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BACKGROUND: Klebsiella pneumoniae is among the most common Gram-negative bacteria isolated to neonatal intensive care units (NICU) and one of the leading causes of morbidity and mortality. The ceftazidime/avibactam (CAZ-AVI) combination is approved for infections caused by aerobic Gram-negative organisms. It is licensed for use in infants over 3 months old. There are no safety and efficacy data regarding the administration of CAZ-AVI to infants younger than 3 months, except for a few case reports. CASE PRESENTATION: This report describes a severely intoxicated 24-day-old, full-term, male neonate transferred to NICU level III from a secondary maternity hospital due to the deterioration of his general condition. On day four of admission, blood culture revealed the pan-drug-resistant (PDR) K. pneumoniae ss. pneumoniae, susceptible only to CAZ-AVI, which thus represented the only treatment option. Off-label CAZ-AVI was administered intravenously as a salvage therapy. CONCLUSIONS: In healthcare settings, treating resistant K. pneumoniae presents serious challenges, especially in NICU patients. The off-label treatment with CAZ-AVI for 17 days was safe and effective in this one-month-old patient. A year later, the patient was healthy with normal cognitive development.
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Probiotics possibly affect local and systemic immune reactions and maintain the intestinal immune homeostasis in healthy individuals and patients with diseases such as irritable bowel syndrome (IBS). In this single-center, blinded trial, we enrolled 40 individuals (20 patients with IBS and 20 healthy individuals) whose blood and fecal samples were collected before and after a 21-day administration of a product comprising Lactobacillus spp., larch arabinogalactan, and colostrum. The percentage of HLA-DR+ natural killer (NK) cells was higher in healthy individuals (p = 0.03) than in patients with IBS after product supplementation. In the fecal samples of patients with IBS, we observed a decline in IL-6, IFN-γ, TNF-α, and secretory IgA levels and, simultaneously, an increase in IL-10 and IL-17A levels after supplementation, although non-significant, whereas, in healthy individuals, we observed a significant decline in IL-6 and IFN-γ levels after supplementation (p < 0.001). Nevertheless, we observed a clinical improvement of symptoms in 65-75% of patients with IBS and the complete resolution of the initial symptoms in five of the 20 patients. We also observed a possible prophylactic effect by the inducing system antiviral impact accompanied by a trend for local immune tolerance in the gut in healthy individuals, where it is the desirable state.
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Calostro/fisiología , Suplementos Dietéticos , Galactanos/administración & dosificación , Tolerancia Inmunológica , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Síndrome del Colon Irritable/dietoterapia , Síndrome del Colon Irritable/inmunología , Células Asesinas Naturales/inmunología , Lactobacillus , Larix/química , Activación de Linfocitos/inmunología , Adulto , Citocinas/metabolismo , Femenino , Galactanos/aislamiento & purificación , Voluntarios Sanos , Homeostasis , Humanos , Inmunoglobulina A Secretora/metabolismo , Síndrome del Colon Irritable/prevención & control , Masculino , Persona de Mediana Edad , Resultado del Tratamiento , Adulto JovenRESUMEN
Surface layer (S-layer) proteins have been identified in the cell envelope of many organisms, such as bacteria and archaea. They self-assemble, forming monomolecular crystalline arrays. Isolated S-layer proteins are able to recrystallize into regular lattices, which proved useful in biotechnology. Here we investigate the structure and thermal unfolding of the S-layer protein isolated from Lactobacillus salivarius 16 strain of human origin. Using circular dichroism (CD) spectroscopy, and the software CDSSTR from DICHROWEB, CONTINLL from CDPro, as well as CDNN, we assess the fractions of the protein's secondary structural elements at temperatures ranging between 10 and 90 °C, and predict the tertiary class of the protein. To study the thermal unfolding of the protein, we analyze the temperature dependence of the CD signal in the far- and near-UV domains. Fitting the experimental data by two- and three-state models of thermal unfolding, we infer the midpoint temperatures, the temperature dependence of the changes in Gibbs free energy, enthalpy, and entropy of the unfolding transitions in standard conditions, and the temperature dependence of the equilibrium constant. We also estimate the changes in heat capacity at constant pressure in standard conditions. The results indicate that the thermal unfolding of the S-layer protein from L. salivarius is highly cooperative, since changes in the secondary and tertiary structures occur simultaneously. The thermodynamic analysis predicts a "cold" transition, at about -3 °C, of both the secondary and tertiary structures. Our findings may be important for the use of S-layer proteins in biotechnology and in biomedical applications.
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Ligilactobacillus salivarius , Glicoproteínas de Membrana/química , Desplegamiento Proteico , Temperatura , Estructura Secundaria de ProteínaRESUMEN
Barley stress response to ultraviolet radiation (UV) has been intensively studied at both the physiological and morphological level. However, the ability of barley genome to repair UV-induced lesions at the DNA level is far less characterized. In this study, we have investigated the relative contribution of light-dependent and dark DNA repair pathways for the efficient elimination of cyclobutane pyrimidine dimers (CPDs) from the genomic DNA of barley leaf seedlings. The transcriptional activity of barley CPD photolyase gene in respect to the light-growth conditions and UV-C irradiation of the plants has also been analyzed. Our results show that CPDs induced in the primary barley leaf at frequencies potentially damaging DNA at the single-gene level are removed efficiently and exclusively by photorepair pathway, whereas dark repair is hardly detectable, even at higher CPD frequency. A decrease of initially induced CPDs under dark is observed but only after prolonged incubation, suggesting the activation of light-independent DNA damage repair and/or tolerance mechanisms. The green barley seedlings possess greater capacity for CPD photorepair than the etiolated ones, with efficiency of CPD removal dependent on the intensity and quality of recovering light. The higher repair rate of CPDs measured in the green leaves correlates with the higher transcriptional activity of barley CPD photolyase gene. Visible light and UV-C radiation affect differentially the expression of CPD photolyase gene particularly in the etiolated leaves. We propose that the CPD repair potential of barley young seedlings may influence their response to UV-stress.
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Reparación del ADN , Desoxirribodipirimidina Fotoliasa/metabolismo , Regulación de la Expresión Génica de las Plantas , Hordeum/genética , Dímeros de Pirimidina/genética , Daño del ADN , Desoxirribodipirimidina Fotoliasa/genética , Hordeum/fisiología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantones/genética , Plantones/fisiología , Rayos UltravioletaRESUMEN
Antimicrobial activity and antibiotic susceptibility were tested for 23 Lactobacillus and three Bifidobacterium strains isolated from different ecological niches. Agar-well diffusion method was used to test the antagonistic effect (against Staphylococcus aureus, Escherichia coli, Bacillus cereus and Candida albicans) of acid and neutralized (pH 5.5) lyophilized concentrated supernatants (cell-free supernatant; CFS) and whey (cell-free whey fractions; CFW) from de Man-Rogosa-Sharpe/trypticase-phytone-yeast broth and skim milk. Acid CFS and CFW showed high acidification rate-dependent bacterial inhibition; five strains were active against C. albicans. Neutralized CFS/CFW assays showed six strains active against S. aureus (L. acidophilus L-1, L. brevis 1, L. fermentum 1, B. animalis subsp. lactis L-3), E. coli (L. bulgaricus 6) or B. cereus (L. plantarum 24-4Ð). Inhibition of two pathogens with neutralized CFS (L. bulgaricus 6, L. helveticus 3, L. plantarum 24-2L, L. fermentum 1)/CFW (L. plantarum 24-5D, L. plantarum 24-4Ð) was detected. Some strains maintained activity after pH neutralization, indicating presence of active substances. The antibiotics minimum inhibitory concentrations (MICs) were determined by the Epsilometer test method. All strains were susceptible to ampicillin, gentamicin, erythromycin and tetracycline. Four lactobacilli were resistant to one antibiotic (L. rhamnosus Lio 1 to streptomycin) or two antibiotics (L. acidophilus L-1 and L. brevis 1 to kanamycin and clindamycin; L. casei L-4 to clindamycin and chloramphenicol). Vancomycin MICs > 256 µg/mL indicated intrinsic resistance for all heterofermentative lactobacilli. The antimicrobially active strains do not cause concerns about antibiotic resistance transfer and could be used as natural biopreservatives in food and therapeutic formulations.
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A family with 2 siblings with severe spinal muscular atrophy with respiratory distress 1 (SMARD1) was genetically proved to be caused by mutations in IGHMBP2 gene. Both patients developed progressive muscular weakness and respiratory distress and died before 6 months of age. One novel deletion, c.780delG;p.(Gln260Hisfs*24), inherited from the father and a nonsense mutation, c.1488C>A;p.(Cys496*), inherited from the mother were detected. An attempt was made to correlate the genetic-clinical data available in the literature. The clinical case presented in this study might be considered as the most severe form of spinal muscular atrophy respiratory distress 1 reported so far, presumably because of the total absence of IGHMBP2 enzyme activity.
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Proteínas de Unión al ADN/genética , Atrofia Muscular Espinal/genética , Mutación/genética , Síndrome de Dificultad Respiratoria del Recién Nacido/genética , Factores de Transcripción/genética , Salud de la Familia , Humanos , Recién Nacido , Examen Neurológico , Radiografía Torácica , Tórax/patologíaRESUMEN
The theoretical and experimental studies of the particles' electric dipole moments in the microscopic and submicroscopic size range show that in the case of polar and conductive media the interfacial components of the dipole moments are of greatest importance. While in the range of manometer's sizes there seems to be no important problems in the identification and in the estimation of the values of the dipole moments at present, in the micrometer range there are serious problems. In this communication these problems are considered and illustrated by electro-optic investigations of Escherichia coli HB 101.
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Electrones , Escherichia coli/química , Técnicas Electroquímicas , Electroquímica , Escherichia coli/ultraestructura , Microscopía de Fuerza AtómicaRESUMEN
Lactobacillus plantarum strains were isolated from fully ripened, white brined Bulgarian home-made cheeses. Strains were derived from phenotypically homogenous Lactobacillus group and were identified as L. plantarum based on both phenotypic and molecular identification (species-specific and multiplex PCR) methods. Heterogeneity of L. plantarum isolates was evaluated by Rep-PCR analysis. Further antimicrobial activity, antibiotic susceptibility and transit tolerance of the strains were evaluated. Most of them showed broad spectrum of activity against Gram-negative bacteria (including human pathogens) independent on the presence of organic acids or hydrogen peroxide. All strains were sensitive to amoxicillin and sulfamethoxazole/trimethoprim, resistant to clinically relevant beta -lactame antibiotics (penicillin and ampicillin) and to nalidixic acid, ciprofloxacin, streptomycin, and vancomycin. Significant variability in the sensitivity to tetracycline was noted. The tolerance to low pH, bile salts, pepsin and pancreatin at concentrations similar to those in the gastrointestinal tract was strain-dependent. The exposure to bile salts was less destructive than exposure to pH 2.0 for all tested strains. Based on their combined responses to the above selection criteria, four L. plantarum strains, RL29, RL34, RL36 and RL37, were selected as potential probiotics for in vivo studies.
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Biodiversidad , Queso/microbiología , Lactobacillus plantarum/clasificación , Lactobacillus plantarum/aislamiento & purificación , Antibacterianos/biosíntesis , Antibacterianos/farmacología , Antibiosis , Técnicas de Tipificación Bacteriana , Bulgaria , Análisis por Conglomerados , Dermatoglifia del ADN , ADN Bacteriano/genética , Farmacorresistencia Bacteriana , Genotipo , Bacterias Gramnegativas/efectos de los fármacos , Lactobacillus plantarum/genética , Lactobacillus plantarum/fisiología , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana , Reacción en Cadena de la Polimerasa/métodosRESUMEN
BACKGROUND AND PURPOSE: Twenty lactobacilli isolated from vaginal samples of healthy volunteers were characterized according to polyphasic taxonomy. A broad spectrum of activity and protective properties of these vaginal isolates has been reported. METHODS: Phenotypic and genotypic methods such as random amplified polymorphic DNA (RAPD), species-specific and BOX polymerase chain reaction (PCR), amplified ribosomal DNA restriction analysis (ARDRA), and 16S rRNA sequence analyses were applied for Lactobacillus spp. identification. RESULTS: On the basis of carbohydrate utilization profiles using API 50 CHL kits (fermentation pattern), the strains were divided into 7 groups. RAPD- and BOX-PCR analyses revealed heterogeneity within the established phenotypic groups and discriminated successfully between the vaginal strains. The combination of species-specific PCR, ARDRA, and 16S rRNA sequence analyses allowed the species identification of 7 strains as Lactobacillus fermentum, 2 as Lactobacillus gasseri, 1 as Lactobacillus brevis, and 1 as Lactobacillus salivarius. These results were not in agreement with data obtained by classical and molecular methods for some clinical isolates. CONCLUSION: This is the first polyphasic taxonomy study on vaginal lactobacilli from Bulgarian women of reproductive age. These data add to the limited information existing in Bulgaria on the true identity of Lactobacillus spp. in the vaginal microbiota.