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1.
BMJ Case Rep ; 14(2)2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33526521

RESUMO

Vertebral osteomyelitis is an infection of the vertebrae that can lead to spinal degeneration, most commonly caused by Staphylococcus aureus Here, we report an unusual case of pyogenic osteomyelitis caused by Gardnerella vaginalis and Streptococcus parasanguinis in a 61-year-old postmenopausal woman. The patient presented with a 2-week history of worsening lower back pain and fever and a recent episode of cystitis following re-engagement of sexual activity. Imaging revealed a deterioration of vertebrae discs and spinal canal stenosis at the L3-L4 levels with a formation of abscess in the right psoas muscle. Needle aspiration of the abscess identified G. vaginalis and S. parasanguinis and the patient was successfully treated with a 6-week course of ceftriaxone and metronidazole. This case describes an unusual coinfection of two pathogens that normally reside in the urogenital tract and oral cavity, respectively, and highlights the risk posed when these organisms breach the body's normal barriers.


Assuntos
Discite/microbiologia , Gardnerella vaginalis/isolamento & purificação , Vértebras Lombares , Osteomielite/microbiologia , Abscesso do Psoas/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus/isolamento & purificação , Antibacterianos/uso terapêutico , Técnicas de Cultura , Discite/diagnóstico por imagem , Discite/tratamento farmacológico , Discite/fisiopatologia , Feminino , Infecções por Bactérias Gram-Positivas/diagnóstico , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Imagem por Ressonância Magnética , Pessoa de Meia-Idade , Osteomielite/diagnóstico por imagem , Osteomielite/tratamento farmacológico , Osteomielite/fisiopatologia , Pós-Menopausa , Abscesso do Psoas/diagnóstico por imagem , Abscesso do Psoas/tratamento farmacológico , Estenose Espinal/fisiopatologia , Infecções Estreptocócicas/diagnóstico , Infecções Estreptocócicas/tratamento farmacológico
2.
Cancer Radiother ; 25(2): 126-134, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33431297

RESUMO

BACKGROUND: To determine the effects of concurrent irradiation and T-DM1 on HER2-positive breast cancer cell lines. METHODS: Five human breast cancer cell lines (in vitro study) presenting various levels of HER2 expression were used to determine the potential therapeutic effect of T-DM1 combined with radiation. The toxicity of T-DM1 was assessed using viability assay and cell cycle analysis was performed by flow cytometry after BrdU incorporation. HER2 cells were irradiated at different dose levels after exposure to T-DM1. Survival curves were determined by cell survival assays (after 5 population doubling times). RESULTS: The results revealed that T-DM1 induced significant lethality due to the intracellular action of DM1 on the cell cycle with significant G2/M phase blocking. Even after a short time incubation, the potency of T-DM1 was maintained and even enhanced over time, with a higher rate of cell death. After irradiation alone, the D10 (dose required to achieve 10% cell survival) was significantly higher for high HER2-expressing cell lines than for low HER2-expressing cells, with a linearly increasing relationship. In combination with irradiation, using conditions that allow cell survival, T-DM1 does not induce a radiosensitivity. CONCLUSIONS: Although there is a linear correlation between intrinsic HER2 expression and radioresistance, the results indicated that T-DM1 is not a radiation-sensitizer under the experimental conditions of this study that allowed cell survival. However, further investigations are needed, in particular in vivo studies before reaching a final conclusion.


Assuntos
Ado-Trastuzumab Emtansina/uso terapêutico , Antineoplásicos Imunológicos/uso terapêutico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/terapia , Quimiorradioterapia/métodos , Receptor ErbB-2/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Técnicas de Cultura , Feminino , Citometria de Fluxo/métodos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos da radiação , Humanos , Pontos de Checagem da Fase M do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem da Fase M do Ciclo Celular/efeitos da radiação , Tolerância a Radiação/efeitos dos fármacos , Fatores de Tempo
3.
Medicine (Baltimore) ; 100(2): e24090, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33466172

RESUMO

OBJECTIVE: To understood the pathogen detection status and clinical characteristics of suspected pertussis in children and to observe the drug sensitivity and drug resistance genes of Bordetella pertussis (B. pertussis). METHODS: Three hundred fifty-one cases were collected and their nasopharyngeal swab samples were analyzed by culture and fluorescent quantitative polymerase chain reaction. The susceptibility to erythromycin, clindamycin, ampicillin, levofloxacin, and sulfamethoxazole-trimethoprim were tested by E-test for the positive strains, and the susceptibility to erythromycin was also tested for the KB disk diffusion method. The 23S rRNA gene of the positive strains was amplified and sequenced, and statistical analysis was performed in conjunction with clinical data. RESULTS: The positive rate of bacterial culture was 16.8% (59/351), and the positive rate of PCR was 62.4% (219/351). Two cases were positive about bacterial culture and negative for PCR. There were 221 confirmed cases of pertussis. The E-test results showed that the rate of the sensitivity of the 55 strains of pertussis to erythromycin and clindamycin was 50.9% (28/55), the minimum antibiotic concentration50 (MIC50) and MIC90 values were 0.094/>256 and 0.75/>256 mg/L, respectively, and the MIC50/MIC90 to ampicillin, levofloxacin, and sulfamethoxazole were 0.125/0.19, 0.38/0.5, and 0.125/0.25 mg/L, respectively. The KB disk diffusion method showed 27 of the 55 strains 49.1% (27/55) was resistant to erythromycin; all of the resistant strains had the 23S rRNA gene A2047G mutation, and their MIC of erythromycin was >256 mg/L. CONCLUSION: The diagnosis of pertussis by a fluorescent quantitative polymerase chain reaction method is more sensitive than that of bacterial culture. The resistance of B. pertussis to erythromycin was prominent. All of the strains of B. pertussis resistant to erythromycin in our center had the A2047G mutation of the 23S rRNA gene.


Assuntos
Bordetella pertussis/efeitos dos fármacos , Bordetella pertussis/genética , Farmacorresistência Bacteriana/genética , Testes de Sensibilidade Microbiana/métodos , Coqueluche/microbiologia , Antibacterianos/farmacologia , Criança , Pré-Escolar , China , Técnicas de Cultura , Eritromicina/farmacologia , Feminino , Humanos , Lactente , Masculino , Tipagem Molecular , Mutação , Reação em Cadeia da Polimerase , RNA Ribossômico 23S/genética , Coqueluche/diagnóstico
4.
BMJ Case Rep ; 14(1)2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33500298

RESUMO

A 47-year-old man was referred for ongoing workup of an enlarging lung mass. Extensive workup of the mass had been unrevealing for several months until cultures grew Nocardia beijingensis He was successfully treated with trimethoprim/sulfamethoxazole and then doxycycline with near-complete resolution of the mass on follow-up. This case presents a rare species of N. beijingensis It highlights the importance of considering nocardiosis in immunocompetent adults and the challenge in initiating targeted treatment due to delayed culture results.


Assuntos
Abscesso/diagnóstico por imagem , Mediastinite/diagnóstico por imagem , Nocardiose/diagnóstico por imagem , Nocardia , Pneumonia Bacteriana/diagnóstico por imagem , Abscesso/complicações , Abscesso/tratamento farmacológico , Antibacterianos/uso terapêutico , Biópsia , Biópsia com Agulha de Grande Calibre , Sedimentação Sanguínea , Broncoscopia , Proteína C-Reativa , Técnicas de Cultura , Fístula Esofágica/diagnóstico , Fístula Esofágica/etiologia , Humanos , Imunocompetência , Masculino , Doenças do Mediastino/diagnóstico , Doenças do Mediastino/etiologia , Mediastinite/complicações , Mediastinite/tratamento farmacológico , Pessoa de Meia-Idade , Nocardiose/complicações , Nocardiose/tratamento farmacológico , Nocardiose/patologia , Pneumonia Bacteriana/complicações , Pneumonia Bacteriana/tratamento farmacológico , Pneumonia Bacteriana/patologia , Tomografia por Emissão de Pósitrons , Cirurgia Torácica Vídeoassistida , Tomografia Computadorizada por Raios X
5.
Methods Mol Biol ; 2232: 53-60, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33161539

RESUMO

Beneficial plant-microbe interactions are important and desirable for sustainable intensification of agriculture. Here, we describe methods to isolate microbes from the roots of field-grown wheat plants. This includes the rhizosphere and rhizoplane soil, as well as the root endosphere. We also describe a method to test for endosphere competence of putative endophytes.


Assuntos
Agricultura/métodos , Técnicas de Cultura/métodos , Raízes de Plantas/microbiologia , Triticum/microbiologia , Endófitos/genética , Microbiota/genética , Raízes de Plantas/genética , Triticum/genética
7.
Discov Med ; 29(157): 129-137, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33002409

RESUMO

Sepsis is a life-threatening clinical condition demanding accurate and rapid diagnosis of the culprit pathogen, thereby to improve prognosis. Pathogen determination through blood culture is the gold standard for diagnosis but has limitations due to low sensitivity. Recently, circulating DNAs derived from pathogenic organisms were found in the plasma of patients with sepsis and were further proved to be more sensitive biomarkers for the diagnosis of the pathogen origin in sepsis. However, the fundamental molecular characteristics of circulating DNA in patients with sepsis remain unclear. Here, we used specific PCR and Sanger sequencing to verify the microbiology culture results via the corresponding plasma circulating DNA. We analyzed the composition and molecular characteristics of circulating DNA in septic patients using next-generation sequencing technology. We showed the presence of pathogen-derived circulating DNA in the plasma of patients with sepsis. The sizes of circulating DNA fragments derived from pathogenic bacteria showed a skewed unimodal distribution, while those derived from host cells showed a normal unimodal distribution. Lengths of fragments at peak concentration for both origins ranged from 150 bp to 200 bp, and reads mapping to pathogenic bacteria genome distributed uniformly on the reference. Our findings have improved our understanding of microbial circulating DNA in patients with sepsis as a potential methodology for the accurate diagnosis of sepsis, especially in light of an urgent need for such a diagnosis associated with the COVID-19 infection.


Assuntos
Infecções Bacterianas/microbiologia , Ácidos Nucleicos Livres/sangue , DNA Bacteriano/sangue , Sepse/microbiologia , Adulto , Idoso , Infecções Bacterianas/complicações , Infecções Bacterianas/diagnóstico , Betacoronavirus , Ácidos Nucleicos Livres/análise , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Técnicas de Cultura , DNA Bacteriano/análise , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/complicações , Pandemias , Pneumonia Viral , Reação em Cadeia da Polimerase , Sepse/complicações , Sepse/diagnóstico , Análise de Sequência de DNA
8.
PLoS One ; 15(9): e0238769, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32925929

RESUMO

Myxobacteria are one of the most promising secondary metabolites producers. However, they are difficult to isolate and cultivate. To obtain more myxobacteria and know the effects of environmental factors on myxobacterial community, we characterized myxobacterial communities in Dinghushan acidic forest soils of pH 3.6-4.5 with culture-dependent and -independent techniques, and analyzed environmental factors shaping myxobacterial communities. A total of 21 myxobacteria were isolated using standard cultivation methods, including eleven isolates of Corallococcus, nine isolates of Myxococcus and one isolate of Archangium, and contained three potential novel species. In addition, a total of 67 unknown myxobacterial operational taxonomic units (OTUs) were obtained using high-throughput sequencing method. The abundance of Myxococcales account for 0.9-2.2% of bacterial communities, and Sorangium is the most abundant genus (60.1%) in Myxococcales. Correlation analysis demonstrated that bacterial diversity and soil pH are the key factors shaping myxobacterial community. These results revealed an abundant myxobacterial community which is shaped by other bacteria and pH in Dinghushan acidic forest soils.


Assuntos
Técnicas de Cultura/métodos , Myxococcales/crescimento & desenvolvimento , Myxococcales/isolamento & purificação , Microbiologia do Solo , Solo/química , Florestas , Sequenciamento de Nucleotídeos em Larga Escala , Concentração de Íons de Hidrogênio , Myxococcales/genética , RNA Ribossômico 16S/genética
9.
Indian J Dent Res ; 31(3): 420-425, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32769277

RESUMO

Background and Aims: Although Streptococcus mutans has been implicated as the major etiologic agent in the development of dental caries, however, this organism has not been found to be present in all children with caries. Thus it remains to be elucidated whether a single or specific consortium of bacteria is responsible for the caries process. The aim of this study is to evaluate the oral microflora of Indian children suffering from dental caries and to compare the same in children with no caries. Methods: The study was carried out on 67 out-patient 2-14 years old children who reported to the department of Paediatric Dentistry. Dental plaque samples from superficial and deep carious lesions and caries free surfaces in caries active children (n = 35) were collected using a sterile excavator in storage vials and subjected to various conventional and molecular microbial techniques. Caries free children (n = 32), who did not have any carious lesion served as controls. The data obtained was subjected to Pearson's Chi Square/Fischer's Exact tests to determine the statistical difference between the microflora of groups. Results: Main organisms isolated were: Streptococcal species mainly S anginosus, S salivarius, S gordonii: Lactobacilli spp; Coagulase negative Staphylococci, Staph epidermidis, Staph aureus. Other organisms such as Klebsiella spp, Acinetobacter, Enterococcus, E coli could also be found. There was a statistical difference in the frequency of isolation of non-mutans Streptococcal species (P = 0.008) and Lactobacilli species (P = 0.0001) in the two groups. Conclusions: The study suggests that caries activity in this population of children is associated with a diverse microbial flora without detectable S mutans. Main organisms associated with dental caries in this population are: Gram-positive cocci and bacilli mainly the non-mutans Streptococcus and Lactobacilli. Frequent consumption of sugar containing food promotes the presence and growth of cariogenic organisms.


Assuntos
Cárie Dentária , Placa Dentária , Adolescente , Criança , Pré-Escolar , Contagem de Colônia Microbiana , Técnicas de Cultura , Escherichia coli , Humanos , Reação em Cadeia da Polimerase , Streptococcus mutans
10.
PLoS Biol ; 18(7): e3000794, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32730242

RESUMO

The precision and repeatability of in vivo biological studies is predicated upon methods for isolating a targeted subsystem from external sources of noise and variability. However, in many experimental frameworks, this is made challenging by nonstatic environments during host cell growth, as well as variability introduced by manual sampling and measurement protocols. To address these challenges, we developed Chi.Bio, a parallelised open-source platform that represents a new experimental paradigm in which all measurement and control actions can be applied to a bulk culture in situ. In addition to continuous-culturing capabilities, it incorporates tunable light outputs, spectrometry, and advanced automation features. We demonstrate its application to studies of cell growth and biofilm formation, automated in silico control of optogenetic systems, and readout of multiple orthogonal fluorescent proteins in situ. By integrating precise measurement and actuation hardware into a single low-cost platform, Chi.Bio facilitates novel experimental methods for synthetic, systems, and evolutionary biology and broadens access to cutting-edge research capabilities.


Assuntos
Reatores Biológicos , Técnicas de Cultura/instrumentação , Optogenética/instrumentação , Automação , Biofilmes , Proliferação de Células , Simulação por Computador , Software
11.
Eur J Protistol ; 75: 125718, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32604041

RESUMO

Cryptosporidium is a genus of apicomplexan parasites that inhabit the respiratory and gastrointestinal tracts of vertebrates. Research of these parasites is limited by a lack of model hosts. This study aimed to determine the extent to which infection at the embryo stage can enhance the propagation of Cryptosporidium oocysts in chickens. Nine-day-old chicken embryos and one-day-old chickens were experimentally infected with different doses of Cryptosporidium baileyi and Cryptosporidium parvum oocysts. Post hatching, all chickens had demonstrable infections, and the infection dose had no effect on the course of infection. Chickens infected as embryos shed oocysts immediately after hatching and shed significantly more oocysts over the course of the infection than chickens infected as one-day-olds. In chickens infected as embryos, C. baileyi was found in all organs except the brain whereas, C. parvum was only found in the gastrointestinal tract and trachea. In chickens infected as one-day-olds, C. baileyi was only found in the gastrointestinal tract and trachea. Chickens infected as embryos with C. baileyi died within 16 days of hatching. All other chickens cleared the infection. Infection of chickens as embryos could be used as an effective and simple model for the propagation of C. baileyi and C. parvum.


Assuntos
Cryptosporidium parvum/crescimento & desenvolvimento , Cryptosporidium/crescimento & desenvolvimento , Técnicas de Cultura , Oocistos/crescimento & desenvolvimento , Animais , Embrião de Galinha , Galinhas , Criptosporidiose/parasitologia
12.
Rev Bras Parasitol Vet ; 29(2): e022819, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32609247

RESUMO

The aim of this study was to evaluate the efficiency of different substrates for larval development of Ctenocephalides felis felis during its biological cycle. Eight hundred eggs of C. felis felis from a flea maintenance colony were used. Different diets were formulated, in which the main substrates were meat flour, powdered milk, sugar, lyophilized bovine blood, tick metabolites and lyophilized egg. The flea eggs were placed in test tubes (10 per tube) and approximately 2 g of the diet to be tested was added to each tube. There were 10 replicates for each substrate. After 28 days, each tube was evaluated individually for the presence of pupae and emerged adults. The following percentages of the larvae completed the cycle to the adult stage: 67% in diets containing tick metabolites; 55%, meat flour; 39%, dehydrated bovine blood; 14%, powdered milk; and less than 1% in diets containing sugar, lyophilized bovine blood, lyophilized egg or wheat bran. It was concluded that among the diets tested, the one constituted by tick metabolites as the substrate was shown to be the most satisfactory for maintaining a laboratory colony of C. felis felis, followed by the one containing meat flour.


Assuntos
Ctenocephalides , Técnicas de Cultura , Animais , Gatos , Ctenocephalides/crescimento & desenvolvimento , Técnicas de Cultura/métodos , Larva/crescimento & desenvolvimento
13.
J Dairy Sci ; 103(8): 7124-7140, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32600762

RESUMO

Within the rumen, nitrate can serve as an alternative sink for aqueous hydrogen [H2(aq)] accumulating during fermentation, producing nitrite, which ideally is further reduced to ammonium but can accumulate under conditions not yet explained. Defaunation has also been associated with decreased methanogenesis in meta-analyses because protozoa contribute significantly to H2 production. In the present study, we applied a 2 × 2 factorial treatment arrangement in a 4 × 4 Latin square design to dual-flow continuous culture fermentors (n = 4). Treatments were control without nitrate (-NO3-) versus with nitrate (+NO3-; 1.5% of diet dry matter), factorialized with normal protozoa (faunated, FAUN) versus defaunation (DEF) by decreasing the temperature moderately and changing filters over the first 4 d of incubation. We detected no main effects of DEF or interaction of faunation status with +NO3-. The main effect of +NO3- increased H2(aq) by 11.0 µM (+117%) compared with -NO3-. The main effect of +NO3- also decreased daily CH4 production by 8.17 mmol CH4/d (31%) compared with -NO3-. Because there were no treatment effects on neutral detergent fiber digestibility, the main effect of +NO3- also decreased CH4 production by 1.43 mmol of CH4/g of neutral detergent fiber degraded compared with -NO3-. There were no effects of treatment on other nutrient digestibilities, N flow, or microbial N flow per gram of nutrient digested. The spike in H2(aq) after feeding NO3- provides evidence that methanogenesis is inhibited by substrate access rather than concentration, regardless of defaunation, or by direct inhibition of NO2-. Methanogens were not decreased by defaunation, suggesting a compensatory increase in non-protozoa-associated methanogens or an insignificant contribution of protozoa-associated methanogens. Despite adaptive reduction of NO3- to NH4+ and methane inhibition in continuous culture, practical considerations such as potential to depress dry matter intake and on-farm ration variability should be addressed before considering NO3- as an avenue for greater sustainability of greenhouse gas emissions in US dairy production.


Assuntos
Fibras na Dieta/metabolismo , Eucariotos/metabolismo , Hidrogênio/metabolismo , Metano/metabolismo , Microbiota , Nitratos/farmacologia , Compostos de Amônio/metabolismo , Animais , Bactérias/metabolismo , Reatores Biológicos , Técnicas de Cultura , Dieta/veterinária , Ingestão de Alimentos , Ácidos Graxos Voláteis/metabolismo , Fermentação/efeitos dos fármacos , Hidrogênio/análise , Nitritos/metabolismo , Nitrogênio/metabolismo , Rúmen/metabolismo
14.
Nat Protoc ; 15(9): 2867-2890, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32709990

RESUMO

The world is on the cusp of a post-antibiotic era, but researchers and medical doctors have found a way forward-by looking back at how infections were treated before the advent of antibiotics, namely using phage therapy. Although bacteriophages (phages) continue to lack drug approval in Western medicine, an increasing number of patients are being treated on an expanded-access emergency investigational new drug basis. To streamline the production of high-quality and clinically safe phage preparations, we developed a systematic procedure for medicinal phage isolation, liter-scale cultivation, concentration and purification. The 16- to 21-day procedure described in this protocol uses a combination of modified classic techniques, modern membrane filtration processes and no organic solvents to yield on average 23 mL of 1011 plaque-forming units (PFUs) per milliliter for Pseudomonas, Klebsiella, and Serratia phages tested. Thus, a single production run can produce up to 64,000 treatment doses at 109 PFUs, which would be sufficient for most expanded-access phage therapy cases and potentially for clinical phase I/II applications. The protocol focuses on removing endotoxins early by conducting multiple low-speed centrifugations, microfiltration, and cross-flow ultrafiltration, which reduced endotoxins by up to 106-fold in phage preparations. Implementation of a standardized phage cultivation and purification across research laboratories participating in phage production for expanded-access phage therapy might be pivotal to reintroduce phage therapy to Western medicine.


Assuntos
Bacteriófagos/crescimento & desenvolvimento , Bacteriófagos/isolamento & purificação , Técnicas de Cultura/normas , Terapia por Fagos , Bacteriófagos/química , Bacteriófagos/fisiologia , Medicina de Precisão , Controle de Qualidade , Padrões de Referência , Proteínas Virais/análise
15.
PLoS One ; 15(6): e0234747, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32584845

RESUMO

SUMMARY OF BACKGROUND DATA: The vascular buds in the vertebral endplate (VEP) are the structural foundation of nutrient exchange in the intervertebral disc (IVD). VEGF is closely related to angiogenesis in the endplate and intervertebral disc degeneration (IDD). OBJECTIVE: To investigate the effects of static load on vascular buds and VEGF expression in the VEP and to further clarify the relation between IDD and VEGF. METHODS: IVD motion segments were harvested from rabbit lumbar spines and cultured under no-loading conditions (controls) or in custom-made apparatuses under a constant compressive load (0.5 MPa) for up to 14 days. Tissue integrity and the number of vascular buds were determined, and the concentrations and expression of Aggrecan, COL2a1, and VEGFA in the VEPs were assessed after 3, 7, and 14 days of culturing and then compared with those of fresh tissues. RESULTS: Under the constant compression, the morphological integrity of the VEPs was gradually disrupted, and immunohistochemistry results showed a significant decrease in the levels of Agg and COL2a1. During the static load, the number of vascular buds in the VEPs was gradually reduced from the early stage of culture, and ELISA showed that the constant compressive load caused a significant decrease in the VEGFA and VEGFR2 protein concentrations, which were consistent with the immunohistochemistry results. Western blot and RT-PCR results also showed that the loading state caused a significant decrease in VEGFA expression compared with that of fresh and control samples. CONCLUSIONS: Constant compression caused degeneration of the VEP as well as a decreased number of vascular buds, thereby accelerating disc degeneration. VEGFA is involved in this process. We anticipate that regulating the expression of VEGFA may improve the condition of the lesions to the vascular buds in the endplates, thus enhancing the nutritional supply function in IVD and providing new therapeutic targets and strategies for the effective prevention and treatment of IDD.


Assuntos
Vasos Sanguíneos/fisiologia , Regulação da Expressão Gênica , Disco Intervertebral/irrigação sanguínea , Disco Intervertebral/metabolismo , Estresse Mecânico , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Técnicas de Cultura , Masculino , Coelhos
16.
Am Fam Physician ; 101(12): 721-729, 2020 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-32538597

RESUMO

Despite dramatic reductions in the rates of bacteremia and meningitis since the 1980s, febrile illness in children younger than 36 months continues to be a concern with potentially serious consequences. Factors that suggest serious infection include age younger than one month, poor arousability, petechial rash, delayed capillary refill, increased respiratory effort, and overall physician assessment. Urinary tract infections are the most common serious bacterial infection in children younger than three years, so evaluation for such infections should be performed in those with unexplained fever. Abnormal white blood cell counts have poor sensitivity for invasive bacterial infections; procalcitonin and C-reactive protein levels, when available, are more informative. Chest radiography is rarely recommended for children older than 28 days in the absence of localizing signs. Lumbar puncture is not recommended for children older than three months without localizing signs; it may also be considered for those from one to three months of age with abnormal laboratory test results. Protocols such as Step-by-Step, Laboratory Score, or the Rochester algorithms may be helpful in identifying low-risk patients. Rapid influenza testing and tests for coronavirus disease 2019 (COVID-19) may be of value when those diseases are circulating. When empiric treatment is appropriate, suggested antibiotics include ceftriaxone or cefotaxime for infants one to three months of age and ampicillin with gentamicin or with cefotaxime for neonates. For children three months to three years of age, azithromycin or amoxicillin is recommended if pneumonia is suspected; for urinary infections, suggested antibiotics are cefixime, amoxicillin/clavulanate, or trimethoprim/sulfamethoxazole. Choice of antibiotics should reflect local patterns of microbial resistance.


Assuntos
Tomada de Decisão Clínica , Febre/etiologia , Influenza Humana/diagnóstico , Pneumonia Bacteriana/diagnóstico , Infecções Urinárias/diagnóstico , Algoritmos , Combinação Amoxicilina e Clavulanato de Potássio , Antibacterianos/uso terapêutico , Betacoronavirus , Hemocultura , Proteína C-Reativa/metabolismo , Pré-Escolar , Técnicas de Laboratório Clínico , Infecções por Coronavirus/complicações , Infecções por Coronavirus/diagnóstico , Técnicas de Cultura , Humanos , Lactente , Recém-Nascido , Influenza Humana/complicações , Influenza Humana/tratamento farmacológico , Influenza Humana/epidemiologia , Contagem de Leucócitos , Pandemias , Pneumonia Bacteriana/complicações , Pneumonia Bacteriana/tratamento farmacológico , Pneumonia Bacteriana/epidemiologia , Pneumonia Viral , Pró-Calcitonina/metabolismo , Radiografia Torácica , Punção Espinal , Combinação Trimetoprima e Sulfametoxazol , Urinálise , Infecções Urinárias/complicações , Infecções Urinárias/tratamento farmacológico , Infecções Urinárias/epidemiologia
17.
J Oleo Sci ; 69(5): 467-477, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32378550

RESUMO

Esterases catalyze the hydrolysis of ester bonds in fatty acid esters with short-chain acyl groups. In the present study, thirty-seven bacterial isolates were isolated from soil contaminated with waste cooking oil, dairy waste etc. from Shimla and Solan district of H.P. Out of 37 isolates, the isolate RL-1, which gave maximum activity, was identified as Bacillus licheniformis MH061919. The optimization of various production parameters resulted in maximum activity at inoculum age of 24 h and inoculum size of 1.5% (v/v). Esterase gave considerable activity in production medium containing sodium chloride (0.5 % w/v), galactose (1%, w/v), coconut oil (2.0%, v/v) and beef extract (0.3%, w/v) at a temperature of 45℃ and pH 8.5.The enzyme production was enhanced by 3-fold after optimization of production parameters. Further, on optimizing reaction conditions, enzyme gave maximum activity at a temperature of 45℃ and pH 8.5. The para-nitrophenyl acetate (p-NPA) was found to be optimum substrate and metal ions and detergents have inhibitory effect on esterase activity.


Assuntos
Bacillus/enzimologia , Meios de Cultura/química , Técnicas de Cultura/métodos , Esterases/metabolismo , Bacillus/isolamento & purificação , Óleo de Coco , Galactose , Concentração de Íons de Hidrogênio , Nitrofenóis/metabolismo , Carne Vermelha , Cloreto de Sódio , Temperatura , Extratos de Tecidos
18.
Am J Trop Med Hyg ; 103(1): 325-333, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32431272

RESUMO

Schistosomiasis is a parasitic infection highly prevalent in Central Africa where it is co-endemic with many other parasitic infections, including soil-transmitted helminths (STHs). For its optimal control, there is a need of descriptive epidemiological data for each endemic region. The objective of the present study was to determine the epidemiological situation around schistosomiasis in Lambaréné, Gabon. A cross-sectional study was conducted among schoolchildren. One urine sample per day was collected on three consecutive days for the diagnosis of schistosomiasis using a urine filtration technique. One stool sample was collected for the detection of Schistosoma spp. and STH spp. eggs using the Kato-Katz technique, and for larvae, using the coproculture technique. A total of 614 schoolchildren were included in the analysis. The overall prevalence of schistosomiasis and STH infections was 26% (159/614) and 15% (70/473), respectively. Human-freshwater contact was the main risk factor for schistosomiasis in the area (relative risk (RR) = 2.96 [2.20-4.00], P < 0.001). Hematuria (RR = 5.53 [4.30-7.10], P < 0.001) and proteinuria (RR = 2.12 [1.63-2.75], P < 0.001) as well as infection with Trichuris trichiura (RR = 1.86 [1.33-2.61], P = 0.002) and Ascaris lumbricoides (RR = 1.96 [1.19-3.21], P = 0.039) were associated with an increased risk of schistosomiasis. Trichuris trichiura was the highest prevalent STH species in the area. Our study reports a moderate prevalence for schistosomiasis with human-water contact as the main risk factor, whereas the prevalence of STH infections appears to be low. Our results stress the need for the implementation of WHO recommendations for schistosomiasis control.


Assuntos
Ascaríase/epidemiologia , Esquistossomose Urinária/epidemiologia , Tricuríase/epidemiologia , Adolescente , Albendazol/uso terapêutico , Anti-Helmínticos/uso terapêutico , Ascaríase/tratamento farmacológico , Criança , Coinfecção/epidemiologia , Estudos Transversais , Técnicas de Cultura , Fezes/parasitologia , Feminino , Gabão/epidemiologia , Hematúria/epidemiologia , Infecções por Uncinaria/tratamento farmacológico , Infecções por Uncinaria/epidemiologia , Humanos , Masculino , Praziquantel/uso terapêutico , Prevalência , Proteinúria/epidemiologia , Fatores de Risco , Esquistossomose Urinária/tratamento farmacológico , Estrongiloidíase/tratamento farmacológico , Estrongiloidíase/epidemiologia , Tricuríase/tratamento farmacológico
19.
Trends Parasitol ; 36(6): 512-519, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32360314

RESUMO

Trager and Jensen established a method for culturing Plasmodium falciparum, a breakthrough for malaria research worldwide. Since then, multiple attempts to establish Plasmodium vivax in continuous culture have failed. Unlike P. falciparum, which can invade all aged erythrocytes, P. vivax is restricted to reticulocytes. Thus, a constant supply of reticulocytes is considered critical for continuous P. vivax growth in vitro. A critical question remains why P. vivax selectively invades reticulocytes? What do reticulocytes offer to P. vivax that is not present in mature erythrocytes? One possibility is protection from oxidative stress by glucose-6-phosphate dehydrogenase (G6PD). Here, we also suggest supplements to the media and procedures that may reduce oxidative stress and, as a result, establish a system for the continuous culture of P. vivax.


Assuntos
Técnicas de Cultura/normas , Estágios do Ciclo de Vida/fisiologia , Plasmodium vivax/crescimento & desenvolvimento , Reticulócitos/parasitologia , Técnicas de Cultura/tendências , Eritrócitos/enzimologia , Eritrócitos/parasitologia , Interações Hospedeiro-Parasita , Humanos , Estresse Oxidativo , Reticulócitos/enzimologia
20.
Am J Trop Med Hyg ; 103(2): 696-703, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32458777

RESUMO

Despite the implementation of effective conjugate vaccines against the three main bacterial pathogens that cause meningitis, Streptococcus pneumoniae, Haemophilus influenzae type b (Hib), and Neisseria meningitidis serogroup A, the burden of meningitis in West Africa remains high. The relative importance of other bacterial, viral, and parasitic pathogens in central nervous system infections is poorly characterized. Cerebrospinal fluid (CSF) specimens were collected from children younger than 5 years with suspected meningitis, presenting at pediatric teaching hospitals across West Africa in five countries including Senegal, Ghana, Togo, Nigeria, and Niger. Cerebrospinal fluid specimens were initially tested using bacteriologic culture and a triplex real-time polymerase chain reaction (PCR) assay for N. meningitidis, S. pneumoniae, and H. influenzae used in routine meningitis surveillance. A custom TaqMan Array Card (TAC) assay was later used to detect 35 pathogens including 15 bacteria, 17 viruses, one fungus, and two protozoans. Among 711 CSF specimens tested, the pathogen positivity rates were 2% and 20% by the triplex real-time PCR (three pathogens) and TAC (35 pathogens), respectively. TAC detected 10 bacterial pathogens, eight viral pathogens, and Plasmodium. Overall, Escherichia coli was the most prevalent (4.8%), followed by S. pneumoniae (3.5%) and Plasmodium (3.5%). Multiple pathogens were detected in 4.4% of the specimens. Children with human immunodeficiency virus (HIV) and Plasmodium detected in CSF had high mortality. Among 220 neonates, 17% had at least one pathogen detected, dominated by gram-negative bacteria. The meningitis TAC enhanced the detection of pathogens in children with meningitis and may be useful for case-based meningitis surveillance.


Assuntos
Infecções por Escherichia coli/epidemiologia , Malária Cerebral/epidemiologia , Meningite Pneumocócica/epidemiologia , Meningite/epidemiologia , Meningite/microbiologia , África Ocidental/epidemiologia , Pré-Escolar , Técnicas de Cultura , Infecções por Citomegalovirus/líquido cefalorraquidiano , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/epidemiologia , Infecções por Escherichia coli/líquido cefalorraquidiano , Infecções por Escherichia coli/diagnóstico , Feminino , Gana/epidemiologia , Infecções por HIV/líquido cefalorraquidiano , Infecções por HIV/diagnóstico , Infecções por HIV/epidemiologia , Vacinas Anti-Haemophilus/uso terapêutico , Humanos , Lactente , Recém-Nascido , Infecções por Klebsiella/líquido cefalorraquidiano , Infecções por Klebsiella/diagnóstico , Infecções por Klebsiella/epidemiologia , Malária Cerebral/líquido cefalorraquidiano , Malária Cerebral/diagnóstico , Masculino , Meningite/líquido cefalorraquidiano , Meningite/diagnóstico , Meningite por Haemophilus/líquido cefalorraquidiano , Meningite por Haemophilus/epidemiologia , Meningite por Haemophilus/prevenção & controle , Meningite Meningocócica/líquido cefalorraquidiano , Meningite Meningocócica/epidemiologia , Meningite Meningocócica/prevenção & controle , Meningite Pneumocócica/líquido cefalorraquidiano , Meningite Pneumocócica/prevenção & controle , Vacinas Meningocócicas/uso terapêutico , Técnicas de Diagnóstico Molecular , Mortalidade , Reação em Cadeia da Polimerase Multiplex , Níger/epidemiologia , Nigéria/epidemiologia , Vacinas Pneumocócicas/uso terapêutico , Reação em Cadeia da Polimerase em Tempo Real , Infecções por Roseolovirus/líquido cefalorraquidiano , Infecções por Roseolovirus/diagnóstico , Infecções por Roseolovirus/epidemiologia , Senegal/epidemiologia , Infecções Estafilocócicas/líquido cefalorraquidiano , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/epidemiologia , Togo/epidemiologia
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