Real time PCR-based evaluation of live attenuated lumpy skin disease virus vaccines for immunogenicity and efficacy.

Introduction: Burn wounds are highly susceptible to bacterial infections, particularly from organisms that produce biofilms and exhibit multidrug resistance. These biofilms make infections more difficult to treat by protecting bacteria from antibiotics and slowing the healing process. Identifying the microorganisms involved and understanding their resistance patterns are essential for guiding treatment and improving outcomes in burn patients. Aim: To investigate the microbial profile, antibiotic susceptibility, and biofilm production in bacterial isolates from burn wound infections in patients at a tertiary care hospital. Materials and Methods: This cross-sectional study was conducted in the Department of Microbiology, Shri M.P. Shah Government Medical College, Jamnagar, Gujarat, India, over a one-year period (October 2018-September 2019). A total of 100 samples were collected from patients with burn wounds, yielding 48 bacterial isolates. Swab samples were cultured for bacterial growth, identified using standard biochemical methods, and tested for antibiotic susceptibility using the Kirby-Bauer disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Biofilm production was assessed using the Tissue Culture Plate (TCP), tube adherence, and Congo Red Agar (CRA) methods. Demographic details including age, sex, type, and extent of burns were recorded. Data were analysed using Statistical Package for the Social Sciences (SPSS) version 24.0, applying the Chi-square test, with p-values <0.05 considered statistically significant. Results: Of the 100 burn wound specimens, 40 (40.0%) showed positive cultures, yielding a total of 48 bacterial isolates. The predominant pathogen was Pseudomonas aeruginosa (17; 35.4%), followed by Staphylococcus aureus (12; 25.0%), with methicillin resistance identified in 7 (58.3%) of the S. aureus isolates. Biofilm formation by the TCP method was detected in 38 (79.2%) isolates. Multidrug resistance (MDR) was observed in 34 (70.8%) isolates, of which 33 (97.1%) were biofilm producers, compared with 3 (21.4%) among non Multidrug Resistant Organisms (MDROs) (χ2 =30.25, p-value <0.001). Deep burns accounted for 25 (52.1%) culture-positive cases and superficial burns for 23 (47.9%), with biofilm positivity rates of 20 (80.0%) and 18 (78.3%), respectively. Conclusion: Pseudomonas aeruginosa and Staphylococcus aureus were the most common bacterial isolates from burn wound infections, with a notable prevalence of MDR strains. Biofilm production was frequently observed, particularly among Pseudomonas aeruginosa isolates, and showed a strong association with multidrug resistance. These findings highlight the importance of considering biofilm formation when managing burn wound infections, as it plays a critical role in antimicrobial resistance and may contribute to treatment failure.

Vitex negundo Linn.: A decade of advances in phytochemistry, pharmacological activities, and biotechnological interventions (2015-2025) - A comprehensive review.

Regulation of heat tolerance by iron chlorine E6 in “84 K” poplar tissue culture seedlings: A study coupled with metabolomic and transcriptomic analysis

Meta-topolin riboside (mTR) has shown superior effects over benzyladenine (BA) in promoting root and shoot development in passion fruit tissue culture, yet its underlying mechanisms remain unclear. Understanding the metabolic differences between mTR and BA treatments can inform optimized propagation strategies for high-quality planting material in this economically important crop. Nodal buds of Passiflora edulis "Tainung No. 1" were cultured on BA- or mTR-supplemented medium, then analyzed using ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) in SWATH acquisition mode. Metabolite features were extracted, statistically filtered (VIP > 1, FC > 1.5 or < 0.667, p < 0.05), identified via multiple databases, and subjected to enrichment and pathway analysis. SWATH-MS detected 2823 ions in positive and 1637 in negative mode, with 21 significant metabolites identified in each mode. mTR treatment upregulated metabolites linked to root development (e.g., 5,10-methylenetetrahydrofolate, daidzin, and hesperidin) and stem elongation (amygdalin), while BA treatment had higher levels of kinetin, gibberellin A4, and lignans. Pathway analysis highlighted folate metabolism as significantly enriched in mTR samples. mTR treatment altered phytohormone, flavonoid and phenolic profiles in ways that likely promote rooting, shoot elongation and oxidative stress resilience, explaining its superior growth performance over BA. These insights can guide refined cytokinin use in micropropagation and broader applications in horticultural biotechnology.

Tissue cultivation conditions and transcriptome analysis of Tremella fuciformis primordia for industrial production purposes.

Currently, there is limited knowledge on the impact of immunity to hemagglutinin (HA) and/or neuraminidase (NA) on the transmission of influenza viruses. Therefore, using intramuscular vaccination, intranasal vaccination, or infection with reassortant viruses, we induced immunity to each antigen alone or both antigens combined in ferrets. We then assessed transmission of the 2009 pandemic H1N1 virus from these ferrets to naïve respiratory contacts. For all strategies used to induce immunity, combined immunity to HA and NA resulted in the largest reductions in transmission. Moreover, immunity to HA and NA conferred additive rather than synergistic reductions in transmission. No escape variants emerged in our transmission studies, and logistical regression showed that the probability of transmission was less than 50% when viral titers in donors were reduced to 101.5 and 102 median tissue culture infectious dose per ml on days 1 and 3 postinfection, respectively. These studies define the relationship between immunity to HA and NA on transmission and identify a threshold titer indicative of decreased transmission in ferrets.

Virus-induced genome editing: toward crop breeding applications.

Background: Primary cell culture closely matches the physiological and biochemical features of an in vivo system and it serves as a representative model for studying many key issues of disease, reproduction, genetics and biotechnology. The present study aimed to develop a primary culture system from the Gill, Liver and Kidney tissues of Osteobrama belangeri, which is an important candidate species for aquaculture. Methods: The primary culture was established from tissues through the explant method. The cultured cells were maintained in Leibovitz’s L-15 Medium supplemented with FBS (Fetal Bovine Serum) and Antibiotic Antimycotic Solution 100X Liquid. The growth optimization of cultured cells was performed at different incubation temperatures (25°C, 28°C, 30°C) and FBS concentrations (5%, 10% and 15%). Result: The maximum growth rate of cultured gill cells was recorded at 30°C temperature and 15% FBS. The cultured cells were characterized for species authentication using DNA barcoding. The sequence analysis reveals and confirms the species of origin. This primary culture of different tissues provides a foundational in vitro platform for conservation genetics, disease diagnostics and biotechnological applications in O. belangeri.

To select and breed superior varieties of Dendrobium officinale with high quality and strong resistance to adverse conditions, a systematic selection process was employed to screen for outstanding strains, complemented by tissue culture for seed propagation. Following screening and self-purification, a new variety, ‘Tiefeng No.1’, was developed. Between 2019 and 2023, a comprehensive assessment of its characteristics, regional product ratio tests, and productivity research was meticulously conducted. The results indicated that the new variety of Dendrobium officinale is stable and of excellent quality. The polysaccharide content ranged from 44.35% to 58.55%, and the mannose content varied from 14.03% to 22.38%, both of which meet the standards set by the Chinese Pharmacopoeia (2020). The anthocyanidin content was measured at 94.76 to 115.43 μg/g, which is double that of the Yueqing landraces. It exhibits good frost resistance and disease resistance. Through production verification and demonstration, ‘Tiefeng No.1’ has proven to possess excellent quality and high yield, showcasing significant potential for promotion in the primary growing regions.

Goatpox is a severe viral disease in goats, and vaccination with live attenuated strains remains the most effective strategy for control. Conventional vaccine production relies on primary or Vero cells, but these substrates are limited by unstable supply, high cost, and suboptimal virus yields. This study aimed to establish a scalable suspension culture system using immortalized lamb testis (iLT) cells to support efficient replication of the goatpox virus (GTPV) AV41 strain for vaccine production. Adherent iLT cells were adapted to suspension by stepwise passaging under reduced serum. Eight commercial suspension media were screened, and four were selected for growth evaluation in 125-mL shaker flasks (20-mL working volume; 1 × 106 cells/mL), with stability assessed up to the 40th passage. For virus production, suspension cultures at 2-3 × 106 cells/mL were inoculated with GTPV-AV41 at 5-10% (v/v), samples were collected at 72-144 h post-inoculation, and viral replication was quantified by real-time quantitative PCR (RT-qPCR) using a cycle threshold (Ct) -50% tissue culture infectious dose (TCID50) calibration curve, with infection confirmed by indirect immunofluorescence. Suspension-adapted iLT cells maintained robust growth for over 40 continuous passages, reaching densities exceeding 1.2 × 107 cells/mL with >95% viability.Following GTPV-AV41 inoculation, the suspension culture system supported efficient viral replication, yielding a maximum viral titer of 108.00TCID50 equivalents/mL in cell lysates at 72 h post-inoculation. Efficient viral entry and replication were further confirmed by indirect immunofluorescence assays. Suspension-cultured iLT cells represent a promising platform for scalable goatpox vaccine production, combining high growth performance with efficient viral replication. This study presents a scalable suspension-culture platform by adapting iLT cells under serum-reduced conditions, enabling stable high-density growth and efficient replication of goatpox virus for cost-effective vaccine manufacturing.

Acute and chronic wounds are a major clinical burden, with persistent inflammation, impaired fibroblast function, defective angiogenesis, and disordered extracellular matrix deposition. The translational potential of existing in vitro models is limited by their poor durability and physiological relevance. The present paper aims to develop a robust in vitro organotypic model to simulate the early phases of both acute and chronic wounds and to validate it by testing the biocompatibility of clinically available wound dressings. Human fibroblasts and vascular endothelial cell lines were cultured at a ratio of 1:1 for 48 h, either on uncoated tissue culture plastic or on tissue culture plastic coated with a synthetic substrate (PhenoDrive-Y) that biomimics the extracellular matrix and promotes cell organization into tissue-like structures on a 2D plane (i.e., angiogenesis sprouting and fibroblast organization around it). Wound conditions were then created by damaging the formed structures using a conventional scratch procedure and introducing U937 human macrophage cells to the model to simulate either the onset of an acute wound or that of a chronic wound through the simultaneous spiking of the culture with relevant cytokines, i.e., IL-6 and TNF-α. The formation of new tissue-like structures in the scratch area was quantified by the extent of scratch closure after a further 24 h of incubation. Morphological analysis of wound healing was performed by light microscopy, while angiogenesis was assessed by CD31 immunostaining by confocal microscopy. The deposition of components of the extracellular matrix was determined both qualitatively and quantitatively by Picrosirius Red staining for collagen production and by Alcian Blue staining for glycosoaminoglycan synthesis on the adhering cells and their supernatants. Macrophage polarization into either M1 or M2 phenotype was studied by immunostaining with iNOS (M1) and CD206 (M2) antibodies by confocal microscopy. The model was validated by studying the gap closure areas in simulated acute and chronic wound-like conditions when incubated with clinically available wound dressings, N-A Ultra and Kaltostat. PhenoDrive-Y allowed the formation of tissue-like structures on the 2D tissue culture plane as opposed to the formation of cell monolayers on the uncoated tissue culture plastic. Upon mechanical damage, cell migration was significantly different; uncoated control co-cultures achieved complete closure as an indistinct monolayer by 24 h, while the organotypic wound models showed a slower percentage of damage closure. A further delay in the closure of the damaged area was observed when chronic wound-like conditions were simulated. Angiogenesis in chronic wound conditions was considerably impaired compared to the acute conditions. The analysis of the extracellular matrix component synthesis, specifically collagen and polysaccharides, revealed the deposition of dense, organized collagen fibers in the acute wound model, in contrast to the thin, fragmented collagen fibers and intracellular polysaccharides observed under chronic wound-like conditions. This corresponded to a statistically significant increase in the levels of both collagen and polysaccharides detected as soluble molecules in the supernatants. Macrophage polarization showed no statistically significant differences in the acute and chronic wound models, though iNOS did significantly decrease after N-A application in acute and chronic models. However, acute wound-like conditions showed a restoration of the vascularized tissue-like structures after treatment with these types of dressings, albeit through different organizational pathways, whereas only minimal improvement was noted under chronic wound conditions, particularly in the case of the N-A dressing. The organotypic dermis model for the onsets of acute and chronic wounds emerges as a highly versatile tool to understand healing mechanisms in the absence or presence of co-morbidities and to assess the biocompatibility of wound dressings as well as the safety, efficacy and dosage of drugs.

Mesenchymal stem/stromal cells (MSCs) are known to secrete wound healing factors. However, the delivery of the MSCs and their secretomes remains a barrier to the development of a successful regenerative medicine. Therefore, we examined the content and paracrine activity of serum free MSC conditioned media (MSC CM) generated in routine tissue culture and when cultured on a plasma polymerized membrane previously used clinically in cell therapies for skin wounds. Serum free MSC CM from murine and human MSC cultures were subjected to proteomic analysis by mass spectrometry and quantitative immunoassays and their paracrine effects were tested on dermal fibroblasts in vitro. MSC CM contained multiple wound healing factors, including extracellular matrix proteins and soluble factors. MSC CM harvested from flask cultures and plasma polymerized membrane cultures significantly increased dermal fibroblast adhesion, proliferation, and scratch wound closure (for mouse MSC CM) compared to control media. This study has: (i) shown that MSC secreted factors increased dermal fibroblast activities seen in wound healing; (ii) identified target factors potentially responsible for these paracrine effects; (iii) shown that MSC secretomes generated on a plasma polymerized membrane used in skin cell therapies also have wound healing paracrine effects.

Background: Cardava banana (Musa spp.) production in the Philippines is constrained by slow multiplication rates and disease transmission associated with traditional sucker propagation, as well as the high cost and limited accessibility of tissue culture. Macropropagation offers a low-cost alternative, particularly when combined with locally available farm waste substrates. This study evaluated the effects of five farm waste-based media on shoot emergence and early growth of macropropagated and traditionally propagated Cardava bananas under greenhouse conditions, assessed field growth and yield performance and examined the educational outcomes of integrating macropropagation activities into science instruction. Method: A factorial experiment in a Completely Randomized Design compared soil (control), soil amended with coconut husk, compost, mung bean pod hull and rice hull. Field trials compared the growth and yield of macropropagated and traditionally propagated plants. Educational outcomes were evaluated using a quasi-experimental pretest–posttest design involving hands-on propagation activities. Results: Macropropagated plants exhibited significantly faster shoot emergence and greater early growth than traditionally propagated plants across all media (p less than 0.001). Among substrates, soil amended with mung bean pod hulls produced the fastest emergence and greatest plant height, followed by compost- and coconut husk–amended soils, while rice hull–amended soil performed the poorest. Field evaluation showed significantly higher yields in macropropagated plants, with an average increase of 7.10 kg per plant (p less than 0.001). Educational results indicated substantial improvements in posttest scores, student engagement, interest in agriculture and knowledge retention compared with theory-based instruction. The findings demonstrate that macropropagation supported by farm waste-based substrates provides a flexible and sustainable approach to improving Cardava banana production while enhancing experiential learning in agricultural education.

Molecular characterization of non-polio enterovirus in children with acute flaccid paralysis in Iraq.

Background: Tuberculous pleuritis (TBP) presents a diagnostic challenge due to its paucibacillary nature. The gold standard for diagnosing TBP requires the identification of Mycobacterium tuberculosis in tissue culture and evidence of granulomatous inflammation in pleural specimens. We aimed to determine the diagnostic yield and accuracy of medical thoracoscopy, as well as its utility in early diagnosis and treatment in a low-prevalence region. Methods: This is a retrospective study of patients with suspected TBP between 2020 and 2024 in a large hospital system in New York City. A subgroup of patients underwent medical thoracoscopy with pleural biopsy for histopathological and microbiological evaluation, and their results were compared to those of a cohort initiated on RIPE treatment without pleural tissue analysis. Results: Of 129 patients included, medical thoracoscopy was performed in 46 cases, with 31 (67.4%) diagnosed with TBP, 4 (8.7%) with nontuberculous pleuritis, and 11 (23.9%) with chronic idiopathic pleuritis. The diagnostic yield for tuberculosis was 67.4% in a low-prevalent region, with an overall accuracy of 89.1%. Out of 83 patients who were initiated on RIPE therapy based on clinical suspicion of TBP, 18 (21.7%) had no microbiological evidence of tuberculosis. The time to RIPE initiation was shorter in the medical thoracoscopy group (3.0 [IQR, 1.0–5.0] days vs. 5.0 [IQR, 3.0–13.0] days, P = 0.006). Conclusions: Medical thoracoscopy offers high accuracy and a diagnostic yield in a low-prevalent region, facilitating early diagnosis and leading to more timely and appropriate treatment initiation. Diagnosis of TBP in areas of low prevalence may help reduce drug resistance and inappropriate treatment regimens.

Rapid genetic transformation of herbaceous peony without tissue culture via Agrobacterium rhizogenes: Optimization using rhizomes, stems, roots, and seedlings.

The decline of proteostasis is a central hallmark of aging, the earliest manifestations of which have remained difficult to capture in human tissues with conventional model systems. The skin is a continuously renewing and environmentally exposed organ offering a uniquely accessible window into aging biology. Skin organoid technologies allow for long-term culturing of human epidermal and full-thickness skin-like tissues that accurately recapitulate important aspects of cellular heterogeneity, spatial organization, and stem cell dynamics. In this perspective, we discuss how skin organoids are beginning to reveal early proteostasis alterations-encompassing impaired protein folding, reduced proteasomal activity, and autophagy dysfunction-that precede overt structural and functional hallmarks of skin aging, with particular emphasis on underexplored regulators- sebaceous gland and sebocyte-specific proteostasis, autophagy, and inflammaging. We also highlight emerging insights, conceptual challenges, and experimental limitations, and outline future directions for integrating skin organoids with skin-on-a-chip, single-cell proteomics, and stress-reporting approaches to advance proteostasis-targeted interventions in skin aging.

Spinal cord reactive-antibodies identified by serological antigen selection show prognostic value in traumatic spinal cord injury patients.

Extended invitro culture (EIVC) of porcine embryos beyond Day 7 is critical for understanding peri-implantation development but remains technically challenging due to the rapid degeneration of blastocysts. While defined serum-free supplements, such as B27 and growth factor cocktails (FGF2, LIF, IGF1; FLI), have successfully supported EIVC in bovine and murine models, their efficacy in porcine embryos has not been systematically evaluated against fetal bovine serum (FBS). This study investigated the effects of these supplements on the long-term viability and molecular integrity of porcine blastocysts. Porcine blastocysts produced invitro were cultured from Day 7 to Day 11 in Porcine Zygote Medium (PZM) supplemented with either 10% FBS, FLI cocktail, B27 or no supplement (Control). Contrary to findings in other species, supplementation with FLI or B27 failed to support porcine blastocyst survival, resulting in complete degeneration by Day 11, similar to the control group. In contrast, FBS supplementation significantly extended blastocyst viability and maintained structural integrity up to Day 11. Although blastocyst expansion plateaued after Day 9, the FBS group retained a significantly higher total cell number and preserved the expression of pluripotency (OCT4, SOX2) and lineage (CDX2, GATA6) markers compared to other groups. Furthermore, FBS significantly suppressed apoptosis, as evidenced by a lower BAX/BCL2 ratio. These findings demonstrate a distinct species-specific requirement for porcine EIVC; defined supplements effective in other mammals are insufficient for pigs, suggesting that while FBS is essential for preventing rapid degeneration and maintaining cellular viability, further optimisation is required to fully support extended developmental progression comparable to invivo peri-implantation elongation.