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1.
Emerg Infect Dis ; 30(4): 795-799, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38526241

RESUMO

Uncommon Salmonella Infantis variants displaying only flagellar antigens phenotypically showed identical incomplete antigenic formula but differed by molecular serotyping. Although most formed rough colonies, all shared antimicrobial resistances and the presence of usg gene with wild-type Salmonella Infantis. Moreover, they were undistinguishable wild-type Salmonella Infantis by whole-genome sequencing.


Assuntos
Cadeia Alimentar , Aves Domésticas , Animais , Itália/epidemiologia , Salmonella/genética , Sorotipagem
2.
Clin Genet ; 2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38840272

RESUMO

The current genetic diagnostic workup of congenital cataract (CC) is mainly based on NGS panels, whereas exome sequencing (ES) has occasionally been employed. In this multicentre study, we investigated by ES the detection yield, mutational spectrum and genotype-phenotype correlations in a CC cohort recruited between 2020 and mid-2022. The cohort consisted of 67 affected individuals from 51 unrelated families and included both non-syndromic (75%) and syndromic (25%) phenotypes, with extra-CC ocular/visual features present in both groups (48% and 76%, respectively). The functional effect of variants was predicted by 3D modelling and hydropathy properties changes. Variant clustering was used for the in-depth assessment of genotype-phenotype correlations. A diagnostic (pathogenic or likely pathogenic) variant was identified in 19 out of 51 probands/families (~37%). In a further 14 probands/families a candidate variant was identified: in 12 families a VUS was detected, of which 9 were considered plausibly pathogenic (i.e., 4 or 5 points according to ACMG criteria), while in 2 probands ES identified a single variant in an autosomal recessive gene associated with CC. Eighteen probands/families, manifesting primarily non-syndromic CC (15/18, 83%), remained unsolved. The identified variants (8 P, 12 LP, 10 VUS-PP, and 5 VUS), half of which were unreported in the literature, affected five functional categories of genes involved in transcription/splicing, lens formation/homeostasis (i.e., crystallin genes), membrane signalling, cell-cell interaction, and immune response. A phenotype-specific variant clustering was observed in four genes (KIF1A, MAF, PAX6, SPTAN1), whereas variable expressivity and potential phenotypic expansion in two (BCOR, NHS) and five genes (CWC27, KIF1A, IFIH1, PAX6, SPTAN1), respectively. Finally, ES allowed to detect variants in six genes not commonly included in commercial CC panels. These findings broaden the genotype-phenotype correlations in one of the largest CC cohorts tested by ES, providing novel insights into the underlying pathogenetic mechanisms and emphasising the power of ES as first-tier test.

3.
Plasmid ; 114: 102566, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33582117

RESUMO

Salmonella enterica subsp. enterica serovar Napoli (S. Napoli) ranks among the top serovars causing human infections in Italy, although not common in other European countries. Isolates are generally pan-susceptible or resistant to aminoglycosides only, however data on antimicrobial resistance genes in strains of S. Napoli are limited. Recently an isolate encoding resistance to third generation cephalosporins was reported. This study aimed to characterize plasmid-encoded cephalosporin resistance due to the blaCTX-M-15 gene in a human S. Napoli isolate in Italy, and to investigate plasmid stability over time. S. Napoli 16/174478 was confirmed to be ESBL-producing. The blaCTX-M-15 gene was shown to be located on an IncI1α plasmid of 90,272 bp (50.03 GC%) encoding for 107 coding sequences (CDS). The plasmid was successfully transferred by conjugation to an E. coli 1816 recipient strain (conjugation frequency 3.9 × 10-2 transconjugants per donor). Transconjugants were confirmed to carry the IncI1α plasmid, and to be ESBL-producing strains as well. Moreover, transconjugant colonies maintained the plasmid for up to 10 passages. The identification of S. Napoli isolates able to produce ESBLs is of great concern, as this pathogen is frequently associated with invasive infections and a higher risk of bacteraemia, and its reservoir has not yet been clearly identified.


Assuntos
Escherichia coli , Salmonella , Antibacterianos/farmacologia , Escherichia coli/genética , Humanos , Itália , Plasmídeos/genética , Salmonella/genética , Sorogrupo , beta-Lactamases/genética
4.
Fish Shellfish Immunol ; 119: 384-396, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34687879

RESUMO

Ascidians are marine invertebrate chordates belonging to the earliest branch (Tunicata) in the chordate phylum, therefore, they are of interest for studying the evolution of immune systems. Due to the known genome, the non-colonial Ciona robusta, previously considered to be C. intestinalis type A, is a model species for the study of inflammatory response. The internal defense of ascidians mainly relies on hemocytes circulating in the hemolymph and pharynx. Hemocytes can be in vivo challenged by LPS injection and various granulocyte and vacuolated cell populations differentiated to produce and release inflammatory factors. Molecular biology and gene expression studies revealed complex defense mechanisms involving different inflammatory hemocytes. Furthermore, cloning procedures allowed sequence analyses and molecular studies disclose immune-related gene families including TOLL-like receptors, galectins, C-type lectins, collectins, interlectins, pentraxine-like, peroxinectins, complement factors-like, TNFα-like, IL-17-like, TGF-like, MIF-like. These genes are promptly upregulated by the inflammatory stimulus and show a time course of transcription similar to each other. Domains sequence similarity and phylogenetic relationships with the vertebrate counterparts are shedding some light on immune-related gene evolution. Selective bioassays as well as bioinformatic approaches have allowed the characterization of antimicrobial peptides and the identification of post transcriptional molecular mechanisms able of influencing dynamics of gene regulation are described. In synthesis, the purpose of this article is to further explore the topic of hemocyte and molecules related to internal defence of ascidians involved in the inflammatory reaction, as well as to discuss current and future study options through a detailed literature review.


Assuntos
Ciona intestinalis , Animais , Peptídeos Antimicrobianos , Ciona intestinalis/imunologia , Hemócitos , Lipopolissacarídeos , Filogenia
5.
BMC Genomics ; 21(1): 202, 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32131727

RESUMO

BACKGROUND: Salmonella enterica subsp. enterica serovar Napoli (S. Napoli) is among the top serovars causing human infections in Italy, although it is relatively uncommon in other European countries; it is mainly isolated from humans and the environment, but neither the reservoir nor its route of infection are clearly defined. This serovar is characterized by high genomic diversity, and molecular evidences revealed important similarities with typhoidal serovars. RESULTS: 179 S. Napoli genomes as well as 239 genomes of typhoidal and non-typhoidal serovars were analyzed in a comparative genomic study. Phylogenetic analysis and draft genome characterization in terms of Multi Locus Sequence Typing (MLST), plasmid replicons, Salmonella Pathogenicity Islands (SPIs), antimicrobial resistance genes (ARGs), phages, biocide and metal-tolerance genes confirm the high genetic variability of S. Napoli, also revealing a within-serovar phylogenetic structure more complex than previously known. Our work also confirms genomic similarity of S. Napoli to typhoidal serovars (S. Typhi and S. Paratyphi A), with S. Napoli samples clustering primarily according to ST, each being characterized by specific genomic traits. Moreover, two major subclades of S. Napoli can be clearly identified, with ST-474 being biphyletic. All STs span among isolation sources and years of isolation, highlighting the challenge this serovar poses to define its epidemiology and evolution. Altogether, S. Napoli strains carry less SPIs and less ARGs than other non-typhoidal serovars and seldom acquire plasmids. However, we here report the second case of an extended-spectrum ß-lactamases (ESBLs) producing S. Napoli strain and the first cases of multidrug resistant (MDR) S. Napoli strains, all isolated from humans. CONCLUSIONS: Our results provide evidence of genomic plasticity of S. Napoli, highlighting genomic similarity with typhoidal serovars and genomic features typical of non-typhoidal serovars, supporting the possibility of survival in different niches, both enteric and non-enteric. Presence of horizontally acquired ARGs and MDR profiles rises concerns regarding possible selective pressure exerted by human environment on this pathogen.


Assuntos
DNA Bacteriano/genética , Infecções por Salmonella/microbiologia , Salmonella enterica/classificação , Sequenciamento Completo do Genoma/métodos , Farmacorresistência Bacteriana Múltipla , Ilhas Genômicas , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Itália , Filogenia , Plasmídeos/genética , Salmonella enterica/genética , Salmonella enterica/imunologia , Salmonella enterica/isolamento & purificação , Sorogrupo , Febre Tifoide/microbiologia , Resistência beta-Lactâmica
6.
BMC Vet Res ; 16(1): 455, 2020 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-33228646

RESUMO

BACKGROUND: Surgical sterilization is the most effective method of contraception for dogs. It also prevents pyometra and reduces the risk of mammary tumour development. However, this procedure also has negative effects, such as urinary incontinence. Steroid hormone deprivation following gonadectomy could also affect canine vaginal mucosa conditions and the microbial community colonizing the vaginal tract. This hypothesis was tested by comparing the vaginal cytology and microbial community of two groups of bitches, including 11 in anoestrus and 10 sterilized bitches (post-pubertal sterilization in the last 4 years). Bacteria were identified through metataxonomic analysis, amplifying the V3-V4 regions of 16S rRNA gene, and culturing methods. RESULTS: Vaginal mucosa cytology was suggestive of dystrophic conditions in sterilized bitches, whereas a typical anoestrus pattern with parabasal and intermediate cells was appreciable in anoestrous animals. Metataxonomic analysis revealed large inter-individual variability. Salmonella, Mycoplasma and Staphylococcus were present in moderate quantities in almost all the samples in both groups. Mollicutes (class level) and Tenericutes (phylum level) were commonly present in moderate quantities in anoestrus samples, whereas these microbes were present at high levels in a single sample from the sterilized group. Based on culturing, a higher number of different species were isolated from the anoestrous bitches, and Mycoplasma canis was exclusively identified in an anoestrous bitch. Staphylococcus spp. was the most frequently isolated genus in both groups, followed by Streptococcus spp., and, among gram-negative bacteria, Escherichia spp. and Haemophilus spp. A comparison of the numbers of the most frequently isolated genera of bacteria from vaginal cultures of bitches revealed that Pasteurella and Proteus were the most frequently identified in sterilized animals based on metataxonomic analysis (p-value = 0.0497 and 0.0382, respectively), whereas Streptococcus was significantly and most frequently isolated from anoestrous bitches using culture methods (p value = 0.0436). CONCLUSIONS: In this preliminary investigation, no global patterns of the vaginal bacteria community were noted that characterized the condition of the bitches; however, cytology suggested local modifications. Sterilization after puberty caused minimal alterations in the vaginal microbial community of bitches within 4 years after surgery.


Assuntos
Microbiota , Esterilização Reprodutiva/veterinária , Vagina/microbiologia , Anestro , Animais , Bactérias/classificação , Cães , Feminino , Histerectomia/veterinária , Mucosa/citologia , Ovariectomia/veterinária , Projetos Piloto , Vagina/citologia
7.
Mar Drugs ; 18(4)2020 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-32290587

RESUMO

BACKGROUND: Previously published work has demonstrated that the LPS injection of Ciona robusta leads to the overexpression of a truncated form of an immune-related mRNA (C8short) by means of Ciona robusta (CR) alternative polyadenylation (APA) (CR-APA). METHODS: The 3D structure of the C8short-derived Ciona robusta chemo-attractive peptide (CrCP) was evaluated by homology modeling. The biological activity of the CrCP was studied in vitro using a primary human dermal cell line (HuDe). Real-Time PCR was used to investigate the expression levels of genes involved in cell motility. NF-κB signaling was studied by western blotting. RESULTS: In silico modeling showed that CrCP displayed structural characteristics already reported for a short domain of the vertebrate CRK gene, suggesting its possible involvement in cell migration mechanisms. In vitro assays demonstrated that CrCP was capable of inducing the motility of HuDe cells in both wound healing and chemo-attractive experiments. qPCR demonstrated the capability of CrCP to modulate the expression of the matrix metalloproteinase-7 (MMP-7) and E-cadherin genes. Finally, western blot analysis demonstrated that treatment with CrCP induced activation of the NF-κB signaling pathway. CONCLUSION: Our results describe the characterization of the 3D structure and chemo-attractive activity of an LPS-induced CrCP peptide from Ciona robusta.


Assuntos
Anti-Inflamatórios/farmacologia , Ciona , Peptídeos/farmacologia , Animais , Linhagem Celular/efeitos dos fármacos , Linhagem Celular/metabolismo , Movimento Celular/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , RNA Mensageiro/metabolismo
8.
Foodborne Pathog Dis ; 16(8): 558-561, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30916593

RESUMO

Salmonella enterica serovar 1,4,[5],12:i:- has emerged over the last two decades as one of the most common serovars causing human salmonellosis in Europe. It is supposed to originate from Salmonella enterica serovar Typhimurium due to antigenic and genotypic similarities between the two serovars. Due to the high level of similarity, the multilocus variable-number tandem repeat analysis (MLVA) protocol designed for Salmonella Typhimurium routine typing is commonly used also for the characterization of S. 1,4,[5],12:i. Nevertheless, the Salmonella Typhimurium-based MLVA protocol often shows poor discriminatory power for S. 1,4,[5],12:i. Indeed, only a limited number of MLVA profiles have been described for S. 1,4,[5],12:i:-. Moreover, based on the MLVA clustering, S. 1,4,[5],12:i:- is supposed to display high clonality. The aim of the present work was to assess whether the five loci of Salmonella Typhimurium investigated by MLVA are sufficiently accurate to correctly assign S. 1,4,[5],12:i:- isolates. For this purpose, 38 epidemiologically unrelated S. 1,4,[5],12:i:- were subjected to whole-genome sequencing. Isolates were selected among a collection of monophasic strains isolated in Italy from different sources over the period 2014-2016 and belonging to the five most commonly detected MLVA profiles. Results confirmed the possible clonality for S. 1,4,[5],12:i:- serovar in the light of the scarce difference observed in terms of single-nucleotide polymorphisms (SNPs) among investigated isolates. Nevertheless, unrelated isolates on the basis of the difference of SNP number were characterized as indistinguishable by MLVA profile, thus suggesting an insufficient resolution of MLVA. Hence, we can conclude that MLVA-based approach does not seem a valuable proxy to deepen into the epidemiological relationship among S. 1,4,[5],12:i:- isolates. These evidences can be useful to avoid incorrect assignment especially when surveillance data are used for outbreak investigations.


Assuntos
Infecções por Salmonella/epidemiologia , Salmonella enterica/isolamento & purificação , Técnicas de Tipagem Bacteriana , DNA Bacteriano/análise , Humanos , Itália/epidemiologia , Repetições Minissatélites , Tipagem de Sequências Multilocus , Infecções por Salmonella/microbiologia , Salmonella enterica/classificação , Salmonella enterica/genética , Sequenciamento Completo do Genoma
9.
J Hum Genet ; 62(2): 277-290, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27734839

RESUMO

Oculocutaneous albinism (OCA) is characterized by hypopigmentation of the skin, hair and eye, and by ophthalmologic abnormalities caused by a deficiency in melanin biosynthesis. In this study we recruited 321 albino patients and screened them for the genes known to cause oculocutaneous albinism (OCA1-4 and OCA6) and ocular albinism (OA1). Our purpose was to detect mutations and genetic frequencies of the main causative genes, offering to albino patients an exhaustive diagnostic assessment within a multidisciplinary approach including ophthalmological, dermatological, audiological and genetic evaluations. We report 70 novel mutations and the frequencies of the major causative OCA genes that are as follows: TYR (44%), OCA2 (17%), TYRP1 (1%), SLC45A2 (7%) and SLC24A5 (<0.5%). An additional 5% of patients had GPR143 mutations. In 19% of cases, a second reliable mutation was not detected, whereas 7% of our patients remain still molecularly undiagnosed. This comprehensive study of a consecutive series of OCA/OA1 patients allowed us to perform a clinical evaluation of the different OCA forms.


Assuntos
Albinismo Oculocutâneo/diagnóstico , Albinismo Oculocutâneo/genética , Antígenos de Neoplasias/genética , Antiporters/genética , Proteínas do Olho/genética , Glicoproteínas de Membrana/genética , Proteínas de Membrana/genética , Proteínas de Membrana Transportadoras/genética , Oxirredutases/genética , Adulto , Idoso , Testes Genéticos , Humanos , Masculino , Melaninas/biossíntese , Pessoa de Meia-Idade
10.
J Hum Genet ; 60(9): 467-71, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26016411

RESUMO

Oculocutaneous albinism (OCA) is characterized by hypopigmentation of the skin, hair and eye, and by ophthalmologic abnormalities caused by a deficiency in melanin biosynthesis. OCA type IV (OCA4) is one of the four commonly recognized forms of albinism, and is determined by mutation in the SLC45A2 gene. Here, we investigated the genetic basis of OCA4 in an Italian child. The mutational screening of the SLC45A2 gene identified two novel potentially pathogenic splicing mutations: a synonymous transition (c.888G>A) involving the last nucleotide of exon 3 and a single-nucleotide insertion (c.1156+2dupT) within the consensus sequence of the donor splice site of intron 5. As computer-assisted analysis for mutant splice-site prediction was not conclusive, we investigated the effects on pre-mRNA splicing of these two variants by using an in vitro minigene approach. Production of mutant transcripts in HeLa cells demonstrated that both mutations cause the almost complete abolishment of the physiologic donor splice site, with the concomitant unmasking of cryptic donor splice sites. To our knowledge, this work represents the first in-depth molecular characterization of splicing defects in a OCA4 patient.


Assuntos
Albinismo Oculocutâneo/genética , Antígenos de Neoplasias/genética , Proteínas de Membrana Transportadoras/genética , Sítios de Splice de RNA/genética , Pré-Escolar , Humanos , Masculino , Mutação de Sentido Incorreto , Splicing de RNA/genética
11.
Molecules ; 20(6): 9879-89, 2015 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-26029857

RESUMO

BC-11 is an easily synthesized simple thiouronium-substituted phenylboronic acid, which has been shown to be cytotoxic on triple negative MDA-MB231 breast cancer cells by inducing a perturbation of cell cycle when administered at a concentration equal to its ED50 at 72 h (117 µM). Exposure of cells to BC-11, either pre-absorbed with a soluble preparation of the N-terminal fragment of urokinase-plasminogen activator (uPa), or in co-treatment with two different EGFR inhibitors, indicated that: (i) BC-11 acts via binding to the N-terminus of the enzyme where uPa- and EGF receptor-recognizing sites are present, thereby abrogating the growth-sustaining effect resulting from receptor binding; and (ii) the co-presence of the EGFR inhibitor PD153035 potentiates BC-11's cytotoxicity. Exposure of cells to a higher concentration of BC-11 corresponding to its ED75 at 72 h (250 µM) caused additional impairment of mitochondrial activity, the production of reactive oxygen species and promotion of apoptosis. Therefore, BC-11 treatment appears to show potential for the development of this class of compounds in the prevention and/or therapy of "aggressive" breast carcinoma.


Assuntos
Antineoplásicos/farmacologia , Ácidos Borônicos/farmacologia , Células Epiteliais/efeitos dos fármacos , Inativadores de Plasminogênio/farmacologia , Quinazolinas/farmacologia , Ativador de Plasminogênio Tipo Uroquinase/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sinergismo Farmacológico , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/genética , Receptores ErbB/metabolismo , Feminino , Expressão Gênica , Humanos , Glândulas Mamárias Humanas/efeitos dos fármacos , Glândulas Mamárias Humanas/metabolismo , Glândulas Mamárias Humanas/patologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/agonistas , Espécies Reativas de Oxigênio/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/genética , Ativador de Plasminogênio Tipo Uroquinase/metabolismo
12.
Biologicals ; 42(5): 294-7, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25132375

RESUMO

In search of novel gene signatures for osteo-differentiation of mesenchymal stem cells (MSCs), we submitted cDNA preparations from undifferentiated and differentiating MSCs to differential display- and semiquantitative-PCR and found down-regulation of COL5A1 in osteo-induced cultures at days 21 and 28, when the mineralized matrix accumulates. We also cultured osteo-differentiating MSCs onto type V collagen substrates and found a decrease in the accumulation of extracellular calcium compared to those grown in uncoated flasks. To our knowledge, this is first evidence that type V collagen might represent a stromal component that impairs osteogenesis.


Assuntos
Colágeno Tipo V/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Osteogênese/genética , Osteogênese/fisiologia , Diferenciação Celular , Células Cultivadas , Colágeno Tipo V/genética , Regulação para Baixo , Humanos , Osteoblastos/citologia , Osteoblastos/metabolismo
14.
Foodborne Pathog Dis ; 11(2): 138-44, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24328499

RESUMO

Salmonella enterica subsp. enterica serovar 4,[5],12:i:- DT193 is recognized as an emerging monophasic variant of Salmonella Typhimurium in many European countries. Resistance to ampicillin, streptomycin, sulphonamides, and tetracycline (R-type ASSuT) is described as one of the most common profiles of resistance within this clone. Recently, strains presenting such features were isolated from two unrelated outbreaks in Italy. Strains were characterized by pulsed-field gel electrophoresis (PFGE), performed with XbaI, BlnI, and SpeI, and multiple-locus variable-number tandem repeat analysis (MLVA). XbaI-PFGE showed strains related to the two outbreaks as indistinguishable. Conversely, both BlnI-PFGE and MLVA characterized the strains related the two outbreaks as different. XbaI-PFGE identified two profiles, differing by one band, within strains isolated from one of the two outbreaks. Also BlnI-PFGE and MLVA generated different profiles among the strains related to that outbreak. Combining the PFGE profiles obtained by XbaI and BlnI and comparing them with the MLVA profiles, the two methods grouped the same isolates based on identity. Moreover, genomic deletions of the genes included in the operon fljAB, the flanking iroB gene, and the closely located STM2757 gene were investigated. For all strains, the same profile of deletion characterized by the absence of fljA, fljB, and hin genes and the presence of STM2757 and iroB genes was identified. This profile of deletion represents a mixture between two profiles of Salmonella 4,[5],12:i:- described as the "Spanish" and the "U.S." clones. This study demonstrated that although strains of Salmonella 4,[5],12:i:- DT193 ASSuT are highly clonal, minor differences between strains may be seen during the same outbreak by using in parallel PFGE with different restriction enzymes, MLVA, and the analysis of molecular markers related to the operon fljAB. The combination of these different molecular approaches was essential to clarify the epidemiological relationship among the strains.


Assuntos
Surtos de Doenças , Doenças Transmitidas por Alimentos/epidemiologia , Produtos da Carne/microbiologia , Salmonella enterica/genética , Salmonella enterica/isolamento & purificação , Animais , Proteínas de Bactérias/genética , Galinhas , Mapeamento Cromossômico , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Fezes/microbiologia , Flagelina/genética , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Itália/epidemiologia , Proteínas Repressoras/genética , Salmonella enterica/classificação , Salmonella typhimurium/genética , Salmonella typhimurium/isolamento & purificação , Suínos
15.
Foodborne Pathog Dis ; 11(6): 497-9, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24666380

RESUMO

Salmonella 4,[5],12:i:- is a variant of Salmonella Typhimurium, which lacks the expression of phase-2 flagellar antigen, generally associated with the deletion of the fljB gene. Additional mechanisms involving the fljAB operon ( fljA, fljB, and hin genes) lead to the lack of expression of phase-2 flagellar antigens also in Salmonella strains harboring the fljB gene. For 20 S. 4,[5],12:i:- strains, defined as "inconsistent" Salmonella Typhimurium variants since they had phenotypically behaved as monophasic, even though the fljB gene was conserved, the fljAB operon was characterized in order to explain the ineffective expression of the phase-2 flagellar antigen. The monophasic phenotype for a first group of strains (9) was likely due to the absence of the hin gene, leading to the inhibited switch between the expression of phase-1 and phase-2 flagellar genes. For a second group of strains (5), the monophasic phenotype could be attributed to nonconservative point mutations identified in fljA and hin genes, which could hamper the proper expression of invertase gene and the fljA, acting as repressor of the phase-1 flagellar gene. Finally, for a last group of inconsistent strains (6), a plausible reason for their monophasic phenotype was not found, since the genes involved in the expression of phase-2 flagellar antigen were fully conserved. Moreover, the collection of inconsistent Salmonella Typhimurium isolates investigated were characterized by distinct molecular profiles, as demonstrated by multiple-locus variable-number tandem repeat analysis, and phenotype variability, as demonstrated by phage-typing. This study highlights the usefulness of investigating the entire fljAB operon when a definitive identification of the monophasic or biphasic status of Salmonella Typhimurium strains is needed (for instance, in the context of epidemiological investigations aimed to identify the relatedness among strains).


Assuntos
Proteínas de Bactérias/metabolismo , Flagelina/metabolismo , Variação Genética , Modelos Biológicos , Óperon , Proteínas Repressoras/metabolismo , Salmonella typhimurium/isolamento & purificação , Animais , Proteínas de Bactérias/genética , Tipagem de Bacteriófagos , Fezes/microbiologia , Flagelina/genética , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Humanos , Itália , Carne/microbiologia , Tipagem Molecular , Tipagem de Sequências Multilocus , Mutação , Proteínas Repressoras/genética , Reprodutibilidade dos Testes , Intoxicação Alimentar por Salmonella/microbiologia , Salmonella typhimurium/classificação , Salmonella typhimurium/genética , Salmonella typhimurium/metabolismo , Especificidade da Espécie
16.
Chemosphere ; 349: 140739, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38000557

RESUMO

2,2',4,4'-tetra-bromodiphenyl ether (BDE-47) is widespread in the environment and biological samples. Its association with health risks is an increasing concern, yet information on BDE-47 immunotoxicity remains limited. This study investigated the impact of BDE-47 on innate and adaptive immune responses through in vitro and in vivo approaches. BDE-47's capacity to directly induce cell responses and modulate responses induced by known stimuli was studied in vitro using the RAW 264.7 murine macrophage cell line and spleen-derived lymphocytes, and in vivo using keyhole limpet hemocyanin (KLH)-immunized BALB/c mice orally administered (28 d) at dose levels (7.5, 15.0 and 30 mg/kg/bw/d) derived from relevant toxicokinetic data from rodent models. RAW 264.7 cells stimulated with lipopolysaccharide (LPS) and exposed to BDE-47 exhibited unchanged cell viability but decreased release of interleukin (IL)-6. Primary splenocytes from naïve mice stimulated with anti-CD3/anti-CD28 antibodies and exposed to BDE-47 showed a significant decrease of IL-17 A and IFNγ production. In vivo data showed that BDE-47 significantly reduced the KLH-specific antibody response. A generally decreasing trend of IFNγ, IL-10 and IL-5 production was observed after in vitro antigen-specific restimulation of spleen cells. Histopathological effects on liver, spleen, small intestine and thyroid were detected at the highest dose in the absence of general toxicity. In addition, the expression of Mm_mir155 and Mm_let7a was induced in livers of exposed mice. The data obtained in this study suggest that exposure to BDE-47 may perturb innate and adaptive immune responses, thus possibly decreasing resistance to bacterial and viral infections.


Assuntos
Imunidade , Interleucina-6 , Camundongos , Animais , Modelos Animais de Doenças , Camundongos Endogâmicos BALB C , Hemocianinas
17.
Biomater Sci ; 12(13): 3374-3388, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38787753

RESUMO

This study details the design, fabrication, clinical trials' evaluation, and analysis after the clinical application of 3D-printed bone reconstruction implants made of nHAp@PLDLLA [nanohydroxyapatite@poly(L-lactide-co-D,L-lactide)] biomaterial. The 3D-printed formulations have been tested as bone reconstruction Cranioimplants in 3 different medical cases, including frontal lobe, mandibular bone, and cleft palate reconstructions. Replacing one of the implants after 6 months provided a unique opportunity to evaluate the post-surgical implant obtained from a human patient. This allowed us to quantify physicochemical changes and develop a spatial map of osseointegration and material degradation kinetics as a function of specific locations. To the best of our knowledge, hydrolytic degradation and variability in the physicochemical and mechanical properties of the biomimetic, 3D-printed implants have not been quantified in the literature after permanent placement in the human body. Such analysis has revealed the constantly changing properties of the implant, which should be considered to optimize the design of patient-specific bone substitutes. Moreover, it has been proven that the obtained composition can produce biomimetic, bioresorbable and bone-forming alloplastic substitutes tailored to each patient, allowing for shorter surgery times and faster patient recovery than currently available methods.


Assuntos
Durapatita , Impressão Tridimensional , Humanos , Durapatita/química , Implantes Absorvíveis , Substitutos Ósseos/química , Crânio/cirurgia , Poliésteres/química , Masculino , Desenho de Prótese , Materiais Biocompatíveis/química , Feminino
18.
Mol Carcinog ; 52(5): 348-58, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-22213077

RESUMO

We previously reported that ductal infiltrating carcinomas (d.i.c.) of the human breast display profound modifications of the stromal architecture, associated with anomalous collagen composition. Among the major alterations observed in the interstitial collagen, the relative increase of type V collagen content was detected. When type V collagen was used as an "in vitro" substrate for 8701-BC d.i.c. cells, it appeared able to restrain cell growth, inhibit cell motility and invasion "in vitro", and modify the expression levels of genes coding for apoptosis factors, caspases and stress response proteins. In the present paper we demonstrate that type V collagen induces the down-regulation of protein kinase C η, an event that may be, at least in part, responsible of the previously-reported modifications of cell morphology and growth rate, and that appears to be involved in the already-observed changes of expression levels of genes encoding for anti- (Bcl-2) and pro-apoptotic factors (Bad, Dapk, Bcl-Xs) and enzymes (caspase 5 and 8).


Assuntos
Neoplasias da Mama/metabolismo , Colágeno Tipo V/metabolismo , Proteína Quinase C/metabolismo , Apoptose/genética , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Caspase 8/genética , Caspase 8/metabolismo , Caspases/genética , Caspases/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Fragmentação do DNA , Regulação para Baixo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Isoenzimas/genética , Isoenzimas/metabolismo , Oligonucleotídeos Antissenso , Proteína Quinase C/genética , Proteína de Morte Celular Associada a bcl/genética , Proteína de Morte Celular Associada a bcl/metabolismo , Proteína bcl-X/genética
19.
Anal Bioanal Chem ; 405(6): 1893-901, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22729357

RESUMO

Emerging evidence indicates that cadmium (Cd) is able to regulate gene expression, drastically affecting the pattern of transcriptional activity in human normal and pathological cells. We have already shown that exposure of MDA-MB231 breast cancer cells to 5 µM CdCl(2) for 96 h, apart from significantly affecting mitochondrial metabolism, induces modifications of the expression level of genes coding for members of stress response-, mitochondrial respiration-, MAP kinase-, NF-κB-, and apoptosis-related pathways. In the present study, we have expanded the knowledge on the biological effects of Cd-breast cancer cell interactions, indicating PLP2 (proteolipid protein-2) as a novel member of the list of Cd-upregulated genes by MDA-MB231 cancer cells and, through the application of transfection techniques with specific antisense oligonucleotides, we have demonstrated that such over-expression may be an upstream event to some of the changes of gene expression levels already observed in Cd-treated cells, thus unveiling new possible molecular relationship between PLP2 and genes linked to the stress and apoptotic responses.


Assuntos
Neoplasias da Mama/genética , Cloreto de Cádmio/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Proteínas com Domínio MARVEL/genética , Oligonucleotídeos Antissenso/genética , Proteolipídeos/genética , RNA Mensageiro/química , Apoptose/efeitos dos fármacos , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Caspases/genética , Caspases/metabolismo , Linhagem Celular Tumoral , Feminino , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Humanos , Proteínas com Domínio MARVEL/antagonistas & inibidores , Proteínas com Domínio MARVEL/metabolismo , Conformação de Ácido Nucleico , Proteolipídeos/antagonistas & inibidores , Proteolipídeos/metabolismo , RNA Mensageiro/genética , Estresse Fisiológico/genética , Transfecção
20.
Microbiol Resour Announc ; 12(5): e0004323, 2023 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-37022181

RESUMO

We report the genome sequence of a Salmonella enterica subsp. enterica serovar Bispebjerg strain that was isolated from a turkey flock in 2011. The genome analysis of the strain, a rare and multihost serovar, revealed its pathogenic potential due to antimicrobial resistance and a plethora of Salmonella pathogenicity islands and virulence factors.

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