Fungal infection in surgical pathology – Our institutional experience

Background: Fungal infection has a worldwide prevalence and it affects almost all age groups and both the sexes. Due to neoteric treatment modalities nowadays, people are enjoying longevity of life. However, on the other hand, immune status is compromised that has resulted in increased prevalence of fungal infection in society. Fungi reported by surgical pathologist, either incidentally or in clinically suspicious cases, greatly improve morbidity and outcome of the patients. Objective: The objective of this study is to study the prevalence of fungal infection in surgical pathology tissue and frequency and spectrum-based analysis in relation to age, gender, and systemic involvement. Materials and Methods: Ethically approved retrospective observational secondary data analysis is done based on histomorphological and cytomorphological evaluation of 17 fungal cases diagnosed at the Department of Pathology, New Civil Hospital, Surat, from January 2015 to January 2017. Results: Out of 17 cases, most common fungus seen was Candida albicans mainly affecting gastrointestinal tract. Common age group affected was 41–60 years and few rare fungi, i.e., mucormycosis and tinea corporis were also reported. Conclusion: Distribution of fungi in tissue section in relation to frequency, age, sex, various organs, and its correlation with clinical symptoms is analyzed in our study and results are found to be yielding that provided outlook in the prevalence of infectious agent.

Antifungal susceptibility and virulence of Aspergillus fumigatus environmental strains from a public tertiary hospital in Metro Manila, Philippines

@#<p><strong>Background and Objectives:</strong> The increase in the number of invasive Aspergillus infections has been observed among immunocompromised and hospitalized patients. In the Philippines to date, no published data focused on the prevalence of Aspergillus species or any other thermotolerant fungal species in a hospital environment. This research served as a primary study to characterize the antifungal susceptibility of environmental strains of Aspergillus fumigatus from a hospital facility against three antifungal agents and to determine the virulence of these isolates on BALB/c mice using an animal survival assay.<br /><strong>Methodology:</strong> Ten environmental strains of A. fumigatus were isolated from three air-conditioned wards in a medical facility using Andersen Air Sampler. The antifungal susceptibility profile of the isolates was determined against Voriconazole, Amphotericin B and Caspofungin. The virulence of these isolates was also tested on BALB/c mice using an animal survival assay. Moreover, the lung tissues of infected BALB/c mice were subjected to histopathological analyses using Gomori Methenamine Silver stain (GMS) and Hematoxylin & Eosin (H&E) stains.<br /><strong>Results:</strong> Etest result for antifungal susceptibility testing showed that two of the ten isolates were resistant to Amphotericin B (AF2-A and AF-3A); one isolate resistant to Voriconazole (AF2-A) and an isolate that manifested non- susceptibility to Caspofungin m(AF2-A). Epidemiological cut-off values were determined for each antifungal following the M38-A2 CLSI guidelines. BALB/c mice median survival analysis revealed that the isolate with the highest Minimum Inhibitory Concentration (MIC= 4.89 ?g/ml) for Voriconazole resulted in the most number of mortality with the least number of observation days. GMS AND H&E histopathology slides showed fungal elements embedded on left lung lobe of mice.<br /><strong>Conclusion:</strong> This study showed that there were strains of Aspergillus fumigatus from a hospital indoor air which were considered as resistant strains to Voriconazole, Amphotericin B, and Caspofungin (AF2-A and AF3-A). Lung tissues of infected mice showed characteristics of bronchopneumonia.</p>

Evaluation of Real-time PCR and Gomori-Methenamine Silver Stain for Diagnosing of Pneumocystis Pneumonia / 现代检验医学杂志

Objective To compare real-time PCR and gomori-methenamine silver stain in the diagnosis of pneumocystis peumonia (PCP).Methods 2 525 unrepeated specimens from suspected PCP patient admitted in Peaking Union Medical College Hospital were collected in 2014.2 492 samples were detected by gomori-methenamine silver stain,33 samples were detected by real-time PCR,and 429 samples were detected by both methods at the meanwhile.With clinical diagnosis as reference standard,the sensitivity,specificity,positive predictive value and negative predictive value of the two methods were analysised.Results Positive rate of gomori-methenamine silver stain was 1.2 ％ (30/2 492).The first three specimen types were sputum,tracheal intubation suction and bronchoalveolar lavage fluid,the positive rate was 0.70 ％ (13/1 845),4.00％ (10/250) and 2.72％ (7/257) respectively.Positive rate of realtime PCR was 34.20％ (158/462),and the positive rate of sputum and bronchoalveolar lavage fluid was 30.61％ (105/343) and 44.54％ (53/119) respectively.The sensitivity were 13.97％ vs 72.07％,specificity were 100％ vs 94.24％,positive predictive value were 100％ vs 92.14％ and negative predictive value were 55.36％ vs 78.26％ for gomori-methenamine silver stain and real-time PCR respectively.All of which were statistically significant analysed by x2 test for paired data.The x2 value and P alue were x2 =68.625,P＜0.01;x2 =4.296,P＜0.05;x2 =6.380,P＜0.01 and x2 =11.873,P＜0.01.Conclusion The real-time PCR had higher sensitivity,fewer interference factors and more clinical diagnostic value,so clinicians should make more use of real-time PCR to diagnose PCP earlier.

Application of special staining techniques in pathological diagnosis of fungi infections in HIV/AIDS patients / 中华临床感染病杂志

Objective To apply special staining techniques in pathological diagnosis of fungal infections in HIV/AIDS patients.Methods Pathological data of 20 HIV/AIDS patients complicated with fungi infections in Shanghai Public Health Clinical Center during February 2010 and November 2013 were retrospectively analyzed.Tissue specimens were stained with hematoxylin and eosin (HE),Periodic acidSchiff (PAS) and methenamine silver nitrate (MSN),and the sections were observed under optical microscope.Results Among 20 HIV/AIDS patients complicated with fungi infections,2 were infected with pulmonary cryptococcosis; 3 were penicillium marneffei infections in skin,lung and abdominal mesenteric lymph nodes; 5 were histoplasma capsulatum infections in epiglottis,neck lymph nodes,oral cavity,abdominal cavity and skin; 4 were aspergillus infections in maxillary sinus,lung and vocal cords,and 3 of them were combined with tuberculous lesions; 6 were candida albicans infections in liver,pharynx,esophagus and stomach.In tissues stained with HE the infiltration of inflammatory cells,granuloma formation and coagulative necrosis were observed,and the shape of fungi needed careful observation to avoid missed diagnosis and misdiagnosis.In tissues stained with PAS,fungal spores and pseudohypha were presented in bright amaranth,and cell nucleus was in purple-blue.In tissues stained with MSN,fungal spores and pseudohypha were identified clearly in brown-black.Conclusion HE plus PAS and MSN staining will help pathological diagnosis of fungi infection.

Recurrent urinary tract infections: evaluation of the prophylactic efficacy of urinary antiseptics methenamine and methylthioninium chloride / Infecções do trato urinário recorrentes: avaliação da eficácia profilática dos anti-sépticos urinários metenamina e cloreto de metiltionínio

Introduction: Urinary tract infection (UTI) is a very common condition in clinical practice, affecting an estimated 50% of all adult women during a lifetime. The most common causative agent is E. coli UTI may also be caused by S. saprophyticus, Enterobacteria (Klebsiella sp and Serratia sp.), Enterococcus sp., and P. aeruginosa. Recurrent UTIs occur at least twice per semester or three times a year. Prophylactic measures to prevent recurrent UTIs include changes in contraception methods, cranberry products, increased fluid intake, urination after intercourse, vaginal estrogen therapy for post-menopausal women, antibiotics, and urinary tract antiseptic agents. Objectives: To evaluate the use of a combination of methenamine and methylthioninium chloride in the prophylaxis of recurrent uncomplicated lower UTIs, with respect to: · Signs and symptoms of UTI· Etiologic agent(s)· Recurrence rates· Need for antibiotic therapy in case of recurrence · Incidence of adverse events associated with the treatment, including any reported alterations of laboratory testsMaterials & methods: A descriptive, analytic, restrospective study was performed at Hospital Universitário Constantino Otaviano - UNIFESO. Medical charts from patients presenting recurrent uncomplicated lower UTI attended from 2001-present were analyzed, including the following information: Demographic data (age, gender, weight, ethnicity, living conditions) medical history/ signs and symptoms of UTI identification of treatment and dosing regimens treatment duration recurrence rates and need for antibiotic therapy in case of recurrence other medications prescribed and records of adverse events. Results: E. coli was identified as etiologic agent in 80% of the patients. Following antibiotic therapy, all patients received prophylactic treatment with the combination of methenamine and methylthioninium chloride. Treatment duration ranged from three to six months. Adverse events were observed in 13/60 patients...

Introdução: A infecção do trato urinário é bastante comum na prática clínica, afetando aproximadamente 50% de todas as mulheres adultas em algum momento da vida. O agente etiológico mais comum é a E. coli. A ITU pode também ser causada por S. saprophyticus, Enterobactérias (Klebsiella sp. e Serratia sp.), Enterococcus sp. e P. aeruginosa. ITUs recorrentes ocorrem ao menos duas vezes por semestre ou três vezes ao ano. Medidas profiláticas para prevenir a recorrência incluem alterações nos métodos contraceptivos, produtos de oxicoco, aumento da ingestão de fluidos, micção após a relação sexual, e terapia vaginal de estrogênio para mulheres em pós-menopausa, bem como antibióticos e anti-sépticos urinários. Objetivos: Avaliar o uso da combinação de de metentamina e cloreto de metiltionínio na profilaxia de ITUs recorrentes não-complicados, com respeito à: · Sinais e sintomas de ITU · Agente etiológico · Número de recorrências · Necessidade de terapia antibiótica em caso de recorrência · Incidência de efeitos adversos do tratamento, inclusive quaisquer alterações de exames laboratoriais. Materiais e métodos: Foi realizado um estudo descritivo, analítico, retrospectivo no Hospital Universitário Constantino Otaviano – UNIFESO. Prontuários médicos disponíveis de pacientes apresentando ITU recorrente não complicada, atendidos de 2001-presente foram analisados, incluindo-se as seguintes informações: dados demográficos (idade, sexo, peso, etnia, condições habitacionais) histórico médico/ sinais e sintomas de ITU identificação do tratamento e regime de tratamento recorrência necessidade de tratamento antibiótico no caso de recorrência outros medicamentos prescritos e registro de efeitos adversos. Resultados: E. coli foi identificado como agente etiológico em 80% dos pacientes. Após terapia antibiótica, todos os pacientes receberam tratamento profilático com a combinação de metenamina e cloreto de metiltionínio. A duração do tratamento variou entre três a seis meses...

The Evaluation of Usefulness of PAS Stain and GMS Stain as Clinical Diagnostic Methods of Tinea unguium / 대한의진균학회지

BACKGROUND: Tinea unguium is a common problem seen in clinical practice. Considering the many differential diagnoses of dystrophic nails, it is important to make a definitive diagnosis of dermatophyte infection before the initiation of antifungal therapy. Potassium hydroxide (KOH) preparation and fungal culture, which are commonly used in the diagnosis of these infections, often yield false-negative results. Recent reports have suggested that nail plate biopsy (Bx) using periodic acid-Schiff (PAS) (Bx/PAS) stain may be a very sensitive technique for the diagnosis of tinea unguium. OBJECTIVE: The purpose of this study was to evaluate the usefulness of PAS and Grocott's methenamine silver (GMS) staining of nail specimen in the diagnosis of tinea unguium as a standard method. METHODS: We evaluated 75 nail specimens from suspected tinea unguium using KOH preparation, biopsy using periodic acid-Schiff stain, and Grocott's methenamine silver stain. RESULTS: Of the 75 nails which were negative on potassium hydroxide mounting, 43 and 39 cases were tested positive respectively on periodic acid-Schiff stain and Grocott's methenamine silver stain. CONCLUSION: Bx/PAS and Bx/GMS are the sensitive methods for the diagnosis of tinea unguium. They are indicated if clinical suspicion of onychomycosis is high and KOH preparation shows no fungal elements.

A method of assessing the thickness of myocardial basement membranes / 中国药理学通报

Aim To observe the thickness changes of cardiomyocyte and microvascular basement membranes(BM)and the expression of collagen Ⅳ in Gk rats of different months and STZ-induced diabetic rats,and to explore the method that operated easily and could assess the thickness of myocardial BM.Methods GK rats of the fourth,fifth,sixth,seventh months,Streptozotocin induced diabetes rats of fifth months and normal Wister rats were respectively divided into different groups with ev eight rats in each group.The myocardial section was stained by methenamine silver and immunohistochemistry was performed for collagen Ⅳ.Left ventricular endocardium was selected as measuring view.Through computer image analysis,the ratio of BM positive staining density(BMPSD)and positive staining density of collagen Ⅳ(CPSD)were calculated separately in all myocardium.Take the ratio to assess the thickness of myocardial BM and the expression of collagen Ⅳ.Results The glycemia,BMPSD and CPSD of the two diabetes rats were higher than those of the normal rats,having highly remarkable difference(P0.05).Conclusion By methenamine silver staining for myocardial section,calculating BMPSD through computer image analysis,can be an index to assess the thickness of myocardial BM.

Immunocytochemical Detection of Pneumocystis Carinii in Bronchoalveolar Lavage / 대한세포병리학회지

Pneumocystis carinii is an established cause of pulmonary infections in immuno- compromised hosts. Several cytological stains, such as Papanicolaou, Gomori methenamine silver(GMS) and Diff-Quik have been used for detection of the organism, but occasionally can be laborious and, due to a degree of nonspecificity, may be misleading. We evaluated the diagnostic utility of immunocytochemical stains that recognize P. carinii in bronchoalveolar lavage from experimentally induced P. carinii pneumonia rats(n=15). In addition to routine stains for diagnosis by morphologic recognition of P. carinii on Papanicolaou, GMS and Diff-Quik stains, bronchoalveolar lavage samples were reacted with immunocytochemical stains using monoclonal antibodies(MAB) 092 and 902. In bronchoalveolar lavage P. carinii organisms were detected in 9 of 10 cases (90%) using each MAB 092 and 902, whereas GMS and Diff-Quik stains demonstrated P. carinii in 13(86%) and 11(73%) of 15 cases respectively. In lung tissue specimens(n=15) P. carinii organisms were well identified on GMS stain and immunohistochemical stains using MAB 092 and 902 in all cases. We believe that the immunocytochemical staining using MAB 092 and/or 902 is a very useful and diagnostic tool in addition to GMS and Diff-Quik stain to detect P. carinii organisms in bronchoalveolar lavage.

ULTRASTRUCTURAL STUDIES ON PARIETAL CELLS OF MOUSE STOMACH USING CL~- LOCALIZATION AND POLYSACCHARIDES STAINING TECHNIQUES / 解剖学报

Cl~- localization and periodic acid-silver methenamine techniques have been used for ultrastructural studies on parietal cells of mouse stomach. Cl~- is located only in the intracellular canaliculi (IC) and intercellular space (IS)of parietal cells. The tubulovesicular system (TS) shows various reactions in periodic acid-silver methenamine staining. The positive reaction was demonstrated in the TS which is located near the apical membranes. This indicates that acid secretion is accompanied by polysaccharides secretion in which the TS takes part, and the polysaccharides secretory activity shows its cyclicity which is characterized by the changes of amounts of secretion. The cycle of parietal cells do not synchronize.