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1.
Parasitol Res ; 123(9): 320, 2024 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-39254766

RESUMO

Cutaneous leishmaniasis (CL) stands out as a significant vector-borne endemic in Pakistan. Despite the rising incidence of CL, the genetic diversity of Leishmania species in the country's endemic regions remains insufficiently explored. This study aims to uncover the genetic diversity and molecular characteristics of Leishmania species in CL-endemic areas of Baluchistan, Khyber Pakhtunkhwa (KPK), and Punjab in Pakistan. Clinical samples from 300 CL patients were put to microscopic examination, real-time ITS-1 PCR, and sequencing. Predominantly affecting males between 16 to 30 years of age, with lesions primarily on hands and faces, the majority presented with nodular and plaque types. Microscopic analysis revealed a positivity rate of 67.8%, while real-time PCR identified 60.98% positive cases, mainly L. tropica, followed by L. infantum and L. major. Leishmania major (p = 0.009) showed substantially greater variation in nucleotide sequences than L. tropica (p = 0.07) and L. infantum (p = 0.03). Nucleotide diversity analysis indicated higher diversity in L. major and L. infantum compared to L. tropica. This study enhances our understanding of CL epidemiology in Pakistan, stressing the crucial role of molecular techniques in accurate species identification. The foundational data provided here emphasizes the necessity for future research to investigate deeper into genetic diversity and its implications for CL control at both individual and community levels.


Assuntos
Variação Genética , Leishmaniose Cutânea , Leishmaniose Cutânea/epidemiologia , Leishmaniose Cutânea/parasitologia , Paquistão/epidemiologia , Humanos , Masculino , Adolescente , Adulto , Feminino , Adulto Jovem , Criança , Pessoa de Meia-Idade , Leishmania/genética , Leishmania/classificação , Leishmania/isolamento & purificação , Pré-Escolar , Análise de Sequência de DNA , Leishmania tropica/genética , Leishmania tropica/isolamento & purificação , Leishmania tropica/classificação , Leishmania major/genética , Leishmania major/classificação , Leishmania major/isolamento & purificação , DNA de Protozoário/genética , Filogenia , Epidemiologia Molecular , Idoso , Reação em Cadeia da Polimerase em Tempo Real
2.
Genes Dis ; 11(6): 101143, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39253579

RESUMO

Pancreatic cancer, a highly fatal malignancy, is predicted to rank as the second leading cause of cancer-related death in the next decade. This highlights the urgent need for new insights into personalized diagnosis and treatment. Although molecular subtypes of pancreatic cancer were well established in genomics and transcriptomics, few known molecular classifications are translated to guide clinical strategies and require a paradigm shift. Notably, chronically developing and continuously improving high-throughput technologies and systems serve as an important driving force to further portray the molecular landscape of pancreatic cancer in terms of epigenomics, proteomics, metabonomics, and metagenomics. Therefore, a more comprehensive understanding of molecular classifications at multiple levels using an integrated multi-omics approach holds great promise to exploit more potential therapeutic options. In this review, we recapitulated the molecular spectrum from different omics levels, discussed various subtypes on multi-omics means to move one step forward towards bench-to-beside translation of pancreatic cancer with clinical impact, and proposed some methodological and scientific challenges in store.

3.
Vet Res Commun ; 2024 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-39264573

RESUMO

A polymerase chain reaction (PCR) survey was performed at an amateur parrot breeding facility in Italy to investigate the presence and molecular characteristics of adenoviruses. Eighty psittacine birds, belonging to seven parrot species, were sampled by cloacal swabs; in addition, 15 livers were collected from specimens that were found dead. Seventy-two out of 95 samples collected were positive for adenoviruses, with a prevalence rate of 75.8%. All seven psittacine species tested positive for at least one genus of the family Adenoviridae; notably, adenoviral infection was found for the first time in the hooded parrot (Psephotellus dissimilis). Based on the sequences and phylogenetic analysis, 57 sequences were psittacine adenovirus 2, seven sequences were duck adenovirus 1 and two sequences were identified as psittacine adenovirus 5. The six remaining sequences showed low nucleotide and amino acid identity with the reference strains of accepted species or types, revealing the presence of novel adenoviruses belonging to the genera Aviadenovirus, Barthadenovirus and Siadenovirus. There were identical adenovirus sequences in both apparently healthy and dead birds suggesting that further investigation into the role and impact of these viruses on the health of psittacine birds is warranted.

4.
Transgenic Res ; 2024 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-39249190

RESUMO

Dehydroascorbate reductase (DHAR), an indispensable enzyme in the production of ascorbic acid (AsA) in plants, is vital for plant tolerance to various stresses. However, there is limited research on the stress tolerance functions of DHAR genes in sweet potato (Ipomoea batatas [L.] Lam). In this study, the full-length IbDHAR1 gene was cloned from the leaves of sweet potato cultivar Xu 18. The IbDHAR1 protein is speculated to be located in both the cytoplasm and the nucleus. As revealed by qRT-PCR, the relative expression level of IbDHAR1 in the proximal storage roots was much greater than in the other tissues, and could be upregulated by high-temperature, salinity, drought, and abscisic acid (ABA) stress. The results of pot experiments indicated that under high salinity and drought stress conditions, transgenic Arabidopsis and sweet potato plants exhibited decreases in H2O2 and MDA levels. Conversely, the levels of antioxidant enzymes APX, SOD, POD, and ACT, and the content of DHAR increased. Additionally, the ratio of AsA/DHA was greater in transgenic lines than in the wild type. The results showed that overexpression of IbDHAR1 intensified the ascorbic acid-glutathione cycle (AsA-GSH) and promoted the activity of the related antioxidant enzyme systems to improve plant stress tolerance and productivity.

5.
Front Bioeng Biotechnol ; 12: 1458362, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39295845

RESUMO

Urinary cell-free DNA (UcfDNA) is gaining recognition as an important biomarker for diagnosing bladder cancer. UcfDNA contains tumor derived DNA sequences, making it a viable candidate for non-invasive early detection, diagnosis, and surveillance of bladder cancer. The quantification and qualification of UcfDNA have demonstrated high sensitivity and specificity in the molecular characterization of bladder cancer. However, precise analysis of UcfDNA for clinical bladder cancer diagnosis remains challenging. This review summarizes the history of UcfDNA discovery, its biological properties, and the quantitative and qualitative evaluations of UcfDNA for its clinical significance and utility in bladder cancer patients, emphasizing the critical role of UcfDNA in bladder cancer diagnosis. Emerging bioactive technologies and materials currently offer promising tools for multiple UcfDNA analysis, aiming to achieve more precise and efficient capture of UcfDNA, thereby significantly enhancing diagnostic accuracy. This review also highlights breakthroughs in detection technologies and substrates with the potential to revolutionize bladder cancer diagnosis in clinic.

6.
One Health ; 19: 100869, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-39220760

RESUMO

Fascioliasis, only foodborne trematodiasis of worldwide distribution, is caused by Fasciola hepatica and F. gigantica, liver flukes transmitted by freshwater snails. Southern and southeastern Asia is an emerging hot spot of F. gigantica, despite its hitherto less involvement in human infection. In Vietnam, increasing cases have been reported since 1995, whereas only sixteen throughout 1800-1994. A database was created to include epidemiological data of fascioliasis patients from the 63 Vietnam provinces throughout 1995-2019. Case profiles were based on serology, symptoms, eosinophilia, imaging techniques, stool egg finding, and post-specific-treatment recovery. Radio broadcasting about symptoms and costless diagnosis/treatment led patients to hospitals after symptom onset. Yearly case numbers were modelled and spatio-temporally analyzed. Missing data and confounders were assessed. The countrywide spread has no precedent. It started in the central coast, including 53,109 patients, mostly adults and females. Seasonality, linked to vegetable consumption, peaks in June, although the intensity of this peak differs according to relief/climatic zones. Incidence data and logistic regression curves are obtained for the first time in human fascioliasis. Fasciolid hybrids accompanying the spreading F. gigantica flukes, and climate change assessed by risk index correlations, are both ruled out as outbreak causes. Human-guided movements of livestock from an original area prove to be the way used by fasciolids and lymnaeid vectors to expand geographically. Radix viridis, a highly efficient transmitting and colonizing vector, played a decisive role in the spread. The use of irrigated crop fields, widely inhabited by R. viridis, for livestock grazing facilitated the transmission and spread of the disease. General physician awareness and diagnostic capacity improvement proved the successful impact of such knowledge transfer in facilitating and increasing patient infection detection. Information, education and communication to the public by radio broadcasting demonstrated to be very helpful. Fasciola gigantica is able to cause epidemic and endemic situations similar to F. hepatica. The magnitude of the human outbreak in Vietnam is a health wake-up call for southern and southeastern countries of Asia which present the highest human population densities with increasing food demands, uncontrolled livestock inter-country exchange, foreign import practices, and monsoon's increasing climate change impact.

7.
BMC Med ; 22(1): 352, 2024 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-39218882

RESUMO

BACKGROUND: The radiogenomic analysis has provided valuable imaging biomarkers with biological insights for gliomas. The radiogenomic markers for molecular profile such as DNA methylation remain to be uncovered to assist the molecular diagnosis and tumor treatment. METHODS: We apply the machine learning approaches to identify the magnetic resonance imaging (MRI) features that are associated with molecular profiles in 146 patients with gliomas, and the fitting models for each molecular feature (MoRad) are developed and validated. To provide radiological annotations for the molecular profiles, we devise two novel approaches called radiomic oncology (RO) and radiomic set enrichment analysis (RSEA). RESULTS: The generated MoRad models perform well for profiling each molecular feature with radiomic features, including mutational, methylation, transcriptional, and protein profiles. Among them, the MoRad models have a remarkable performance in quantitatively mapping global DNA methylation. With RO and RSEA approaches, we find that global DNA methylation could be reflected by the heterogeneity in volumetric and textural features of enhanced regions in T2-weighted MRI. Finally, we demonstrate the associations of global DNA methylation with clinicopathological, molecular, and immunological features, including histological grade, mutations of IDH and ATRX, MGMT methylation, multiple methylation-high subtypes, tumor-infiltrating lymphocytes, and long-term survival outcomes. CONCLUSIONS: Global DNA methylation is highly associated with radiological profiles in glioma. Radiogenomic global methylation is an imaging-based quantitative molecular biomarker that is associated with specific consensus molecular subtypes and immune features.


Assuntos
Neoplasias Encefálicas , Metilação de DNA , Glioma , Imageamento por Ressonância Magnética , Humanos , Glioma/genética , Glioma/imunologia , Metilação de DNA/genética , Feminino , Masculino , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/imunologia , Neoplasias Encefálicas/patologia , Pessoa de Meia-Idade , Adulto , Aprendizado de Máquina , Fenótipo , Idoso , Biomarcadores Tumorais/genética
8.
Acta Parasitol ; 2024 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-39190275

RESUMO

PURPOSE: The identification of the external attaching fish parasitic cymothoid, Nerocila phaiopleura Bleeker 1857, is still based on the brief description of Australian specimens provided by Bruce (1987). The present study aimed to provide a redescription and molecular characterisation of Indian specimens of N. phaiopleura. MATERIALS AND METHODS: Morphological identification was carried out based on microscopic examinations and taxonomic drawings. mitochondrial DNA cox1 was selected as the target gene for sequencing and molecular identification. Nucleotide genetic divergence (p-distance) and base-pair differences among the different species were determined using MEGA11. RESULTS: Nerocila phaiopleura can be well separated from its congeners by the following combination of characteristics: Body about 2.4 times as long as wide, cephalon broadly rounded anteriorly; coxae posteriorly directed, acute and extending beyond their corresponding pereonite; pereonite 7 posterior angle produced, extending to the pleonite 1; pleonites 1 and 2 ventrolateral process posteriorly directed; uropod exopod straight and elongate, 1.7-2.0 times longer than endopod; uropod endopod lateral margin not serrate, no notch on medial margin; pereopods with short ischium; pleotelson triangular. The p-distance among N. phaiopleura and other available Nerocila spp. ranged from 21 to 19%. CONCLUSION: This study represents the first detailed taxonomic redescription of Indian specimens of N. phaiopleura. Key taxonomic features of the life stages and molecular data are provided here to identify the species properly. Interspecific genetic divergence between N. phaiopleura and other Nerocila spp. is assessed for the first time. Studies in cymothoid life histories, genetics, and morphology are necessary to understand one of the least understood parasite families.

9.
ACS Nano ; 18(34): 23301-23309, 2024 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-39151088

RESUMO

Nucleic acid nanoparticles (NANPs) are increasingly used in preclinical investigations as delivery vectors. Tools that can characterize assembly and assess quality will accelerate their development and clinical translation. Standard techniques used to characterize NANPs, like gel electrophoresis, lack the resolution for precise characterization. Here, we introduce the use of charge detection mass spectrometry (CD-MS) to characterize these materials. Using this technique, we determined the mass of NANPs varying in size, shape, and molecular mass, NANPs varying in production quality due to formulations lacking component oligonucleotides, and NANPs functionalized with protein and nucleic acid-based secondary molecules. Based on these demonstrations, CD-MS is a promising tool to precisely characterize NANPs, enabling more precise assessments of the manufacturing and processing of these materials.


Assuntos
Espectrometria de Massas , Nanopartículas , Ácidos Nucleicos , Nanopartículas/química , Ácidos Nucleicos/química , Ácidos Nucleicos/análise , Tamanho da Partícula , DNA/química
10.
Genes (Basel) ; 15(8)2024 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-39202403

RESUMO

MicroRNA482 (miR482) is a conserved microRNA family in plants, playing critical regulatory roles in different biological activities. Though the members of the miR482 gene family have been identified in plants, a systematic study has not been reported yet. In the present research, 140 mature sequences generated by 106 precursors were used for molecular characterization, phylogenetic analysis, and target gene prediction, and the competing endogenous RNA (ceRNA) network mediated by miR482 was summarized. The length of mature sequences ranged from 17 nt to 25 nt, with 22 nt being the most abundant, and the start and end of the mature sequences had a preference for uracil (U). By sequence multiplex comparison, it was found that the mature sequences of 5p were clustered into one group, and others were clustered into the other group. Phylogenetic analysis revealed that the 140 mature sequences were categorized into six groups. Meanwhile, all the precursor sequences formed a stable hairpin structure, and the 106 precursors were divided into five groups. However, the expression of miR482 varied significantly between different species and tissues. In total, 149 target genes were predicted and their functions focused on single-organism process, cellular process, and cell and cell part. The ceRNA network of miR482 in tomato, cotton, and peanut was summarized based on related publications. In conclusion, this research will provide a foundation for further understanding of the miR482 gene family.


Assuntos
Regulação da Expressão Gênica de Plantas , MicroRNAs , Família Multigênica , Filogenia , MicroRNAs/genética , RNA de Plantas/genética , Plantas/genética , Redes Reguladoras de Genes , Solanum lycopersicum/genética
11.
J Appl Microbiol ; 135(8)2024 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-39108074

RESUMO

AIMS: To evaluate the prevalence, molecular characteristics, antimicrobial susceptibility, and epithelial invasion of Streptococcus agalactiae strains isolated from pregnant women and newborns in Rio de Janeiro, Brazil. METHODS AND RESULTS: A total of 67 S. agalactiae isolates, 48 isolates from pregnant women and 19 from neonates, were analyzed. Capsular type Ia and V were predominant (35.8%/each). The multilocus sequence typing analysis revealed the presence of 19 STs grouped into 6 clonal complexes with prevalence of CC17/40.3% and CC23/34.3%. The lmb and iag virulence genes were found in 100% of isolates. Four S. agalactiae strains, belonging to CC17/ST1249 and CC23/ST23, were able to adhere to A549 respiratory epithelial cells. Antimicrobial resistance was verified mainly to tetracycline (85%), erythromycin (70.8%), and clindamycin (58.3%). Four S. agalactiae isolates were multidrug resistant. The resistance genes tested were found in 92.5% of isolates for tetM, 58.2% for ermB, 28.4% for mefAE, and 10.4% for tetO. CONCLUSION: The study showed a high prevalence of virulence and antimicrobial genes in S. agalactiae strains isolated from pregnant women and newborns, supporting the idea that continued surveillance is necessary to identify risk factors and perform long-term follow-up in pregnant women and neonates in Rio de Janeiro.


Assuntos
Antibacterianos , Células Epiteliais , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Infecções Estreptocócicas , Streptococcus agalactiae , Streptococcus agalactiae/genética , Streptococcus agalactiae/efeitos dos fármacos , Streptococcus agalactiae/isolamento & purificação , Feminino , Humanos , Brasil , Gravidez , Infecções Estreptocócicas/microbiologia , Antibacterianos/farmacologia , Recém-Nascido , Células Epiteliais/microbiologia , Farmacorresistência Bacteriana/genética , Adulto , Fatores de Virulência/genética , Complicações Infecciosas na Gravidez/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Virulência/genética
12.
Fish Shellfish Immunol ; 153: 109838, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39151839

RESUMO

IL-26 is a crucial inflammatory cytokine that participates in defending host cells against infections. We initially cloned and identified the cDNA sequences of interleukin (IL)-26 in channel catfish (Ictalurus punctatus). The open reading frame (ORF) of IpIL-26 was 537 bp in length, encoding 178 amino acids (aa). Constitutive expression of IpIL-26 was observed in tested tissues, with the highest level found in the gill and spleen. To explore the function of IpIL-26 in channel catfish, different stimuli were used to act on both channel catfish and channel catfish kidney cells (CCK). The expression of IpIL-26 could be up-regulated by bacteria and viruses in multiple tissues. In vitro, recombinant IpIL-26 (rIpIL-26) could induce the expression levels of inflammatory cytokines such as TNF-α, IL-1ß, IL-6, IL-20, and IL-22 playing vital roles in defending the host against infections. Our results demonstrated that IpIL-26 might be an essential cytokine, significantly affecting the immune defense of channel catfish against pathogen infections.


Assuntos
Sequência de Aminoácidos , Doenças dos Peixes , Proteínas de Peixes , Regulação da Expressão Gênica , Ictaluridae , Imunidade Inata , Interleucinas , Filogenia , Alinhamento de Sequência , Animais , Ictaluridae/imunologia , Ictaluridae/genética , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Proteínas de Peixes/química , Interleucinas/genética , Interleucinas/imunologia , Imunidade Inata/genética , Doenças dos Peixes/imunologia , Alinhamento de Sequência/veterinária , Regulação da Expressão Gênica/imunologia , Perfilação da Expressão Gênica/veterinária , Sequência de Bases , DNA Complementar/genética
13.
Pathogens ; 13(8)2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39204256

RESUMO

The cereal cyst nematode (Heterodera latipons) is becoming an economically important species in global cereal production as it is being identified in many new cereal cultivated areas and causes significant losses. Consequently, understanding its biology becomes crucial for researchers in identifying its vulnerabilities and implementing effective control measures. In the current study, different morphological and biochemical changes of H. latipons cysts containing eggs with infective juveniles from a barley field in Jordan were studied during the summer of 2021, at two sample dates. The first, at the harvest of the cereal crop (June 2021), when the infective second-stage juveniles (J2s) were initiating diapause, and the second, before planting the sequent cereal crop (late October 2021), when the J2s were ending diapause. The studied population was characterized morphologically and molecularly, showing 98.4% molecular similarity to both JOD from Jordan and Syrian "300" isolates of H. latipons. The obtained results and observations revealed that there were dramatic changes in all the investigated features of the cysts and eggs they contained. Morphological changes such as cyst color, sub-crystalline layer, and thickness of the rigid eggshell wall were observed. A slight change in the emergence time of J2s from cysts was observed without any difference in the number of emerged J2s. The results of biochemical changes showed that the total contents of carbohydrates, glycogen, trehalose, glycerol, and protein were higher in cysts collected in October when compared to those cysts collected in June. The SDS-PAGE pattern indicated the presence of a protein with the size of ca. 100 kDa in both sampling dates, whereas another protein (ca. 20 kDa) was present only in the cysts of October. Furthermore, the expression of trehalase (tre) gene was detected only in H. latipons collected in October. The outcomes of this study provide new helpful information that elucidates diapause in H. latipons and may be used for the implementation of new management strategies of cyst nematodes.

14.
J Med Cases ; 15(9): 250-255, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39205695

RESUMO

The identification of a NUP214-ABL1 fusion has been seen in about 6% of patients with T lymphoblastic leukemia (T-ALL). It has been described at a lower frequency in B-lymphoblastic leukemia (B-ALL) patients as well. To our knowledge, this is the first case report documenting a NUP214-ABL1 fusion in a patient with newly diagnosed myelodysplastic syndrome (MDS) as identified by next-generation sequencing (NGS). A case report by Wang et al recently described a case report of the first NUP214-ABL1 fusion in a patient with newly diagnosed acute myeloid leukemia (AML). This shows that this specific translocation is not isolated to lymphoid malignancies, and can be associated with myeloid malignancies as well. The potential use of tyrosine kinase inhibitors (TKIs) as a line of treatment for patients who harbor this translocation makes this finding of particular interest. However, while there have been individual reports noting the effect of TKIs in T-ALLs with NUP214-ABL1 fusions, additional research is needed to fully understand the role of this mutation in myeloid derived malignancies, and its corresponding treatment and prognostic implications.

15.
J Med Virol ; 96(9): e29896, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39206836

RESUMO

Respiratory syncytial virus is a major causative agent of lower respiratory tract infection in children, especially infants with substantial morbidity and mortality implications. The virus undergoes continuous evolution documented by accumulation of mutations in the glycoprotein gene necessitating vigilant surveillance to provide essential data to epidemiologists and researchers involved in development of vaccines. This study was aimed to perform molecular characterization of respiratory syncytial virus (RSV) among children ≤ 5 years admitted in hospital. In the current study we observed RSV-A (2019 (n = 95) and 2021 (n = 61) seasons) and RSV-B (2022 season (n = 68)). Phylogenetic analysis revealed all RSV-A strains (n = 47) to be GA.2.3.5 and RSV-B (n = 22) were classified as GB.5.0.5a. Selection pressure analysis identified one positive (P274L/V) and one negative site (P230T) in RSV-A, while in RSV-B there was only one negatively selected site (S295). This study spanning over three seasons contributes to RSV evolutionary dynamics in India emphasizing the importance of on-going surveillance to inform effective public health strategies and vaccine development efforts.


Assuntos
Filogenia , Infecções por Vírus Respiratório Sincicial , Vírus Sincicial Respiratório Humano , Infecções Respiratórias , Humanos , Infecções por Vírus Respiratório Sincicial/epidemiologia , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/classificação , Vírus Sincicial Respiratório Humano/isolamento & purificação , Lactente , Pré-Escolar , Infecções Respiratórias/virologia , Infecções Respiratórias/epidemiologia , Feminino , Masculino , Índia/epidemiologia , Estações do Ano , Hospitalização/estatística & dados numéricos , Recém-Nascido , Genótipo
16.
Int J Mol Sci ; 25(16)2024 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-39201488

RESUMO

The plerocercoid larva of Spirometra mansoni can cause a parasitic zoonosis-sparganosis. Malate dehydrogenase (MDH) plays a very important role in the life activities of parasites. However, little is known about the MDH family in S. mansoni. We identified eight new MDH members in S. mansoni in this study. Clustering analysis divided SmMDHs into two groups and revealed patterns similar to the conserved motif organization. RT-qPCR suggested that five MDHs were highly expressed in the mature proglottid and that three MDHs were highly expressed in the gravid proglottid. Phylogenetic analysis revealed that SmMDHs contain both conserved family members and members in the process of further diversification. rSmMDH has an NAD binding domain, a dimer interface and a substrate binding domain. Natural SmMDH was immunolocalized in the tissues and follicles around the uterus in the mature or gravid proglottid and eggshells. The maximum forward and reverse reaction activities of rSmMDH were observed at pH 8.5 and 9.0, respectively. The optimum temperature for enzyme activity was 37 °C in the forward reaction and 40 °C in the reverse reaction. These results lay the foundation for studying the molecular functions and mechanisms of MDHs in S. mansoni and related taxa.


Assuntos
Malato Desidrogenase , Filogenia , Spirometra , Animais , Spirometra/genética , Spirometra/enzimologia , Malato Desidrogenase/genética , Malato Desidrogenase/metabolismo , Malato Desidrogenase/química , Proteínas de Helminto/genética , Proteínas de Helminto/química , Proteínas de Helminto/metabolismo , Família Multigênica , Sequência de Aminoácidos
17.
Int J Food Microbiol ; 423: 110845, 2024 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-39079449

RESUMO

The primary objective of this study was to characterize lactic acid bacteria (LAB) strains derived from sourdough for possible utilization as functional starters to produce sourdough and various cereal-based fermented foods. A total of 350 autochthonous LAB strains were isolated from 65 Type I sourdough samples and characterized using six random amplified polymorphic DNA (RAPD) primers at intra- and interspecific levels. Species identification of selected strains representing distinct clusters from RAPD analysis was performed based on the 16S rRNA region. The LAB strains were identified as Companilactobacillus crustorum (n = 135), Levilactobacillus brevis (n = 125), Latilactobacillus curvatus (n = 40), Companilactobacillus paralimentarius (n = 32), and Lactiplantibacillus plantarum (n = 18). A total of 66 LAB strains were selected for technological characterization along with two commercial strains. The characterization involved acidity development, EPS production potential, leavening activity, and growth abilities under harsh conditions. Principle component analysis (PCA) identified 2 Lp. plantarum and 14 Lev. brevis strains as the most relevant technologically. Among them, Lp. plantarum L35.1 and Lev. brevis L37.1 were resistant to tetracycline. Evaluation of probiotic characteristics (survival in pH 2.5 and bile presence, auto aggregation capacity, hydrophobic activity, antioxidant activity, antimicrobial activity) by PCA identified four strains with relevance to Lactobacillus rhamnosus GG (LGG), which were further selected for in vitro digestion assays. Lactiplantibacillus plantarum L7.8, Lev. brevis L55.1, and L62.2 demonstrated similar viability indices to LGG, along with increased auto aggregation capacity and antioxidant activity. These strains are promising as candidate starters for producing sourdough and sourdough-related fermented food products.


Assuntos
Pão , Fermentação , Microbiologia de Alimentos , Técnica de Amplificação ao Acaso de DNA Polimórfico , Pão/microbiologia , RNA Ribossômico 16S/genética , Alimentos Fermentados/microbiologia , Lactobacillales/genética , Lactobacillales/isolamento & purificação , Lactobacillales/classificação , Lactobacillales/metabolismo , Filogenia , Antibacterianos/farmacologia , DNA Bacteriano/genética , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Lactobacillus/classificação , Lactobacillus/metabolismo
18.
Genes (Basel) ; 15(7)2024 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-39062736

RESUMO

The spurge Euphorbia characias is known for its latex, which is rich in antioxidant enzymes and anti-phytopathogen molecules. In this study, we identified a novel polyubiquitin protein in the latex and leaves, leading to the first molecular characterization of its coding gene and expressed protein in E. characias. Using consensus-degenerate hybrid oligonucleotide primers (CODEHOP) and rapid amplification of cDNA ends (5'/3'-RACE), we reconstructed the entire open reading frame (ORF) and noncoding regions. Our analysis revealed that the polyubiquitin gene encodes five tandemly repeated sequences, each coding for a ubiquitin monomer with amino acid variations in four of the five repeats. In silico studies have suggested functional differences among monomers. Gene expression peaked during the summer, correlating with high temperatures and suggesting a role in heat stress response. Western blotting confirmed the presence of polyubiquitin in the latex and leaf tissues, indicating active ubiquitination processes. These findings enhance our understanding of polyubiquitin's regulatory mechanisms and functions in E. characias, highlighting its unique structural and functional features.


Assuntos
Euphorbia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Poliubiquitina , Euphorbia/genética , Poliubiquitina/genética , Poliubiquitina/metabolismo , Proteínas de Plantas/genética , Estações do Ano , Látex/metabolismo , Látex/química , Folhas de Planta/genética , Folhas de Planta/metabolismo , Filogenia
19.
PeerJ ; 12: e17518, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38952990

RESUMO

Potato farming is a vital component of food security and the economic stability especially in the under developing countries but it faces many challenges in production, blackleg disease caused by Pectobacterium atrosepticum (Pa) is one of the main reason for damaging crop yield of the potato. Effective management strategies are essential to control these losses and to get sustainable potato crop yield. This study was focused on characterizing the Pa and the investigating new chemical options for its management. The research was involved a systematic survey across the three district of Punjab, Pakistan (Khanewal, Okara, and Multan) to collect samples exhibiting the black leg symptoms. These samples were analyzed in the laboratory where gram-negative bacteria were isolated and identified through biochemical and pathogenicity tests for Pa. DNA sequencing further confirmed these isolates of Pa strains. Six different chemicals were tested to control blackleg problem in both vitro and vivo at different concentrations. In vitro experiment, Cordate demonstrated the highest efficacy with a maximum inhibition zones of 17.139 mm, followed by Air One (13.778 mm), Profiler (10.167 mm), Blue Copper (7.7778 mm), Spot Fix (7.6689 mm), and Strider (7.0667 mm). In vivo, Cordate maintained its effectiveness with the lowest disease incidence of 14.76%, followed by Blue Copper (17.49%), Air One (16.98%), Spot Fix (20.67%), Profiler (21.45%), Strider (24.99%), and the control group (43.00%). The results highlight Cordate's potential as a most effective chemical against Pa, offering promising role for managing blackleg disease in potato and to improve overall productivity.


Assuntos
Pectobacterium , Doenças das Plantas , Solanum tuberosum , Solanum tuberosum/microbiologia , Pectobacterium/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Paquistão
20.
Biofactors ; 2024 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-38989918

RESUMO

Extracellular vesicles are secreted by all eukaryotic cells and they have an important role in intercellular signaling. Plant extracellular vesicles (PEVs) are a novel area of research that has gained attention due to their potential implications in biomolecule transport and therapeutic applications. PEVs are lipid bilayer-enclosed structures that contain a diverse cargo of biomolecules such as proteins and lipids. Moreover, it is known that PEVs have a noticeable therapeutic potential for various conditions such as inflammation and oxidative stress. However, there are critical problems such as removing the endosomes and plant-derived biomolecules that decrease the standardization and therapeutic efficacy of PEVs. In our study, the aim was to characterize plant cell suspension-derived extracellular vesicles (PCSEVs) obtained from two different plant cell suspension cultures: Stevia rebaudiana and Vaccaria hispanica. These vesicles were isolated using ultrafiltration and characterized with nanoparticle tracking analysis (NTA) and atomic force microscopy (AFM). The molecular composition of PCSEVs was profiled and the cellular uptake assay was performed. Our results demonstrated that PCSEVs have a spherical shape, less than 200 nm. In the fatty acid analysis, the primary components in PCSEVs were palmitic acid, linoleic acid, and cis-vaccenic acid. The protein content of Stevia rebaudiana-derived EVs (SDEVs) was largely associated with proteins involved in extracellular structures and functions. Conversely, Vaccaria hispanica-derived EVs (HDEVs) displayed a higher presence of cytosolic proteins. These findings contribute to the understanding of PCSEVs and open up potential avenues in extracellular vesicle research, pointing to promising prospects for future innovations in various fields.

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