Search Results (230)

Purpose: The adenoid microbiota plays a key role in adenoid hypertrophy (AH). This study explored the molecular epidemiology and antimicrobial resistance of Haemophilus. Influenzae (H. influenzae) strains in pediatric AH patients. Methods: Retrospective analysis of pediatric AH patients undergoing endoscopic adenoidectomy. Adenoid tissue samples were cultured to screen for pathogens. H. influenzae strains were identified by 16S rRNA sequencing and serotyped via q-PCR. Multilocus sequence typing (MLST) and ftsI gene analysis were conducted using PubMLST. β-lactamase genes (bla_TEM-1, bla_ROB-1) were detected by PCR, and antibiotic susceptibility testing (AST) was performed using the Etest method. For imipenem-resistant strains, the acrRAB efflux pump gene cluster and ompP2 porin gene were sequenced and compared with those of the wild-type strain Rd KW20. Results: Over 8 months, 56 non-duplicate H. influenzae strains were isolated from 386 patients. The detection rate was highest in children under 5 years (30.5%) compared to those aged 5–10 years (13.4%) and 10–15 years (8.7%). Of 49 sub-cultured strains, all were non-typeable H. influenzae (NTHi). MLST identified 22 sequence types (STs) and 13 clonal complexes (CCs), with CC11 (26.5%), CC3 (14.3%), and CC107 (14.3%) being predominant. Common STs included ST103 (22.4%), ST57 (10.2%), and ST107 (10.2%). Most strains belonged to the ftsI group III-like+ (57.1%). β-lactamase positivity was 98.0% (48/49), with bla_TEM-1 (95.9%) and bla_ROB-1 (18.4%) detected. AST showed low susceptibility to ampicillin (10.2%), amoxicillin–clavulanate (34.7%), azithromycin (12.2%), and trimethoprim–sulfamethoxazole (14.3%). Among the β-lactamase-positive strains, 44/48 were β-lactamase-positive ampicillin-resistant (BLPAR); none were β-lactamase-negative ampicillin-resistant (BLNAR). Imipenem susceptibility was 91.8% (45/49). No carbapenemases were found in the imipenem-resistant strains, but mutations in acrRAB (88.12–94.94% identity) and ompP2 (77.10–82.94% identity) were observed. Conclusions: BLPAR NTHi strains of CC11 are major epidemic strains in pediatric AH. Imipenem resistance in H. influenzae likely results from porin mutations rather than carbapenemase activity. Enhanced surveillance of H. influenzae’s role in AH and its resistance patterns is warranted. Full article

Multiple myeloma is a hematologic malignancy characterized by the clonal proliferation of plasma cells within the bone marrow. The tumor microenvironment plays a crucial role in multiple myeloma pathogenesis, progression, and drug resistance. Traditional two-dimensional cell culture models have been instrumental in multiple myeloma research. However, they fail to recapitulate the complex in vivo bone marrow microenvironment, leading to limited predictive value for clinical outcomes. Three-dimensional cell culture models emerged as more physiologically relevant systems, offering enhanced insights into multiple myeloma biology. Scaffold-based systems (e.g., hydrogels, collagen, and Matrigel), scaffold-free spheroids, and bioprinted models have been developed to simulate the bone marrow microenvironment, incorporating key components like mesenchymal stromal cells, osteoblasts, endothelial cells, and immune cells. These models enable the functional assessment of cell adhesion-mediated drug resistance, cytokine signaling networks, and hypoxia-induced adaptations, which are often lost in 2D cultures. Moreover, 3D platforms demonstrated improved predictive value in preclinical drug screening, facilitating the evaluation of novel agents and combination therapies in a setting that better mimics the in vivo tumor context. Hence, 3D cultures represent a pivotal step toward bridging the gap between basic myeloma research and translational applications, supporting the development of more effective and patient-specific therapies. Full article

Campylobacter jejuni and Campylobacter coli are the two main campylobacter species that cause foodborne campylobacteriosis. Recent studies have reported that Campylobacter spp. are prone to developing resistance to antibiotics commonly used for their treatment, with many C. coli strains identified as multidrug-resistant. This study presents the results of the whole-genome sequencing analysis of the multidrug-resistant C. coli strain BCT3 isolated in Greece from a stool specimen of a pediatric patient presenting with diarrhea. The strain was isolated using selective culture media and, based on antimicrobial susceptibility tests, was found to be resistant to ciprofloxacin, tetracycline, erythromycin, azithromycin, clarithromycin, and doxycycline. To further characterize it, we performed whole-genome sequencing, which identified strain BCT3 as C. coli. Moreover, multilocus sequence typing assigned the BCT3 to the sequence type (ST) 872, belonging to clonal complex ST-828. The presence of multiple virulence genes revealed its pathogenic potential. The detection of antimicrobial resistance genes and mutated alleles was indicative of its resistance to fluoroquinolones, macrolides, and tetracyclines, supporting the observed phenotype. To our knowledge, this is the first reported clinical case of such a multidrug-resistant C. coli strain in Greece. Full article

Background: During the COVID-19 pandemic, the emergence of multidrug-resistant (MDR) pathogens, driven by heightened antibiotic usage and device-associated infections, has posed significant challenges to healthcare. This study reports an outbreak of Proteus mirabilis producing NDM-5 and CTX-M-15 β-lactamases in a hospital in Buenos Aires, Argentina, from October 2020 to April 2021. To our knowledge, this represents the first documented outbreak of NDM-5-producing P. mirabilis in the country. Methods: A total of 82 isolates were recovered from 40 patients, with 41.5% from blood cultures and 18.3% from respiratory and urinary samples, among others. Antimicrobial susceptibility testing, PCR-based methods, and MALDI-TOF MS cluster analysis were conducted. Whole genome sequencing (WGS) was performed to characterize the MLST, resistome and plasmid content. Biofilm formation assays and in vitro rifampicin susceptibility tests were also conducted. Result: Most isolates exhibited resistance to carbapenems, cephalosporins, aminoglycosides, and fluoroquinolones, while retaining susceptibility to aztreonam. Genetic analysis confirmed the co-presence of the bla_NDM-5 and bla_CTX-M-15 genes. Clonal relationships was supported by PCR-based typing and MALDI-TOF MS cluster analysis. WGS revealed a resistome comprising 25 resistance genes, including rmtB and both β-lactamases, as well as the presence of an incomplete IncQ1 replicon associated with multiple resistance determinants. MLST classified this clone as belonging to ST135. Despite the biofilm-forming capacity observed across strains, rifampicin demonstrated potential for disrupting established biofilms at concentrations ≥32 µg/mL in vitro. The MDR profile of the outbreak strain significantly limited therapeutic options. Conclusions: This study highlights the growing threat of NDM-producing P. mirabilis in Argentina. The absence of surveillance cultures from the index case limits insights into the outbreak’s origin. These findings underscore the importance of integrating genomic surveillance into infection control protocols to mitigate the spread of MDR pathogens. Full article

Callus induction is one of the most important techniques in plant-based industries. Important features in the use of callus induction are the maintenance of pluripotency and the proliferation of cells. Although the importance of callus induction is also understood in ginseng, there are no studies on the genetic modules associated with callus induction and growth regulation. Panax ginseng embryo tissue was wounded and cultured in callus-inducing media, and its time-course physiology was observed. Time-course callus samples were collected for total RNA extraction and RNA-Seq analysis using the Illumina HiSeq X Ten platform. P. ginseng embryo tissue was wounded and treated with varying amounts of gamma radiation in callus-inducing media, and samples were also collected for total RNA extraction and RNA-Seq analysis. A combinatory analysis of various network analyses was used to reveal the regulatory network underlying callus development. We were able to determine the time-course physiology of callus development and the dose-dependent effect of gamma radiation on callus development. Network analysis revealed two networks correlated with callus induction and two networks correlated with callus growth. Our research provides a regulatory network illustrating how callus is induced and growth is regulated in P. ginseng. This result would be helpful in the development of a cell culture system or clonal propagation protocol in P. ginseng. Full article

Re-engineering of CHO cells using genome editing and the overexpression of multiple helper genes is the central track for obtaining better cell lines for the production of biopharmaceuticals. Using two subsequent rounds of genome editing of the CHO S cells, we have developed the cell line CHO 4BGD with four knockouts of two pro-apoptotic genes bak1 and bax, and two common selection markers genes—glul (GS) and dhfr, and additional copies of genes bcl-2 and beclin-1 used for enhancement of macroautophagy. The NGS sequencing of 4BGD cells revealed that all eight targeted alleles were successfully disrupted. Two edited loci out of eight contained large inserts of non-relevant DNA. Further data analysis shows that cells have no off-target DNA editing events, and all known CHO genes are preserved. The cells obtained are completely resistant to the induction of apoptosis, and they are suitable for the generation of stably transfected cell lines with the dhfr selection marker. They also properly undergo the target gene amplification. The 4BGD-derived clonal cell line that secretes the monoclonal antibody retains the ability for prolonged fed-batch culturing. The method of obtaining multiply edited CHO cells using the multiplex CRISPR/Cas9 editing and simultaneous stable transfection of plasmids, coding for the housekeeping genes, is suitable for the rapid generation of massively edited CHO cells. Full article

Latin America offers a unique point of view into the adaptation of viticulture to climate change through its rich diversity of climates, traditional knowledge, and scientific innovation. This review synthesizes the current research and technological developments across major wine-producing countries including Argentina, Brazil, Chile, Uruguay, the Dominican Republic, and Haiti. Argentina shows key adaptation strategies, including high-altitude vineyard relocation, clonal and rootstock selection, canopy and water management, and the conservation of Criolla and other autochthonous grapevine varieties. In Brazil, tropical viticulture and breeding programs led by Embrapa exemplify advancements in disease-resistant and climate-resilient cultivars. Chile’s heroic and southern viticulture highlights the importance of old vines, microclimatic heterogeneity, and territorial identity. Uruguay stands out for its terroir-based research and producer-led adaptation strategies. This review also addresses systemic challenges in scientific publishing, particularly the underrepresentation of Latin American researchers in global vitivinicultural discourse. These disparities underscore the need for inclusive science that values local knowledge and promotes equity in research funding and dissemination. Overall, Latin America stands out not only as a region highly vulnerable to climate change, but as an emerging model of adaptation and innovation, demonstrating how resilient, sustainable, and culturally rooted wine production can thrive under shifting environmental conditions. Full article

Oil palm (Elaeis guineensis Jacq.) is the leading global oil-producing crop due to its high oil yield. Increasing global demands for palm oil require efficient propagation. Conventional breeding is practical but slow, making micropropagation an attractive alternative for rapidly multiplying superior genotypes. However, transitioning from in vitro to ex vitro conditions causes physiological stress, restricting survival and productivity. This study assessed gas exchange and chlorophyll fluorescence dynamics during acclimatization from in vitro conditions to field establishment, comparing the seedlings obtained in vitro with conventional seed-derived palm seedlings to conventional seed-derived palms. A pronounced photosynthetic efficiency decline occurred after transfer from in vitro culture, followed by a gradual recovery. The photosynthetic rate (A) increased from 0.86 µmol m⁻² s⁻¹ early in acclimatization to 15.43 µmol m⁻² s⁻¹ in field-established seedlings. Physiological characterization using CO₂ and light response curves identified the reductions in carboxylation efficiency and overall quantum yield CO₂. These biochemical constraints gradually diminished during acclimatization, facilitating a transition from heterotrophic to autotrophic growth. Chlorophyll fluorescence analysis revealed remarkable photoinhibition during initial ex vitro stages, indicated by a decreased maximum quantum efficiency of photosystem II. However, the seedlings progressively restored photochemical function throughout subsequent acclimatization phases. These findings highlight the importance of carefully regulating environmental parameters—particularly irradiance, humidity, and carbon availability—during early seedling acclimatization. The effective management of growth conditions significantly mitigates physiological stress, ensuring robust photosynthetic activity and optimized stomatal regulation. The improved acclimatization practices, therefore, can substantially enhance seedling survival rates, physiological resilience, and the overall field performance of micropropagated oil palms. Future research should focus on refining acclimatization protocols, emphasizing targeted physiological interventions to maximize the efficiency, commercial viability, and sustainability of oil palm clonal propagation. Full article

The production of influenza A virus (IAV) using Madin-Darby Canine Kidney (MDCK) cells is a key strategy for efficient influenza vaccine manufacturing. However, challenges remain in optimizing cell culture processes for higher yield and efficiency. This study aims to evaluate different process intensification strategies on two distinct clonal MDCK suspension cell lines (C59 and C113) for improved IAV production. A semi-perfusion strategy was used to push cells towards high cell density (HCD), achieving up to 17 × 10⁶ C113 cells/mL and 42 × 10⁶ C59 cells/mL, respectively. Next, a Tangential Flow Depth Filtration (TFDF)-based perfusion process with direct harvest during IAV production was established, resulting in high titers and a 10-fold higher space-time yield for C59 and a 4-fold improvement for C113 compared to batch operation. In addition, the suitability of N-1 perfusion was evaluated for batch and intensified fed-batch processes. Cells taken from the N-1 perfusion showed different cell-specific growth rates, but this had no effect on virus titers except for processes started from oxygen-deprived precultures. Finally, comparable virus titers were obtained when the production bioreactor was directly inoculated from an HCD cryovial. Taken together, seed train intensification and TFDF-based perfusion majorly reduced process times and improved IAV production. Full article

Clinical success in treating complicated urinary tract infections (cUTIs) depends on accurate pathogen detection, given the common occurrence of polymicrobial infections and antimicrobial resistance. This multicenter, randomized, investigator-blinded study compared polymerase chain reaction (PCR)-based diagnostics to conventional culture and sensitivity (C&S) testing in guiding the treatment of cUTIs. PCR identified polymicrobial infections in 43.52% of cases, a significantly higher rate than that observed with C&S (31.95%, p = 0.033). Patients in the C&S arms with undetected polymicrobial infections had a significantly higher clinical failure rate (33.33%, 14/42, p = 0.041) compared to those with concordant polymicrobial infection identification by both methods (22.22%, 12/54). PCR also detected additional pathogens in 54.44% (92/169) of cases in the C&S arm, where clinical failure was significantly higher when C&S missed pathogens (28.26% vs. 14.29%, p = 0.015). Similarly, when C&S failed to detect phenotypic resistance (compared to PCR), clinical failure occurred in 50% (16/42) of cases, compared to 13.22% (21/121, p = 0.001) when resistance detection was concordant (PCR and C&S). To further illustrate the clinical impact, patient-level case analyses are included to demonstrate how PCR-guided therapy improved pathogen detection and enabled more appropriate antimicrobial selection compared to C&S. These findings highlight the limitations of C&S in detecting polymicrobial infections, antimicrobial resistance, and hetero-resistance due to its limited clonal analysis, supporting the integration of PCR for more accurate diagnostics and optimized cUTI management. Full article

Globally, people are cultivating finger millet, an important cereal, to improve food availability and health benefits for humans. However, the biotechnological research on this millet is limited and insufficient in this field. The primary focus of this study is to optimize an efficient regenerated protocol for initiating further plant transformation studies, using the shoot apex as an explant and various growth regulators. For example, three cytokinins (BAP, TDZ, and Kin) at different concentrations were used to induce multiple shoots of finger millet. Among these, TDZ (4.5 µM) provided the maximum number (17.3) of shoots as compared to BAP and Kin. IBA (2.46 µM), along with MS medium, was used for the induction of roots, where 5.6 roots were produced in an individual shoot and the length of the root was longer with a size of 8.2 cm after two weeks of incubation. The clonal fidelity of the in vitro regenerated plantlets of finger millet was confirmed by ISSR primers. Overall, the present work developed a robust and reliable procedure for the establishment of efficient and reproducible regeneration through the shoot apex that will be useful for the genetic improvement of this crop. The genetic enhancement of these millets as well as the successful creation of transgenic plant varieties modified for resistance to biotic and abiotic challenges in the near future would be aided by this study. Full article

Garlic (Allium sativum L.) is the second most significant species within the Allium genus worldwide, widely used in cooking and both traditional and modern medicine due to its beneficial biological and therapeutic properties. In Italy, diverse pedo-climatic conditions and historical–cultural fragmentation have led to the development of various garlic landraces, prized for their unique organoleptic qualities and cultural importance. This study aimed to assess the intra-varietal diversity and uniqueness of two red garlic landraces from the Lazio region in central Italy, “Aglio Rosso di Castelliri” and “Aglio Rosso di Proceno”, using SSR and ISSR molecular markers, along with evaluations of bulb morphological traits, total phenolic content, and antioxidant properties. The molecular analysis included 11 accessions of “Aglio Rosso di Castelliri”, nine of “Aglio Rosso di Proceno”, and 15 control accessions, comprising eight Italian red-type garlic landraces, four Spanish red garlic commercial varieties, two white garlic accessions, and an accession of A. ampeloprasum var. holmense used as an outgroup. SSR and ISSR markers revealed moderate genetic diversity within the collection, with mean PIC values of 0.41 and 0.17, respectively. The molecular data identified four distinct genetic clusters, with the two Lazio landraces forming separate groups, indicating their genetic distinctiveness. The results from the STRUCTURE analysis support the hypothesis that these landraces may have originated from the widely cultivated “Aglio Rosso di Sulmona” or a common ancestral population once prevalent in central Italy. The study also revealed significant intra-population genetic diversity within the two garlic landraces, underscoring the need for in situ conservation and clonal selection. Phenotypic evaluations confirmed the distinctiveness of the two landraces, with “Aglio Rosso di Castelliri” characterized by smaller bulbs and cloves with higher dry matter content and distinct color profiles. Additionally, significant variation in total phenolic content and antioxidant activity was observed by analyzing 13 selected accessions from the two landraces (six from “Aglio Rosso di Proceno” and seven from “Aglio Rosso di Castelliri”) and five red garlic control accessions, with the two Lazio landraces exhibiting higher levels than the control group. This study highlights the importance of integrating molecular, phenotypic, and chemical analyses to understand garlic landrace diversity, with significant implications for their conservation and protection of local agro-food products. Full article

Escherichia albertii is a lactose-negative Escherichia that causes gastritis and enteritis in humans. An analysis of possible sources of infection points out that poultry may be a significant reservoir for this pathogen. The question of whether E. albertii can cause infections in poultry is still unanswered. Our article describes the isolation of E. albertii, for the first time in Russia, from the intestines of birds on a quail farm and a characterization of obtained cultures. We isolated different bacteria from pathological poultry material using bacteriological methods and ruled them out as probable causes for enteritis. The biochemical identification of E. albertii and antibiotic sensitivity were performed using a Vitek-2 Compact instrument. Bacterial identification was carried out using the MALDI-TOF Biotyper instrument. E. albertii-specific genes, virulence factor genes, and microcin genes were detected by real-time PCR. It was concluded that E. albertii isolated from sites of intestinal inflammation are a potential cause of enteritis and high poultry mortality—up to 15% of total livestock for 10- to 20-day-old quails. One of the E. albertii culture differed from the main group of Escherichia by its biochemical properties, and subsequent PCR analysis showed a lack of the intimin gene (eae). We describe the first occasion of infection caused by E. albertii in industrial quails. During the study, it was found that, according to the molecular–genetic and phenotypic properties of isolated strains in quails, there were at least two clonal groups of E. albertii differing in antibiotic resistance, biochemical indices, and presence of the eae (intimin) gene. Full article

For the first time, adventitious and hairy root cultures of Salvia apiana (white sage) have been established and analyzed for the content of secondary metabolites. Non-transformed roots derived from sterile seedlings were maintained on a full-strength IBA-supplemented SH medium. Adventitious roots yielded up to 44.5 mg/g and 18.7 mg/g DW rosmarinic acid when grown in shake flasks and immersion-column bioreactors, respectively. Transformed root cultures were established from S. apiana microshoots, infected with A4 and LBA9402 strains of Rhizobium rhizogenes. The obtained hairy root cultures (three and two clonal lines established using A4 and LBA9402 strains, respectively) were maintained in the PGR-free, full-strength SH medium. The most productive root line, established using A4 strain, accumulated rosmarinic acid at 38.1 and 39.6 mg/g DW when grown in shake flasks and spray bioreactors, respectively. Neither adventitious nor transformed roots of S. apiana produced diterpenoids, identified in roots of the field-grown plants, and instead proved to be a selective source of rosmarinic acid. Full article

In vitro shoot culture and cryopreservation (CP) are techniques essential for the ex situ preservation of genetic resources and the production of plant propagation material of clonally propagated horticultural crops. Changes in plant-associated microbiota diversity and composition induced by in vitro cultivation and CP treatment could have a negative effect on the growth and ex vitro adaptation of the in vitro propagated shoots. Therefore, the aim of the present study was to assess changes in endophytic bacteria diversity in domestic apple tissues induced by in vitro cultivation and CP treatment and to investigate the potential of the bacterial inoculum to improve the rooting and ex vitro acclimatisation of the propagated shoots. Metataxonomic analysis revealed a variation in the endophytic bacteria diversity and taxonomic composition between the field-grown tree dormant bud and the in vitro propagated or CP-treated shoot samples of apple cv. Gala. Whereas Sphingobacteriaceae, Sphingomonadaceae, Pseudomonadaceae, and Beijerinckiaceae families were the most prevalent families in the bud samples, Enterobacteriaceae, Bacillaceae, and Lactobacillaceae were dominant in the in vitro shoots. The bacterial inoculum effect on rooting and ex vitro acclimatisation was assessed using four isolates selected by screening the endophytic isolate collection. Bacillus sp. L3.4, B. toyonensis Nt18, or a combined inoculum resulted in a 21%, 36%, and 59% increase in cumulative root length and a 41%, 46%, and 35% increase in the biomass accumulation of ex vitro acclimatised plantlets, respectively. Root zone microbiota functional diversity analysis implied that growth stimulation was not related to improved nutrient uptake but could involve a pathogen-suppressing effect. The results demonstrate that the application of plant growth-promoting bacteria can potentially improve the performance of the in vitro propagated germplasm. Full article