RESUMEN

Use of tumor-suppressive microRNAs (miRNAs) as anti-cancer agents is hindered by the lack of effective delivery vehicles, entrapment of the miRNA within endocytic compartments, and rapid degradation of miRNA by nucleases. To address these issues, we developed a miRNA delivery strategy that includes (1) a targeting ligand, (2) an endosomal escape agent, nigericin and (3) a chemically modified miRNA. The delivery ligand, DUPA (2-[3-(1,3-dicarboxy propyl) ureido] pentanedioic acid), was selected based on its specificity for prostate-specific membrane antigen (PSMA), a receptor routinely upregulated in prostate cancer-one of the leading causes of cancer death among men. DUPA was conjugated to the tumor suppressive miRNA, miR-34a (DUPA-miR-34a) based on the ability of miR-34a to inhibit prostate cancer cell proliferation. To mediate endosomal escape, nigericin was incorporated into the complex, resulting in DUPA-nigericin-miR-34a. Both DUPA-miR-34a and DUPA-nigericin-miR-34a specifically bound to, and were taken up by, PSMA-expressing cells in vitro and in vivo. And while both DUPA-miR-34a and DUPA-nigericin-miR-34a downregulated miR-34a target genes, only DUPA-nigericin-miR-34a decreased cell proliferation in vitro and delayed tumor growth in vivo. Tumor growth was further reduced using a fully modified version of miR-34a that has significantly increased stability.

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Helicobacter pylori is one of the most common bacteria affecting human societies worldwide, and is mainly associated with gastrointestinal complications due to different virulence factors. This study aimed to investigate some virulence genes of H. pylori in gastric biopsies of patients with gastritis in Sari city, North of Iran. Informed consent forms were obtained from the studied patients, and those who needed endoscopy were included in the study. To evaluate the prevalence of cagA, iceA1, iceA2, vacA, dupA, and oipA genes, gastric biopsies with positive or negative rapid urease test were collected from 50 patients (25 in each group) with gastro-duodenal diseases. The bacterial DNAs were extracted by a specific kit, and the presence of the genes was analyzed by PCR using specific primers. Eighteen (72%) biopsies from 25 H. pylori-positive samples were cagA-positive, while 17 (68%) biopsies contained the vacA gene, and 11 (44%) samples had both vacA and cagA genes. However, 16 (64%), 12 (48%), 13 (52%), and 14 (56%) biopsies contained dupA, iceA1, iceA2, and oipA genes, respectively. Due to the significant role of the studied virulence factors in the pathogenicity of H. pylori, the high prevalence of these factors in biopsies of patients with gastritis is a concern needing to the management in this region.

RESUMEN

Helicobacter pylori is believed to induce gastropathy; however, the exact pathogenic molecules involved in this process have not been elucidated. Duodenal ulcer promoting gene A (DupA) is a virulence factor with a controversial role in gastric inflammation and carcinogenesis. To explore and confirm the function of DupA in gastropathy from the perspective of the microbiome, we investigated the microbial characteristics of 48 gastritis patients through 16S rRNA amplicon sequencing. In addition, we isolated 21 H. pylori strains from these patients and confirmed the expression of dupA using PCR and qRT-PCR. Bioinformatics analysis identified diversity loss and compositional changes as the key features of precancerous lesions in the stomach, and H. pylori was a characteristic microbe present in the stomach of the gastritis patients. Co-occurrence analysis revealed that H. pylori infection inhibits growth of other gastric inhabiting microbes, which weakened the degradation of xenobiotics. Further analysis showed that dupA+ H. pylori were absent in precancerous lesions and were more likely to appear in erosive gastritis, whereas dupA- H. pylori was highly abundant in precancerous lesions. The presence of dupA in H. pylori caused less disturbance to the gastric microbiome, maintaining the relatively richness of gastric microbiome. Overall, our findings suggest that high dupA expression in H. pylori is correlated with a high risk of erosive gastritis and a lower level of disturbance to the gastric microbiome, indicating that DupA should be considered a risk factor of erosive gastritis rather than gastric cancer.

Asunto(s)

Úlcera Duodenal , Gastritis , Microbioma Gastrointestinal , Infecciones por Helicobacter , Helicobacter pylori , Lesiones Precancerosas , Neoplasias Gástricas , Úlcera Gástrica , Humanos , ARN Ribosómico 16S/genética , Neoplasias Gástricas/genética

RESUMEN

BACKGROUND: Cerebrotendinous xanthomatosis (CTX) is a rare autosomal recessive metabolic disease caused by mutations in CYP27A1. It has a low incidence rate, insidious onset, and diverse clinical manifestations. It can be easily misdiagnosed and can go unrecognized by clinicians, leading to delayed treatment and worsened patient outcomes. CASE SUMMARY: A 38-year-old male was admitted to our hospital with a history of unabating unstable posture and difficulty in walking for more than 30 years. Subsequently based on the patient's medical history, clinical symptoms, magnetic resonance imaging and gene sequencing results, he was finally diagnosed with CTX. Due to the low incidence rate of the disease, clinicians have insufficient knowledge of it, which makes the diagnosis process more tortuous and prolongs the diagnosis time. CONCLUSION: Prompt diagnosis and treatment of CTX improve patient outcomes.

RESUMEN

BACKGROUND: Helicobacter pylori infection is one of the most common infectious diseases in humans. Dental plaque is considered as a reservoir of this bacterium, which could play an important role in the development of gastrointestinal problems. Our aim was to investigate the prevalence of H. pylori and its virulence factors in dental plaques in children with and without dental caries. METHODS: Among children aged 6 to 12 years, a total of 72 children were enrolled in the study, including 36 cases with total DMFT/dmft > 3 (case group) and 36 participants with total DMFT/dmft < 1 (control group). After removing supra-gingival plaques from the lower first permanent molar teeth, the samples were examined using PCR method for the presence of H. pylori and some of its virulence factors. Statistical analysis was performed using chi-square, Fisher' exact test, t-tests, and logistic regression. RESULTS: Of 72 participants, 40 cases were male, and 32 cases were female. The minimum and maximum values of total DMFT/dmft indices were zero and ten, respectively, and the mean ± SD value of total DMFT/dmft was 2.78 ± 3.22. Except for vegetable consumption (p = 0.045), there was no significant difference between the two groups regarding gastrointestinal disorders, feeding methods in infancy (p = 0.058), frequency of daily brushing (p = 0.808), frequency of dental visits (p = 0.101), and history of dental scaling (p = 0.246) and professional topical fluoride therapy (p = 0.5). Out of 72 samples, 15 cases were positive for H. pylori DNA (20.8%), and there was no significant association between the presence of this bacterium in dental plaque and dental caries (p = 0.281). The frequency of virulence factors detected in 15 H. pylori cases was as follows: cagA in six cases (40.0%), vacAm1 in three cases (20.0%), and vacAs1 in one case (6.7%). There was no significant difference between the groups regarding the prevalence of virulence factors. CONCLUSION: Our results indicate the presence of H. pylori along with some virulence factors in dental plaques as a reservoir of this bacterium in children in Iran. Although there was no significant association between this bacterium and the incidence of dental caries, dental health in children needs to be seriously taken into consideration.

Asunto(s)

Caries Dental , Placa Dental , Infecciones por Helicobacter , Helicobacter pylori , Antígenos Bacterianos , Proteínas Bacterianas/genética , Estudios de Casos y Controles , Niño , Caries Dental/epidemiología , Placa Dental/epidemiología , Placa Dental/microbiología , Femenino , Genotipo , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/epidemiología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/genética , Humanos , Masculino , Prevalencia , Factores de Virulencia/genética

RESUMEN

Helicobacter pylori is an organism associated with ulcer disease and gastric cancer. The latter is one of the most prevalent malignancies and currently the fourth major cause of cancer-related deaths globally. The pathogen infects about 50% of the world population, and currently, no treatment ensures its total elimination. There has been an increase in our understanding of the pathophysiology and pathogenesis mechanisms of H. pylori over the years. H. pylori can induce several genetic alterations, express numerous virulence factors, and trigger diverse adaptive mechanisms during its adherence and colonization. For successful colonization and infection establishment, several effector proteins/toxins are released by the organism. Evidence is also available reporting spiral to coccoid transition as a unique tactic H. pylori uses to survive in the host's gastrointestinal tract (GIT). Thus, the virulence and pathogenicity of H. pylori are under the control of complex interplay between the virulence factors, host, and environmental factors. Expounding the role of the various virulence factors in H. pylori pathogenesis and clinical outcomes is crucial for vaccine development and in providing and developing a more effective therapeutic intervention. Here we critically reflect on H. pylori infection and delineate what is currently known about the virulence and pathogenesis mechanisms of H. pylori.

Asunto(s)

Gastritis , Infecciones por Helicobacter , Helicobacter pylori , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Gastritis/microbiología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/genética , Humanos , Virulencia/genética , Factores de Virulencia/genética

RESUMEN

Equine gastric ulcer syndrome (EGUS) is a multifactorial disorder and one of the most common diseases in horses. The objective of this research was to detect one of the potential risk factors of equine squamous gastric disease (ESGD), the Helicobacter pylori specific gene, and tracing the presence of the duodenal ulcer-promoting gene (dupA) as a possible virulence marker. Gastric fluid together with faecal samples were collected from twenty rural horses from around Tabriz, Iran. Throughout the endoscopic examinations, the type, numbers, severity, and the location of the lesions were documented. Nine of twenty horses exhibited macroscopic lesions in the squamous mucosa that were later classified into grades 1, 2, 3, and 4. Only three of these horses exhibited H. pylori in their gastric fluid samples, whereas all faecal samples were H. pylori-negative. All the H. pylori-positive cases manifested severe forms of ESGD (grades 3-4). The age and sex were both unrelated to the lesion severity and ESGD status in this study. Research is required to further discuss the virulence aspects of dupA regarding ESGD.

RESUMEN

Helicobacter pylori is the etiological agent of chronic gastritis, peptic ulcer, and gastric cancer. The duodenal ulcer-promoting gene dupA, which is located in the plasticity region of the H. pylori genome, is homologous to the virB gene which encodes a type IV secretion protein in Agrobacterium tumefaciens. Studies have shown associations between H. pylori dupA-positive strains and gastroduodenal diseases. However, whether dupA acts as a risk factor or protective factor in these diseases remains unclear. Therefore, in this study, we aimed to verify the presence of the dupA gene in infectious H. pylori strains in the Brazilian mid-west and to investigate its association with the clinical outcomes of patients with dyspepsia. Additionally, the phylogenetic origin of the strains was determined. Gastric biopsies from 117 patients with dyspepsia were analyzed using histological and molecular techniques. The hpx gene (16S rRNA) was used to screen for H. pylori infection, and positive samples were then subjected to dupA gene detection and sequencing. The estimated prevalence of H. pylori infection was 64.1%, with the dupA gene being detected in a high proportion of infectious strains (70.7%). Furthermore, a risk analysis revealed that for women, a dupA-positive H. pylori infection increased the chance of developing gastritis by twofold. The partial dupA sequences from isolated infectious strains in this work are similar to those of strains isolated in westerns countries. This study provides useful insights for understanding the role of the H. pylori dupA gene in disease development.

Asunto(s)

Proteínas Bacterianas , Infecciones por Helicobacter , Helicobacter pylori , Factores de Virulencia , Proteínas Bacterianas/genética , Brasil/epidemiología , Dispepsia/complicaciones , Dispepsia/epidemiología , Dispepsia/microbiología , Femenino , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/epidemiología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/clasificación , Helicobacter pylori/genética , Helicobacter pylori/patogenicidad , Humanos , Masculino , Filogenia , Factores Protectores , ARN Ribosómico 16S/genética , Factores de Riesgo , Factores de Virulencia/genética

RESUMEN

Common variable immunodeficiency (CVID) is one form of the primary immunodeficiencies (PIDs). CVID is characterized by variable clinical manifestations. Genetic alteration is a cause of the disease in many cases. In the current paper we described Patient N of 45 years old, who have been suffering from frequent various infections and therefore attended an immunologist and clinical geneticist. Immunoglobulins (Ig) A, M, and G deficiency was found in the patient. As a result of medical genetic counselling primary immunodeficiency has been suggested as a diagnosis. Further molecular genetic testing using clinical exome sequencing (Next Generation Sequencing method) revealed a likely-pathogenic variant c.204dupA (p.Leu69ThrfsX12, rs72553875) of TNFRSF13B gene in the patient. The gene variant was found in homozygous state. According to the international medical literature and genomic databases TNFRSF13B gene mutations lead to the CVID development and in some patients are characterized by isolated IgA deficiency and in the other group of patients can lead to decrease of IgA, IgM, and IgG. The patient had a family history of cancer and autoimmune inflammatory bowel disease (erosive-ulcerative enterocolitis). Moreover, one sibling of the patient died at the age of 3 weeks from complications of toxoplasmosis infection. The other sibling of 51 years old have been also suffering from recurrent infectious diseases. Thus, the genetic cause of the disease was identified in the proband. It has been shown that homozygosity for variant c.204dupA of TNFRSF13B gene is characterized by the deficiency of all three classes of Ig. Medical genetic counselling and modern molecular genetic methods application is an important step in management of people with signs of immunodeficiency. Such approach helps to make a diagnosis to the patient, to find an exact molecular reason of the condition, to use effective treatment, and to perform preventive measures in patient`s family.

Asunto(s)

Inmunodeficiencia Variable Común , Adulto , Humanos , Recién Nacido , Persona de Mediana Edad , Inmunodeficiencia Variable Común/diagnóstico , Inmunodeficiencia Variable Común/genética , Inmunodeficiencia Variable Común/complicaciones , Mutación , Inmunoglobulina A/genética , Inmunoglobulina G , Inmunoglobulina M/genética , Proteína Activadora Transmembrana y Interactiva del CAML/genética

RESUMEN

Aim: Ligand-targeted therapeutics are experiencing increasing use for treatment of human diseases due to their ability to concentrate a desired drug at a pathologic site while reducing accumulation in healthy tissues. For many ligand-targeted drug conjugates, a critical aspect of conjugate design lies in engineering release of the therapeutic payload to occur only after its internalization by targeted cells. Because disulfide bond reduction is frequently exploited to ensure intracellular drug release, an understanding of the redox properties of endocytic compartments can be critical to ligand-targeted drug design. While the redox properties of folate receptor trafficking endosomes have been previously reported, little is known about the trafficking of prostate-specific membrane antigen (PSMA), a receptor that is experiencing increasing use for drug targeting in humans. Methods: To obtain this information, we have constructed a PSMA-targeted fluorescence resonance energy transfer pair that reports on disulfide bond reduction by changing fluorescence from red to green. Results: We show here that this reporter exhibits rapid and selective uptake by PSMA-positive cells, and that reduction of its disulfide bond proceeds steadily but incompletely following internalization. The fact that maximal disulfide reduction reaches only ~50%, even after 24 h incubation, suggests that roughly half of the conjugates must traffic through endosomes that display no reducing capacity. Conclusion: As the level of disulfide reduction differs between PSMA trafficked and previously published folate trafficked conjugates, it also follows that not all internalizing receptors are translocated through similar intracellular compartments. Taken together, these data suggest that the efficiency of disulfide bond reduction must be independently analyzed for each receptor trafficking pathway when disulfide bond reduction is exploited for intracellular drug release.

RESUMEN

Gastric cancer constitutes one of the most prevalent malignancies in both sexes; it is currently the fourth major cause of cancer-related deaths worldwide. The pathogenesis of gastric cancer is associated with the interaction between genetic and environmental factors, among which infection by Helicobacter pylori (H. pylori) is of major importance. The invasion, survival, colonization, and stimulation of further inflammation within the gastric mucosa are possible due to several evasive mechanisms induced by the virulence factors that are expressed by the bacterium. The knowledge concerning the mechanisms of H. pylori pathogenicity is crucial to ameliorate eradication strategies preventing the possible induction of carcinogenesis. This review highlights the current state of knowledge and the most recent findings regarding H. pylori virulence factors and their relationship with gastric premalignant lesions and further carcinogenesis.

Asunto(s)

Antígenos Bacterianos/fisiología , Proteínas Bacterianas/fisiología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/patogenicidad , Factores de Virulencia/fisiología , Animales , Carcinogénesis/patología , Mucosa Gástrica/microbiología , Mucosa Gástrica/patología , Humanos

RESUMEN

Functional profiling of CFTR-directed therapeutics offers the potential to provide significant benefits to young people with cystic fibrosis (CF). However, the development of 2D airway epithelial cell models for individual response tests in CF children remains a central task. The objective of this study was to determine the utility of EpiXTM technology for expansion of nasal epithelial cells for use in electrophysiological CFTR function measurements. An initial harvest of as few as 20,000 cells was sufficient to expand up to 50 million cells that were used to generate air-liquid interface (ALI) cultures for ion transport studies with the Ussing assay. CFTR function was assessed by measuring responses to forskolin and the CFTR potentiator VX-770 (ivacaftor) in ALI cultures generated from passage 3 and 4 cells. Short-circuit current (Isc) measurements of blocked CFTR currents (ΔICFTRinh) discriminated CFTR function between healthy control (wild type, WT) and patients with intermediate (F508del/R117H-7T: 56% WT) and severe (F508del/F508del: 12% WT) CF disease. For the mixed genotypes, CFTR activity for F508del/c.850dupA was 12% WT, R334W/406-1G>A was 24% WT, and CFTRdele2,3(21 kb)/CFTRdele2,3(21 kb) was 9% WT. The CFTR correctors VX-809 (lumacaftor) and VX-661 (tezacaftor) significantly increased CFTR currents for F508del/R117H to 73 and 67% WT, respectively. Cultures with the large deletion mutation CFTRdele2,3(21 kb) unexpectedly responded to VX-661 treatment (20% WT). Amiloride-sensitive sodium currents were robust and ranged between 20-80 µA/cm2 depending on the subject. In addition to characterizing the electrophysiological profile of mutant CFTR activity in cultures for five genotypes, our study exemplifies the promising paradigm of bed-to-bench side cooperation and personalized medicine.

RESUMEN

BACKGROUND: Disease severity is tremendously variable in tuberous sclerosis complex (TSC). In contrast with the detailed guidelines available for TSC diagnosis and management, clinical practice lacks adequate tools to evaluate the prognosis, especially in the case of in utero diagnosis. In addition, the correlation between genotypes and phenotypes remains a challenge, in part due to the large number of mutations linked to TSC. In this report, we describe a case of severe TSC diagnosed in utero and associated with a specific mutation in the gene tuberous sclerosis complex 2 (TSC2). CASE PRESENTATION: A mother was referred for a thorough investigation following the observation by ultrasound of cardiac abnormalities in her fetus. The mother was healthy and reported frequent, intense and long-lasting hiccups/spasms in the fetus. The fetus of gestational age 33 weeks and 4 days was found to have multiple cardiac tumors with cardiac ultrasound. Brain magnetic resonance imaging (MRI) performed in utero revealed the presence of sub-ependymal nodules and of abnormal signals disseminated in the white matter, in the cerebral cortex and in the cerebellum. Following diagnosis of definite TSC, pregnancy interruption was chosen by the parents. Genetic testing of the fetus exposed a duplication in exon 41 of TSC2 (c.5169dupA), which was absent in the parents. The autopsy ascertained the high severity of brain damage characterized by an extensive disorganisation of white and grey matter in most cerebral lobes. CONCLUSIONS: This case presentation is the first to depict the association between a de novo TSC2 c.5169dupA and multi-organ manifestation together with indications of a particularly high disease severity. This report can help physicians to perform early clinical diagnosis of TSC and to evaluate the prognosis.

Asunto(s)

Proteína 2 del Complejo de la Esclerosis Tuberosa/genética , Esclerosis Tuberosa/diagnóstico por imagen , Ultrasonografía Prenatal , Adulto , Autopsia , Exones , Femenino , Feto/patología , Pruebas Genéticas , Genotipo , Humanos , Mutación , Fenotipo , Embarazo

RESUMEN

Helicobacter pylori (H. pylori) is a microaerophilic, Gram-negative, human gastric pathogen found usually in the mucous lining of stomach. It infects more than 50% of the world's population and leads to gastroduodenal diseases. The outcome of disease depends on mainly three factors: Host genetics, environment and bacterial factors. Among these, bacterial virulence factors such as cagA, vacA are well known for their role in disease outcomes. However, based on the global epidemiological results, none of the bacterial virulence (gene) factors was found to be associated with particular diseases like duodenal ulcer (DU) in all populations. Hence, substantial importance has been provided for research in strain-specific genes outside the cag pathogenicity island, especially genes located within the plasticity regions. dupA found within the plasticity regions was first demonstrated in 2005 and was proposed for duodenal ulcer development and reduced risk of gastric cancer in certain geographical regions. Due to the discrepancies in report from different parts of the world in DU development related to H. pylori virulence factor, dupA became an interesting area of research in elucidating the role of this gene in the disease progression. In this review, we shed light on the detailed information available on the polymorphisms in dupA and their clinical relevance. We have critically appraised several pertinent studies on dupA and discussed their merits and shortcomings. This review also highlights dupA gene as an important biomarker for DU in certain populations.

Asunto(s)

Úlcera Duodenal , Infecciones por Helicobacter , Helicobacter pylori , Neoplasias Gástricas , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Úlcera Duodenal/diagnóstico , Úlcera Duodenal/epidemiología , Infecciones por Helicobacter/epidemiología , Helicobacter pylori/genética , Humanos , Factores de Virulencia/genética

RESUMEN

Cabazitaxel, a novel tubulin inhibitor with poor affinity for P-glycoprotein, is a second-generation taxane holding great promise for the treatment of metastatic castration-resistant prostate cancer. However, its poor solubility and lack of target-ability limit its therapeutic applications. Herein, we develop a biodegradable, enzyme-responsive, and targeted polymeric micelle for cabazitaxel. The micelle is formed from two amphiphilic block copolymers. The first block copolymer consists of PEG, an enzyme-responsive peptide, and cholesterol; whereas the second block copolymer consists of a targeting ligand, PEG and cholesterol. The enzyme-responsive peptide is cleavable in the presence of matrixmetaloproteinase-2 (MMP-2), which is overexpressed in the tumor microenvironment of prostate cancer. The micelle showed a very low critical micelle concentration (CMC), high drug loading, and high entrapment efficiency. Release of cabazitaxel from the micelle is dependent on the cleavage of the enzyme-responsive peptide. Moreover, the micelle showed dramatically higher cellular uptake in prostate cancer cells compared to free cabazitaxel. Importantly, the ligand-coupled polymeric micelle demonstrated better inhibition of tumor growth in mice bearing prostate cancer xenografts compared to unmodified micelle and free cabazitaxel. Taken together, these findings suggest that the enzyme-responsive cabazitaxel micelle is a potent and promising drug delivery system for advanced prostate cancer therapy. STATEMENT OF SIGNIFICANCE: Herein, we develop a biodegradable, enzyme-responsive, and actively targeted polymer micelle for cabazitaxel, which is a novel tubulin inhibitor with poor affinity for P-glycoprotein. Despite cabazitaxel's great promise for metastatic castration-resistant prostate cancer, its poor solubility, lack of target-ability, and high systemic toxicity limit its therapeutic applications, and therefore a targeted delivery system is highly needed for cabazitaxel. Our results demonstrate the importance of active targeting in targeted prostate cancer therapy. Encapsulating cabazitaxel in the micelle increases its activity and is expected to reduce its systemic toxicity, which is a major hurdle in its clinical applications. Moreover, the polymeric micelle may servers as a promising nanoscale platform for the targeted delivery of other chemotherapeutic agents to prostate cancer.

Asunto(s)

Micelas , Neoplasias de la Próstata , Animales , Línea Celular Tumoral , Humanos , Masculino , Ratones , Polímeros , Neoplasias de la Próstata/tratamiento farmacológico , Taxoides/farmacología , Taxoides/uso terapéutico , Microambiente Tumoral

RESUMEN

INTRODUCTION: Helicobacter pylori is a principal cause of gastric cancer. The aim of this study was to determine the prevalence and contribution of duodenal ulcer promoting gene A (dupA), the plasticity region genes and sigma factors in relation to their pathological expression of H. pylori infections in the Nigerian population. METHODOLOGY: Polymerase Chain Reaction was used to analyze a total of forty-nine H. pylori strains isolated from patients attending various endoscopic units in tertiary hospitals in Nigeria for complete dupA (G27 variant), jhp0917, jhp0918, other plasticity region genes jhp 914/917, jhp0914, jhp0940 and sigma factors. RESULTS: PCR results indicated that the prevalence of complete dupA (G27 variants), jhp0917, jhp0918 and other plasticity region genes jhp0914, jhp0914/0917 and jhp0940 in the H. pylori strains were 4%, 53%, 88%, 73%, 12% and 0% respectively. The prevalence values of the sigma factors were 96%, 92%, 80% for rpoN,  fliA and rpoD respectively. However, the endoscopic findings showed that erosion, normal mucosal, ulcer, hyperaemic stomach, mucosal atrophy and oedematous stomach in the patients where the H. pylori strains were isolated were 40.8%, 32.7%, 10.2%, 8.2%, 2.0% and 6.1% respectively. There was significant association between jhp0917, jhp914/917 and G27 variant and the endoscopic findings, while other plasticity genes showed no association with the endoscopic findings. CONCLUSION: These results suggest that the presence of jhp0917, jhp0914/917 and G27 variant could be used as marker to predict the pathological effect of severity in Nigeria patients with H. pylori infection.

Asunto(s)

Úlcera Duodenal/epidemiología , Infecciones por Helicobacter/epidemiología , Helicobacter pylori/genética , Factor sigma/genética , Factores de Virulencia/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Niño , Femenino , Genes Bacterianos , Helicobacter pylori/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Nigeria , Reacción en Cadena de la Polimerasa , Adulto Joven

RESUMEN

Various glutamate urea ligands have displayed high affinities to prostate specific membrane antigen (PSMA), which is highly overexpressed in prostate and other cancer sites. The multivalent versions of small PSMA-targeted molecules are known to be even more efficiently bound to the receptor. Here, we employ a well-known urea-based ligand, 2-[3-(1,3-dicarboxypropyl)-ureido] pentanedioic acid (DUPA) and triazine dendrimers in order to study the effect of molecular size on multivalent targeting in prostate cancer. The synthetic route starts with the preparation of a dichlorotriazine bearing DUPA in 67% overall yield over five steps. This dichlorotriazine reacts with G1, G3, and G5 triazine dendrimers bearing a 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) group for 64Cu-labeling at the core to afford poly(monochlorotriazine) intermediates. Addition of 4-aminomethylpiperidine (4-AMP) and the following deprotection produce the target compounds, G1-(DUPA)4, G3-(DUPA)16, and G5-(DUPA)64. These targets include 4/16/64 DUPA groups on the surface and a DOTA group at the core, respectively. In vitro cell assay using PC3-PIP (PSMA positive) and PC3-FLU (PSMA negative) cells reveals that G1-(DUPA)4 has the highest PC3-PIP to PC3-FLU uptake ratio (10-fold) through the PSMA-mediated specific uptake. While G5-(DUPA)64 displayed approximately 12 times higher binding affinity (IC50 23.6 nM) to PC3-PIP cells than G1-(DUPA)4 (IC50 282.3 nM) as evaluated in a competitive binding assay, the G5 dendrimer also showed high non-specific binding to PC3-FLU cells. In vivo uptake of the 64Cu-labeled dendrimers was also evaluated in severe combined inmmunodeficient (SCID) mice bearing PC3-PIP and PC3-FLU xenografts on each shoulder, respectively. Interestingly, quantitative imaging analysis of positron emission tomograph (PET) displayed the lowest tumor uptake in PC3-PIP cells for the midsize dendrimer G3-(DUPA)16 (19.4 kDa) (0.66 ± 0.15%ID/g at 1 h. p.i., 0.64 ± 0.11%ID/g at 4 h. p.i., and 0.67 ± 0.08%ID/g at 24 h. p.i.). Through the specific binding of G1-(DUPA)4 to PSMA, the smallest dendrimer (5.1 kDa) demonstrated the highest PC3-PIP to muscle and PC3-PIP to PC3-FLU uptake ratios (17.7 ± 5.5 and 6.7 ± 3.0 at 4 h p.i., respectively). In addition, the enhanced permeability and retention (EPR) effect appeared to be an overwhelming factor for tumor uptake of the largest dendrimer G5-(DUPA)64 as the uptake was at a similar level irrelevant to the PSMA expression.

Asunto(s)

Dendrímeros/química , Dendrímeros/farmacocinética , Glutamato Carboxipeptidasa II/metabolismo , Glicoproteínas de Membrana/metabolismo , Neoplasias de la Próstata/metabolismo , Triazinas/química , Animales , Transporte Biológico , Línea Celular Tumoral , Glutaratos/química , Humanos , Masculino , Ratones , Tomografía de Emisión de Positrones , Neoplasias de la Próstata/diagnóstico por imagen , Urea/análogos & derivados , Urea/química

RESUMEN

Helicobacter pylori is a major human pathogen that causes a wide range of gastrointestinal pathology. Progression of H. pylori induced gastritis to more severe disease has been found to highly correlate with the array of virulence factors expressed by the pathogen. The objective of this study was twofold: first, to characterize the genetic diversity of H. pylori strains isolated from 41 non-atrophic gastritis patients in Switzerland, an issue that has not been investigated to date. And second, to assess the prevalence and sequence variation of H. pylori virulence factors (cagA, vacA, iceA and dupA) and genes encoding outer membrane proteins (OMPs; babA, babB, sabA, sabB, hopZ, hopQ and oipA) by whole genome sequencing (WGS) using an Illumina MiSeq platform. WGS identified high genetic diversity in the analyzed H. pylori strains. Most H. pylori isolates were assigned to hpEurope (95.0%, 39/41), and the remaining ones (5.0%, 2/41) to hpEastAsia, subpopulation hspEAsia. Analysis of virulence factors revealed that 43.9% of the strains were cagA-positive, and the vacA s1 allele was detected in 56.0% of the isolates. The presence of cagA was found to be significantly associated (P < 0.001) with the presence of vacA s1, babA2 and hopQ allele 1 as well as expression of oipA. Moreover, we found an association between the grade of gastritis and H. pylori abundance in the gastric mucosa, respectively and the presence of cagA, vacA s1 and hopQ allele 1. Among our 41 gastritis patients, we identified seven patients infected with H. pylori strains that carried a specific combination of virulence factors (i.e., cagA, vacA s1 allele and babA2 allele), recently implicated in the development of more severe gastrointestinal pathology, like peptic ulcer disease and even gastric cancer. To this end, WGS can be employed for rapid and detailed characterization of virulence determinants in H. pylori, providing valuable insights into the pathogenic capacity of the bacterium. This could ultimately lead to a higher level of personalized treatment and management of patients suffering from H. pylori associated infections.

RESUMEN

BACKGROUND: The global prevalence of H. pylori approaches 50%, with prevalence rates between 20 and 40% in developed countries and up to 90% in Africa and other developing nations of the world. Development of H. pylori-associated diseases is determined by a number of virulence factors. This study aimed at determining the prevalence of H. pylori infections and virulence genes (cagA, dupA, and vacA); the relationship between virulence factors and gastroduodenal diseases among patients. METHODS: Gastric biopsies were obtained from patients and cultured, DNA was extracted from cultured isolates and biopsies for PCR assay after which samples were investigated using standard laboratory procedures. Data of associated risk factors were obtained with the aid of questionnaires. RESULTS: Of the 444 participants, H. pylori was detected in 115 (25.9%) from culture analysis and 217 (48.9%) by direct PCR method. Ninety-eight (85.2%) of the culture-positive patients were also detected by PCR giving an overall prevalence of 52.7% (234/444). The highest number of H. pylori isolates 76.9% (180/234) was obtained from patients suffering from pangastritis. The CagA virulence gene was found in 62% (145/234), dupA in 53.4% (125/234) and vacA in 90.6% (212/234). VacA genotype s1 m1 was the most prevalent [56.4% (132)] followed by s2 m2 [11.5% (27)], s2 m1 [10.3% (24)] and [s1 m2 9.4% (22)]. There was a significant association observed in vacA s1 and peptic ulcer disease, as well as vacA s1/m2 and gastric erosion (P < 0.05). CONCLUSION: The study revealed a significant association between virulence genes and the development of certain forms of gastric infections while the variations in H. pylori detection and the associated risk factors investigated in the study were not significantly related.

Asunto(s)

Gastritis/microbiología , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/epidemiología , Helicobacter pylori/genética , Úlcera Péptica/microbiología , Virulencia/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Biopsia , ADN Bacteriano/análisis , Femenino , Gastritis/diagnóstico , Genotipo , Infecciones por Helicobacter/microbiología , Helicobacter pylori/aislamiento & purificación , Helicobacter pylori/patogenicidad , Hospitales de Enseñanza , Humanos , Masculino , Persona de Mediana Edad , Úlcera Péptica/diagnóstico , Prevalencia , Factores de Riesgo , Factores Socioeconómicos , Sudáfrica/epidemiología , Estómago/patología , Factores de Virulencia/genética , Adulto Joven

RESUMEN

The discovery of Helicobacter pylori in 1983 challenged researchers around the world to identify this pathogen's major virulence factors. The main rationale for this kind of research was to identify a biomarker associated with specific diseases following H. pylori colonization. Among different investigated virulence factors, duodenal ulcer promoting gene A (dupA) has been found to be associated with duodenal ulcer (DU), but its effect was different in various geographical regions. To determine the prevalence of dupA, we applied both classic primer pairs and our newly developed primers producing a highly conserved segment in PCR method. In our survey, 143 (47%) H. pylori isolates were obtained from 304H. pylori-colonized individuals [age range of 19-92; 113 (37%) males with the mean age of 50 and 191 (63%) females with the mean age of 49]. The presence of the dupA gene was investigated by using the different specific primers. The prevalence of the 112 bp segment isolated from H. pylori strains recovered from DU, GU and atrophy groups were significantly higher (81%, p value = .002, 64%, p = .065, 68% and p = .047 38%, respectively) than our control group, where the prevalence of the 112 bp segment was only 38%. Interestingly, a significant relationship was observed between the occurrence of DU and the presence of the 112 bp segment [p = .002; OR: 6.98; (95% CI: 1.94-25.00)]. Taken as a whole, we believe the 112 bp region of H. pylori dupA may serve as the first detected biomarker for the early detection of DU in patients admitted to hospitals.

Asunto(s)

Susceptibilidad a Enfermedades , Úlcera Duodenal/epidemiología , Úlcera Duodenal/etiología , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/microbiología , Helicobacter pylori/genética , Factores de Virulencia/genética , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Medición de Riesgo , Factores de Riesgo