Methenamine Silver Research Articles

Histomorphological features of mucormycosis with rise and fall of COVID-19 pandemic

IntroductionDue to Corona Virus disease -19, India saw a surge of mucormycosis cases, associated with high death rate. India, during the month of May to July 2021 saw a surge of mucormycosis from all states, with close to 50,000 cases just in a span of 3 months. ObjectiveTo examine the histopathological appearances of rhino-orbital/rhino-maxillary/sino-nasal mucormycosis in the backdrop of the ongoing COVID 19 pandemic. Material and methodsThe study involved analysis of 60 biopsy samples of suspected rhino-maxillary /rhino-orbital mucormycosis received from post-COVID-19 patients. A preliminary review of the slides showing hyphal forms of fungal organisms with un-doubtful tissue / mucosal invasion was included. All samples were examined under Hematoxylin and Eosin stains along with special fungal stains. Data thus obtained were analyzed statistically. Special stains for fungus namely Periodic Acidic Schiff (PAS) and Gomori Methenamine silver (GMS) were utilized to confirm and/or to differentiate the fungal organisms and to highlight the cell wall of the fungus. ResultsThe mean age of the patients with mucormycosis was 51.68 years and 72 (83.33%) of them were males. Acute type of inflammation was noted in 44 (73.33%), granulomatous inflammation in 14 (23.33%) of cases. Bony invasion and perineural invasion was observed in 5 (8.33%) and 55 (91.67%) cases, respectively. The dominant fungus were mucorales in 58 (96.67%), aspergillous, along with mucorales in 12 (20%) and combination of mucorales and candida identified in 8 (13.33%) cases. ConclusionBesides all the histological appearance of angioinvasion, bone, and soft tissue invasion, a notable aspect was the shift in inflammatory pattern, which was more granulomatous in nature, with a decrease in fungal load correlating with the drop of COVID second wave. This proves that as immunity develops, the host's response to secondary opportunistic infections changes.

Alternate Special Stains for the Detection of Mycotic Organisms in Oral Cyto-Smears-A Histomorphometric Study.

In the wake of the COVID-19 pandemic, fungal infections of the maxillofacial region have become prevalent, making their accurate diagnosis vital. Histopathological staining remains a simple, cost-effective technique for differentiation and diagnosis of the causative fungal organisms. The present study aims to evaluate the staining efficacy of Periodic Acid-Schiff (PAS), Alcian Blue, Safranin-O and Gomori’s Methenamine Silver (GMS) on fungal smears. This research work also attempts to study the morphometric characteristics of Candida albicans, Aspergillus flavus, Rhizopus oryzae. Candida albicans, Aspergillus flavus and Rhizopus oryzae, 10 smears each, were stained using PAS, Alcian Blue, Safranin-O and GMS. The morphological characteristics and staining efficacy were examined, and semi-quantitative scoring was performed. Candida albicans, Aspergillus flavus and Rhizopus oryzae were stained for the first time with Safranin-O. The morphometric traits were then analyzed using an image analysis software. Safranin-O provided the most reliable staining efficacy amongst the stains and optimum morphological definition for all three organisms. Safranin-O was found to be superior to PAS and GMS, ensuring detection of even the most minute mycotic colonies. The hyphae of Aspergillus flavus to be the largest, and the spores and fruiting body of Rhizopus oryzae were found to be the largest amongst the three organisms compared. Early and accurate diagnosis of fungal infections can significantly reduce morbidity in orofacial fungal infections.

A Rare Presentation of Sarcoidosis: Dysphagia, Pancytopenia, and Acute Renal Failure

Sarcoidosis is a multisystem inflammatory chronic disorder that can virtually affect any organ system in the body. Most commonly affected organs are the intrathoracic structures with 90% of the reported cases involving the lungs. Esophageal involvement in sarcoidosis is extremely rare. Involvement of the esophagus and kidney along with hematological involvement is extremely uncommon in the same patient. Here, we present a case of a 58-year-old gentleman with a similar rare presentation. The patient presented with shortness of breath, productive cough, fatigue, and difficulty in swallowing, along with a weight loss of 20-30 pounds over three months. Laboratory workup was significant for leukopenia (2900 K/UL), serum creatinine level of 2.7 mg/dL (baseline: 1.2-1.7), and raised angiotensin-converting enzyme level at 187 nmol/ml/min. Chest X-ray showed bilateral widespread fine reticulonodular opacities, chest CT showed extensive bilateral reticulonodular opacities throughout the lung parenchyma, and fine-needle aspiration cytology of the right lung showed noncaseating granulomas. No fungal or acid-fast organisms were identified, and no evidence of malignancy was seen. Special stains for fungal (Grocott's methenamine silver and periodic acid-Schiff) and acid-fast organisms (acid-fast bacilli (AFB) and fluorescent AFB) were negative. Esophagogastroduodenoscopy (EGD) with gastric biopsy showed acute and chronic inflammation and no intestinal metaplasia, dysplasia, or malignancy was identified. Bronchoalveolar lavage was done, which showed macrophages (74%), neutrophils (6%), eosinophils (3%), and lymphocytes (17%), and was negative for malignant cells. QuantiFERON and AFB sputum/Mycobacterium tuberculosis polymerase chain reaction were negative. The patient was initially started on intravenous fluids and calcitonin, which significantly improved renal function and the calcium status of the body. Then prednisone 40 mg daily was started, which improved swallowing and breathing. After a week, prednisone was changed to 20 mg daily and was continued at the time of discharge.

Heavy Metal Enhancement Technique for Diaminobenzidine in Immunohistochemistry Enables Ultrastructural Observation by Low-vacuum Scanning Electron Microscopy.

Low-vacuum scanning electron microscopy (LV-SEM) is a powerful tool that allows to observe light microscopic specimens with periodic acid-silver methenamine (PAM) staining at a higher magnification, simply by removing the coverslip. However, it is not suitable for observation of immunohistochemistry (IHC) using 3,3'-diaminobenzidine (DAB) due to insufficient backscattered electron image. Traditional heavy metal enhancement techniques for DAB in IHC, (1) osmium tetroxide and iron, (2) cobalt, (3) methenamine silver (Ag), (4) gold chloride (Gold), and (5) both Ag and Gold (Ag + Gold), were examined by LV-SEM. Tissue specimens from Thy1.1 glomerulonephritis rat kidney stained with α-smooth muscle actin and visualized with DAB were enhanced by each of these enhancement methods. We found, in light microscopic and LV-SEM, that the enhancement with Ag, Gold, or Ag + Gold had better intensity and contrast than others. At a higher magnification, Ag + Gold enhancement showed high intensity and low background, although only Ag or Gold enhancement had nonspecific background. Even after observation by LV-SEM, the quality of specimens was maintained after remounting the coverslip. It was also confirmed that Ag + Gold enhancement could be useful for IHC using clinical human renal biopsy. These findings indicate that Ag + Gold provided an adequate enhancement in IHC for both LM and LV SEM observation.

Biliary candidiasis mimicking IgG4‐related disease in a patient with normal immunity

AbstractPurposeTo report a rare case of biliary candidiasis in a host with normal immunity.MethodsLaboratory tests indicated an elevated G level of 1000 pg/ml. Microscopic examination of pathological sections demonstrated multiple, narrow base, budding yeast cells following Grocott's methenamine silver staining and periodic acid‐Schiff staining. Subsequent sequence analysis supported the diagnosis of biliary candidiasis.ResultsA 31‐year‐old previously healthy man developed obstructive jaundice under no obvious inducement. He experienced cholecystectomy and biliary drainage. Surgical specimens revealed granulomatous chronic inflammation in the gallbladder, liver, bile ducts and mesenteric lymph nodes. He was misdiagnosed with tuberculosis and received antituberculosis treatment. However, his condition did not improve. He was finally diagnosed with biliary candidiasis and administered the antifungal treatment with fluconazole. His symptoms improved after such treatment.ConclusionsWe reported a case of biliary candidiasis that mimicked IgG4‐RD in a host with normal immunity. Our findings highlighted the need for pathological diagnosis in patients with symptoms resembling IgG4‐RD.

Acute Abdominal Distention in a Premature Low-birthweight Infant.

Acute Abdominal Distention in a Premature Low-birthweight Infant.

Grocott Methenamine Silver Staining Is the Optimal Approach to Histological Diagnosis of Pulmonary Cryptococcosis.

BackgroundHistological staining methods for Cryptococcus identification vary in accuracy. This study aimed to investigate the clinical value of Grocott methenamine silver (GMS), periodic acid-Schiff (PAS), and Alcian blue (AB) staining in the diagnosis of pulmonary cryptococcosis (PC).MethodsFrom April 2004 to June 2021, the clinical and pathological data of 152 patients with PC were collected from the Department of Pathology, Sun Yat-sen University Cancer Center. The sensitivity and identifiability of GMS, PAS, and AB staining for histological diagnosis were systematically evaluated using statistical methods combined with the microscopic characteristics of PC cases.ResultsStatistical analysis showed that the detection rates of GMS, PAS, and AB staining were 100.0% (152/152), 94.7% (144/152), and 81.6% (124/152), respectively. McNemar’s test showed that the sensitivity of GMS was significantly higher than those of PAS (P = 0.008) and AB stains (P < 0.001). Both PAS and AB stains had obvious non-specific staining, which interfered with the detection of Cryptococcus, and increased diagnostic difficulties. In contrast, in GMS staining, Cryptococcus spores were prominent with a clean background and were clearly observed at low or medium power magnification, with the identifiability significantly better than those of PAS or AB staining.ConclusionGMS staining had sensitivity up to 100%, and identifiability that was better than those of PAS and AB staining. GMS is the best method for histological diagnosis of PC.

Microscopic Imaging and Labeling Dataset for the Detection of Pneumocystis jirovecii Using Methenamine Silver Staining Method

Pneumocystis jirovecii pneumonia is one of the diseases that most affects immunocompromised patients today, and under certain circumstances, it can be fatal. On the other hand, more and more automatic tools based on artificial intelligence are required every day to help diagnose diseases and thus optimize the resources of the healthcare system. It is therefore important to develop techniques and mechanisms that enable early diagnosis. One of the most widely used techniques in diagnostic laboratories for the detection of its etiological agent, Pneumocystis jirovecii, is optical microscopy. Therefore, an image dataset of 29 different patients is presented in this work, which can be used to detect whether a patient is positive or negative for this fungi. These images were taken in at least four random positions on the specimen holder. The dataset consists of a total of 137 RGB images. Likewise, it contains realistic, annotated, and high-quality microscope images. In addition, we provide image segmentation and labeling that can also be used in numerous studies based on artificial intelligence implementation. The labeling was also validated by an expert, allowing it to be used as a reference in the training of automatic algorithms with supervised learning methods and thus to develop diagnostic assistance systems. Therefore, the dataset will open new opportunities for researchers working in image segmentation, detection, and classification problems related to Pneumocystis jirovecii pneumonia diagnosis.

Invasive Pulmonary Aspergillosis Diagnosis via Peripheral Blood Metagenomic Next-Generation Sequencing.

Invasive pulmonary aspergillosis (IPA) is one of the major causes of morbidity and mortality in immunocompromised patients such as hematological malignancies, hematopoietic stem cell transplantation, and solid organ transplantation. The diagnosis of IPA in these patients is still difficult because it has no obvious specificity in clinical symptoms, signs and imaging, and test sensitivity of blood 1,3-β-d-glucan test, galactomannan are low. Therefore, we still need to explore more diagnostic methods. In our study, via peripheral blood metagenomic next-generation sequencing (mNGS), five patients were tested positive for Aspergillus DNA and then quickly diagnosed as IPA. Out of the 5 cases, 1 was proven and 4 were probable IPA. The underlying diseases of the 5 patients were myelodysplastic syndrome (2 cases), acute myeloid leukemia (2 cases), and renal transplantation (1 case). Then they were diagnosed as IPA using other methods such as lung histopathology, bronchoalveolar lavage fluid (BALF) mNGS, and sputum culture or sputum mNGS. In case 1, sputum culture suggested Aspergillus flavus. In case 2, both Grocott methenamine silver (GMS) stain of lung histopathology and lung tissue mNGS suggested Aspergillus infection. In cases 3 and 4, BALF-mNGS suggested Aspergillus infection. In case 5, sputum mNGS suggested Aspergillus infection. In conclusion, detecting the cfDNA of Aspergillus via peripheral blood mNGS can be used to diagnose IPA and is a rapid and non-invasive diagnosis method.

Pneumocystis Jirovecii Pneumonia Diagnosis via Metagenomic Next-Generation Sequencing.

The incidence of non-HIV-infected Pneumocystis Jirovecii Pneumonia (PJP) is increasing. The prognosis for non-HIV PJP is poor and diagnostic tests are of lower sensitivity in non-HIV patients. Metagenomic next-generation sequencing (mNGS) was compared with routine detection assays, including Gomori methenamine silver (GMS) stain and polymerase chain reaction (PCR) technique. Specimens of 4 bronchoalveolar lavages (BAL) and 1 lung tissue samples were obtained from 4 non-HIV patients from our hospitals. Although both GMS and mNGS were positive for P. jirovecii with PCR as positive control, the testing time of mNGS was obviously shorter than GMS. Compared with the traditional GMS method, mNGS has absolute advantages. However, the issue with PJP presentations having atypical symptoms and ambiguous imaging features persists. Hence, the disease can easily be ignored. Secondly, PJP progresses rapidly in non-HIV-infected patients and can cause severe respiratory failure with unfavorable prognosis. This study affirms that mNGS can be used to quickly and accurately diagnose PJP, but a combination of clinical judgement of symptoms, laboratory testing, and imaging examination is required to make a comprehensive judgment along with mNGS test results.

Characteristics and Prognostic Factors of Non-HIV Immunocompromised Patients With Pneumocystis Pneumonia Diagnosed by Metagenomics Next-Generation Sequencing.

ObjectiveThe aim of this study was to evaluate the potential of metagenomic next-generation sequencing (mNGS) for the diagnosis of pneumocystis pneumonia (PCP) in patients with non-human immunodeficiency virus-infection and to discuss the clinical characteristics and identify prognostic factors associated with patients with non-HIV PCP.MethodsForty-six patients with PCP who were admitted in respiratory intensive care unit (ICU) between May 2018 and May 2020 were retrospectively reviewed. The subjects were divided into survivor and non-survivor groups according to the patients' outcome. Conventional methods and mNGS for detecting Pneumocystis jirovecii (P. jirovecii) were analyzed. The patients' demographics, comorbidities, laboratory parameters, and treatments were compared and evaluated in both groups to identify risk factors for mortality by using univariate and multivariate logistic regression.ResultsMetagenomic next-generation sequencing (mNGS) showed a satisfying diagnostic performance of 100% positive of detecting P. jirovecii from bronchoalveolar lavage (BAL) specimens in forty-six patients with non-HIV PCP, compared to only 15.2% for Gomori Methenamine silver (GMS) staining and 84.8% for Serum 1,3-beta-D-glucan (BDG). Among them, the mean age was 46.4-year-old (range 18–79-year-old) and mortality rate was 43.5%. The dominant underlying conditions were connective tissue diseases (34.8%), autoimmune kidney diseases (30.4%), followed by hematologic malignancies (10.9%), and solid organ transplantation (6.5%). A total of 38 cases (82.6%) received glucocorticoid and 19 cases (41.3%) used immunosuppressant within 3 months before diagnosed PCP. Multiple infections were very common, over two thirds' cases had mixed infections. Compared with survivors, non-survivors had a higher acute physiology and chronic health evaluation II (APACHE II) score (14.4 ± 4.8 vs. 10 ± 3.4), Procalcitonin (PCT) [ng/ml: 0.737 (0.122–1.6) vs. 0.23 (0.095–0.35)], lactic dehydrogenase (LDH) [U/L: 1372 (825.5–2150) vs. 739 (490.5–956)], and neutrophil-lymphocyte ratio (NLR) [21.6 (15.67–38.2) vs. 11.75 (5.1–15.52)], but had a lower PaO2/FiO2 ratio (mmHg:108.8 ± 42.4 vs. 150.5 ± 47.5), lymphocytes [×109/L: 0.33 (0.135–0.615) vs. 0.69 (0.325–1.07)] and CD4+ T cells [cell/μl: 112 (53.5–264) vs. 255 (145–303.5)], all P < 0.05. Furthermore, we found non-survivors' PaO2/FiO2 ratio of day 3 and day 7 had not improved when compared with that of day one, and platelet level and NLR became worse. Multivariate analysis showed that other pathogens' co-infection (OR = 9.011, 95% CI was 1.052–77.161, P = 0.045) and NLR (OR = 1.283, 95% CI was 1.046–1.547, P = 0.017) were the independent risk factors of poor prognosis.ConclusionmNGS is a very sensitive diagnostic tool for identifying P. jirovecii in patients who are non-HIV immunocompromised. PCP in patients who are non-HIV infected is associated with a high rate of multiple infections and severe condition. Mixed infection and elevation of NLR were the independent risk factors of poor prognosis.

Comparison of Antioxidant Effect of Crocus Sativus and Vitamin E on Amakicin Induced Nephrotoxicity in Albino Rats

ABSTRACT&#x0D; Objectives: The present study evaluated the protective effect of Crocus Sativus against Amikacin-induced Nephrotoxicity.&#x0D; Methods: The prospective descriptive study was conducted at the department of orthopedic surgery, Dr. Ruth KM Pfau Adult laboratory albino rats of the Wistar strain were used in this experimental study. In this study, 75 albino rats were subdivided into 3 groups, including experimental groups B and C. The control group (A) received normal saline intraperitoneally, group (B) received Amikacin 30mg/kg body weight intraperitoneally and Group (C) received Crocus sativus 50 mg/kg body weight orally and vitamin E orally in a dose of 50mg/kg of body weight. Kidneys of rats obtained after dissection were processed, sectioned, and stained in PAS Hematoxylin and Gomoris Methenamine Silver stains. The present study was conducted at Dow University of Health Sciences in the Institute of Biomedical Sciences and Animal House of Dow University and Dow Diagnostic Research and Reference Laboratory. The duration of the study was Three months. Inclusion criteria were Albino rats of both sexes, kidneys of either side (right &amp; left), rats with an approximate weight of 140 to 180 grams were included in the study. Exclusion criteria were overweight rats and pregnant rats.&#x0D; Results: A significant increase in kidney weight was observed in group B. A considerable addition was observed in proximal tubular in groups B and C. A considerable increase was also observed in the number of intact proximal tubule nuclei and the nuclear diameter of groups B &amp; C.&#x0D; Conclusion: The study provides evidence that Crocus Sativus has protective effects in Amikacin-induced nephrotoxicity, comparable to widely used Antioxidant Vitamin E. The study recommends the addition of Crocus Sativus in dietary supplements after further evaluation in human subjects.

Covid-19 associated mucormycosis

: Corona virus disease-2019 (COVID-19) is associated with various opportunistic, bacterial, and fungal infections. High risk groups include people with diabetes especially diabetic ketoacidosis, solid organ transplantation, long-term systemic corticosteroid use, and iron overload. Many cases of mucormycosis were reported worldwide. Mucormycosis is an invasive fungal infection with high morbidity and mortality. Early diagnosis and treatment with appropriate anti-fungals lead to improved outcomes. The aim of this study is to establish the factors associated with mucormycosis in a COVID-19 setting, like comorbidities and treatment protocols for treatment of COVID-19. The histological patterns and tissue reactions to mucormycosis also were studied. In patients with ongoing COVID-19 infection and in a post COVID-19 scenario, we studied the biopsy findings of mucormycosis in various sites like rhino-orbital, lung, gastric and trachea. Material for this study is from a tertiary care hospital in South India. Patient age ranged from 30 years to 74 years. Mean age of the patients was 51 years. Male to female ratio was 1:1.1. Tissue from sino- nasal mucosa, peri orbital tissue, exenterated eyeball, lung tissue and tissue from rare sites like gastric and tracheal mucosal lesions were also included in this study. Tissue was fixed in 10% buffered formalin. Routine Haematoxylin and Eosin(H&amp;E) stains were done. Gomori’s Methenamine silver (GMS) stains were done on all cases. Tissue was submitted for fungal cultures in all the cases.Total of twenty-three cases were diagnosed as mucormycosis based on the morphology and special stains in this analysis. Histology revealed areas of infarction in all cases with neutrophilic infiltration. Granulomatous reaction was seen in seven cases and melanin pigment was seen in two cases. 100% of patients were diabetics. There was neutrophilia in 100% of cases. Lymphopenia was seen in 85.7%, C-Reactive Protein (CRP) was elevated in 100% of cases.Ferritin was done in 14 patients and D-dimer was done in 17 patients and in all patients, these were elevated. All patients were treated with steroids according to the treatment protocol for COVID-19, Remdesivir was given in 72.7% of cases and second immunomodulator drugs like Tocilizumab in four cases and Baricitinib in one case.Mucormycosis is an emerging problem with COVID-19. It is important to carefully monitor blood glucose levels and take into account underlying medical conditions of patients before initiation of steroid therapy. Early recognition of symptoms and early diagnosis has a better outcome in patients with mucormycosis associated with COVID-19 infection.

Nocardiosis in Free-Ranging Cetaceans from the Central-Eastern Atlantic Ocean and Contiguous Mediterranean Sea.

Simple SummaryCharacterization, description, and geographical location of harmful bacterial agents in cetaceans are important for population surveillance and health monitoring around the world. This research compiles the pathologic features of nocardiosis in five free-ranging delphinids from the Canary Islands and Andalusia. All examined animals showed a disseminated pattern of infection with characteristic suppurative to pyogranulomatous lesions with thromboembolism in two or more organs. The obtained results provide the first record of N. otitidiscaviarum in cetaceans, the first account of N. farcinica in free-ranging dolphins, and confirmation of nocardiosis in central eastern Atlantic Ocean.We report the pathologic features of nocardiosis in five free-ranging delphinids from the Canary Islands and Andalusia, namely four striped dolphins (Stenella coerulealba) and one bottlenose dolphin (Tursiops truncatus). All animals had a multiorgan (disseminated) pattern of infection involving suppurative to pyogranulomatous and thromboembolic lesions in two or more organs. Most affected organs were (by decreasing order) lung, pulmonary lymph nodes, liver, kidney, adrenal glands, and central nervous system. Typical intralesional and intravascular branched and filamentous bacteria were highlighted by Grocott’s methenamine silver and Gram stains. Bacterial analysis including 16S rRNA gene sequencing identified Nocardia farcinica in two striped dolphins and Nocardia otitidiscaviarum in one striped dolphin and the bottlenose dolphin. All dolphins tested (n = 4) for cetacean morbillivirus were negative; one dolphin had concurrent cutaneous herpesvirosis. These results provide the first record of N. otitidiscaviarum in cetaceans, the first account of N. farcinica in free-ranging dolphins, and confirmation of nocardiosis in central eastern Atlantic Ocean. These results expand the known geographic range of nocardiosis in cetaceans.

Aerossacolitis and Pneumonia in an Indian Peafowl Caused by Lactobacillus agilis

Background: The peafowl is an ornamental bird that has the habit of eating directly from the earthy soil, which makes this bird more susceptible to endoparasites. One important endoparasite is Eucoleus contortus, which leads to inflammatory processes that alter the local microbiota, potentializing disease. By the other way, a member of the bird’s microbiota there is the genus Lactobacillus, but when occurs some imbalance, these bacteria can overgrowth and even cause some infection. This report describes the pathological and microbiological findings of chronic necrotizing pneumonia and aerossacolitis caused by Lactobacillus agilis in a peafowl, associated with parasitism by E. contortus. Case: A peafowl (Pavo cristatus), adult, male, who lived on a farm with contact with other species of animal, was submitted to post-mortem examination due to sudden death. This animal lived in an extensive system on the property and was the only one of its species. During the gross evaluation, the air sacs were filled with solid yellowish crumbly material. The same material was observed forming well-defined nodules that occupied &gt; 50% of the lung parenchyma. Histological analysis showed multiple parabronchi dilated and filled with caseous necrosis, characterized by abundant cellular debris and fibrin deposition. These areas were surrounded by the proliferation of fibrous connective tissue and inflammatory infiltrate of macrophages, giant cells, lymphocytes, and plasma cells. The air sacs parenchyma showed fibrin deposition and mixed inflammatory infiltrate. Multiple gram-positive bacilli were observed within the caseous foci in Gram-stained slides. In the crop and esophageal mucosa, cross-sections of filiform nematodes morphologically compatible with E. contortus were associated with chronic inflammatory infiltrate and epidermal hyperkeratosis. A lung section was submitted to Gram-Brown-Hopps and Ziehl-Neelsen (ZN) stains for bacterial investigation, and Grocott's methenamine silver (GMS) stain for fungal investigation. Short gram-positive bacilli bacteria are observed within the caseous foci in Gram-stained slides. By the other way, no agents were identified on the ZN and GMS stains. Following the analysis, lung fragments were cultivated at aerobic and microaerophilic conditions on sheep blood agar and McConkey agar. All the microbial cultures were incubated at 37°C to 48 h. Pure culture, in microaerophilic condition, of Gram-positive bacilli, was observed. The isolated bacterium was identified by MALDI-TOF MS as L. agilis.Discussion: Although uncommon, L. agilis was the single bacterium identified and therefore, associated as a primary cause of necrotic pneumonia and aerossacolitis in the studied peafowl. The presence of E. contortus could induce the aspiration of regurgitated of little amount of material from the gastrointestinal tract with sufficient bacterial load to initiate an infection, but not enough to smother the animal. The gradual aspiration can induce a chronic inflammatory condition. Infections by bacteria from the host microbiota have been observed in animals and humans with immunodeficiency. In summary, both the parasitosis and the inflammation could be resulted by the parasite and the aspiration of gastric product, which probably interfered in the immune response and allowed the overgrowth of L. agilis. In the current case report, based on macroscopic, microscopic and bacteriological results, we have provided insights to understand how the parasitosis made possible a pneumonia from a bacterium from the host's microbiota. Finally, to the best of our knowledge, this is the first report of L. agilis as causal agent of fatal necrotic pneumonia and aerossacolitis in peafowl.Keywords: respiratory infection, ornamental birds, secondary infection, parasitosis.

Cytopathological analysis of bronchoalveolar lavage fluid in patients with and without HIV infection

BackgroundHuman immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) infection can lead to a broad spectrum of lung diseases, including infectious diseases and tumors. Recently, with the wide application of bronchoscopes and cytopathology of bronchoalveolar lavage fluid (BALF), the diagnostic efficiency of lung diseases has improved. The present study focuses on analyzing the cytopathologic characteristics of BALF in the diagnosis of HIV/AIDS-related lung disease and comparing the lung disease spectrum between HIV and HIV-uninfected patients.MethodsBALF specimens were collected from 2211 patients. Using ThinPrep liquid-based technology, the cytologic smears were prepared by staining with Hematoxylin and Eosin (HE), Gomori's methenamine silver (GMS), and Periodic Acid Schiff (PAS), acid-fast and immunocytochemical (ICC) staining. Real-time PCR was used to detect cytomegalovirus (CMV) and Mycobacterium tuberculosis (M. tuberculosis) in the remaining BALF. PCR-reverse dot hybridization was used for mycobacterial species identification.ResultsFrom the 2211 BALF specimens, 1768 (79.96%) were specimens from HIV-infected patients, and 443 (20.04%) were speciments from HIV-uninfected patients. The HIV-infected patients with a median age of 38.5 ± 11.3 years were markedly younger than the HIV-uninfected patients (52.9 ± 14.9 years) (p < 0.01). We found that 1635 (92.5%) HIV-infected patients were males, showing a prominently higher proportion than those without HIV infection (71.1%) (p < 0.01). Meanwhile, 1045 specific lesions were found in 1768 HIV-infected patients (59.1%), including 1034 cases of infectious diseases and 11 neoplastic lesions, also exhibiting a distinctly higher proportion compared to the HIV-uninfected patients (12.2%) (p < 0.001). For the HIV-infected group, a distinctly higher proportion of single infection lesions (724/1768, 41%) was noted than the HIV-uninfected group (14/443, 3.2%) (p < 0.001). Among single infection lesions, the most common was Cytomegalovirus(CMV) infection (20.9%) for the HIV-infected group, followed by Pneumocystis jiroveci(PJ) (13.0%), Fungal (3.5%), and Mycobacterial infections (3.4%), of which M. tuberculosis infection accounted for 3.1%. Double infections (300/1768, 17.0%) and Triple infections (10/1768, 0.6%) were found only among the patients with HIV. The malignancies among HIV-infected patients included adenocarcinomas (0.22%), small cell carcinomas (0.2%), squamous cell carcinomas (0.1%), and diffuse large B-cell lymphoma (0.1%). HIV-infected patients exhibited a significantly lower incidence of neoplastic lesions (0.6% vs. 9.0%) than the HIV-uninfected patients (p < 0.001).ConclusionsThere was a significant difference in the spectrum of lung diseases between HIV-infected and non-infected patients diagnosed by BALF cytopathology.

Abstract WP237: Uric Acid Expressed Within Carotid Atherosclerotic Plaque And Serum Uric Acid Are Associated With Cerebrovascular Events

Introduction: Uric acid (UA) level within carotid plaque specimens and its association with cerebrovascular events has not been detected or quantified. Systemically, serum uric acid (SUA) is a marker of inflammation and risk factor for atherosclerosis. However, its association with carotid plaque instability and subsequent cerebrovascular events remains unclear. Hypothesis: We hypothesized that UA may play a role in carotid artery atherosclerosis and stroke pathogenesis. In patients undergoing carotid endarterectomy (CEA), we aimed to determine whether UA is present differentially in carotid plaques of symptomatic versus asymptomatic patients and whether SUA is associated with cerebrovascular symptoms before surgery and major adverse cardiovascular events (MACE) in the follow-up to CEA. Methods: We prospectively collected 32 carotid plaques during CEA. We qualitatively assessed the presence of UA by using Gomori methenamine silver (GMS) staining and immunohistochemistry with anti-UA antibodies. We extracted and measured the quantity of UA in carotid plaques by using an enzymatic colorimetric assay. A retrospective analysis of a clinical database of 534 consecutive patients who underwent CEA was performed. We defined as symptomatic those patients with history of stroke, TIA and amaurosis fugax. Results: UA positive staining was detected in higher number of carotid plaques from symptomatic compared to asymptomatic patients on both staining, GMS [20 (86.9%) vs 2 (22.2%); p=0.001] and immunohistochemistry anti-UA [16 (69.5%) vs 1 (11.1%); p=0.004]. Moreover, we found a significantly higher concentration of UA in carotid plaque from symptomatic compared to asymptomatic patients [25.1 (9.5) μg/g vs 17.9 (3.8) μg/g; p=0.021]. SUA level in patients prior to CEA was significantly higher in the symptomatic compared to the asymptomatic group [6.0 (4.8-7.1) mg/dL vs 5.2 (4.4-6.3) mg/dL; p&lt;0.001] and it was significantly higher in patients who developed MACE after CEA versus those that did not [6.0 (4.8-7.1) mg/dL vs 5.4 (4.5-6.6) mg/dL; p=0.021]. Conclusions: UA may provide a mechanistic explanation as systemic biomarker and potential tissue contributor for carotid plaque instability and subsequent cerebrovascular and cardiovascular symptoms.

Nonhealing Ulcer at a Surgical Site

A man in his 80s was evaluated for a 4.5 cm necrotic ulcer with a violaceous border at a previous surgical site (Figure 1). The nonhealing ulcer despite treatment was concerning for squamous cell carcinoma (Marjolin ulcer). Punch biopsy from the ulcer border revealed epidermal hyperplasia with intraepidermal acantholysis, dermal hemorrhage, and lymphoplasmacytic inflammation (Figure 2). Tissue cultures, Grocott methenamine silver, Gram, and acid-fast bacillus stains and immunohistochemistry for herpes simplex virus were unremarkable.

Study of mucormycosis in COVID-19 patients in a tertiary care center, Raichur Institute of Medical Sciences, Raichur.

Coronavirus disease-2019 (COVID-19), caused by severe acute respiratory syndrome (SARS-CoV-2), has become an emergency global public health issue. Disease pattern of COVID-19 ranges from mild to life-threatening pneumonia with associated bacterial and fungal coinfections. Due to associated comorbidities (e.g., diabetes mellitus and chronic obstructive pulmonary disease) and immunocompromised conditions (e.g., corticosteroid therapy), these patients are prone to develop severe opportunistic infections. The aim of this study was to investigate the incidence of fungal coinfection of hospitalized patients with COVID-19 infection (SARS-CoV-2) in RIMS, Raichur. Our study included 94 patients with invasive fungal infection who presented with either COVID-19 infection or had recovered from COVID-19 infection. Tissue samples from suspected sites were received in formalin for histopathological examination, and these were stained stains with hematoxylin and eosin and with periodic acid-Schiff/Gomori's methenamine silver wherever required. A total of 94 biopsies were received with clinical suspicion of mucormycosis. Out of these, 52 (55.3%) were positive for mucormycosis and 42 (44.7%) were negative. Out of 52 positive cases, 46 (88.4%) were males and 6 (11.6%) were females. The highest number of cases was seen in age group of 51-60 years (26.92%). Out of the 52 positive cases, KOH mount was positive for fungal elements in 8 cases and negative for fungal elements in 29 cases. Uncontrolled diabetes and overzealous use of steroids are two of the main factors aggravating the incidence of mucormycosis in COVID-19 patients. Early surgical intervention, histopathological confirmation and rapid and judicious intravenous antifungal treatment should be initiated for better patient outcomes.

Частота встречаемости гломерулопатий при новообразованиях почек в Красноярске

Introduction. Renal malignancies accounted for 3.9% of all malignant neoplasms in Russia in 2019 and were among the top ten most common. Paraneoplastic syndrome can occur in malignant neoplasms and manifest as paraneoplastic nephropathies and glomerulopathies. The aim of the study was to analyze archival renal specimens to assess the incidence of glomerulopathies in renal neoplasms. Materials and methods. We analyzed 122 kidney samples with a tumor after nephrectomy in Krasnoyarsk regional clinical oncologic hospital named after A.I. Kryzhanovsky. To assess the cellular composition of the glomeruli and basement membrane structure, the following histochemical stains were used: PAS reaction, Masson’s trichrome stain, methenamine silver, and congo red stain. Results. The age of the patients ranged from 28 to 84 years. Malignant neoplasms were diagnosed in 114 cases (93.4%), whereas benign neoplasms were detected in 8 patients (6.6%). Fifty-six cases (45.9%) showed glomerulopathies. In 55 cases, glomerulopathies were found in malignant neoplasms (98.2%) and one case was found in a patient with a benign neoplasm (1.8%). Diabetic nephropathy was detected in 43 cases, ac-counting for 76.8%. Glomerulonephritis was diagnosed in 7 patients (12.5%) and focal segmental glomerulosclerosis was revealed in 6 cases (10.7%). Conclusion. Glomerulopathies in renal tumors are rather common and therefore their diagnosis is important in the study of the operative material. Keywords: kidney, tumour, glomerulopathies, paraneoplastic syndrome, diabetic nephropathy, glomerulonephritis