Warthin-Starry Stain Research Articles

P30 Exposure to particulate matter induces structural and pigmentary changes to human skin

Abstract Introduction and aims The majority of the world’s population reside in places with high levels of air pollution. The impact of air pollution on human health has been extensively studied and demonstrates a significant correlation between particulate matter (PM) and risk of cardiovascular and respiratory diseases. However, little is known about the impact of air pollution on human skin health. Here, we develop an ex vivo skin model to investigate cutaneous changes induced by the topical application of PM. Methods PM (0, 0.5, 3, 6, 10, 20 and 40 μg mL−1) was topically applied to Genoskin® NativeSkin Access® (Genoskin, Salem, MA, USA) human ex vivo skin samples of White European ancestry (n = 3) for 8 days, with reapplication on day 4. Resultant samples were assessed using haematoxylin and eosin staining (for morphometric analyses), immunofluorescence for Ki67 and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) (to determine cellular turnover), while pigmentary changes were analysed by colourimetry, Warthin–Starry (WS) staining and immunohistochemistry for the melanin markers tyrosinase (TYR), gp100 and c-kit. Results Morphological assessment revealed that epidermal thickness was significantly reduced following exposure to higher doses of PM (r = −0.9163, P < 0.01). In addition, there was reduced epidermal cell proliferation (Ki67) and increased apoptosis (TUNEL) at higher doses of PM. Exposure to PM resulted in a dose-dependent decrease in individual typology angle, which is indicative of skin colour becoming darker (r = −0.7895, P < 0.05). Furthermore, WS staining demonstrated significantly increased epidermal melanin (r = 0.9583, P < 0.01) and significantly increased expression of melanin markers TYR (P < 0.01), gp100 (P < 0.001) and c-kit (P < 0.01) in response to PM exposure. Conclusions This study confirms that PM induces pigmentary changes to ex vivo human skin by increasing epidermal melanin. Increased expression of c-kit in response to PM exposure suggests that increased pigmentation occurs owing to the upregulation of the microphthalmia-associated transcription factor signalling pathway. Further investigation of the mechanistic drivers of increased pigmentation is warranted.

Immunohistochemical Demonstration of Mast Cells in Bovine Papillomatous Digital Dermatitis of Holstein-Friesian Dairy Cattle

Mast cells (MC)are unique members of immune system; their location and functions are of great importance in health and disease status. This study aims to evaluate MCs distribution and heterogenity in healthy and digital dermatitis lesions of dairy cattle for the first time. A total of 50 skin samples, 25 healthy and 25 with digital dermatitis (DD) lesions were sampled in Holstein-Friesian dairy cattle. All samples were stained with hematoxylin-eosin (H&E) for microscopic examination and also stained with Toluidine blue for MCs demonstration. Epidermal acanthosis and hyperkeratosis with dermal inflammatory cell infiltrations were in consistent with digital dermatitis lesions. Spirochetal agents were successfully demonstrated with Warthin-Starry staining in 22 out of 25 DD samples. Increased number of mast cell were observed in digital dermatitis samples when compared with healthy skin samples. The average number of intact MCs were 4.6 ± 2 and 9.3 ± 1 in healthy and digital dermatitis samples, respectively. Two-fold increase in the number of intact MCs in digital dermatitis samples was observed. Degranulated mast cell numbers in tissue sections were also higher in digital dermatitis samples and 4.25-fold increase was recorded in affected skin samples. Immunophenotype of MCs in skin samples were identified by immmunohistochemical stainings with anti-tryptase and chymase antibodies. Only tryptase positive MCs were observed in healthy and DD samples. In the statistical analysis, differences in the mean intact and degranulated MCs were found to be significant (mean intact: p<0.05, mean degranulated: p<0.01). Our results possibly suggest that MCs may have important roles in the pathogenesis of bovine digital dermatitis.

Canine Gallbladder Erosion/Ulcer and Hemocholecyst: Clinicopathological Characteristics of 14 Cases.

(1) Background: Gallbladder mucosal erosion and/or ulceration are illnesses associated with unexpected gallbladder intra-cystic bleeding (hemocholecyst), an under-reported problem in dogs. (2) Methods: Clinicopathological characteristics of 14 dogs with gallbladder erosion/ulcer were investigated in this single-center retrospective study using clinical data and archived gallbladder tissues of client-owned dogs. (3) Results: Canine gallbladder erosion/ulcer tends to occur in older, neutered dogs of various breeds. Vomiting, lethargy, and anorexia are common. Concurrent gallbladder rupture occurred in 5/14 cases (35.7%), while rupture was absent in 6/14 cases (42.8%) and undetermined in 3/14 (21.4%) cases. Histologically, the gallbladder wall was markedly thickened due to mucosal hyperplasia, inflammatory infiltrates, fibrosis, edema, hemorrhage, and smooth muscle hyperplasia/hypertrophy. Twelve out of fourteen cases (85.7%) had concurrent cholecystitis of varying severity. Bacteria were detected by Giemsa or Warthin-Starry stain in 8/14 (57.1%) cases. Bacterial rods immunoreactive to the anti-Helicobacter antibody were present in one case. Mucosal epithelial cells of the gallbladder erosion/ulcer cohort were immunopositive for the cyclooxygenases COX-1 or COX-2 in only 5/14 (35.7%) cases. In contrast, COX-1 and COX-2 were more frequently expressed in a reference pool of cases of gallbladder mucocele (n = 5) and chronic cholecystitis (n = 5). COX-1 was expressed in 9/10 cases (90.0%) of gallbladder mucocele and chronic cholecystitis and in 10/10 cases (100%) for COX-2. (4) Conclusions: Canine gallbladder erosion/ulcer is an under-reported condition which requires active clinical intervention. Based on the clinicopathological information reported in this study in addition to the COX-1 and COX-2 IHC results, we suggest that canine gallbladder erosion/ulcer may be related to decreased cytoprotection physiologically provided by arachidonic acid, but which is decreased or absent due to reduced COX expression because of yet undetermined etiologies.

Leptospirosis in Wild Mice: An Ongoing Concern for the Azorean Islands

Background: Leptospirosis is a worldwide zoonotic infectious disease that affects Human and animals worldwide with increased risk in regions with temperate climates. Such is the case of the Azorean islands (a Portuguese archipelago with 9 islands) where this disease is considered endemic. Rodents of these islands have been considered the reservoir species for Leptospira and regulating measurements have been previously developed to control and prevent this disease in the archipelago. This study aims to determine a more recent infection rate of leptospirosis in the house-mouse (Mus musculus). Methodology & Findings: This retrospective study covering a period of 8 years (2011 to 2018), was performed by live capturing Mus musculus in 6 locations of S. Miguel Island (the most populated island of the archipelago). After euthanasia and necropsy of the mice, kidneys were surgically removed and processed in standard histological routine, and slides were stained with modified Warthin-Starry silver stain for spirochetes and scanned for the presence of Leptospires. It was possible to determine that from the 133 wild mice captured, 89 were positive for leptospirosis, representing a 66.91% infection rate. Conclusion: This study reveals that leptospirosis still represents an increased risk to Human lives and an uncalculated economical loss to the dairy farms of the Azorean islands.

Upper Lip Nodule as the First Manifestation of Sarcoidosis: A Case Report

Introduction: Sarcoidosis is a chronic disorder characterized by the presence of multiple granules, which could affect different systems and tissues. These granules are histologically characterized by non-caseous granulomatous inflammation; however, their etiology is still unknown. Previous reports state that different antigens may be involved. Sarcoidosis has been rarely reported in the Brazilian population. The objective of this study was to report a clinical case of sarcoidosis presenting with oral manifestations as the first sign and symptom. Case Presentation: An 85-year-old, nonsmoking, white woman was referred to the Department of Stomatology of our clinic in 2019, complaining of a persistently growing nodule on the upper lip and in the right arm subcutaneous tissue lasting for three months. Both lesions were painless and non-mobile/cohesive nodules with borrachoid consistency on palpation, well-delimited margins, and overlying normal-colored mucosa/skin. The oral nodule was surgically excised and sent for routine histopathological analysis, revealing granulomatous inflammation, the presence of central lobules of activated macrophages, epithelioid histiocytes, multinucleated Langerhans cells, and a peripheral cuff of mononuclear cells (lymphocytes). There was no evidence of central necrotic areas; however, asteroid bodies (i.e., starry inclusions) were rarely interspersed within multinucleated Langerhans cells. Warthin-Starry, Fite, and PAS special staining did not show any evidence of granulomatous pathogens, which was compatible with non-infectious granulomatosis. Complementary tests revealed hypercalcemia and elevated serum levels of alkaline phosphatase and angiotensin-converting enzyme. Conclusions: Sarcoidosis can manifest itself in several ways, so the diagnostic process needs to be thorough and comprehensive to confirm the disease. In addition, disease progression should be regularly monitored in patients.

Melanin-dependent tissue interactions induced by a 755-nm picosecond-domain laser: complementary visualization by optical imaging and histology

Fractional picosecond-domain lasers (PSL) induce optical breakdown, which correlates histologically to vacuolization in the epidermis and dermis. In this ex vivo porcine study, we sought to establish a framework for the investigation of laser-tissue interactions and their dependence on melanin density. Light- (melanin index: 24.5 [0–100]), medium- (58.7), and dark-pigmented (> 98) porcine skin samples were exposed to a 755-nm fractional PSL and examined with dermoscopy, line-field confocal optical coherence tomography (LC-OCT), conventional OCT, and subsequently biopsied for digitally stained ex vivo confocal microscopy (EVCM) and histology, using hematoxylin and eosin (HE) and Warthin-Starry (WS) melanin staining. Dermoscopy showed focal whitening in medium- and dark-pigmented skin. Similarly, LC-OCT and OCT visualized melanin-dependent differences in PSL-induced tissue alterations. Vacuoles were located superficially in the epidermis in dark-pigmented skin but at or below the dermal–epidermal junction in medium-pigmented skin; in light-pigmented skin, no vacuoles were observed. Histology confirmed the presence of vacuoles surrounded by areas void of WS staining and disrupted stratum corneum in darker skin. The combined use of optical imaging for multiplanar visualization and histological techniques for examination of all skin layers may mitigate the effect of common artifacts and attain a nuanced understanding of melanin-dependent laser-tissue interactions.

Multi-hospital retrospective study comparing gastric adenocarcinoma in Hispanic versus non-Hispanic patients.

e16027 Background: Gastric cancer is one of the most prevalent malignancies in the United States and carries a particularly high rate of mortality. In the United States, the overall gastric cancer rate continues to decline amongst Caucasian and African American patients; however, it remains static amongst Hispanic patients. While numerous studies have been conducted to better understand the underlying causes leading to the disproportionate incidence of gastric cancer among Hispanics, it is still not well understood. This retrospective study sought to compare the clinicopathological features and overall survival of Hispanic patients with gastric adenocarcinoma with non-Hispanic patients treated in three safety-net county hospitals in Southern California. Methods: This is a retrospective cohort study of 123 patients with newly diagnosed gastric adenocarcinoma who were treated at three safety-net county hospitals in Southern California between 2010 and 2022. The subjects were categorized as Hispanic and non-Hispanic. Clinical, pathological, and survival data were obtained from the patient chart and compared. Results: Of the 123 patients included in the study, 87 (71%) were Hispanic. At time of diagnosis, Hispanic patients were found to be significantly younger than non-Hispanics (55.4 +/- 13.5 vs 60.8 +/- 12.2, p-value 0.03). They were also more likely to be misdiagnosed (defined as greater than one prior encounter of misdiagnosis within six months) relative to non-Hispanics (26% vs 14%, p-value 0.007) despite no differences in insurance/PCP status. Hispanic patients were more likely to present with metastatic disease (65% vs 30%, p-value <0.001), and were found to have a higher rate of peritoneal carcinomatosis (30% vs 8%, p-value 0.01). Hispanics were also more likely to present with disease in the gastric antrum (47% vs 19%, p-value 0.004) and have a higher rate of diffuse subtype histology (32% vs 22%, p-value 0.05). Similar rates of Helicobacter pylori infection were found within the tissue biopsy sample using Warthin Starry stain amongst Hispanic and non-Hispanic patients (17% vs 17%). Lastly, the mortality rate was higher in Hispanics relative to non-Hispanics at time of last contact (65% vs 35%, p-value <0.001). Conclusions: Upon comparing clinicopathological features and mortality among gastric cancer patients who were treated in multiple safety net county hospitals, there were significant differences noticed in Hispanic American patients. Relative to non-Hispanics, Hispanic patients were more likely to be misdiagnosed, more likely to present with metastatic disease, and have higher rates of mortality despite no disparities in insurance and PCP status. Further investigation is warranted regarding possible risk factors including exposures to possible environmental triggers or if there is a genetic predisposition in Hispanic populations.

An observational study to assess the Helicobacter pylori infection rates in patients with cholelithiasis

Background: Cholelithiasis and chronic cholecystitis cholelithiasis is fairly common all around the world. Since chronic inflammation causes repeated trauma to the gallbladder mucosa and DNA damage led by high or abnormal bile acid exposure, gallstones are closely related to the development of hepatobiliary cancers. After cholecystectomy, some patients experience ongoing upper abdomen pain, which is distressing for the surgeons and raises the possibility of a concomitant upper gastrointestinal issue, which in most cases turned out to be peptic ulcer disease. Numerous studies have found various Helicobacter species in the gallbladder tissue, gallstones, and bile taken from the gallbladder. We conducted this study to assess the relationship between cholelithiasis and H. pylori infection in the stomach. Ascending infection of H. pylori from the stomach and duodenum to biliary tree may have function in development of gall stone production and hepatobiliary carcinomas. Methods: Hospital based cross sectional observational study on patients admitted in surgical wards of SMS Hospital Jaipur for laparoscopic cholecystectomy. All patients underwent UGI and gastric antral biopsy followed by laparoscopic cholecystectomy, gastric mucosa and GB mucosa were subjected to Warthin starry silver stain for detection of H. pylori. Results: Out of 116 patients who underwent upper GI endoscopy followed by laparoscopic cholecystectomy, 29 (25%) patients were positive for H. pylori infection. Conclusions: This study shows significant positive co-relation between H. pylori infection in stomach, gall bladder to chronic cholecystitis and cholelithiasis, which signifies gastric colonisation of H. pylori has it’s role in biliary pathologies through ascending infection, the effectiveness of H. pylori eradication therapy in preventing these pathologies need to be studied further.

Immunohistochemical detection of Treponema pallidum in skin samples with clinical and histopathological correlations and Warthin-Starry staining critical analysis

BackgroundSyphilis in its different phases may be a difficult diagnosis in clinical and histopathological grounds. ObjectivesThe present study objectives were to evaluate the detection and tissue distribution of Treponema pallidum in skin lesions of syphilis. MethodsA blinded diagnostic accuracy study was performed with immunohistochemistry and Warthin-Starry silver staining in skin samples from patients with syphilis and other diseases. Patients attended two tertiary hospitals between 2000 and 2019. Prevalence ratios (PR) and 95% confidence intervals (95% CI) were calculated for the association between immunohistochemistry positivity and clinical-histopathological variables. ResultsThirty-eight patients with syphilis and their 40 biopsy specimens were included in the study. Thirty-six skin samples were used as non-syphilis controls. The Warthin-Starry technique was unable to accurately demonstrate bacteria in all samples. Immunohistochemistry showed spirochetes only in skin samples from patients with syphilis (24/40) with 60% sensitivity (95% CI 44.8‒75.2). Specificity was 100% and accuracy, 78.9% (95% CI 69.8‒88.1). Most cases had spirochetes in both dermis and epidermis and there was a high bacterial load. Study limitationsCorrelation between immunohistochemistry and clinical or histopathological characteristics was observed but was limited statistically due to the small sample size. ConclusionsSpirochetes were promptly seen in an immunohistochemistry protocol, which can contribute to the diagnosis of syphilis in skin biopsy samples. On the other hand, the Warthin-Starry technique showed to be of no practical value.

A147 “ALREADY GLUTEN-FREE”: A CASE REPORT AND INTERNAL REVIEW OF CHALLENGES IN THE DIAGNOSTIC WORK-UP FOR CHILDREN WITH CELIAC DISEASE

Abstract Background A single positive tissue transglutaminase IgA (TTG) result does not diagnose celiac disease (CeD). Starting a gluten free diet (GFD) without the proper approach to diagnosis can result in misdiagnosis and inappropriate lifelong gluten restriction. Helicobacter pylori (H. pylori) infection can be a mimicker of CeD symptomatically, biochemically, and histologically. Purpose We report a case of a patient referred for CeD with a false-positive TTG and severe iron deficiency anemia who had started a GFD at the time of referral. Method In addition to the case review, we reviewed referrals for possible CeD from June 1 to September 1, 2022 at McMaster Children’s Hospital to identify the proportion of patients started on a partial or full GFD before their initial gastroenterology consultation. Result(s) Case Report: A 10-year-old female was referred for a positive TTG result 6x the upper limit of normal (72 U/mL; ULN 12U/mL) and iron deficiency anemia with a hemoglobin (Hb) of 85 and MCV of 62. She had been informed her blood work was diagnostic for CeD and to start a GFD prior to referral to Pediatric Gastroenterology. At the time of consultation she had symptoms of GERD, diarrhea, and epigastric pain. Following her consultation, she started a 6-month gluten challenge to proceed to endoscopy. During this time her TTG normalized (2.7 U/ml), however her iron deficiency anemia persisted and was refractory to iron supplementation (Hb 90, MCV 65.9). Endoscopy revealed flattened mucosa in D2 but was otherwise normal. Villous blunting in D1 with severe lymphoplasmacytic gastritis in the body with microabscesses was seen on biopsies. H. pylori was negative on Warthin-Starry staining and the pathologist reported a Marsh stage of 3a. Despite the villous blunting, due to the normalized TTG and ongoing epigastric pain, anemia, and presence of lymphoid follicles, a carbon-14 urea breath test was completed which was positive for H. pylori (count rate 286 dpm, normal < 50 dpm). The patient received treatment for H. pylori infection for 14 days with 8 week follow-up revealing a negative breath test. There was complete resolution of symptoms, normalization of laboratory results (Hb 143, MCV 82), and negative TTG at 6 months. Quality improvement review On internal review of the patients seen in celiac clinic as new referrals from June 1 to September 1 2022, 22/55 patients (42%) had already started a full or partial GFD, therefore requiring gluten challenge and delay in diagnostic investigations. Conclusion(s) This case demonstrates the importance of following confirmational testing pathways for CeD. It highlights the role of considering potential causes of false positive TTG and the differential diagnosis of CeD such as H. pylori, NSAIDs, and Crohn’s disease. The identification of 42% of patients referred for suspected CeD who were already on a GFD, suggests a need for improved education of primary care providers to prevent misdiagnosis and unnecessary life-long gluten elimination diets in children. Please acknowledge all funding agencies by checking the applicable boxes below None Disclosure of Interest None Declared

Investigación con PCR en Tiempo Real e Histopatología sobre la presencia de H. felis, H. heilmannii y H. pylori en Perros

Helicobacter species such as H. heilmannii, H. pylori, H. felis, H. bizzozeronii and H. salomonis have been identified in cats and dogs, and research suggesting that these species may be zoonotic agents and has been studied intensified in recent years. The aim of this study was to reveal the presence, comparing the histopathological findings and Real-time PCR results of H. felis, H. heilmannii, and H. pylori in the stomach and liver tissues taken during the necropsies of owned, stray or shelter dogs. The material of the study consisted of stomach and liver tissues taken from 35 dogs that died for different reasons and were brought for necropsy. DNA copies of H. heilmannii were detected by Real-time PCR in the liver samples of 30 dogs using H. heilmannii-specific primers. In the case of gastric samples, Real-time PCR detected H. heilmannii in 13 cases, H. pylori in 3 cases, both H. heilmannii and H. pylori in 13 cases, and H. felis, H. heilmannii and H. pylori in 3 cases. Microscopically, neutrophil leukocyte infiltration, epithelial degeneration, fibrosis and oedema in the lamina propia, and lymphoplasmacytic cell infiltration were determined in the stomachs. In the Hemotoxylin Eosin staining of the sections, 5 cases and 14 cases in the Warthin–Starry staining were found positive for Helicobacter-like microorganisms. Microscopically, dissociation of the remark cords and hydropic degeneration in hepatocytes, and focal mononuclear cell infiltrations in some sections were determined in the livers. In conclusion, with this study, it was understood that Real-time PCR analyzes are very useful in the diagnosis of H. felis, H. heilmannii, and H. pylori. However, histopathological examinations are necessary to associate the presence of bacteria with the development of the disease.

Painful facial, oral, and genital ulcers

Painful facial, oral, and genital ulcers

IL-8 Links NF-κB and Wnt/β-Catenin Pathways in Persistent Inflammatory Response Induced by Chronic Helicobacter pylori Infection

Helicobacter pylori (H. pylori) infection can cause persistent inflammatory response in human gastric mucosal epithelial cells, which may result in the occurrence of cancer. However, the underlying mechanism of carcinogenesis has not been elucidated yet. Herein, we established the models of chronic H. pylori infection in GES-1 cells and C57BL/6J mice. Interleukin 8 (IL-8) level was detected by ELISA. The expression of NF-κB p65, IL-8, Wnt2 and β-catenin mRNA and proteins was evaluated by real-time PCR, Western blotting, immunofluorescence staining, and immunohistochemistry. The infection of H. pylori in mice was evaluated by rapid urease test, H&E staining and Warthin-Starry silver staining. The morphological changes of gastric mucosa were observed by electron microscopy. Our results showed that in H. pylori infected gastric mucosal cells along with activation of NF-κB signaling pathway and increase of IL-8 level, the expression of Wnt2 was also increased significantly, which preliminarily indicates that IL-8 can positively regulate the expression of Wnt2. Studies in chronic H. pylori infected C57BL/6J mice models showed that there was an increased incidence of premalignant lesions in the gastric mucosa tissue. Through comparing changes of gastric mucosal cell ultrastructure and analyzing the relationship between NF-κB signaling pathway and Wnt2 expression, we found that H. pylori infection activated NF-κB signal pathways, and the massive release of IL-8 was positively correlated with the high expression of Wnt2 protein. Subsequently, the activated Wnt/β-catenin signal pathways may be involved in the malignant transformation of gastric mucosal cells. Collectively, H. pylori chronic infection may continuously lead to persistent inflammatory response: activate NF-κB pathway, promote IL-8 release and thereby activate Wnt/β-catenin pathway. IL-8 probably plays an important role of a linker in coupling these two signal pathways.

Spirochaetosis of the gallbladder – a case report

Spirochaetosis is an infection caused by Brachyspira which are Gram-negative bacteria. The disease classically affects the large bowel and appendix and has not been previously described in the gallbladder. Spirochaetosis is diagnosed on histology and has a characteristic microscopic appearance. We report a case of spirochaetosis of the gallbladder, found incidentally in a cholecystectomy specimen, in a 65-year-old male with acute-on-chronic cholecystitis and cholelithiasis. The gallbladder epithelium showed the presence of a thick fuzzy brush border, and a Warthin-Starry stain highlighted the presence of spirochaetes.

Métodos de diagnóstico convencionais e imunoistoquímica na detecção de espécies de Helicobacter gástricas em cães com gastropatia crônica

ABSTRACT An accurate diagnostic test for Helicobacter spp. infection in dogs is necessary but no gold standard diagnostic method has yet been established. The purpose of this study was to assess the sensitivity of conventional diagnostic methods as opposed to immunohistochemistry (IHC) in the detection of Helicobacter spp. in gastric samples of dogs with chronic gastropathy. Samples of gastric fundus, body and antrum were collected by gastroscopy from 13 domestic dogs presenting chronic vomit and submitted to the rapid urease test (RUT), cytopathology, histopathology with Hematoxylin-Eosin (HE) and Warthin-Starry (WS) stain, and IHC. Cohen's kappa coefficient was carried out to determine the agreement between techniques compared to IHC. Prevalence of colonization detected by IHC was 92% and was higher in the gastric fundus. The sensitivity of each technique in detecting Helicobacter spp. in the gastric fundus, body, and antrum, respectively, were as follows: RUT 50%, 70%, 17%; cytology 58%; 70%, 50%; HE 42%; 70%, 50%; WS 67%; 80%, 50%. We found that IHC was the most sensitive method for Helicobacter spp. diagnosis revealing that gastric fundus samples have higher presence of bacteria. Squash cytology enhances sensitivity and ancillary staining such as WS should be taken into consideration towards diagnosis.

Bacillary Angiomatosis of Lymph Nodes: Unusual Case Presentation in an Immunocompetent Adolescent

Bacillary angiomatosis is a neovascular proliferative condition caused by the Bartonella group of opportunistic bacteria, commonly occurring in patients with advanced Human Immunodeficiency Virus (HIV) and other immunocompromised conditions. However, cases have also been reported to occur rarely in immunocompetent individuals. Skin lesions such as nodules and papules are the most common manifestations, although cases have been rarely reported in other sites such as the oral cavity, respiratory tract, gastrointestinal tract, and bones. Isolated lymph node involvement or systemic manifestations such as fever, chills, anorexia, and weight loss are very rare. Since Bartonella is a fastidious Gram-negative bacterium that is difficult to grow and isolate in culture, diagnosis relies on histological examination of the affected organ/tissue and demonstration of the causative bacteria using special stains such as Warthin-Starry, Giemsa, Gram’s, and silver stains. Herein, the authors presented a case of bacillary angiomatosis in a 14-year-old immunocompetent adolescent male who presented with recurrent fever and painful enlargement of the axillary lymph nodes. The case is notable for its unusual clinical manifestations of systemic symptoms, specifically anorexia and significant weight loss, leading to suspicion of malignancy. Following the establishment of the diagnosis, the boy experienced complete recovery, with regained appetite and weight, following treatment with a long duration of antibiotics.

Ezrin immunoexpression in gastric cells of domestic cats infected with Helicobacter spp

The aim of this study was to evaluate de immunoexpression of ezrin in gastric cells of domestic cats infected with Helicobacter spp. and with chronic gastritis. Twenty paraffin-embedded gastric samples were selected based on previous positive results for Helicobacter spp. in the Rapid Urease Test, Warthin-Starry staining and cytology. Haematoxylin-eosin stained sections was done to evaluate inflammatory cell infiltrates. Immunohistochemical analysis was done using anti-Helicobacter pylori and anti-Ezrin antibodies. The analysis of inflammatory infiltrates revealed 8/20 (40%) in score 0, 11/20 (55%) in score 1 and 1/20 (5%) in score 2. The labelling observed in the immunohistochemical analysis using anti-Helicobacter spp. antibody showed no samples with score 0; 4/20 (20%) with score 1; 7/20 35% with score 2 and 9/20 (45%) with score 3. Ezrin overexpression on the cytoplasm of parietal cells was revealed in 18 out of 20 samples (90%). Of these, 10 cases (45%) achieved the score 1; 6 cases (30%) the score 2 and 2 cases (10%) the score 3. On the surface and pit cells there was an increase in Ezrin immnoexpression in 12 out of the 20 samples (60%), of which 8 samples (40%) achieved the score 1 and 4 samples (20%) the score 2. No sample were classified in score 3. Statistically significant differences (p = 0.026) were observed between the inflammatory infiltrate in the gastric mucosa and the immunoexpression of Ezrin in the cytoplasm of parietal cells. It was concluded that ezrin had an increased immunoexpression in the gastric mucosa of cats with chronic gastritis.

Coptis, Pinellia, and Scutellaria as a promising new drug combination for treatment of Helicobacter pylori infection.

Helicobacter pylori (H. pylori) is the most important infectious agent and plays an important role in the progression of chronic gastritis and the development of gastric cancer. To identify efficient therapeutic agents or strategies that can treat H. pylori infection. We performed literature analysis, experimental validation, and network pharmacology. First, traditional Chinese medicine (TCM) prescriptions for the treatment of H. pylori infection were obtained from the China National Knowledge Infrastructure, China Biology Medicine, China Science and Technology Journal Database, and WanFang databases. In addition, we conducted a relevant search by Reference Citation Analysis (RCA) (https://www.referencecitationanalysis.com). Next, we used TCM Inheritance Support System V2.5 to identify core drug combinations in the TCM prescriptions. Then, an H. pylori-associated chronic mouse model of gastritis was established. The antibacterial properties and anti-inflammatory potential of the core drug combination were evaluated by the rapid urease test, modified Warthin-Starry silver staining, histopathological analysis, and enzyme linked immunosorbent assay. Finally, the active compounds, hub targets, and potential signaling pathways associated with the core drug combination were analyzed by network pharmacology. The TCM treatment of H. pylori was mainly based on reinforcing the healthy Qi and eliminating pathogenic factors by simultaneously applying pungent dispersing, bitter descending, cold and warm drugs. The combination of Coptis, Pinellia, and Scutellaria (CPS) was identified as the core drug combination from 207 prescriptions and 168 herbs. This drug combination eradicated H. pylori, alleviated the gastric pathology induced by H. pylori infection, and reduced the expression levels of tumor necrosis factor-α (P = 0.024) and interleukin-1β (P = 0.001). Moreover, a total of 35 compounds and 2807 targets of CPS were identified using online databases. Nine key compounds (tenaxin I, neobaicalein, norwogonin, skullcapflavone II, baicalein, 5,8,2'-trihydroxy-7-methoxyflavone, acacetin, panicolin, and wogonin) and nine hub target proteins (EGFR, PTGS2, STAT3, MAPK3, MAPK8, HSP90AA1, MAPK1, MMP9, and MTOR) were further explored. Seventy-seven signaling pathways were correlated with H. pylori-induced inflammation and carcinogenesis. In summary, we showed that CPS is the core drug combination for treating H. pylori infection. Animal experiments demonstrated that CPS has bacteriostatic properties and can reduce the release of inflammatory cytokines in the gastric mucosa. Network pharmacology predictions further revealed that CPS showed complex chemical compositions with multi-target and multi-pathway regulatory mechanisms. Although the results derived from network pharmacology are not necessarily comprehensive, they still expand our understanding of CPS for treating H. pylori infection.

The correlation between reflux esophagitis and Helicobacter pylori infection based on natural population

Objective: Reflux esophagitis (RE) may be negatively correlated with Helicobacter pylori (H. pylori) infection, but the conclusion and relevant mechanism is still controversial. This study proposed to explore the correlation between RE and H. pylori infection based on natural population. Methods: From July 2013 to December 2014, 3 940 residents aged 40-69 years were recruited in Linqu County of Shandong Province and Hua County of Henan Province by the whole sampling method. All the subjects underwent gastroscopy, and gastric mucosa biopsy specimens were collected for pathological diagnosis and Warthin-Starry (WS) staining to identify H. pylori infection. Venous blood samples of some subjects were collected for H. pylori immunoglobulin G (H. pylori-IgG) detection. Also, demographic and sociological data were collected. Chi-square test and logistic regression were used to analyze the correlation between RE and H. pylori infection. Results: A total of 359 cases of RE were detected. Excluding RE and other upper gastrointestinal organic diseases, 3 382 cases were considered as controls. Chi-square test showed that WS staining positive rate in RE group was significantly lower than that in control group (P=0.023), but there was no significant difference in the positive rate of H. pylori-IgG between the two groups (P=0.281). There were significant differences between RE group and control group in gender composition, age, body mass index (BMI), smoking, alcohol consumption, education level and mucosal active inflammation. Multivariate regression analysis showed that RE was negatively correlated with gastric mucosa active inflammation [OR=0.754 (95%CI 0.600-0.949), P=0.016], and positively correlated with male [OR=4.231 (95%CI 3.263-5.486), P<0.001], age ≥60 years, BMI≥24 kg/m2 [OR=1.540 (95%CI 1.220-1.945), P<0.001]. Compared to those aged 40-49 years and 50-59 years, the odds ratio (OR) of RE in these aged ≥60 years were 1.566 (95%CI 1.144-2.143, P=0.005) and 1.405 (95%CI 1.093-1.805, P=0.008). Conclusion: RE is more closely related to H. pylori present infection. Multivariate analysis showed that RE is negatively correlated with active inflammation of gastric mucosa caused by H. pylori infection, and positively correlated with male, overweight and aged ≥60 years.

S2199 A Rare Case of Intestinal Spirochetosis in an Asymptomatic Immunocompetent Patient

Introduction: Human Intestinal spirochetosis is defined as presence of spirochetes attached to the surface of the intestinal mucosa. Intestinal spirochetes comprise of a heterogeneous group of bacteria. In humans, Brachyspira aalborgi and Brachyspira pilosicoli predominate. Intestinal spirochetosis is primarily reported in developing countries and rare in places with high living standards. Immunocompromised patients and homosexuals are at higher risk for colonization and invasive human intestinal spirochetosis. Patients can be asymptomatic or present with diarrhea, abdominal pain, hematochezia and lower GI bleeding. We present a rare case of asymptomatic human intestinal spirochetosis with no risk factors. Case Description/Methods: A 66-year-old White heterosexual male with past medical history of nicotine use disorder, alcohol dependence, and hypertension was referred to the gastroenterology clinic for persistently elevated liver enzymes and abnormal abdominal imaging results. He denied having any gastrointestinal or systemic symptoms except occasional diarrhea and was sexually active with only one female partner for last 40 years. CT scan of the abdomen & pelvis showed hepatic steatosis, sigmoid diverticulosis with sigmoid wall thickening and a fistulous communication between the sigmoid colon and bladder dome. An active open fistula between bowel and bladder was considered. Further evaluation with colonoscopy showed a total of 2 large polyps and multiple sigmoid diverticula but no evidence of fistula. Histopathology of biopsies showed hyperplastic, adenomatous polyps and spirochetes. Warthin-Starry stain was positive for Barchyspira spirochetes from all the tissue specimens. Patient was referred to an infectious disease specialist. HIV and Rapid Plasma Regain (RPR) were negative. The decision was made to monitor the clinical condition and avoid antibiotics for time being, given the absence of symptoms and concurrent alcohol use. A surgical evaluation was planned to look into a possible colo-vesical fistula, but patient was lost to follow-up (Figure). Discussion: Intestinal spirochetosis is known to cause havoc in veterinary medicine, especially in the swine population (Our patient lived in a trailer park and worked with goats, chickens, dogs, and cats). Intestinal spirochetosis in humans and its clinical significance in an asymptomatic immunocompetent patient is poorly understood. Antibiotics are not recommended unless the patient is symptomatic. If recommended metronidazole is the choice of treatment.Figure 1.: Intestinal Spirochetosis.