Anne Sofie Frølunde, Jan Brink Valentin, Lise Kristensen
et al.
Dermatophyte infections are common in general practice and occur more often in individuals with type 2 diabetes (T2D), but whether they signal undiagnosed T2D remains unclear. We conducted a register-based cohort study including positive PCR tests for dermatophyte infection from the feet or nails, matched 1:3 to individuals from the same geographic area in Denmark. Those with known diabetes, type 1 diabetes or aged under 20 were excluded. Incidence rates (IRs) and incidence rate ratios (IRRs) for new-onset T2D were estimated using Poisson regression. The final cohort comprised 78,140 individuals, with a median age of 51 years, and 60.8% were male. The IR for T2D was 9.23 per 100 person-years in the exposed group and 9.00 in the unexposed group, with an adjusted IRR of 1.00 (0.91–1.11, p=0.94), indicating no significant association. In a sensitivity analysis excluding unexposed individuals with prior topical antifungal treatment, the IRR increased to 1.15 (1.08–1.23, p=0.001). While the primary analysis showed no significant association, the sensitivity analysis suggested a modest increased risk when exposure misclassification was reduced, supporting dermatophyte infection as a possible early signal of undiagnosed T2D in selected populations.
ABSTRACT Flavobacteriaceae is the major participant in the degradation of algal polysaccharides. With diverse polysaccharide utilization loci (PULs) and specific carbohydrate-active enzymes (CAZymes), Flavobacteriaceae strains appear to have different abilities in algal polysaccharide degradation and therefore change their roles in the bacterial community. Here, we identified two novel isolates as two novel species of genus Flagellimonas with the names Flagellimonas alginolytica sp. nov. and Flagellimonas cixiensis sp. nov. Furthermore, the comprehensive genomic comparison of 41 Flagellimonas genomes revealed that Flagellimonas strains were diverse in the CAZymes and PUL profiles and exhibited a preference for polysaccharides derived from brown algae. The evolutionary analysis of alginate utilization loci (AUL) in this genus illuminated that the function genes in AULs, that is, PL7 and PL17, were more reliant on the stable inheritance from ancestors associated with gene duplication and loss rather than horizontal gene transfer (HGT) from outside, and the AUL structures exhibited a trend of simplification which resulted in the incidental decrease in alginate degradation ability. This study highlights the important role of vertical inheritance in the evolution of AULs and proves that the discrepancy in AUL structure can arouse phenotypic differences, providing a new perspective on the evolution of AUL and the niche adaptation mechanism of Flavobacteriaceae strains.IMPORTANCEFlavobacteriaceae play an important role in the marine carbon cycle with their noteworthy ability in algal polysaccharides degradation, which is primarily reliant on diverse polysaccharide utilization loci (PULs). Our study highlights the crucial role of vertical inheritance in the evolution of alginate utilization loci (AUL) in Flagellimonas strains and reveals the AUL structural simplification found in Flagellimonas strains that will lead to the reduction of alginate degradation ability. These insights advance understanding of niche adaptation strategy and related evolutionary mechanisms of Flavobacteriaceae strains.
IntroductionPantoea ananatis has emerged as a significant plant pathogen affecting various crops worldwide, causing substantial economic losses. Bacteriophages and their endolysins offer promising alternatives for controlling bacterial infections, addressing the growing concerns of antibiotic resistance.MethodsThis study isolated and characterized the Pantoea phage PA1 and investigated the role of PA1-LRP in directly damaging bacteria and assisting endolysin PA1-Lys in cell lysis, comparing its effect to exogenous transmembrane domains following the identification and analysis of the PA1-Lys and the PA1-LRP based on whole genome analysis of phage PA1. Additionally, this study also explored how hydrophobic region of PA1-LRP (HPP) contributes to bacterial killing when combined with PA1-Lys and examined the stability and lytic spectrum of PA1-Lys under various conditions.Results and discussionPhage PA1 belonging to the Chaseviridae family exhibited a broad host range against P. ananatis strains, with a latent period of 40 minutes and a burst size of 17.17 phages per infected cell. PA1-Lys remained stable at pH 6-10 and temperatures of 20-50°C and showed lytic activity against various Gram-negative bacteria, while PA1-Lys alone could not directly lyse bacteria, its lytic activity was enhanced in the presence of EDTA. Surprisingly, PA1-LRP inhibited bacterial growth when expressed alone. After 24 h of incubation, the OD600 value of pET28a-LRP decreased by 0.164 compared to pET28a. Furthermore, the lytic effect of co-expressed PA1-LRP and PA1-Lys was significantly stronger than each separately. After 24 h of incubation, compared to pET28a-LRP, the OD600 value of pET28a-Lys-LRP decreased by 0.444, while the OD420 value increased by 3.121. Live/dead cell staining, and flow cytometry experiments showed that the fusion expression of PA1-LRP and PA1-Lys resulted in 41.29% cell death, with bacterial morphology changing from rod-shaped to filamentous. Notably, PA1-LRP provided stronger support for endolysin-mediated cell lysis than exogenous transmembrane domains. Additionally, our results demonstrated that the HPP fused with PA1-Lys, led to 40.60% cell death, with bacteria changing from rod-shaped to spherical and exhibiting vacuolation. Taken together, this study provides insights into the lysis mechanisms of Pantoea phages and identifies a novel lysis-related protein, PA1-LRP, which could have potential applications in phage therapy and bacterial disease control.
Abstract Background The β-galactosidase from Paenibacillus wynnii (β-gal-Pw) is a promising candidate for lactose hydrolysis in milk and dairy products, as it has a higher affinity for the substrate lactose (low K M value) compared to industrially used β-galactosidases and is not inhibited by the hydrolysis-generated product D-galactose. However, β-gal-Pw must firstly be produced cost-effectively for any potential industrial application. Accordingly, the yeast Komagataella phaffii was chosen to investigate its feasibility to recombinantly produce β-gal-Pw since it is approved for the regulated production of food enzymes. The aim of this study was to find the most suitable way to produce the β-gal-Pw in K. phaffii either extracellularly or intracellularly. Results Firstly, 11 different signal peptides were tested for extracellular production of β-gal-Pw by K. phaffii under the control of the constitutive GAP promoter. None of the signal peptides resulted in a secretion of β-gal-Pw, indicating problems within the secretory pathway of this enzyme. Therefore, intracellular β-gal-Pw production was investigated using the GAP or methanol-inducible AOX1 promoter. A four-fold higher volumetric β-galactosidase activity of 7537 ± 66 µkat oNPGal/Lculture was achieved by the K. phaffii clone 27 using the AOX1 promoter in fed-batch bioreactor cultivations, compared to the clone 5 using the GAP promoter. However, a two-fold higher specific productivity of 3.14 ± 0.05 µkat oNPGal/gDCW/h was achieved when using the GAP promoter for β-gal-Pw production compared to the AOX1 promoter. After partial purification, a β-gal-Pw enzyme preparation with a total β-galactosidase activity of 3082 ± 98 µkat oNPGal was obtained from 1 L of recombinant K. phaffii culture (using AOX1 promoter). Conclusion This study showed that the β-gal-Pw was produced intracellularly by K. phaffii, but the secretion was not achieved with the signal peptides chosen. Nevertheless, a straightforward approach to improve the intracellular β-gal-Pw production with K. phaffii by using either the GAP or AOX1 promoter in bioreactor cultivations was demonstrated, offering insights into alternative production methods for this enzyme.
Evgeny V. Mavrodiev, Andrey K. Sytin, Alexey P. Laktionov
et al.
The necessity of the typification of the <i>Apocynum</i> L. s.l. (incl. <i>Poacynum</i> Baill. and <i>Trachomitum</i> Woodson) (Apocynaceae) species is crucial for future dogbane diversity estimations. Still, the original material of many taxa of the genus <i>Apocynum</i> s.l. is doubtful or remains to be discovered. This study’s topic is resolving a long-term taxonomic homonymy around the widely used binomial “<i>A. sibiricum</i>”. The misusing of the name “<i>A. sibiricum</i>” became a reason for long-term misunderstanding of the meaning of the binomial <i>Poacynum sarmatiense</i> (Woodson) Mavrodiev, Laktionov & Yu.E.Alexeev (<i>A. sarmatiense</i> (Woodson) Wissjul.; <i>T. sarmatiense</i> Woodson) and therefore for the misestimation of the <i>Apocynum</i> s.l. diversity in Russian and other Eurasian floras. Resolving this issue, here, we designate the lectotype of <i>A. sibiricum</i> Jacq. and the name “<i>A. sibiricum</i> Pall.” was validated within <i>Poacynum</i> as <i>P. pallasianum</i> Mavrodiev, Sytin, Laktionov & Vasjukov nom. nov. (<i>Apocynum sibiricum</i> auct., non Jacq.) with the lectotype selected from the original collections of Peter Simon Pallas.
The paper presents a clinical and morphological justification for the use of glycyrrhizic acid in poultry farming. It was found that the use of glycyrrhizic acid in broiler chickens enhances the histological picture of the thymus with a pronounced image and differentiation of the cortical layer. Increased mitotic activity of thymocytes and improved blood supply to the organ were determined. Revealed, Fabricius’s bag of broiler chickens, which were given glycyrrhizic acid, has a fairly high level of functional dynamics. Installed, the thickness of the cortical layer of the thymus lobule in broiler chickens in the control group was 1.2 times lower than after the use of glycyrrhizic acid in broiler chickens in the experimental group, and the cerebral layer was 1.8 times less than in the experimental group. It was revealed that glycyrrhizic acid has a beneficial effect on the body of birds.
Manuka honey is known for its unique antibacterial activity, which is due to methylglyoxal (MGO). After establishing a suitable assay for measuring the bacteriostatic effect in a liquid culture with a time dependent and continuous measurement of the optical density, we were able to show that honey differs in its growth retardingeffect on <i>Bacillus subtilis</i> despite the same content of MGO, indicating the presence of potentially synergistic compounds. In model studies using artificial honey with varying amounts of MGO and 3-phenyllactic acid (3-PLA), it was shown that 3-PLA in concentrations above 500 mg/kg enhances the bacteriostatic effect of the model honeys containing 250 mg/kg MGO or more. It has been shown that the effect correlates with the contents of 3-PLA and polyphenols in commercial manuka honey samples. Additionally, yet unknown substances further enhance the antibacterial effect of MGO in manuka honey. The results contribute to the understanding of the antibacterial effect of MGO in honey.
Bernhard A. Huber, Guanliang Meng, Jiří Král
et al.
The southern South American genus Guaranita includes tiny spiders (body length ~1 mm) that lead reclusive lives under ground-objects and run rapidly when disturbed. As a result, they have been poorly collected and studied. Here we report on a recent collection of Guaranita spiders from Argentina, describing one new species (G. auadae Huber sp. nov.) and the previously unknown female of G. dobby Torres et al., 2016. In addition, we provide CO1 barcodes for all (now five) known species, first SEM data, and first chromosome data for the genus. The diploid number of Guaranita goloboffi Huber, 2000 (2n♂ = 11) is among the lowest in araneomorph spiders with monocentric chromosome structure.
Soil microbial diversity, composition, and function are sensitive to global change factors. It has been predicted that the temperature and precipitation will increase in northern China. Although many studies have been carried out to reveal how global change factors affect soil microbial biomass and composition in terrestrial ecosystems, it is still unexplored how soil microbial diversity and composition, especially in microbial functional genes, respond to increasing precipitation and warming in a semiarid grassland of northern China. A field experiment was established to simulate warming and increasing precipitation in a temperate semiarid grassland of the Horqin region. Soil bacterial (16S) and fungal (ITS1) diversity, composition, and functional genes were analyzed after two growing seasons. The result showed that warming exerted negative effects on soil microbial diversity, composition, and predicted functional genes associated with carbon and nitrogen cycles. Increasing precipitation did not change soil microbial diversity, but it weakened the negative effects of simulated warming on soil microbial diversity. Bacterial and fungal diversities respond consistently to the global change scenario in semiarid sandy grassland, but the reasons were different for bacteria and fungi. The co-occurrence of warming and increasing precipitation will alleviate the negative effects of global change on biodiversity loss and ecosystem degradation under a predicted climate change scenario in a semiarid grassland. Our results provide evidence that soil microbial diversity, composition, and function changed under climate change conditions, and it will improve the predictive models of the ecological changes of temperate grassland in future climate change scenarios.
Priyanka Sharma, Ilaria Dando, Raffaele Strippoli
et al.
Autophagy is an evolutionary conserved physiological process with a fundamental role during development, differentiation, and survival of eukaryotic cells. On the other hand, autophagy dysregulation is observed in many pathological conditions, including cancer. In particular, tumor growth and progression are accompanied and promoted by increased autophagy that allows cancer cells to escape apoptosis and to proliferate also in harsh microenvironments. It is, therefore, clear that the impairment of the autophagic process may represent a valid strategy to inhibit or reduce cancer growth and progression. Among the plethora of molecular players controlling cancer growth, a group of small endogenous noncoding RNAs called microRNAs (miRNAs) has recently emerged. In fact, miRNAs can act as either oncogenes or oncosuppressors depending on their target genes. Moreover, among miRNAs, miRNA-34a has been connected with both tumor repression and autophagy regulation, and its expression is frequently lost in many cancers. Therefore, enforced expression of miRNA-34a in cancer cells may represent a valid strategy to reduce cancer growth. However, such strategy is limited by the fast biodegradation and short half-life of miRNA-34a and by the lack of an efficient intracellular delivery system. The following review describes the autophagic process and its role in cancer as well as the role of miRNAs in general and miRNA-34a in particular in regulating tumor growth by modulating autophagy. Finally, we describe the use of nanoparticles as a promising strategy to selectively deliver miRNA-34a to tumor cells for therapeutic and diagnostic purposes.
Luca Sgarra, Alessandro Santo Bortone, Maria Assunta Potenza
et al.
We explored the significance of the L-Arginine/asymmetric dimethylarginine (L-Arg/ADMA) ratio as a biomarker of endothelial dysfunction in stroke patients. To this aim, we evaluated the correlation, in terms of severity, between the degree of endothelial dysfunction (by L-Arg/ADMA ratio), the methylene tetrahydrofolate reductase (MTHFR) genotype, and the interatrial septum (IAS) phenotype in subject with a history of stroke. <b>Methods and Results:</b> L-Arg, ADMA, and MTHFR genotypes were evaluated; the IAS phenotype was assessed by transesophageal echocardiography. Patients were grouped according to the severity of IAS defects and the residual enzymatic activity of MTHFR-mutated variants, and values of L-Arg/ADMA ratio were measured in each subgroup. Of 57 patients, 10 had a septum integrum (SI), 38 a patent foramen ovale (PFO), and 9 an ostium secundum (OS). The L-Arg/ADMA ratio differed across septum phenotypes (<i>p</i> ≤ 0.01), and was higher in SI than in PFO or OS patients (<i>p</i> ≤ 0.05, <i>p</i> ≤ 0.01, respectively). In the PFO subgroup a negative correlation was found between the L-Arg/ADMA ratio and PFO tunnel length/height ratio (<i>p</i> ≤ 0.05; r = − 0.37; R2 = 0.14). Interestingly, the L-Arg/ADMA ratio varied across MTHFR genotypes (<i>p</i> ≤ 0.0001) and was lower in subgroups carrying the most impaired enzyme with respect to patients carrying the conservative MTHFR (<i>p</i> ≤ 0.0001, <i>p</i> ≤ 0.05, respectively). Consistently, OS patients carried the most dysfunctional MTHFR genotypes, whereas SI patients the least ones. <b>Conclusions:</b> A low L-Arg/ADMA ratio correlates with impaired activity of MTHFR and with the jeopardized IAS phenotype along a severity spectrum encompassing OS, PFO with long/tight tunnel, PFO with short/large tunnel, and SI. This infers that genetic MTHFR defects may underlie endothelial dysfunction-related IAS abnormalities, and predispose to a cryptogenic stroke. Our findings emphasize the role of the L-Arg/ADMA ratio as a reliable marker of stroke susceptibility in carriers of IAS abnormalities, and suggest its potential use both as a diagnostic tool and as a decision aid for therapy.
Zyryanov S.K., Chenkurov M.S., Ivzhits M.A.
et al.
Objective.
To investigate the mortality rate, comorbidity prevalence, and etiology of community-acquired pneumonia (CAP) in elderly patient population.
Materials and Methods.
Hospitalized elderly patients with CAP were distributed into the following age groups: 65–74 years (group I), 75–84 years (group II) and 85–94 years (group III). The patients’ medical records were used for determining comorbidities and mortality rate. In order to determine etiology of CAP, sputum or BAL samples were collected. A total of 171 isolates were identified using MALDI-TOF MS.
Results.
The mortality rates were 27.1%, 31.5% and 45.7% in age groups I, II, III, respectively. The most common concomitant diseases in all age groups were arterial hypertension (47.4%, 54.6%, and 62.8% for groups I, II, and III, respectively), chronic heart failure (45.7%, 50.9%, and 60.0%, respectively), and coronary heart disease (15.2%, 25.9%, and 24.3%, respectively). The most frequently isolated bacteria by age group were the following: group I – non-fermenting Gram-negative bacteria (NFGNB) (7.4%), Enterobacterales (6.6%), S. aureus (6.6%); group II – Enterobacterales (13.9%), S. aureus (5.6%), Enterococcus spp. (5.6%), NFGNB (2.8%); group III – NFGNB (15.4%), S. aureus (7.7%), Enterococcus spp. (7.7%), Enerobacterales (7.7%).
Conclusions.
The mortality rates in elderly patients with CAP were high and varied from 27.1% in 65–74 years old patients to 45.7% in 85–94 years old patients. The most common comorbidities in all age groups were arterial hypertension (up to 62.8%), chronic heart failure (up to 60%), and coronary heart disease (up to 25.9%). The main pathogens causing CAP in elderly patient population were Enterobacterales and non-fermenting Gram-negative bacteria.
Savita Chib, Farhan Ali, Aswin Sai Narain Seshasayee
ABSTRACT Prolonged stationary phase is an approximation of natural environments presenting a range of stresses. Survival in prolonged stationary phase requires alternative metabolic pathways for survival. This study describes the repertoire of mutations accumulating in starving Escherichia coli populations in lysogeny broth. A wide range of mutations accumulates over the course of 1 month in stationary phase. Single nucleotide polymorphisms (SNPs) constitute 64% of all mutations. A majority of these mutations are nonsynonymous and are located at conserved loci. There is an increase in genetic diversity in the evolving populations over time. Computer simulations of evolution in stationary phase suggest that the maximum frequency of mutations observed in our experimental populations cannot be explained by neutral drift. Moreover, there is frequent genetic parallelism across populations, suggesting that these mutations are under positive selection. Finally, functional analysis of mutations suggests that regulatory mutations are frequent targets of selection. IMPORTANCE Prolonged stationary phase in bacteria, contrary to its name, is highly dynamic, with extreme nutrient limitation as a predominant stress. Stationary-phase cultures adapt by rapidly selecting a mutation(s) that confers a growth advantage in stationary phase (GASP). The phenotypic diversity of starving E. coli populations has been studied in detail; however, only a few mutations that accumulate in prolonged stationary phase have been described. This study documented the spectrum of mutations appearing in Escherichia coli during 28 days of prolonged starvation. The genetic diversity of the population increases over time in stationary phase to an extent that cannot be explained by random, neutral drift. This suggests that prolonged stationary phase offers a great model system to study adaptive evolution by natural selection.
Fahimeh Azizi, Mohammad B. Habibi Najafi, Mohammad R. Edalatian Dovom
Abstract Lactobacilli, as the largest group of lactic acid bacteria, produce large amounts of antimicrobial metabolites such as organic acids, fatty acids, ammonia, hydrogen peroxide, diacetyl and bacteriocin, which inhibit the growth of pathogenic bacteria and increase shelf life of food. The aim of this study was to identify the Lactobacillus spp. isolated from Iranian raw milk Motal cheese and to detect the presence of bacteriocin genes in the isolated Lactobacillus strains exhibiting antimicrobial activity. For this purpose, 6 Motal cheese samples from Dasht-e-Moghan region, Iran, were subjected to microbial characterization. Nineteen Lactobacillus spp. were isolated and subsequently identified based on biochemical and molecular methods. According to the sequencing of isolates, Lactobacillus spp. consisted primarily of Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei and Lactobacillus buchneri. The identified isolates were then evaluated for antimicrobial activity against Escherichia coli ATCC 25922, Listeria innocua ATCC 33090 and Staphylococcus aureus ATCC 25923. The results of PCR analysis using specific primers of genes encoding Bacteriocin, revealed the presence of Plantaricin A and Plantaricin EF in all Lactobacillus plantarum isolates and Brevicin 174A in 5 of Lactobacillus brevis isolates, whereas the gene encoding Pediocin PA-1 was not observed in any of examined isolates. It is therefore concluded that bacteriocinogenic isolates could be recommended as suitable candidates to be used as starter, adjunct-starter or antimicrobial agents for production of fermented and non-fermented products.
Patrice Bazongo, Imaël Henri Nestor Bassolé, Søren Nielsen
et al.
The proximate composition of seeds and main physicochemical properties and thermal stability of oil extracted from Lannea microcarpa seeds were evaluated. The percentage composition of the seeds was: ash (3.11%), crude oil (64.90%), protein (21.14%), total carbohydrate (10.85%) and moisture (3.24%). Physicochemical properties of the oil were: refractive index, 1.473; melting point, 22.60°C; saponification value, 194.23 mg of KOH/g of oil; iodine value, 61.33 g of I2/100 g of oil; acid value, 1.21 mg of KOH/g of oil; peroxide value, 1.48 meq of O2/kg of oil and oxidative stability index, 43.20 h. Oleic (43.45%), palmitic (34.45%), linoleic (11.20%) and stearic (8.35%) acids were the most dominant fatty acids. Triacylglycerols with equivalent carbon number (ECN) 48 and ECN 46 were dominant (46.96% and 37.31%, respectively). The major triacylglycerol constituents were palmitoyl diolein (POO) (21.23%), followed by dipalmitoyl olein (POP) (16.47%), palmitoyl linoleyl olein (PLO) (12.03%), dipalmitoyl linolein (PLP) (10.85%) and dioleoyl linolein (LOO) (9.30%). The total polyphenol and tocopherol contents were 1.39 mg GAE g−1 DW and 578.56 ppm, respectively. γ-Tocopherol was the major tocopherol (437.23 ppm). These analytical results indicated that the L. microcarpa seed oil could be used as a frying oil and in the cosmetic industry.
Ajay K. Pandey, Chunling Yang, Chunquan Zhang
et al.
Asian soybean rust is an aggressive foliar disease caused by the obligate biotrophic fungus Phakopsora pachyrhizi. On susceptible plants, the pathogen penetrates and colonizes leaf tissue, resulting in the formation of necrotic lesions and the development of numerous uredinia. The soybean Rpp2 gene confers resistance to specific isolates of P. pachyrhizi. Rpp2-mediated resistance limits the growth of the pathogen and is characterized by the formation of reddish-brown lesions and few uredinia. Using virus-induced gene silencing, we screened 140 candidate genes to identify those that play a role in Rpp2 resistance toward P. pachyrhizi. Candidate genes included putative orthologs to known defense-signaling genes, transcription factors, and genes previously found to be upregulated during the Rpp2 resistance response. We identified 11 genes that compromised Rpp2-mediated resistance when silenced, including GmEDS1, GmNPR1, GmPAD4, GmPAL1, five predicted transcription factors, an O-methyl transferase, and a cytochrome P450 monooxygenase. Together, our results provide new insight into the signaling and biochemical pathways required for resistance against P. pachyrhizi.