Skin tropism during Usutu virus and West Nile virus infection: an amplifying and immunological role.

Usutu virus (USUV) and West Nile virus (WNV) are closely related emerging arboviruses belonging to the Flavivirus genus and posing global public health concerns. Although human infection by these viruses is mainly asymptomatic, both have been associated with neurological disorders such as encephalitis and meningoencephalitis. Since USUV and WNV are transmitted through the bite of an infected mosquito, the skin represents the initial site of virus inoculation and provides the first line of host defense. Although some data on the early stages of WNV skin infection are available, very little is known about USUV. Herein, USUV-skin resident cell interactions were characterized. Using primary human keratinocytes and fibroblasts, an early replication of USUV during the first 24 hours was shown in both skin cells. In human skin explants, a high viral tropism for keratinocytes was observed. USUV infection of these models induced type I and III interferon responses associated with upregulated expression of various interferon-stimulated genes as well as pro-inflammatory cytokine and chemokine genes. Among the four USUV lineages studied, the Europe 2 strain replicated more efficiently in skin cells and induced a higher innate immune response. In vivo, USUV and WNV disseminated quickly from the inoculation site to distal cutaneous tissues. In addition, viral replication and persistence in skin cells were associated with an antiviral response. Taken together, these results provide a better understanding of the pathophysiology of the early steps of USUV infection and suggest that the skin constitutes a major amplifying organ for USUV and WNV infection.IMPORTANCEUsutu virus (USUV) and West Nile virus (WNV) are closely related emerging Flaviviruses transmitted through the bite of an infected mosquito. Since they are directly inoculated within the upper skin layers, the interactions between the virus and skin cells are critical in the pathophysiology of USUV and WNV infection. Here, during the early steps of infection, we showed that USUV can efficiently infect two human resident skin cell types at the inoculation site: the epidermal keratinocytes and the dermal fibroblasts, leading to the induction of an antiviral innate immune response. Moreover, following cutaneous inoculation, we demonstrated that both viruses can rapidly spread, replicate, and persist in all distal cutaneous tissues in mice, a phenomenon associated with a generalized skin inflammatory response. These results highlight the key amplifying and immunological role of the skin during USUV and WNV infection.

One year of SARS-CoV-2 circulation in the Nouvelle-Aquitaine region, February 2021-2022, France.

Background: Since 2021, 3 variants of concern (VOC) have spread to France, causing successive epidemic waves. Objectives: To describe the features of Alpha, Delta and Omicron VOC circulation in the Nouvelle-Aquitaine region, France, between February 2021 and February 2022. Study design: Data from the three university hospitals (UH) of Nouvelle-Aquitaine were used to describe regional SARS-CoV-2 circulation (RT-PCR positive rates and identified VOC) as well as its consequences (total number of hospitalizations and admissions in intensive care unit). They were analyzed according to the predominant variant and compared with national data. Results: A total of 611,106 SARS-CoV-2 RT-PCR tests were performed in the 3 Nouvelle-Aquitaine UH during the study period. The 37,750 positive samples were analyzed by variant-specific RT-PCR or whole-genome sequencing. In 2021, Alpha VOC was detected from week 5 until week 35. Delta became the most prevalent variant (77.3%) in week 26, reaching 100% in week 35. It was replaced by Omicron, which was initially detected week 48, represented 77% of positive samples in week 52 and was still predominant in February 2022. The RT-PCR positive rates were 4.3, 4.2, and 21.9% during the Alpha, Delta and Omicron waves, respectively. The ratio between intensive care unit admissions and total hospitalizations was lower during the Omicron wave than during the two previous waves due to the Alpha and Delta variants. Conclusion: This study highlighted the need for strong regional cooperation to achieve effective SARS-CoV-2 epidemiological surveillance, in close association with the public health authorities.

Anti-SARS-CoV-2 Activity of Polysaccharides Extracted from Halymenia floresii and Solieria chordalis (Rhodophyta).

Even after hundreds of clinical trials, the search for new antivirals to treat COVID-19 is still relevant. Carrageenans are seaweed sulfated polysaccharides displaying antiviral activity against a wide range of respiratory viruses. The objective of this work was to study the antiviral properties of Halymenia floresii and Solieria chordalis carrageenans against SARS-CoV-2. Six polysaccharide fractions obtained from H. floresii and S. chordalis by Enzyme-Assisted Extraction (EAE) or Hot Water Extraction (HWE) were tested. The effect of carrageenan on viral replication was assessed during infection of human airway epithelial cells with a clinical strain of SARS-CoV-2. The addition of carrageenans at different times of the infection helped to determine their mechanism of antiviral action. The four polysaccharide fractions isolated from H. floresii displayed antiviral properties while the S. chordalis fractions did not. EAE-purified fractions caused a stronger reduction in viral RNA concentration. Their antiviral action is likely related to an inhibition of the virus attachment to the cell surface. This study confirms that carrageenans could be used as first-line treatment in the respiratory mucosa to inhibit the infection and transmission of SARS-CoV-2. Low production costs, low cytotoxicity, and a broad spectrum of antiviral properties constitute the main strengths of these natural molecules.

Effect of High Temperature and UV Radiation on the Insecticidal Capacity of a Spodoptera frugiperda Nucleopolyhedrovirus Microencapsulated in a Matrix Based on Oxidized Corn Starch.

A multiple nucleopolyhedrovirus native isolate (SfCH32) of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) was encapsulated by spray-drying in a matrix based on oxidized corn starch without and with a fluorescent brightener. The microcapsules were exposed to UV radiation (365 nm) for 0, 2, 4, and 8 h at 25 °C or temperatures of 35, 40, and 45 °C for 8 h. The data obtained with temperatures 35, 40, and 45 °C were contrasted with those obtained at 25 °C. The microcapsules were evaluated for size, shape, and insecticidal capacity against third instar S. frugiperda larvae under laboratory conditions. The 82-84.2% of the encapsulating matrix, in a dry-weight basis, was recovered as NPV microcapsules of heterogeneous shape and size. The exposure to UV radiation and temperatures reduced significantly the insecticidal capacity of tested viruses; however, such capacity was higher for microencapsulated than for non-microencapsulated viruses. The non-encapsulated virus that had been exposed to 45 °C or maintained at UV radiation for 8 h showed the lowest insecticidal activity at 5th day post-inoculation, with a larvae mortality of 25.3 and 16%, respectively. The fluorescent brightener increased significantly the insecticidal capacity of encapsulated and non-encapsulated viruses, causing a mortality of 100% at that time point, and decreased the median lethal time independently of the incubation temperature and exposure time to radiation. The findings suggested that an encapsulating matrix based on oxidized corn starch might protect the insecticidal capacity of NPV under field conditions.

Diversity and genetic structure of natural populations of the palm tree Euterpe precatoria (Arecaceae) / Diversidad y estructura genética de poblaciones naturales de la palmera Euterpe precatoria (Arecaceae)

Abstract Introduction: The natural ecosystems of northern Mato Grosso, Brazil, are in process of fragmentation, mainly due to population growth and the expansion of agriculture. This endangers the palm Euterpe precatoria (locally known as açaí), used for construction, palm hearts, juices and ice cream. Objective: To evaluate the local diversity and genetic structure in native populations of E. precatoria. Methods: We collected leaves from 106 fruiting palms from five populations in Mato Grosso State, for analysis of microsatellite markers with Polymerase Chain Reaction. Results: The five SSR loci revealed a total of 30 alleles, ranging from 5 (EE23 and EE43) to 7 (EE2 and EE15), with an average of 6 alleles per locus. The mean PIC was 0.74 and confirmed low heterozygosity and inbreeding. The UPGMA dendrogram produced two groups and molecular variance revealed greater genetic differentiation within populations. The high levels of homozygous microsatellite loci indicate low genetic diversity. Conclusions: These populations have low gene diversity, high average number of alleles per locus, and rare and exclusive alleles. We recommend the establishment of permanent conservation units with corridors among them.

Resumen Introducción: Los ecosistemas naturales del norte de Mato Grosso, Brasil, están en proceso de fragmentación, principalmente debido al crecimiento de la población y la expansión de la agricultura. Esto pone en peligro la palma Euterpe precatoria (localmente conocida como açaí), utilizada para la construcción, extracción de palmito, preparación de jugos y helados. Objetivo: Evaluar la diversidad local y estructura genética en poblaciones nativas de E. precatoria. Métodos: Recolectamos hojas de 106 palmas fructíferas de cinco poblaciones en el estado de Mato Grosso, para análisis de marcadores microsatélites con el método de Reacción en Cadena de la Polimerasa (PCR). Resultados: Los cinco loci SSR revelaron un total de 30 alelos, que van desde 5 (EE23 y EE43) hasta 7 (EE2 y EE15), con un promedio de 6 alelos por locus. El PIC medio fue de 0.74 y confirmó baja heterocigosidad y endogamia en las poblaciones. El dendrograma UPGMA produjo dos grupos y la varianza molecular reveló una mayor diferenciación genética dentro de las poblaciones. Los loci de microsatélites presentaron un alto nivel de homocigotos lo que indica una baja diversidad genética. Conclusiones: Estas poblaciones tienen baja diversidad genética, alto promedio de alelos por locus y alelos raros y únicos. Recomendamos el establecimiento de unidades de conservación permanentes con corredores entre ellas.

Pooling Rectal, Pharyngeal, and Urine Samples to Detect Neisseria gonorrhoeae, Chlamydia trachomatis, and Mycoplasma genitalium Using Multiplex Polymerase Chain Reaction Is as Effective as Single-Site Testing for Men Who Have Sex With Men.

Background: Screening for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) at pharyngeal, urogenital, and anorectal sites is recommended for men who have sex with men (MSM). Pooling samples is a promising technique, but no data are available when pooled screening also includes Mycoplasma genitalium (MG). The main objective of this study was to examine the sensitivity of pooled samples for detecting CT, NG, and MG in MSM using nucleic acid amplification versus single-site testing. Methods: In this multicenter study, MSM with a positive result for CT, NG, or MG were recalled to the clinic for treatment and were asked to participate in this study. Separate samples were sent to a central virological department that proceeded to form the pooled samples. Testing was performed using the multiplex real-time polymerase chain reaction Allplex STI Essential Assay (Seegene, Seoul, Korea), which can simultaneously detect 7 pathogens. Results: A total of 130 MSM with at least 1 positive test for CT, NG, or MG were included. A total of 25.4% had a coinfection. The sensitivities of pooled-sample testing were 94.8% for CT, 97.0% for NG, and 92.3% for MG. Pooling failed to detect 8 infections, but pooled-sample analysis missed detecting only samples with a low bacterial load (cycle threshold >35). Conclusions: Pooling samples from MSM to detect CT, NG, and MG is as sensitive as individual-site testing for these 3 pathogens using the Allplex assay. Missed infections with a very low bacterial load could have a low impact on further transmission. Clinical Trials Registration. NCT03568695.

Efficacy and Safety of Doravirine-Based Regimens in Real Life: A Prospective Monocentric French Study.

Doravirine (DOR) efficacy and safety have been evaluated in adult naive or treated patients starting a DOR-based regimen between September 15, 2019, and December 31, 2020. Medical history and examination, laboratory results, and tolerance were assessed during the 48 weeks of follow-up. Among the 77 patients included, virological control (VC) was noticed for 66 patients at baseline. Median age was 51 years, and 62% were men. The most common reason for initiating a DOR-based therapy was toxicity (44; 67%) and, especially, weight gain. A virological suppression (VS) was maintained in 55 (83%) patients of the VC group and noticed in 9 (82%) of the non-VC patients at week 48, by intention-to-treat analysis. On treatment analysis, 98% and 100% patients achieved VS in the VC and non-VC groups, respectively. The renal and metabolic tolerance were good. DOR-based regimens appear to be a safe and relevant strategy to circumvent drug interactions and drugs with a poor metabolic tolerance profile.

Antiviral Effect of hBD-3 and LL-37 during Human Primary Keratinocyte Infection with West Nile Virus.

West Nile virus (WNV) is an emerging flavivirus transmitted through mosquito bites and responsible for a wide range of clinical manifestations. Following their inoculation within the skin, flaviviruses replicate in keratinocytes of the epidermis, inducing an innate immune response including the production of antimicrobial peptides (AMPs). Among them, the cathelicidin LL-37 and the human beta-defensin (hBD)-3 are known for their antimicrobial and immunomodulatory properties. We assessed their role during WNV infection of human primary keratinocytes. LL-37 reduced the viral load in the supernatant of infected keratinocytes and of the titer of a viral inoculum incubated in the presence of the peptide, suggesting a direct antiviral effect of this AMP. Conversely, WNV replication was not inhibited by hBD-3. The two peptides then demonstrated immunomodulatory properties whether in the context of keratinocyte stimulation by poly(I:C) or infection by WNV, but not alone. This study demonstrates the immunostimulatory properties of these two skin AMPs at the initial site of WNV replication and the ability of LL-37 to directly inactivate West Nile viral infectious particles. The results provide new information on the multiple functions of these two peptides and underline the potential of AMPs as new antiviral strategies in the fight against flaviviral infections.

Functional and morphological analysis of isolates of phylloplane and rhizoplane endophytic bacteria interacting in different cocoa production systems in the Amazon.

Endophytic bacteria colonize different internal tissues of plants without damaging their cells. They can establish themselves in the same niche as other microorganisms and develop antagonistic activities against phytopathogens. There is little research on the functional and morphological characterization of these bacteria in production systems in the Amazon. Thus, the objective of this work was to functionally and morphologically characterize endophytic bacteria isolated from cocoa trees (Theobroma cacao L.) and evaluate their antagonistic potential against phytopathogens. A total of 197 endophytic bacteria isolates were obtained from leaves and roots of cocoa plants with different production systems and at different times of the year. The characterization of functional groups consisted of proteolytic, amylolytic and cellulolytic activity and ability to fix nitrogen and solubilize phosphate. Morphological diversity was evaluated mainly according to the following parameters: shape, color, size and elevation of the colony. Thirteen isolates of endophytic bacteria, selected by cluster analysis, were used to evaluate the antagonistic potential in paired trials against four species of phytopathogenic fungi. The largest amount of endophytic bacteria was isolated from the root (95.9%), in the dry season. The most expressive activities with regards to the enzyme index were amylolytic (71.9%), proteolytic (70.2%) and nitrogen fixing (38.6%), respectively. The similarity analysis formed two clusters with isolates CS R 2.4 and CS R 2.25 exhibiting 100% similarity. Five isolates displayed inhibitory activity against phytopathogenic fungi, most notably isolate TS R 2.19, which exhibited antagonistic activity against all fungi and mycelial growth inhibition rates between 25.7% and 50.7%. Understanding the interaction between endophytes in cocoa plants is important as a possible additional tool in biological control. Our studies are incipient and the first to be carried out in different cocoa production systems in the state of Pará, Brazil.

Phenol-soluble modulins α are major virulence factors of Staphylococcus aureus secretome promoting inflammatory response in human epidermis.

Staphylococcus aureus is a skin commensal microorganism commonly colonizing healthy humans. Nevertheless, S. aureus can also be responsible for cutaneous infections and contribute to flare-up of inflammatory skin diseases such as atopic dermatitis (AD), which is characterized by dysbiosis of the skin microbiota with S. aureus as the predominant species. However, the role of major virulence factors of this pathogen such as phenol-soluble modulin (PSM) toxins in epidermal inflammation remains poorly understood. Stimulation of primary human keratinocytes with sublytic concentrations of synthetic and purified PSM α3 resulted in upregulation of a large panel of pro-inflammatory chemokine and cytokine gene expression, including CXCL1, CXCL2, CXCL3, CXCL5, CXCL8, CCL20, IL-1α, IL-1ß, IL-6, IL-36γ and TNF-α, while inducing the release of CXCL8, CCL20, TNF-α and IL-6. In addition, using S. aureus culture supernatant from mutants deleted from genes encoding either α-type PSMs or all PSM production, PSMs were shown to be the main factors of S. aureus secretome responsible for pro-inflammatory mediator induction in human keratinocytes. On the other hand, α-type PSM-containing supernatant triggered an intense induction of pro-inflammatory mediator expression and secretion during both topical and basal layer stimulation of an ex vivo model of human skin explants, a physiologically relevant model of pluristratified epidermis. Taken together, the results of this study show that PSMs and more specifically α-type PSMs are major virulence factors of S. aureus inducing a potent inflammatory response during infection of the human epidermis and could thereby contribute to AD flare-up through exacerbation of skin inflammation.

Characterisation of Antiviral Activity of Cathelicidins from Naked Mole Rat and Python bivittatus on Human Herpes Simplex Virus 1.

Hg-CATH and Pb-CATH4 are cathelicidins from Heterocephalus glaber and Python bivittatus that have been previously identified as potent antibacterial peptides. However, their antiviral properties were not previously investigated. In this study, their activity against the herpes simplex virus (HSV)-1 was evaluated during primary human keratinocyte infection. Both of them significantly reduced HSV-1 DNA replication and production of infectious viral particles in keratinocytes at noncytotoxic concentrations, with the stronger activity of Pb-CATH4. These peptides did not show direct virucidal activity and did not exhibit significant immunomodulatory properties, except for Pb-CATH4, which exerted a moderate proinflammatory action. All in all, our results suggest that Hg-CATH and Pb-CATH4 could be potent candidates for the development of new therapies against HSV-1.

Necesidad de equipos de salud mental para pacientes hospitalizados por SARS-CoV-2 y personal de salud de primera línea / Need of mental health teams for patients hospitalized due to SARS-CoV-2 and front-line health personnel

La población con mayor riesgo para desarrollar trastornos mentales durante la pandemia por COVID-19 es la de los pacientes en aislamiento y la del personal médico de primera línea; los malestares que más frecuentemente han sido reportados incluyen: insomnio, ansiedad y depresión. Debido a que el personal de salud mental se considera no esencial para la atención de los pacientes en aislamiento, se desaconseja su ingreso a áreas COVID, por lo que el personal médico de primera línea debe proporcionar intervenciones psicológicas sin saber cómo mitigar la angustia de los pacientes. El personal de salud tampoco recibe atención adecuada debido a algunas barreras para solicitar ayuda, como la falta de tiempo, el estigma y el temor a una intervención no deseada. Las herramientas para la atención a distancia como la telepsiquiatría y el apoyo psicológico en línea no permiten la obtención de datos importantes, como la exploración física. En China se publicaron principios rectores para la atención de la población, los cuales recomendaron la creación de equipos de salud mental. En el presente trabajo se presentan los datos obtenidos por un equipo de salud mental de una unidad del Instituto Mexicano del Seguro Social reconvertida por COVID-19, que muestra diferencias en el malestar emocional y los diagnósticos de trastornos mentales reportados internacionalmente en pacientes en aislamiento y en personal de salud de primera línea, que sugiere la necesidad de equipos integrados por personal de psiquiatría y psicología con evaluaciones presenciales.

People vulnerable to mental health problems during the COVID-19 outbreak include hospitalized patients and frontline health professionals; mostly reported discomforts include: insomnia, anxiety and depression. Under strict infection measures, non-essential personnel such as clinical psychiatrists and psychologists are strongly discouraged from entering isolation wards; therefore, frontline healthcare workers become the main personnel providing psychological interventions to patients in hospitals without knowing how to mitigate patient distress. Health personnel do not receive adequate care for due to lack of time, stigma associated with using mental health services and fear of unwanted intervention. Remote care tools such as tele-psychiatry and online psychological intervention have disadvantages for conditions requiring physical examination. The National Health Commission of China released guiding principles to reduce the psychosocial effects of the COVID-19 outbreak recommending the creation of mental health teams. In this article we present the data obtained by a mental health team in a COVID-19 hospital of the Mexican Institute of Social Security, showing differences in emotional discomfort and diagnoses of mental disorders in patients in isolation and in frontline health personnel, suggesting the need for teams made up of psychiatry and psychology staff with face-to-face on site assessments.

Post-herpetic encephalitis cerebral abscess: Viral reactivation or latency site within central nervous system?

Herpetic encephalitis results from central nervous system invasion by herpes simplex virus. We report the case of a man who developed a cerebral abscess fifteen months after initial Herpetic encephalitis. Retrospectively, antiviral should not have been associated with antibiotics during abscess episode, as transcriptomic analysis reported no viral reactivation.

The Antiviral Activities of Poly-ADP-Ribose Polymerases.

The poly-adenosine diphosphate (ADP)-ribose polymerases (PARPs) are responsible for ADP-ribosylation, a reversible post-translational modification involved in many cellular processes including DNA damage repair, chromatin remodeling, regulation of translation and cell death. In addition to these physiological functions, recent studies have highlighted the role of PARPs in host defenses against viruses, either by direct antiviral activity, targeting certain steps of virus replication cycle, or indirect antiviral activity, via modulation of the innate immune response. This review focuses on the antiviral activity of PARPs, as well as strategies developed by viruses to escape their action.

Distinct genetic basis for root responses to lipo-chitooligosaccharide signal molecules from different microbial origins.

Lipo-chitooligosaccharides (LCOs) were originally found as symbiotic signals called Nod Factors (Nod-LCOs) controlling the nodulation of legumes by rhizobia. More recently, LCOs were also found in symbiotic fungi and, more surprisingly, very widely in the kingdom Fungi, including in saprophytic and pathogenic fungi. The LCO-V(C18:1, fucosylated/methyl fucosylated), hereafter called Fung-LCOs, are the LCO structures most commonly found in fungi. This raises the question of how legume plants such as Medicago truncatula can discriminate between Nod-LCOs and Fung-LCOs. To address this question, we performed a genome-wide association study on 173 natural accessions of M. truncatula, using a root branching phenotype and a newly developed local score approach. Both Nod-LCOs and Fung-LCOs stimulated root branching in most accessions, but the root responses to these two types of LCO molecules were not correlated. In addition, the heritability of the root response was higher for Nod-LCOs than for Fung-LCOs. We identified 123 loci for Nod-LCO and 71 for Fung-LCO responses, of which only one was common. This suggests that Nod-LCOs and Fung-LCOs both control root branching but use different molecular mechanisms. The tighter genetic constraint of the root response to Fung-LCOs possibly reflects the ancestral origin of the biological activity of these molecules.

[Need of mental health teams for patients hospitalized due to SARS-CoV-2 and front-line health personnel]. / Necesidad de equipos de salud mental para pacientes hospitalizados por SARS-CoV-2 y personal de salud de primera línea.

People vulnerable to mental health problems during the COVID-19 outbreak include hospitalized patients and frontline health professionals; mostly reported discomforts include: insomnia, anxiety and depression. Under strict infection measures, non-essential personnel such as clinical psychiatrists and psychologists are strongly discouraged from entering isolation wards; therefore, frontline health-care workers become the main personnel providing psychological interventions to patients in hospitals without knowing how to mitigate patient distress. Health personnel do not receive adequate care for due to lack of time, stigma associated with using mental health services and fear of unwanted intervention. Remote care tools such as tele-psychiatry and online psychological intervention have disadvantages for conditions requiring physical examination. The National Health Commission of China released guiding principles to reduce the psychosocial effects of the COVID-19 outbreak recommending the creation of mental health teams. In this article we present the data obtained by a mental health team in a COVID-19 hospital of the Mexican Institute of Social Security, showing differences in emotional discomfort and diagnoses of mental disorders in patients in isolation and in frontline health personnel, suggesting the need for teams made up of psychiatry and psychology staff with face-to-face on-site assessments.

La población con mayor riesgo para desarrollar trastornos mentales durante la pandemia por COVID-19 es la de los pacientes en aislamiento y la del personal médico de primera línea; los malestares que más frecuentemente han sido reportados incluyen: insomnio, ansiedad y depresión. Debido a que el personal de salud mental se considera no esencial para la atención de los pacientes en aislamiento, se desaconseja su ingreso a áreas COVID, por lo que el personal médico de primera línea debe proporcionar intervenciones psicológicas sin saber cómo mitigar la angustia de los pacientes. El personal de salud tampoco recibe atención adecuada debido a algunas barreras para solicitar ayuda, como la falta de tiempo, el estigma y el temor a una intervención no deseada. Las herramientas para la atención a distancia como la telepsiquiatría y el apoyo psicológico en línea no permiten la obtención de datos importantes, como la exploración física. En China se publicaron principios rectores para la atención de la población, los cuales recomendaron la creación de equipos de salud mental. En el presente trabajo se presentan los datos obtenidos por un equipo de salud mental de una unidad del Instituto Mexicano del Seguro Social reconvertida por COVID-19, que muestra diferencias en el malestar emocional y los diagnósticos de trastornos mentales reportados internacionalmente en pacientes en aislamiento y en personal de salud de primera línea, que sugiere la necesidad de equipos integrados por personal de psiquiatría y psicología con evaluaciones presenciales.

Where to Biopsy to Detect Helicobacter pylori and How Many Biopsies Are Needed to Detect Antibiotic Resistance in a Human Stomach.

This study aims to determine the gastric distribution, density, and diversity of Helicobacter pylori infection. Subtotal resection of the stomachs of three H. pylori-infected and asymptomatic obese patients were collected after a sleeve gastrectomy. Distribution and density of H. pylori were determined using culture and RT-PCR on multiple gastric sites (88, 176, and 101 biopsies per patient). Diversity of H. pylori strains was studied using antibiotic susceptibility testing, random amplified polymorphism DNA (RAPD) typing and cagA gene detection on single-colony isolates (44, 96, and 49 isolates per patient). H. pylori was detected in nearly all analyzed sites (354/365 biopsies, 97%). Antral density was higher in one patient only. The three stomachs were almost exclusively infected by an antibiotic-susceptible strain. One clarithromycin-resistant isolate in one biopsy was detected in two stomachs (1/44 and 1/49 isolates), while in the third one, eight (8/96 isolates) metronidazole-resistant isolates were detected. DNA typing showed infection with cagA-negative strains for one patient, cagA-positive strains for a second patient and the third patient was infected with two different strains of distinct cagA genotypes. Infection with H. pylori is shown to spread to the whole surface of the stomach, but a possibility of minor sub-population of antibiotic-resistant clones, undetectable in routine practice.