken), and the lowest score was 0.65 . The least scores were phenylalanine: 0.34 (exotic chicken), phenylalanine and serine: 0.36 , leucine and aspartic acid: 0.41 (turkey), methionine: 0.31 (quail), and glutamic acid: 0.33 (guinea fowl). Also, guinea fowl egg had the highest total essential amino acid (TEAA) (49.6 g/100 g protein), i.e., % TEAA (55.1%), while exotic chicken egg had the lowest (41.1%), but the highest % NEAA (58.9%).  Guinea fowl eggs had the highest EAA and TAA content. Its consumption should particularly be encouraged for children as this can significantly reduce the risk of protein-energy malnutrition and prevent protein deficiencies. Diets and nutritious meals are necessary for sustaining good health and avoiding sickness. Recent global increases in malnutrition have prompted many studies on the impact of different cuisines on human health and well-being. Depending on their nutritional composition, certain foods are deemed healthier than others . “The nuFoods may be grouped broadly into cereal grains, eggs, fat replacers, fats and oils, fish, fruits, herbs and spices, legumes, meat and poultry, milk and milk products, nuts and seeds, roots and tubers, sweeteners, and vegetables . Each caAmino acids (AA) are the building blocks of all critical organs, muscles (particularly heart muscles), hormones, and biological fluids such as blood. A steady supply of high-quality protein is essential for development and other bodily activities since the human body cannot store protein . Hence, Protein quality assessment aims to establish a protein's capacity to satisfy maintenance demands and particular needs for development, pregnancy, and lactation . For theProtein quality may be described as the capacity of a food protein to fulfil the body's metabolic requirements for AA and nitrogen (N). “It is determined by the AA composition and digestibility of the protein and the bioavailability of the individual AA” . The amiin vivo protein digestibility ,” “in vitro protein digestibility,” “the protein efficiency ratio (PER),” “estimated PER,” “maximum PER,” “net protein ratio (or retention) (NPR),” “protein rating (PR),” “net protein utilisation (NPU),” “biological value (BV) ,” and “the protein digestibility corrected amino acid score (PDCAAS).” “AAS is the ratio of the AA content in 1 g of a target protein to that of a reference protein or requirement” .Amount of amino: the overall amino acid score for a given egg sample was calculated by adding the individual amino acid scores and getting the average.p < 0.05.The mean and standard error were computed and analyzed using the one-way ANOVA using SPSS version 20 at a significance level of 5%; LSD was used for multiple comparisons. Results were reported as means and standard errors as in Eridiong et al. . Signifip < 0.05), with guinea fowl egg having the most protein content (20.3% 0.15%) and turkey egg having the lowest (11.5% ± 0.44%). Similar trends were detected across the boiled egg samples, with guinea fowl eggs having the most protein content (26.6% ± 0.06%), which was considerably more remarkable than the others (p < 0.05), and turkey eggs having the lowest .This is summarised in a column of The essential amino acid and nonessential amino acid profiles are presented in Tables p < 0.05) higher values for all the EAA except for valine which was highest in the exotic chicken egg (6.99 g/100 g protein). Comparing the raw against boiled samples, the concentrations of the EAA were slightly higher in the boiled eggs. Most EAA concentrations varied significantly (p < 0.05) among the different egg species, with a few statistically similar values. The raw exotic and local chicken eggs had statistically (p > 0.05) similar content of tryptophan . In contrast, both species in boiled forms also had statistically (p > 0.05) similar content of histidine  and phenylalanine .All the nine essential amino acids were present in appreciable amounts in raw and boiled samples of the five egg species. Among the EAA, isoleucine and valine had the highest concentrations, while tryptophan had the lowest concentrations. Among both raw and boiled samples, guinea fowl egg had significantly (p < 0.05) different within the various groups, with only a few exceptions. The contents of cystine in both raw (2.52 and 2.43 g/100 g protein) and boiled (2.75 and 2.61 g/100 g protein) exotic and local chicken eggs, respectively, were statistically (p > 0.05) similar. Similarly, exotic chicken and quail egg had statistically (p > 0.05) similar content of alanine .Nine NEAA were found in appreciable amounts in the five egg species; the NEAA results are presented in The total EAA (TEAA), total NEAA (TNEAA), TAA, percentage of the essential amino acid (% EAA), and percentage of the nonessential amino acid (% NEAA) are all presented in All the five egg species in this study contained all the essential amino acids in significant quantities, hence the statement that “egg protein is a complete protein.” Out of the 20 naturally occurring amino acids, 18 were present in the eggs—the 9 EAA and 9 NEAA. Two “neutral” amino acids—glutamine and asparagine —were not found in eggs, and this is confirmed by other similar studies such as Ismail et al.  and LewiGuinea fowl eggs had the highest TAA and TEAA content. As a result, it also had the highest AAS making its protein of the highest quality among the five egg species in this study. The values of the TEAA of the eggs were lower than the value of 45.2 g/100 g protein of the egg reference protein  except fArchachatina marginata [However, most animal proteins are low in cystine (Cys), for example %: 35.5% in arginata ; 27.3%–3arginata ; 23.8%–3arginata ; 26.0–26arginata . The prearginata . This mearginata . Cystinearginata , 30.The three branched-chain amino acids (BCAA), leucine, isoleucine, and valine, were also present in reasonable amounts, as seen in the amino acid scores. Guinea fowl eggs had significantly higher amounts of all three BCAA, and their AAS were relatively high. The BCAA are essential for muscle protein synthesis and can also be oxidized in the muscles during exercise for energy. A study by Blomstrand et al.  showed tFurthermore, results from similar studies state that “the difference in protein and AA patterns of various eggs could be attributed to the impact of the diet composition on the level of crude protein and amino acids” , 33, 34.Biochemically, variations in the amino acid patterns of egg proteins may be ascribed to the kind and proportion of protein in the egg white and egg yolks, such as ovalbumin, ovotransferrin, lysozyme, immunoglobulin Y, and ovomucin . More thThis research showed that the five egg species under study had nine essential amino acids in significant amounts and nine nonessential amino acids. Guinea fowl egg had higher amino acid score than the standard reference (hen's egg). In terms of heating, it was observed that boiling the eggs for just 10 mins under standard conditions did not reduce the amino acid content of the eggs; instead, a slight increase was observed in almost all the AA values of the boiled samples. In addition, the results show that not all eggs are the same in protein quality and certain varieties may be better recommended for frequent consumption by children who have higher protein requirements for growth and development. Consumption of less popular varieties such as guinea fowl and quail eggs should also be promoted due to their relatively higher nutritional value.


Myasthenia gravis (MG) is a rare autoimmune disease with clinical symptoms of fluctuating muscle weakness. Due to the side effects of current therapies, there is an urgent need for a new medication for MG treatment. Nux vomica is a traditional Chinese medicine used in various diseases. However, the mechanism of action of Nux vomica against MG remains unclear. in vivo experiments in mice.  Network pharmacology was used to explore the underlying mechanisms of Nux vomica in MG treatment, which was validated using molecular docking and In vivo experiments revealed that Nux vomica can improve the symptoms of MG.  Twelve bioactive compounds and 72 targets in Nux vomica were screened. Seventy-nine myasthenia-related targets were obtained from the GENECARD and DisGeNET databases. PPI networks of Nux vomica- and myasthenia-related targets were constructed using Bisogenet, and these two networks were subsequently merged into an intersection to establish a core-target PPI network that consisted of 204 nodes and 4,668 edges. KEGG enrichment analysis indicated that 132 pathways were enriched in 204 core targets. In addition, we obtained 50 docking pairs via molecular docking.  Nux vomica is involved in the pathogenesis of MG through the “multicomponent-target-pathway” mechanism. Myasthenia gravis (MG) is an autoantibody-mediated neuromuscular disease that causes fluctuating muscle weakness predominantly in the ocular, bulbar, and proximal limb muscles . The preα-dependent signaling pathway, which restored muscle energy supply and ameliorated gastrocnemius muscle contractility in experimental autoimmune MG rats [L, belonging to the genus Strychnos of the family Loganiaceae. Studies have illustrated that Nux vomica has numerous biological activities, such as analgesic, anti-inflammatory, antitumor, and immunomodulatory effects [Traditional Chinese medicines (TCMs) have received increasing attention because of their high efficacy, safety, and few adverse effects. According to the TCM theory, MG is classified as “Weizheng” and diagnosed as “flaccidity syndrome” and “Qi-deficiency pattern” . Thus, Q MG rats . Another MG rats . Nux vom effects . A previ effects . HoweverNetwork pharmacology, first introduced by British pharmacologist Hopkins, has proven to be an effective strategy for revealing the complicated relationship between TCMs and diseases , 12. TCMhttps://www.genecards.org/) and DisGeNET [https://www.disgenet.org/) databases. Candidate targets were collected after the deletion of duplicates.Therapeutic targets for MG were obtained from GeneCards  (https:/DisGeNET  (https:/https://tcmspw.com/tcmsp.php) is an online platform for integrating pharmacokinetics, medicinal chemistry, therapeutic targets, and drug-target-disease networks [https://www.swissadme.ch/index.php). The therapeutic targets of strychnine were then predicted using the SwissTargetPrediction online tool (https://swisstargetprediction.ch/).TCMSP , Biomolecular Interaction Network Database (BIND), Biological General Repository for Interaction Datasets (BioGRID), Database of Interacting Proteins (DIP), Molecular INTeraction Database (MINT), and Human Protein Reference Database (HPRD) . PPI netCentral network evaluation is considered a primary method for core protein screening in PPI networks. First, the PPI networks of the Nux vomica-related targets and MG-related targets were merged into an intersection. Next, the Cytoscape plugin CytoNCA was applied to assess the intersection from the following six centrality measures: betweenness centrality (BC), closeness centrality (CC), degree centrality (DC), eigenvector centrality (EC), network centrality (NC), and local average connectivity-based method (LAC) . In addihttps://geneontology.org/) and KEGG (https://www.genome.jp/kegg/).Gene ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed based on the public database GO  (https://www.bioconductor.org/packages/release/bioc/html/clusterProfiler.html) was adopted to perform the GO and KEGG pathway enrichment analyses with P values <0.01, adjusted by the Bonferroni correction.Here, https://zinc15.docking.org) was used to download two-dimensional (2D) structures of bioactive compounds [https://www.rcsb.org/) [The zinc database (ompounds . AutoDocompounds . The 3D sb.org/) . The hubsb.org/) . Next, tsb.org/)  online psb.org/) . FinallyFemale C57BL/6J mice (aged 7–8 weeks) were purchased from the Animal Resources Center . All mice were housed in a controlled environment under a 12-h light/dark cycle with 21°C and 45–65% humidity and had free access to food and water.μg MuSK in 100 μL phosphate-buffered saline (PBS) and 100 μL complete Freund's adjuvant , which were intraperitoneally injected into the hind foot. The control group was injected with 100 μL of PBS and 100 μL of CFA. The mice in both groups were injected with cyclophosphamide monohydrate  within 24 h to suppress immune resistance. The aforementioned operation was repeated after 30 days. The body weight of the mice was measured weekly, and a clinical examination was performed every 2 weeks to determine the model-building cycle. After the MG mouse model was successfully constructed, the mice were randomly divided into five groups (n = 8). The mice in the low-, mid-, and high-dose Nux vomica groups were administered low (4 mg/kg), mid (8 mg/kg), and high (16 mg/kg) doses of Nux vomica. The mice in the control and model groups were intragastrically administered equivalent doses of saline. The body weight of the mice was measured weekly, and a clinical examination was performed every 2 weeks. After 30 days, mice were sacrificed by cervical dislocation after injecting sodium pentobarbital (50 mg/kg), and tendon tissues were removed for subsequent experiments.After anaesthesia, mice in the model group were emulsified with 40 For clinical examination, mice were left for 3 min on a flat platform and observed for signs of MG. The clinical muscle weakness was graded as follows: Grade 0.5, the mouse exhibited slightly lower activity than normal after the standard exercise state, and the hanging time was >60 s; Grade 1, the mouse exhibited normal movement, lying prone, staying still, and/or hanging time <60 s; Grade 2, before and after exercise, the mouse was lying prone, staying still, and/or hanging test <30 s; Grade 3, severe weakness, dehydration and paralysis, weight loss >15%, and/or hanging time 0 s; Grade 4, the mouse died. The grip of the mice was observed every 2 weeks. First, the tail of each mouse was gently grabbed and the mouse was suspended above a metal grid. When the mouse grasped the metal grid, the mouse was lifted and the tension was measured 20 times. One end was fixed on the tension device, and the front end of the force measurement device was made into a grid shape. The tail of the mouse was grasped, the front end of the force measuring device was grasped, and the tail of the mouse was pulled forcefully. When the mouse was not strong enough to grasp the front end of the force measuring device, the force measuring device showed the grip force at this time. In addition, clinical progress was assessed using an inverted screening test every 2 weeks. In this test, the mouse was placed in the center of the grid and immediately rotated to the upside-down position, and stably maintained 50 cm above the soft cushion. The time when the mouse fell was recorded and the observation time was limited to 300 s.−ΔΔCt method. Primer sequences for qRT-PCR are listed in Total RNA was extracted from the tendon tissues of the mice using TRIzol® Reagent (Ambion™) , and 500 ng of RNA was reverse-transcribed into cDNA using TaqMan® Reverse Transcription Reagents . The expression of target genes was examined using ABI Prism SDS 7000 (Thermo Fisher Scientific) at 95°C for 1 min, followed by 40 cycles of 95°C for 15 s, and 60°C for 1 min, and then calculated using the 2The predicted target protein expression was detected by western blotting. Briefly, total proteins were extracted from tendon tissue using RIPA lysis buffer . Then, the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a polyvinylidene fluoride membrane . The membranes were blocked with 5% skimmed milk and incubated with primary antibodies anti-EGFR, anti-TP53, anti-AKT1, anti-MYC, and anti-GAPDH  at 4°C overnight. After washing with PBS, the membranes were incubated with a secondary antibody . Enhanced chemiluminescent reagents  were used to detect the protein bands. GAPDH was used as an internal control.P < 0.05.Data are shown as the mean ± standard deviation (SD) and analyzed by one-way ANOVA followed by Tukey's test using SPSS 27.0 software . Differences were considered significant at The TCMSP database was used to search for the bioactive compounds of Nux vomica with the criteria of OB ≥ 30% and DL ≥ 0.18. The TCMSP database was also used to search for the targets of Nux vomica. As a result, 11 active compounds with the targets were found in Nux vomica. Previous studies have shown that strychnine is one of the major bioactive and principal toxic compounds of Nux vomica . TherefoP < 0.01). The GO analysis with the top 15 markedly enriched MF, BP, and CC terms is shown in Figures P < 0.01). The top 15 KEGG pathways in which the targets of Nux vomica bioactive compounds were enriched are presented in To further investigate the 72 targets of the Nux vomica bioactive compounds, GO enrichment analysis was performed using biological processes (BP), molecular function (MF), and cellular component (CC) terms. A total of 103 MFs, 454 BPs, and 48 CCs were found to be enriched . The top 15 significant GO terms, including MF, BP, and CC terms, are shown in Figures P < 0.01). The top 15 KEGG pathways enriched for the targets of bioactive compounds from Nux vomica are shown in In the GO enrichment analysis, a total of 144 molecular functions, 1,570 biological processes, and 151 cell components were identified in 204 core targets of Nux vomica following MG treatment -stylopine, brucine-N-oxide, and isostrychnineN-oxide (II)) are shown in P < 0.01). Clinical examinations were performed every 2 weeks. After 42 days, compared to the control group, the clinical scores were increased, the grip strengths were weakened, and the inverted screen times were shortened in the model group (P < 0.01).As shown in After treatment with Nux vomica, the clinical examination indicators of the MG model  significantly improved. After 28 days of treatment, the weight of the MG mice increased significantly, clinical scores reduced, grip strength increased, and the inverted screen times were increased with the increased dose of Nux vomica .To further verify the effect of Nux vomica against MG, we detected the expression of hub targets by qRT-PCR and western blotting. The results showed that compared to the model group, Nux vomica treatment significantly reduced EGFR and TP53 expression and increased MYC expression in a dose-dependent manner. Moreover, HSPA8 expression was not significantly changed, while AKT1 expression was significantly reduced, and the effect was particularly pronounced in the high-dose group. The qRT-PCR results also showed that the mRNA expression of RPS27, UBC, HSP90AA1, and EP300 was not significantly different between the groups .in vivo experiments.MG is an autoimmune disease of the neuromuscular junction, which is mainly mediated by acetylcholine receptor (AChR) antibody and muscle-specific kinase (MuSK) , 28. PreBased on the public databases, 12 bioactive compounds of Nux vomica were collected by setting OB ≥ 30% and DL ≥ 0.18 . StudiesE infection was relevant to MG, at least in China, and may contribute to MG by inducing replication in the thymus, bystander activation, and stimulation of antigen-specific T cells [in vivo, and results showed that Nux vomica treatment downregulated the expression of EGFR, TP53, and AKT1, while upregulated the MYC level in MG model mice. These findings indicated that EGFR, TP53, AKT1, and MYC may be the key targets for Nux vomica treating MG. In addition, the pathways of Nux vomica against MG were related to EBV infection, hepatitis B, hepatitis C, and other viral infections based on KEGG analysis of the core genes. We also identified the miRNA pathway in cancer. An increasing number of studies have suggested that miRNAs are involved in the occurrence and development of MG [MG occurrence is closely related to continuous viral infections, genetic factors, and abnormal immune responses. Growing evidence has demonstrated that EBV infection is correlated with the occurrence of MG , 37. For T cells . Target nt of MG . The miRnt of MG . In anotnt of MG . It was in vivo experiments. The findings showed that the bioactive compounds of Nux vomica exerted therapeutic effects on MG by acting on multiple targets and multiple pathways. Based on the network pharmacology, this study confirmed the therapeutic effects of Nux vomica on MG probably via downregulating the expression of EGFR, TP53, and AKT1 and upregulating MYC level. Our data may provide a valuable theoretical basis for future MG diagnosis and treatment and also serve as a reference for further mechanistic research on Nux vomica treating MG. However, we also found that the mechanisms of Nux vomica against are involved in the PI3K-Akt signaling pathway and miRNAs, so further experiments are needed to prove these predictions. Besides, the therapeutic efficacy of Nux vomica on MG is needed to be verified in a clinical trial.Our study systematically analyzed the potential effects of Nux vomica in the treatment of MG based on network pharmacology and validated its effects in combination with


Kawasaki disease (KD) is an acute idiopathic systemic vasculitis with a self- limiting course that predominantly affects children under 5 years old, particularly in the East Asian countries. Nevertheless, to date, the data on KD in Malaysia are limited. This study aimed to evaluate the epidemiology, clinical features, treatment, and outcomes of KD among the pediatric patients admitted to Hospital Canselor Tunku Muhriz (HCTM), Kuala Lumpur, Malaysia.A retrospective cohort study of 66,500 pediatric patients presented at HCTM from the year 2004 to 2021 was conducted.p < 0.05). Interestingly, abnormal liver parameters (p < 0.05) and incomplete KD (p < 0.05) were significantly related to IVIG resistance.62 KD cases out of 66,500 pediatric admissions were reported, with a male-to-female ratio of 1.58 to 1. Majority of KD patients (95.0%) were younger than 5 years old. Prior infection was reported in 5 KD patients (8.1%). Apart from the classical features, manifestations of various organ systems including cardiovascular (16.1%), gastrointestinal (43.5%), neurological (1.61%), musculoskeletal (1.61%), and genitourinary (17.7%) systems were observed. There was a significant association between sterile pyuria and coronary artery aneurysm (CAA) (The presence of family history, immunological disorder, and previous infection in our KD patients suggested that there is a possibility of genetic, immunological, and infectious roles in the pathophysiology of KD. IVIG resistance is more likely to occur in KD patients with hepatic dysfunction or incomplete KD presentation. These findings highlighted the significant contribution of laboratory parameters to the prognosis of KD, prompting more in-depth research on the KD scoring systems and their relevance in this country. Kawasaki Disease (KD) is an acute systemic febrile vasculitis of unknown etiology mainly affecting children younger than 5 years old . KD was This study was carried out at Hospital Canselor Tuanku Muhriz (HCTM). HCTM is a tertiary medical center and is one of the university hospitals in Malaysia. It is located in Bandar Tun Razak, Kuala Lumpur and is administered by Universiti Kebangsaan Malaysia. The study was conducted from October 2021 until October 2022.In this retrospective study, patients’ data were retrieved from the HCTM Case Mix system using the International Classification of Diseases (ICD) code, ICD-10 (M30.3) Mucocutaneous lymph node syndrome [Kawasaki]. A total of 103 patients who attended HCTM with the diagnosis of KD from 2004 until 2021 were included. 41 patients were excluded due to misdiagnosis [24], missing records [10], and duplicated medical records [7]. The remaining 62 patients were enrolled in the study and classified into 2 groups: complete KD and incomplete KD. The medical records of the patients were reviewed and analyzed. This includes demographic data, clinical features, laboratory results at the time of diagnosis, echocardiographic findings, treatment, and clinical outcomes. These data were collected from December 2021 until May 2022.All registered data of patients who were admitted to HCTM between 2004 and 2021 with the diagnosis of KD based on the health information system were included in this study. With this approach, there were two methodological limitations. The first limitation was type I error, which happened when non-KD patients were coded as KD in the system. These patients did not meet the criteria to be diagnosed as complete or incomplete KD and were excluded from this study. The second limitation was type II error, which happened when KD patients were not coded as KD in the system. These patients' data could not be traced and subsequently not included in this study.Meanwhile, the exclusion criteria of this study are (i) patients' data with repeated names and reference numbers, (ii) patients' data that could not be accessed at all due to loss of information or patient's file. Repetitions of data were considered as a single entry. However, any incomplete dataset was accepted and reported as it is.A data collection form was formed through extensive literature review and focused group discussion with the pediatric specialists. The form was constructed using Google Form to ease the data collection process of KD patients. Patients' information that was extracted were (i) demographic data; (ii) predisposing factors; (iii) clinical features; (iv) laboratory results; (v) echocardiographic findings; (vi) IVIG treatment; (vii) secondary treatment, and (viii) clinical outcome.Complete and incomplete KD were diagnosed according to the AHA guideline. In this guideline, complete KD is diagnosed based on the definition of fever occurrence of ≥5 days and the presence of ≥4 out of 5 principal clinical criteria. These clinical criteria consisted of [i] oral mucosal changes such as strawberry tongue; [ii] bilateral non-exudative conjunctival injection; [iii] polymorphous rash; [iv] changes of extremities such as erythema, oedema, or desquamation; [v] cervical lymphadenopathy. Incomplete KD is diagnosed when the patient only fulfills less than 4 of the principal clinical criteria or if there is any supportive laboratory or echocardiographic findings . LaboratX2 test for categorical data were used to identify any significant differences between the compared variables. The level of statistical significance was set at p < 0.05.SPSS version 28 was used to analyze the data. Frequencies and percentages were used to describe the categorical variables while continuous data were presented as median . Descriptive statistics were used to describe the demographic backgrounds of our KD patients. Meanwhile, parametric tests such as student T-test for normally distributed continuous data, Mann Whitney U test for continuous data that were not normally distributed and non-parametric tests such as the n = 42) had the highest occurrence rate compared to Chinese  while other races including Burmese, mixed Chinese Japanese, and Indian were less than 5%.In our study, there were 62 patients with KD, with a male-to-female ratio of 1.5 to 1 with 20 patients (32.0%) diagnosed with incomplete KD and 42 patients (68.0%) diagnosed with complete KD. There are 59 patients (95.0%) who were less than 5 years old, and within this age cohort there were 42 patients (71.0%) who were less than 2 years old. Among all the races, Malay , upper respiratory tract infection virus , varicella zoster virus  and, Group A streptococcus . In regard to breastfeeding being suggested as a protective factor from KD  had a positive family history of KD. Another patient (1.6%) had a positive family history of immune thrombocytopenic purpura (ITP). In terms of infection, there were 5 patients who presented with history of infections within 4 weeks before the diagnosis of KD including rotavirus  had different initial diagnoses before they were diagnosed with KD. Out of the 5 classical criteria for KD apart from fever, the most common feature was polymorphous rash which was presented in 91.9% (n = 57) of the patients. This was followed by cervical lymphadenopathy , conjunctival injection , erythematous oral cavity , and palmar erythema .All 62 patients had fever upon admission.  The avern = 27). Cardiovascular manifestation only presented in 16.1% (n = 10) of KD patients during presentation with 6 of them having CAA upon admission. Joint manifestation and aseptic meningitis were only present in 1 patient each respectively in this study. Other common feature was flaring of the Bacillus Calmette-Guerin (BCG) scar which was observed in 38.7% of the patients (n = 24). The details on the frequency of each clinical feature are documented in Apart from the classical features, other systemic involvements were also considered. The most commonly involved was the gastrointestinal system, with vomiting and diarrhea as the most common symptoms , with neutrophilia in 66.1% (n = 41) of the patients. Anemia was reported in 40.3% (n = 25) of the patients with a median hemoglobin level of 10.8 g/dl. As for the platelet count, 38.7% (n = 24) of children with KD had thrombocytosis. Interestingly, patients with complete KD had a significantly higher platelet count than those with incomplete KD (p = 0.03) (n = 58) and erythrocyte sedimentation rate (ESR)  due to the inflammatory nature of KD. Nevertheless, there were no significant difference of hemoglobin level (p = 0.38), CRP level (p = 0.54), and sodium level (p = 0.22) between complete KD and incomplete KD patients.Evaluation of the hematological parameters showed elevated white cell count in 48.4% ( = 0.03) . A signin = 29) of the patients had increased alanine aminotransferase (ALT) level ranging from 23 to 1435 U/l while hypoalbuminemia was observed in 90.9% (n = 50) of the KD patients. Only 18.2% (n = 10) of the KD patients had raised bilirubin. None of the patients had elevated gamma-glutamyl transferase (GGT). Among 42 children who developed hyponatremia, those younger than 2 years old were predominantly affected . Hypokalemia was only found in 9.7% (n = 6) of the KD patients.Liver abnormalities were also documented in our patients. Out of 56 KD patients who underwent liver profile investigations, 51.8% (n = 57) of KD patients received IVIG and aspirin as the first line of treatment. 4.8% (n = 3) of KD patients were treated with aspirin alone due to late presentation while one patient was given IVIG only. 10.3% (n = 6) of KD patients who were given IVIG treatment were IVIG resistant and required a second dose of IVIG. None of the patients received third dose of IVIG, steroids or any other immunosuppressive therapy. Of note, deranged hepatic parameters such as raised ALT level and hypoalbuminemia were documented in all IVIG-resistant patients (p = 0.03). Meanwhile, there was a significant association between incomplete KD and IVIG resistance in which 83.3% (n = 5) of the IVIG-resistant patients had incomplete KD (p < 0.05). Side effects of IVIG were observed in 2 (3.5%) of the KD patients on IVIG treatment, with one suspected to have autoimmune hemolytic anemia (AIHA) while another patient developed anaphylactic reactions.A total of 91.9% (n = 10) of the KD patients, mainly presented with CAA , of which 4 (66.7%) of them presented with small aneurysm and 2 (33.3%) with medium aneurysm. Younger children were identified to be complicated with CAA with a median age of 12.50 ± 7.89 months compared to older children (21.52 ± 20.22 months) (p = 0.03). However, there were no significant correlation between: IVIG-resistance (p = 1.00); gender (p = 1.00); timing of diagnosis (p = 0.07); with CAA in this cohort. Echocardiographic follow-up after at least one month showed CAA regression in 3 (75%) of the KD patients, with one (25%) had persistent aneurysm, while the other two defaulted the follow up. On the other hand, valvular insufficiency  was noted in 2 (3.2%) and pericardial effusion in 3 (4.8%) KD patients, respectively. The duration of follow-up for our KD patients ranged from 2 months to 10 years (median: 2 months), with longer duration particularly noted in those with cardiovascular sequelae. No death cases were reported.Cardiovascular complications were detected in 16.1% ( months) . We alsoKD incidence was on a rising trend in HCTM, Malaysia for the past 17 years, which was consistent with other Asian countries including Japan, China, South Korea, India, Iran and Australia. , 26–28. n = 20) were diagnosed with incomplete KD with 19 of them are less than 5 years old. Similarly, a study in Switzerland reported 29% of incomplete KD cases, indicating increasing challenges to diagnose KD  and was significantly higher than a previous study in southern Malaysia (6%) , 28. TheIn this study, our KD patients had leukocytosis (48.4%), anemia (40.3%) and thrombocytosis (38.7%) which were in line with most reported literature , 62. We n = 57) received IVIG as the first line of treatment, which aimed to halt immunological responses to reduce cardiac sequelae in KD patients  of them presented with cardiac complication, mainly presenting with CAA (n = 6). This finding was higher than that of a wide-scale study done in Japan (9%) and another study in Canada (2.4%), which probably attributed to delayed diagnosis and initiation of treatment in our population (p > 0.05) which was contradictory to other studies  guidelines was not used in this study due to lacking data on height and weight in some of our KD patients .KD incidence in our center showed an increasing trend which was similarly observed in other studies. Nevertheless, it is still lower compared to other East Asian countries particularly Japan and Korea. Children younger than 5 years old and males were predominantly affected by KD. Our study further revealed the presence of sterile pyuria to be associated with CAA, and IVIG resistance in KD is likely among those with hepatic dysfunction or incomplete KD diagnosis. These findings highlighted the significant contribution of laboratory parameters to the prognosis of KD, prompting more in-depth research on the KD scoring systems and the relevance in this country.


The first group (CTR) served as control; the second group (IVM) received subcutaneous injection of IVM (0.2 mg/kg body weight); the third group (ZnNPs) fed on zinc nanoparticles (60 mg/kg diet); and the fourth group (ZnNPs + IVM) were administered IVM and zinc nanoparticles at the same doses. The experiment lasted for 9 weeks. Results revealed that IVM-intoxicated rabbits showed impaired growth performance parameters, including body weight, total body weight gain (TBWG), total feed intake (TFI), and feed conversion ratio (FCR). Moreover, carcass characteristic and fertility parameters  were also impaired after IVM administration. Additionally, testicular malondialdehyde (MDA) and antioxidant  levels as well as the histopathology and immunohistochemical expression of caspase 3 and PCNA in the testes and epididymis were detrimentally affected. On the contrary, ZnNP administration efficiently improved most of these parameters in IVM-intoxicated rabbits. In conclusion, ZnNPs exhibited promising ability for improving the growth and fertility status of rabbits and reducing the deleterious effects of IVM possibly through the suppression of apoptotic and oxidative pathways.Ivermectin isthe medication of choice for treating human onchocerciasis and is used in veterinary medicine to treat a variety of ectoparasites and endoparasites. This study was designed to investigate the effects of zinc nanoparticles (ZnNPs) on the fertility of male rabbits exposed to experimental ivermectin (IVM) intoxication. A total of 72 mature male rabbits were equally divided into 4 groups ( Commercial rabbits have recently gained considerable attention due to their high prolificacy and rapid growth rate compared to broiler chicken  had better growth, reproduction, and immunity compared to those who were not. To date, however, no available studies have investigated the impact of ZnNPs on the side effects of IVM. Therefore, the present study aimed to determine the possible impact of ZnNPs on the growth and fertility of male rabbits receiving IVM treatment, focusing specifically on the possible mechanisms of action.The Institutional Animal Care and Use Committee of University of Alexandria approved the experimental protocol used in this study (Permit #2021/013/97).Zn oxide 99.99%  was purchased as a commercial product from Sigma Company, Egypt. Nano-Zn oxide 97%  was purchased as a commercial product from Sigma Company, Egypt. The size of Zn oxide nanoparticles was ˂ 50 nm according to the manufacturer company. IVM was purchased from Alfasan International BV Company, the Netherlands.A total number of 72 V-line male rabbits (3 months old) were used in this experiment. Rabbits were individually reared in batteries  of galvanized wire net, equipped with an automatic drinker and a manual feeder. Rabbits were reared in an open house system . The temperature was adjusted to 19–23 °C. Relative humidity was nearly 60% with a 16-h light and 8-h dark cycle. Fresh tap water was continuously available for consumption via stainless steel nipples located inside each cage. Rabbits were acclimated for 2 weeks before the beginning of experimental procedures.After acclimatation, animals were divided into four groups (18 bucks/group), and each group was subdivided into three replicates (6 bucks/replicate). Bucks were fed a balanced basal diet containing all the required nutrients according to NRC  . Total weight gain and total feed intake were recorded, after which the average feed conversion ratio, that is, the amount of total feed intake/total body gain, was calculated.For the assessment of carcass characteristics, nine rabbits from each treatment were randomly selected. Rabbits were weighed in a fasted state before slaughtering to determine the live body weight. After the slaughtered rabbits were bled, the skin, genitals, urinary bladder, gastrointestinal tract, and the distal part of the legs were removed. Hot carcasses  were weighed. The dressing percentage and the ratio of thigh, skin (with head skin), skin (without head skin), head with skin, head without skin, heart, lung, liver, spleen, abdominal fat, and kidney relative to the live BW were calculated.Organ dimensions  were measured after slaughter. Circumference was measured using a measuring tape, whereas testicular length was measured using an obstetrical pelvimeter.6) at 40 × magnification using light microscopy . The percentages of progressive motility and viability were evaluated for at least 200 spermatozoa from each buck. Assessment of motile sperm at the warm stage showed progressive forward movement under 100 × magnification using a light microscope. Assessment of live and dead sperm was performed by counting 200 sperm cells using an eosin-nigrosin staining mixture. Complete or partial purple-stained sperm cells were considered non-viable, whereas non-stained sperm cells were considered viable.Immediately after slaughtering, the animals were dissected and the epididymis was collected as quickly as possible and placed in a clean Petri plate. Thereafter, the cauda epididymis was separated from the whole epididymis, cut into several pieces, immersed in 3 mL pre-warmed phosphate buffer saline (PBS) solution, and incubated for 10 min at 37 °C to allow for sperm release from the epididymal lumen , plasma testosterone levels were estimated following the methods described by Tietz .At the end of the experimental period, rabbits were slaughtered and dissected. The testes were carefully removed, cut into small pieces, and preserved at − 20 °C for further analysis of Zn, reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA). Testicular Zn , SOD , and CAT  were assessed using spectrophotometric procedures  with commercially available kits  according to the manufacturer’s instructions. GSH  and MDA  testicular contents were examined using the colorimetric method according to Beutler et al.  and OkhaImmediately after slaughtering, specimens were collected from testes of control and treated bucks. The collected samples were washed and immersed for 48 h in a 10% neutral-buffered formalin solution for fixation. Fixed samples were prepared using the routine paraffin-embedding technique Bancroft . Briefly2O2 0.3% (v/v) (PBS). Thereafter, sections were incubated for 1 h with 10% (v/v) normal goat serum to block non-specific immunologic reagent binding, and tissue sections were incubated overnight at 4 °C with anti-Caspase-3 antibody rabbit monoclonal [EPR18297]  and Anti-PCNA Mouse monoclonal antibody [24/PCNA] . Afterwards, sections were washed in PBS and treated for 60 min with biotin-conjugated goat anti-rabbit IgG antiserum . The sections were then rewashed in PBS and treated for 30 min with streptavidin-peroxidase conjugate . The streptavidin–biotin complex was observed for 3 min with a 3,3′-diaminobenzidine tetrahydrochloride (DAB)-H2O2 solution. Finally, Mayer’s hematoxylin solution was used to counterstain the sections. Several original micrographs were obtained from five high-power fields/sections/organs at random and used for quantitative histomorphometric examination of immunostaining. Caspase-3 and PCNA positive brown color cells were counted in each micrograph  using manual computer-assisted cell counting (ImageJ plug-in-cell counter.jar) with ImageJ  , and endogenous peroxidase activity was quenched in phosphate-buffered saline with HP < 0.05. All values were expressed as mean ± standard error. Histomorphometric analysis of caspase-3 and PCNA immune expressions were analyzed using the non-parametric analysis using Kruskal–Wallis test to assess the significance between mean scores obtained from Wilcoxon rank-sum test.Statistical analyses of the data were performed using SAS software SAS . One-wayP < 0.0001) reduction in live BW at 18, 20, and 21 weeks old compared to the control group. In addition, IVM-treated rabbits had significantly lower total BW gain (TBWG) and total feed intake (TFI) (P < 0.0001) compared to control group, which caused worst FCR. Conversely, rabbits in the ZnNPs group had significantly higher TFI (P < 0.0001) compared to control rabbits. Interestingly, rabbits in the ZnNPs + IVM group showed significant greater improvement in live BW at 16, 18, and 20 weeks old and TFI, with a subsequent improvement in FCR, compared to the IVM group.The effects of dietary inorganic Zn replacement through ZnNPs with or without IVM injection on the growth performance of buck rabbits are presented in Table P < 0.0001) in the ZnNPs group and significantly lower (P < 0.0001) in the IVM group compared to the control group. However, the relative weight of the thigh was significantly improved (P < 0.0001) in the ZnNPs group compared to the control group. Conversely, skin (with or without head skin) and abdominal fat showed no significant change (P < 0.0001) compared to the control group. In addition, the relative weights of the head with skin, heart, and lungs were significantly lower (P < 0.0001) compared to the control group. The relative weights of the head without skin and spleen were significantly decreased in IVM group (P < 0.0001) compared to the control group. Additionally, the relative weights of the liver were significantly greater (P < 0.0001) in the IVM group and significantly lower (P < 0.0001) in ZnNPs and ZnNPs + IVM groups compared to control group. The relative weight of the kidney was significantly lower (P < 0.0001) in the IVM and ZnNPs + IVM groups compared to the control group.The effects of dietary ZnNP supplementation with or without IVM injection on dressing percentage and carcass quality of buck rabbits are shown in Table P < 0.0001) in ZnNPs group compared to the control group. However, the penile length of the different groups differed significantly (P < 0.0001) compared to that of the control group, with ZnNPs group and CTR group having the greatest and least values . In addition, testicular circumference and length were significantly lower (P < 0.0001) in all treatment groups compared to the control group. However, testicular width was significantly higher (P < 0.0001) in ZnNPs + IVM compared to control group.Data in Table P < 0.0001) in ZnNPs and ZnNPs + IVM group compared to the control group  higher in ZnNPs group compared to the control group. Meanwhile, serum testosterone concentration at 18 weeks old was significantly higher (P < 0.0001) in ZnNPs and ZnNPs + IVM groups than in the control group. At 20 weeks old, serum testosterone concentrations did not significantly differ (P < 0.0001) among the groups.In Table P < 0.0066) than the other groups. Meanwhile, the CTR and IVM groups reached puberty at 19 and 20 weeks old, respectively  significantly earlier (ly Table .As shown in Table Our findings showed that the level of testicular MDA (an index of oxidative process) was significantly improved in the IVM group and significantly lower in ZnNPs group compared to the control group. Meanwhile, rabbits in ZnNPs + IVM group showed no significant difference in MDA levels compared to the control group.As shown in Fig. P < 0.05) declined in IVM group compared to control rats. However, oral administration of ZnNPs in ZnNPs + IVM group significantly (P < 0.05) improved the score of spermatogenesis compared to rats in IVM group . Our results are consistent with those presented in Tag-El Din , which sP < 0.05) decreased the TFI due to the decreased in appetite, which significantly decreased the BW and total gain. Fortunately, the present study revealed that replacement of inorganic Zn with nano-Zn could overcome the negative effects of repeated IVM injections on the growth and FI. Hence, the co-administration of nano-Zn had ameliorative effects by reversing the negative effects of IVM on the BW, gain, and feed consumption in rabbits.On the contrary, our findings revealed that weekly IVM injections significantly deteriorated the growth and feed intake among rabbits receiving dietary inorganic Zn. Similarly, El-Shobokshy et al.  found thP < 0.0001) relative weights of the heart, lungs, kidneys, and livers compared to those receiving inorganic Zn (CTR). Consistent with our results, Tag-El Din  improved the dressing percentage and reversed the negative effects of IVM injection on organ relative weights, especially of the spleen and liver. Similarly, previous studies by Khaldoun-Oularbi et al.  compared to the inorganic Zn groups. Moreover, similar findings in Japanese quails were reported by Reda et al. (P < 0.001) enhanced serum SOD and glutathione peroxidase (GPX) but reduced MDA levels. Moreover, Kamel et al. (In line with this, the present study revealed that dietary nano-Zn replacement significantly enhanced Zn concentrations in the testes of ZnNP-treated group. Our results were consistent with those obtained by Zhao et al.  who founa et al. . Given ta et al. . MDA is a et al. . Our resa et al.  who founl et al.  stated tl et al.  reportedIn contrast, rabbits in the IVM groups showed lower enzymatic activity of GSH, SOD, and CAT and higher MDA levels. This result highlights the harmful and immunosuppressive effects of IVM injection in rabbits. Omshi et al.  stated tThe effect of IVM on genital organ dimensions was demonstrated in the IVM group, especially with regard to the size and weight of the testis plus epididymis and weight of accessory glands. This result was in agreement with that reported by El-Far  who demoIVM affected semen parameters, such that the IVM group had significantly lower semen motility, livability, and concentration than the other groups. This result was in agreement with those reported by Elzoghby et al.  who founReplacement of inorganic Zn with Zn nanoparticles enhanced the BW, weight gain, and FCR of male rabbits. In addition, fertility and oxidative parameters, as well as histopathologic findings, were also improved in ZnNP-treated rabbits. Dietary supplementation of ZnNPs for IVM-intoxicated rabbit ameliorated the negative impact of IVM and improved the performance of male rabbits potentially via its antioxidant and antiapoptotic pathways. Hence, we recommend including ZnNPs in the diets of rabbit exposed to IVM injections.


Students were confined to their homes due to the national closure of educational institutions during the COVID 19 pandemic, thus presenting an unprecedented risk to children’s education, protection, and wellbeing.This study aimed to understand the determinants of subjective wellbeing of adolescents and youth (aged 11–21 years) during the COVID-19 pandemic in India.A cross-sectional web-based survey was adapted, pre-tested, and finalized to obtain the participant’s responses from schools and colleges. Participants aged 11–17 years were engaged through schools. Consent procedures were followed. The survey link was disseminated through social media for the participants aged 18–21 years. The survey was made available in English and Hindi. The data was collected from March-June, 2021.p < 0.01) difference in the level of student’s life satisfaction before and during the COVID-19 pandemic. Of the students who were dissatisfied with their general life during the pandemic, nearly 63.4% felt sadness followed by other feelings, i.e., boredom (around 60.5%), loneliness (63.7%), and anxiety (62.2%).Overall, 1,596 students completed the survey. Out of 1,596 students, 1252 (78%) were below 18 years and 344 (21.5%) participants were 18 years and above. Results suggest a statistically significant (This study highlights the need for innovative strategies for adolescents and parents to adopt and promote overall subjective wellbeing, especially during public health crises such as the COVID-19 pandemic. The COVID-19 pandemic has created unprecedented challenges for people all around the world. In the current scenario, the main measure to contain the COVID-19 infection spread includes self-isolation, quarantine, mask-wearing, hand washing, and social distancing . ChildreIn India, the new academic year (2020–2021) was initiated virtually in April 2020 in New Delhi and in June 2020 in other parts of the country. While this shift to virtual learning has been able to provide some semblance of normalcy for students, who are otherwise holed up in the safety of their homes, it has also exposed widespread social and economic disparities in society. This is reflected by the inequitable distribution and capacity to access digital platforms due to the family’s absence of multiple electronic devices and limited financial capacity to afford electronic devices . FurtherThe primary objective of this study is to explore the determinants of subjective wellbeing of adolescents (11–17 years) and youth (18–21 years) during the COVID-19 pandemic. The secondary objective is to understand the changes in the subjective wellbeing of adolescents before and during the pandemic.A cross-sectional survey was conducted using an online platform to understand the general life satisfaction of school/college students before and during the COVID-19 lockdown across 15 Indian states. The survey responses were self-reported. The survey used in one of the studies conducted by the University of Luxemburg  was adopThe study included participants from schools and colleges in the age group of 11–21 years. The participants under study belonged to the selected schools across 15 states/union territories  of India. Overall, 1,596 students completed the survey.The data was collected through web-based forms, which were circulated through social media, such as Facebook and WhatsApp, for the participants aged 18–21 years. The online survey form included nearly 83 questions including socio-demographic characteristics of the respondents. The survey link was open for participants aged 18 and above. Participants aged 11–17 years were engaged through schools. Consent procedures were followed. The survey link was shared with the students by the school authority following school consent. Informed assent was obtained from the participants below 18 years and consent was obtained from participants above 18 years. The study was approved by PHFI IEC (TRC-IEC 446.1/20). The data was collected from March–June, 2021.The explanatory variables used in this study were: age, gender, father/mother’s education, father/mother’s occupation, wealth index, physical activity, and psycho-social feelings (sad/worried/bored/lonely) during COVID-19. The wealth of the adolescent’s households was assessed by asking them questions about various material assets .The wealth index (a composite measure of a household’s cumulative living standard) was created based on the suggested guideline of DHS . We have considered responses on households’ material possessions  to create a wealth index using principal component analysis (PCA), scoring then converting into “5 quintiles,” and further (as suggested by DHS).We used 14 items-each representing an individual response  on a distinct household item. The questions to assess the household’s assets were: Does your household have-electricity/a fixed telephone/television/radio/refrigerator/car/moped/scooter/motorcycle/washing machine/Internet/computer/smart phone/tablet.The PCA was used to generate scores using responses on household assets. The score was further converted into 5 quintiles using the function “xtile” in Stata software. In this way, we created the variable “wion” with five quintiles. These five categories were labeled as poorest = 1, poorer = 2, 3 = medium, 4 = richer, and 5 = richest, as per the guidelines suggested by the DHS on the creation of wealth index/wealth quintile.The responses (dichotomous) on these households’ assets were used to create the variable “wealth index” using PCA. The scores generated through PCA were converted into 5 quintiles. The 5 quintiles were then further divided into three categories: poor, medium, and rich. The variable “wealth index” was used to understand the socio-economic status (SES) of adolescents.The dependent variable in this study was “general life satisfaction during the pandemic,” which was assessed by the question “How satisfied are you generally with your life during the time of the coronavirus?” This question included four response categories, i.e., very dissatisfied, not satisfied, satisfied, and very satisfied. We created the variable “gen_life_satis” by dichotomizing the variable as 0 = satisfied (satisfied/very satisfied) or 1 = dissatisfied (very dissatisfied/not satisfied) for analysis purposes.The variables “father_occupation,” “wealth_index,” “Psysc,” and “Phy_active” were considered the main outcome variables in the logistic regression modeling. The variable “father_occupation” was assessed by the question What is your father’s occupation?The question had seven closed-ended options: Professional ; Semi-Professional ; clerical, shop owner, or farmer; Skilled worker ; Semi-skilled worker ; Unskilled worker (Laborer); and Unemployed. We have recategorized this variable into four categories: 1 = professional , 2 = skilled (skilled/semi-skilled worker), 3 = unemployed, and 4 = other . Similarly, the variable “wealth_index” was created by recategorizing the variable “wion” into 1 = poor (poorest/poorer), 2-medium, and 3-rich (richer/richest). The independent variable “Psysc” was created through the responses of four items (sad/worried/bored/lonely) which were dichotomized  for analysis purposes. We also created a variable named “Phy_active” by simply using the dichotomous response to the question Did you perform PT/drill during COVID-19 lock-down?Descriptive analyses including exploratory graphs and percentage distribution of categorical variables were done to understand the distribution of data. Bivariate analysis using non-parametric tests, i.e., Chi-square at a 5% level of significance, was used to understand the association between outcome and predictors. The absolute changes were calculated to understand the changes in the wellbeing of adolescents before and during the pandemic. The negative effects (sad/worried/bored/lonely) during COVID-19 were assessed with a four-item scale, adapted from the International Survey of Children’s Well-Being. Adolescents were asked to rate the frequency with which they experienced specific feelings along a 4-point scale ranging from “almost never (0)” to “very often .” The reThe variables “age” and “gender” were treated as the co-variates in the predictive modeling. We also estimated the predicted score, margin, and interaction effect of covariates, which was illustrated through a “marginal plot.”Out of 1,596 students, 1252 (78%) were below 18 years and 344 (21.5%) participants were 18 years and above. Around 61.4% of female and 38.6% of male participants responded to the survey, as shown in p = 0.000) difference in the level of student’s life satisfaction before and during the COVID-19 pandemic. The value of absolute change reveals that the level of student’s life satisfaction has decreased due to the pandemic, which resulted in increasing dissatisfaction in general lifestyle, including a longer stay at home and online schooling during COVID-19 (More than three quarters of the students (84.0%) were satisfied/very satisfied with their general school life before the COVID-19 pandemic. This study found that the proportion of dissatisfaction among students increased during COVID-19. More than half of the students (57.5%) felt dissatisfied with their life at home during lockdown . The resCOVID-19 .p = 0.000), indoor games (p = 0.000), and PT/drill (p = 0.000) for before and during COVID pandemic lockdown. While no statistically significant difference was found for yoga (p = 0.210) before and during the COVID period (Most of the students played (88%) outdoor sports  and did PT/drill (83.1%) before COVID-19, while relatively fewer students played outdoor sports (12%) due to lockdown caused by COVID-19 in their respective areas. This study found that the lockdown has changed the regular playing habits of the students. The proportion of students playing indoor games has increased from 36.1% (before COVID-19) to 63.9% during COVID-19 restrictions . BinomiaD period .This study found that the overall proportion of students who were dissatisfied with their general lives during the pandemic increased from 16.0% (before COVID-19) to 57.5% (during COVID-19).p = 0.000), gender (0.007), occupation (0.002), and wealth index (p = 0.020), were significantly associated with the outcome variable . Also, this study found a significant increase in dissatisfaction levels among students during the COVID-19 lockdown in their respective areas (The respondents who felt dissatisfied during the pandemic who were younger than 18 years (61.4%) were more than those over 18 years (43.0%). While the proportion of dissatisfied female students with their general lives during the pandemic was about 60%, which was relatively higher than their male counterparts (53.2%). During the pandemic, more than half of the students with unemployed fathers felt dissatisfied with their general lives at home. The proportion of students who belonged to the middle (53%) and richer (56%) sections of the society was relatively lesser than those who had a poor SES (61.5%). This study found that the education of parents was not significantly associated with the level of dissatisfaction among students during the pandemic, while other variables, such as age group , loneliness (63.7%), and anxiety (62.2%) .The results of the logistic regression revealed that the students with unemployed fathers  were significantly more likely to feel dissatisfied with their general lives during the pandemic, compared to the students whose fathers were professional workers. The odds of dissatisfaction were lower among the students who belonged to middle socioeconomic status  or richer compared to those who belonged to poor sections of the society. Students who felt sad/bored/lonely/anxious were 2 times more  likely to be dissatisfied with their general lives compared to those who did not feel such negative feelings. The students who did PT/drill were less likely to feel dissatisfied than those who did not do any physical activity during the pandemic .The age and sex interaction modeling and margin plot provided us with the probability of dissatisfaction among students by age and gender. On average, the probability of dissatisfaction among male students below 18 years is 0.58 (58%), while it is 0.62 (62%) among female students in the same age group .p = 0.01) difference in the level of student’s life satisfaction before and during the COVID-19 pandemic. There are statistically significant differences between proportions of students playing outdoor sports (p = 0.000), indoor games (p = 0.000), and PT/drill (p = 0.000) before and during the COVID pandemic lockdown. Of the students who were dissatisfied with their general life during the pandemic, nearly 63.4% felt sadness followed by other feelings, i.e., boredom (around 60.5%), loneliness (63.7%), and anxiety (62.2%). Students who felt sad/bored/lonely/anxious were 2 times more  likely to be dissatisfied with their general lives compared to those who did not feel such negative feelings.The findings of our study revealed that there is a statistically significant  variables like age group, gender, occupation, and wealth index were the determinants of general life satisfaction, (ii) students with unemployed fathers and students from poor economic strata of the society were dissatisfied more, (iii) the level of dissatisfaction among female students below 18 years was more compared with the male students in the same age group, and (iv) students who did some sort of physical activity felt less dissatisfaction in general. A similar study was conducted to investigate the correlates of different dimensions of subjective wellbeing in adolescents (10–16 years) from Luxembourg, Germany, and Brazil, which revealed that adolescents from low-income homes may be more vulnerable to the negative impacts of COVID-19 that can affect mental health , to be better prepared for future developments . The curThe raw data supporting the conclusions of this article will be made available by the authors on request.The studies involving human participants were reviewed and approved by Public Health Foundation of India-Institutional Ethics Committee (PHFI-IEC) (TRC-IEC 446.1/20). Written informed consent for participation was not provided by the participants’ legal guardians/next of kin because: Adolescents and youth (aged 11–21 years) were invited to participate and complete the web-based online survey during COVID-19 pandemic. Informed assent was completed by all the participants below 18 years of age and informed consent was obtained from participants aged 18 and above. Adolescents below 18 years of age were invited through schools after obtaining consent from school authorities.MA, RM, KS, and TR conceptualized the study design and provided guidance to implement the study. TR collected, organized, and wrote the manuscript. VM supported the analysis of the data with TR under the guidance of MA, RM, and KS. MA, RM, KS, SS, MK, and SG revised the manuscript critically for intellectual contents. All authors approved the final manuscript.