Methenamine Silver Nitrate Research Articles

The Vital Role of Special Stains in Immunocompromised Cases: A Focus on Histoplasmosis Screenings for Enhanced Kaposi`s Sarcoma Diagnosis Beyond H&E Morphology

Abstract Introduction/Objective This case report emphasizes the critical importance of thorough pathology-based investigation before making a definitive diagnosis of Kaposi’s Sarcoma (KS). In particular, we demonstrate the importance of using special stains in addition to H&E staining to determine if the lesion is KS or a fungal infection, specifically cutaneous Histoplasmosis. This highlights the necessity of comprehensive diagnostics to ensure accurate and timely identification of underlying conditions, guiding appropriate treatment strategies. Methods/Case Report In Kampala Uganda, a 56-year-old male with a history of HIV/AIDS presented with skin lesions on the lower extremities. The lesions were initially suspected to be Kaposi’s Sarcoma due to their appearance and the patient’s immunocompromised status. Skin biopsy results supported the initial suspicion, showing spindle cell proliferation in the dermis, a common feature of KS. Given his compromised immune system, it was imperative to conduct a thorough investigation to rule out potential etiologies, including opportunistic infections. Additional tests, including a Grocott Methenamine-Silver Nitrate stain, revealed the presence of Histoplasmosis. The patient did not exhibit typical symptoms of Disseminated Histoplasmosis, such as respiratory distress or fever. Resource constraints hindered comprehensive follow-up, impacting long-term outcome assessment. Results (if a Case Study enter NA) NA Conclusion Histoplasmosis is widespread in Africa, accounting for 61% of global cases, with 87% concentrated in Western and Central Africa. The coexistence of KS and Histoplasmosis poses diagnostic challenges due to overlapping clinical manifestations. KS primarily affects the skin, while Histoplasmosis causes invasive vascular and pulmonary infections. However, in Africa, notably in immunocompromised individuals, histoplasmosis can cause cutaneous infections. Failure to recognize and address fungal infections may result in delayed or inappropriate treatment, exacerbating the patient’s condition. This case report underscores the importance of maintaining a high index of suspicion for opportunistic infections in this population for timely intervention.

Modification of Grocott's staining procedure with heat treatment and oxidation by periodic acid for mucormycosis in tissue: a method to detect Mucor spp.

The sensitivity of the Grocott-modified Gomori's methenamine-silver nitrate technique for the detection of fungi is sometimes low, especially for Mucor spp. We modified the Grocott technique by replacing chromic acid with periodic acid in the oxidation step. The use of periodic acid instead of chromic acid enhanced the detectability of Mucor spp. in histopathological sections. Other parameters should be assessed with a high number of cases under different conditions. We propose our protocol as one of the options in practice, especially in cases suspected of Mucor spp. infection.

Painful facial, oral, and genital ulcers

Painful facial, oral, and genital ulcers

Rinite em caprino por Exserohilum rostratum (Setosphaeria rostrata)

ABSTRACT: We described a case of rhinitis case caused by Exserohilum rostratum in a 3-year-old, mixed breed, afemale goat, presenting with decreased appetite, dyspnea, and face deformity, with evolution of approximately 2 months. Necropsy revealed an extensive brownish yellow, irregular, ulcerated, and friable focal lesion in the nasal cavity. Purulent secretion drained from the lesion, affecting the nasal vestibule and extending the caudal portion of the dorsal concha, associated with nasal meatus obstruction, destruction of the nasal septum, and nasal bone palatine and vomer. Histologically, the lesion was characterized as rhinitis and pyogranulomatous and necrotizing osteomyelitis (multifocal to coalescent, sharp, and chronic) and associated with a myriad of fungal structures. Morphologically, the hyphae were characterized by thin, slightly tortuous, and rarely septate walls (2-6 μm diameter), multiple chains of individual conidia or in groups (8-10 μm diameter), and pigmented chlamydoconidia (7.5-15 μm diameter). They showed positive results for the special histochemical techniques such as Fontana Masson, methenamine silver nitrate by Grocott, and periodic acid by Schiff. E. rostratum was identified as the causative agent using microbiological isolation associated with molecular identification. Pyrogranulomatous rhinitis caused by E. rostratum in goats is unprecedented, being the first isolation of the agent in animals in Brazil and the first case of nasal phaeohyphomycosis in the goats.

Strawberry gingivitis – First sign of Wegener's granulomatosis

Wegener's Granulomatosis (WG) is an immunologically mediated rare multisystem disease characterized by necrotizing granulomatous inflammation affecting the upper and lower respiratory tracts, disseminated vasculitis and glomerulonephritis. WG oral lesions are reported to occur in 6%–13% of patients; they were an initial presenting feature in only 2% of cases. A case of strawberry gingivitis as the first sign of WG presented with erythematous and painful localized gingival enlargement affecting the maxillary anterior region of jaw in a 17-year-old boy is reported here. Correlation of histopathology with routine hematoxylin and eosin and special stains such as Grocott-Gomori's methenamine-silver nitrate and periodic acid–Schiff, peripheral blood smear, chest radiographs, Mantoux test and clinical presentation were established in diagnosing this rare entity. By the above-mentioned procedures, we arrived at the diagnosis of WG. The aim of reporting this case was to emphasize that, the dental surgeon often being the first person to examine the oral cavity, should be familiar with the typical appearance of gingiva as “strawberry gingivitis,” its clinical course, diagnostic parameters and adequate management of WG.

Esophageal Leukoplakia.

CASE REPORT A 64-year-old white man with a history of tobacco use and stable weight presented to the gastroenterology clinic due to an 11-month history of solid-food esophageal dysphagia. On physical examination, patient was alert and in no acute distress. His vital signs were within normal limits. His head and neck examination revealed normal oropharyngeal mucosa. No thyromegaly, lymphadenopathy, or masses in his neck were noted on palpation. Abdominal examination presented normal bowel sounds, without distension or tenderness. Upper endoscopy demonstrated white plaques at the gastroesophageal junction (Figure 1).Figure 1.: Upper endoscopy showing white plaques at the gastroesophageal junction.Biopsies obtained from this abnormality showed esophageal epidermoid metaplasia consistent with leukoplakia and mild patchy active chronic inflammation. Gomori methenamine–silver nitrate stain for fungal organisms was negative. Given the potential for malignant transformation of leukoplakia, endoscopic resection was suggested for the patient. An endoscopic submucosal dissection was planned, due to the lesion size to ensure compete removal. Chromoscopy with methylene blue was used to mark the borders of the lesion, and endoscopic submucosal dissection was accomplished without adverse events (Figure 2). Histologic examination confirmed hyperplastic squamous esophageal mucosa with marked hyperkeratosis without dysplasia and negative margins. During the 2-year follow-up, the patient reported complete resolution of his dysphagia. An upper endoscopy was ordered for surveillance.Figure 2.: Chromoscopy with methylene blue was used to mark the borders of the lesion.Epidermal metaplasia is a histological correlate to a white patch on the surface of a mucous membrane that cannot be rubbed off, known as leukoplakia.1 It is a common finding in oral mucosa, but rare in esophageal mucosa.1,2 The etiology of this condition in the esophagus is unclear and thought to be associated with esophageal tobacco and alcohol exposure or possibly an unusual response to chronic acid reflux.1,3,4 Although generally asymptomatic, symptom presentation can include dysphagia.3 Endoscopic features include slightly elevated whitish surface, which can be translucent or cobblestone-like in appearance.2,4 The natural history is unknown. As adjacent dysplasia and possible squamous cell carcinoma of the esophagus has been reported, and it is suggested that there is a potential for malignant transformation and therefore strict surveillance or endoscopic resection or ablation is warranted.1,5 DISCLOSURES Author contributions: PV Hernandez drafted the manuscript. D. Snyder, A. Kahn, KK Wang, DA Katzka, JL Horsley-Silva revised the manuscript. JL Horsley-Silva is the article guarantor. Financial disclosure: None. Previous presentation: This case report was presented as a poster at the 2019 ACG Annual Meeting; October 25-30, 2019; San Antonio, TX. Informed consent was obtained for this case report.

Editing the genome of Aphanomyces invadans using CRISPR/Cas9

BackgroundThe clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is increasingly being used for genome editing experiments. It is a system to add, delete and/or replace parts of a gene in situ in a time- and cost-efficient manner. The genome of many organisms has been edited using this system. We tested the CRISPR/Cas9 system in Aphanomyces invadans, an oomycete, which is the causative agent of epizootic ulcerative syndrome (EUS) in many fish species. Extracellular proteases produced by this oomycete are believed to play a role in EUS virulence.MethodsWe designed three single guide-RNAs (gRNA) to target A. invadans serine protease gene. These gRNAs were individually combined with the Cas9 to form ribo-nucleo-protein (RNP) complex. A. invadans protoplasts were then transfected with RNP complexes. After the transfection, the target gene was amplified and subjected to sequencing. Zoospores of A. invadans were also transfected with the RNP complex. Three groups of dwarf gourami (Trichogaster lalius) were then experimentally inoculated with (i) non-treated A. invadans zoospores; (ii) RNP-treated A. invadans zoospores; and (iii) autoclaved pond water as negative control, to investigate the effect of edited serine protease gene on the virulence of A. invadans in vivo.ResultsFluorescence microscopy showed sub-cellular localization of RNP complex in A. invadans protoplasts and zoospores. Sequencing results from the protoplast DNA revealed a point mutation in the target gene. A matching mutation was also detected in zoospores after similar treatment with the same RNP complex. In vivo results showed that the CRISPR/Cas9-treated A. invadans zoospores did not produce EUS clinical signs in the fish. These results were then confirmed by histopathological staining of the muscle sections using Gomori’s methenamine silver nitrate and hematoxylin and eosin stains.ConclusionsResults obtained in this study indicate that the RNP complex caused effective mutation in the target gene. This hindered the production of serine protease, which ultimately impeded the manifestation of EUS in the fish. Our methods thus establish a promising approach for functional genomics studies in A. invadans and provide novel avenues to develop effective strategies to control this pathogen.

INVESTIGATION OF CYTOSTATIC ACTIVITY OF THE METHENAMINE SILVER IN VITRO AND IN VIVO

Background: Developing of novel pharmaceutic agents is an actual topic for chemotherapy. Heavy metals possess a significant cytotoxic properties and some of them are widly introduced in medical practice, e.g. cisplatin.Aim and Objective of this study was to investigate the cytostatic effects of prospective silver-based preparation on viral bovine leukemia model in vivo and lymphoblast cells in vitro.Material and Methods: Silver salt of methenamine was suggested as prospective substance and tested was tested on black-motley cattle with bovine leukemia to reveal influence on hematological parameters. Preparation was applied in form of 0.5% solution in different doses and was administered as single intravenous injection. Blood parameters were determined by triple hematological monitoring. Investigation of cytostatic activity was conducted on the leukemic cells of HUT-102 line in Vitro in concentration range of 0.03-3.0 mg/ml.Results: The results showed normalization of blood parameters in dose-dependent manner after intravenous injection of methenamine silver salt. It was revealed a pronounced inhibitory effect of silver-based drug on the leukemic cells both in vitro and in vivo. Control blood test one month after experimental therapy confirmed the stability of the achieved results.Conclusion: The tested drug showed a moderate cytostatic effect. Methenamine silver nitrate can be considered as prospective pharmacological agent. Bovine leukemia was shown as a convenient model for the development of new treatment of blood diseases. Continuation of the work is of interest for both veterinary and medical pharmacotherapy.Keywords: methenamine silver nitrate, HUT 102 cells; lymphocytes, leucosis, cytostatic activity, bovine leukemia, silver hexamethylenetetramine.

Bronchopulmonary Aspergillosis Associated with Acute Myocardial Infarction in Dog

Background: Fungi of the genus Aspergillus are ubiquitous microorganisms of opportunistic character, of which animals and humans are continually exposed. Are described three main forms of aspergillosis in dogs: nasal, disseminated and bronchopulmonary. This form which is restricted to the lung parenchyma is considered rare in dogs. The aim of this paper is to report a case of pulmonary aspergillosis associated to acute myocardial infarction in a dog, focusing on epidemiological, clinical, pathological and immunohistochemical characteristics. Case: A male dog, 4-year-old, Pinscher, clinically presented dyspnea, anorexia, vomiting, concentrated urine and dark and mollifed faeces. On physical examination, there were congested oral and conjunctival mucosas, dehydration and petechiae in the outer ear. The animal died fve days after the clinical attendance and was sent for necropsy. The macroscopic lesions were restricted to the lung, heart and liver. There were multifocal areas of consolidation in the lung parenchyma and yellowish content in the trachea and bronchi lumen. The heart was rounded and with pale areas in the myocardium. Liver was discreet accentuation of the lobular pattern. Histologically the lesion was characterized by pyogranulomatous and necrotizing bronchopneumonia, multifocal to coalescing, moderate, associated with myriads of intralesional hyphae consistente with Aspergillus spp., associated with acute myocardial infarction. The hyphae weakly stained basophilic by hematoxylin and eosin were strongly impregnated by Grocott’s methenamine silver nitrate and showed strong immunolabelling of the cytoplasm, which were evidenced in brown. There were areas of acute myocardial infarction and hepatocellular degeneration and congestion in the liver, although fungal hyphae were not observed in these tissues. Discussion: The diagnosis of pulmonary aspergillosis was established based on the morphological and tinctorial characteristics of the agent and confrmed by immunohistochemistry. In this case, is not included in the clinical history any conditions that could promote immunosuppression, neither were observed morphological changes in the pathological examination which suggested other intercurrent diseases. It is likely that the dog has been exposed to the inhalation of large amounts of conidia for a long period of time, condition that favors the occurrence of the disease, even in immunocompetent animals. Probably were not evidenced severe respiratory clinical signs, that characterize the pulmonary impairment, due to the shortcourse of the disease. The pyogranulomatous and necrotizing inflammatory reaction is typical of fungal infection and was restricted to the lung parenchyma. The areas of coagulation necrosis observed in the myocardium are probably secondary to vascular injury caused by the fungi, with subsequent thromboembolism, ischemia and infarction. In Brazil, there are few reports of aspergillosis in dogs, have been reported the nasal and systemic forms. In the world literature, there are few reports about this bronchopulmonary form of the disease. Although uncommon, pulmonary aspergillosis should be considered in the differential diagnosis of diseases that affect the lower respiratory tract of dogs, as well as it’s systemic complications resulting from angioinvasive characteristic of the fungus, which in this case was determinant to the death of the animal. Keywords: Aspergillus, fungal disease, respiratory tract

Investigation of Pneumocystis jirovecii colonization in patients with chronic pulmonary diseases in the People's Republic of China.

BackgroundThe detection of Pneumocystis jirovecii DNA in respiratory specimen from individuals who do not have signs or symptoms of pneumonia has been defined as colonization. The role of P. jirovecii colonization in the development or progression of various lung diseases has been reported, but little information about P. jirovecii colonization in patients is available in the People’s Republic of China.ObjectiveTo determine the prevalence of P. jirovecii colonization in patients with various pulmonary diseases, including the acute and stable stage of COPD, interstitial lung diseases, cystic fibrosis, and chronic bronchiectasis.Materials and methodsA loop-mediated isothermal amplification (LAMP) and a conventional polymerase chain reaction (PCR) method for detecting P. jirovecii were developed. Ninety-eight HIV-negative patients who were followed-up and who had undergone bronchoscopy for diagnosis of various underlying respiratory diseases were included in the study. Sputa of these patients were analyzed with LAMP amplification of P. jirovecii gene. In addition, conventional PCR, Giemsa and Gomori’s methenamine silver nitrate staining assays were applied to all specimens.ResultsThe sensitivity and specificity test showed that there was no cross-reaction with other fungi or bacteria in detecting the specific gene of P. jirovecii by LAMP, and the minimum detection limits by LAMP was 50 copies/mL. P. jirovecii DNA was detected in 62 of 98 (63.3%) sputa specimens by LAMP assay and 22.45% (22/98) by conventional PCR. However, no P. jirovecii cysts were found by Giemsa and Gomori’s methenamine silver nitrate in all of gene-positive specimens.ConclusionThe results of our study showed that prevalence of P. jirovecii colonization is particularly high in patients with chronic pulmonary diseases in the People’s Republic of China, and the LAMP method is better for evaluation of the colonization of P. jirovecii in sputum specimen than conventional PCR.

Cutaneous Invasive Aspergillosis: Retrospective Multicenter Study of the French Invasive-Aspergillosis Registry and Literature Review.

Invasive aspergillosis (IA) has poor prognosis in immunocompromised patients. Skin manifestations, when present, should contribute to an early diagnosis. The authors aimed to provide prevalence data and a clinical and histologic description of cutaneous manifestations of primary cutaneous IA (PCIA) and secondary CIA (SCIA) in a unique clinical series of IA and present the results of an exhaustive literature review of CIA. Cases of proven and probable IA with cutaneous manifestations were retrospectively extracted from those registered between 2005 and 2010 in a prospective multicenter aspergillosis database held by the National Reference Center for Invasive Mycoses and Antifungals, Pasteur Institute, France. Patients were classified as having PCIA (i.e., CIA without extracutaneous manifestations) or SCIA (i.e., disseminated IA). Among the 1,410 patients with proven or probable IA, 15 had CIA (1.06%), 5 PCIA, and 10 SCIA. Hematological malignancies were the main underlying condition (12/15). Patients with PCIA presented infiltrated and/or suppurative lesions of various localizations not related to a catheter site (4/5), whereas SCIA was mainly characterized by disseminated papules and nodules but sometimes isolated nodules or cellulitis. Histologic data were available for 11 patients, and for 9, similar for PCIA and SCIA, showed a dense dermal polymorphic inflammatory infiltrate, with the epidermis altered in PCIA only. Periodic acid Schiff and Gomori-Grocott methenamine silver nitrate staining for all but 2 biopsies revealed hyphae compatible with Aspergillus. Aspergillus flavus was isolated in all cases of PCIA, with Aspergillus fumigatus being the most frequent species (6/10) in SCIA. Two out 5 PCIA cases were treated surgically. The 3-month survival rate was 100% and 30% for PCIA and SCIA, respectively. Our study is the largest adult series of CIA and provides complete clinical and histologic data for the disease. Primary cutaneous IA should be recognized early, and cases of extensive necrosis should be treated surgically; its prognosis markedly differs from that for SCIA. Any suppurative, necrotic, papulonodular, or infiltrated skin lesion in an immunocompromised patient should lead to immediate biopsy for histologic analysis and mycological skin direct examination and culture.

Low incidence of Pneumocystis jirovecii pneumonia in an unprophylaxed liver transplant cohort

Liver transplant recipients are managed with a range of immunosuppressive regimens that place them at heightened risk of life-threatening opportunistic infections such as Pneumocystis jirovecii pneumonia (PJP). No routine PJP prophylaxis is used at out institution. We reviewed the incidence of PJP in this cohort of unprophylaxed liver transplant recipients. We examined all liver transplants performed between January 2000 and January 2012 in Ireland's National Liver Transplant Centre, St. Vincent's University Hospital, Dublin. Cases were identified through a computerized database and through the histopathology and microbiology registration system. The diagnosis of PJP was confirmed by identification of Pneumocystis cysts in bronchoalveolar lavage (BAL) fluid or on autopsy specimens using Grocott-Gomori methenamine-silver nitrate or modified Wright-Giemsa staining methods. During the study period, 687 liver transplants were performed. We found 7 cases of PJP with an incidence rate of 0.84 per 1000 person transplant years. Five cases occurred within 12months of transplant with 2 cases occurring at 56 and 60months, respectively. Two cases were diagnosed at postmortem; 1 previously had negative cytology from BAL, while the other could not be bronchoscoped because of rapid deterioration in the clinical condition. Three of the 5 treated patients died. The incidence of PJP in this cohort was very low, but the case fatality rate was high. Two cases occurred well after the usual recommended period of prophylaxis. In institutions with a very low risk of infection, targeted rather than universal prophylaxis may be reasonable.

Hybridization quality and bond strength of adhesive systems according to interaction with dentin.

Objective:To evaluate the hybridization quality and bond strength of adhesives to dentin.Materials and Methods:Ten human molars were ground to expose the dentin and then sectioned in four tooth-quarters. They were randomly divided into 5 groups according to the adhesive used: Two single-step self-etch adhesives – Adper Prompt (ADP) and Xeno III (XE), two two-step self-etching primer systems – Clearfil SE Bond (SE) and Adhe SE (ADSE), and one one-step etch-and-rinse system – Adper Single Bond (SB). Resin composite (Filtek Z250) crown buildups were made on the bonded surfaces and incrementally light-cured for 20 s. The restored tooth-quarters were stored in water at 37°C for 24 h and then sectioned into beams (0.8 mm2 in cross-section). Maximal microtensile bond strength (μ-TBS) was recorded (0.5 mm/min in crosshead speed). The results were submitted to one-way ANOVA and Tukey's test (α = 0.05). Thirty additional teeth were used to investigate the hybridization quality by SEM using silver methenamine or ammoniacal silver nitrate dyes.Results:SE reached significantly higher μ-TBS (P < 0.05); no significance was found between ADSE and XE (P > 0.05), and between SB and ADP (P > 0.05); ADSE and XE were significantly higher than SB and ADP (P < 0.05). The bonding interface of SB showed the most intense silver uptake. SE and ADSE showed more favorable hybridization quality than that observed for ADP and XE.Conclusions:The bond strength and hybridization quality were affected by the interaction form of the adhesives with dentin. The hybridization quality was essential to improve the immediate μ-TBS to dentin.

Pathology Quiz Case 1

A 61-YEAR-OLD MAN PRESENTED WITH SEvere otalgia and otorrhea. He had a 35year history of right-sided intermittent otorrhea, aural fullness, and tinnitus and a 3-month history of decreased balance. Topical and systemic antibiotic therapy was initiated, but his symptoms persisted, and new symptoms of severe facial pain and fever developed. He had no history of immunocompromise. Physical examination revealed an edematous, mildly erythematous right external auditory canal, with purulent discharge obscuring the tympanic membrane. There was no pain with movement of the pinna. Temperature, light touch, and sharp and dull stimuli were reduced in the right trigeminal distribution. Audiometric testing showed a down-sloping, moderate to profound mixed hearing loss in the patient’s right ear. Computed tomography of the temporal bone with intravenous contrast showed opacification of the right middle ear, sclerotic mastoid with erosion of the carotid canal adjacent to the eustachian tube, and a peripherally enhancing low-density petrous apex collection (Figure1). Gadolinium-enhanced magnetic resonance imaging also demonstrated a low-signal focus in the right petrous apex, with enhancement and thickening of the cisternal portion of the right trigeminal nerve and meninges of the posterior clivus and sphenoid wing (Figure 2). Hematoxylin-eosinstainingofbiopsyspecimensofgranulomatous tissue from the protympanum and petrous apex demonstratedacuteandchronicallyinflamedciliatedcolumnarepitheliumandgranulation tissue(Figure3).GrocottGomori methenamine-silver nitrate staining (Figure 4) showedaggregatesofgram-positive filamentousbacteria in entangledmasses focallybreakingup intococcoidandbacillary forms.Cultures identifiedProteusmirabilis,Enterococcus avium, and multiple strains of Corynebacterium species with sensitivity to ciprofloxacin. What is your diagnosis?

′Unstained′ and ′de stained′ sections in the diagnosis of chromoblastomycosis: A clinico-pathological study

Diagnosis of chromoblastomycosis is frequently missed for many reasons: (1) rarity of the lesion (2) requirement of careful search for diagnostic 'sclerotic' bodies which are often sparse in clinical material such as tissue and exudates (3) often they elicit tissue reactions such as verrucous lesion and micro abscesses, misleading the diagnosis (4) lack of 'clinical suspicion'. (1) To compare the feasibility of 'unstained', 'stained' and 'de stained' sections in identification of the diagnostic ' sclerotic' bodies (2) To study histopathological features of chromoblastomycosis, and (3) To highlight the importance of clinical suspicion in the diagnosis of chromoblastomycosis. This is a retrospective study conducted on six clinically unsuspected, histopathologically diagnosed cases of chromoblastomycosis. Most common clinical presentation was verrucous lesion over extremities affecting adult men engaged in outdoor works. Sclerotic bodies and their characteristic features were easily identified in both 'unstained' and 'de stained' sections. Special stains such as Fontana Masson and Gomori's methenamine silver nitrate are useful for demonstrating hyphal forms in keratinized layers and for illustration purposes. Both 'unstained' and 'de stained' sections can be used for rapid screening of sclerotic bodies. 'De stained' sections can be used as a suitable substitute for special stains for confirmation and for retrospective review of all verrucous lesions to diagnose the missed lesion. Clinical suspicion is very important in the diagnosis of chromoblastomycosis as it helps pathologist to screen for diagnostic sclerotic bodies.

Rhino-orbito-cerebral infection due to Rhizopus in a diabetic patient

A 55-year-old diabetic female presented with persistent headache, bilateral nasal blockage, common cold and nasal discharge since 10 days. The patient also complained of bilateral nasal bleeding, anosmia and ptosis of the eye since the last five days. Clinical diagnosis of rhino-orbito-cerebral was confirmed by computed tomography (CT) scan of paranasal sinuses and brain. The diagnosis of fungal infection due to Rhizopus species was confirmed by Potassium hydroxide (KOH) examination, Hematoxylin and Eosin staining and Gomori's Methenamine Silver Nitrate staining on mucosal biopsy, followed by culture on Sabouraud's Dextrose agar. Surgical debridement was done and intravenous Amphotericin B was administered to the patient, who responded well to the treatment.

Morphologic Organization of Pulmonary Granulomas in Mice Infected with Paracoccidioides brasiliensis

A morphologic study of the lungs was carried out in Swiss mice infected with yeast isolated from Paracoccidioides brasiliensis (Pb18). The lung was processed 1, 2, 4, and 8 weeks after inoculation for histologic staining with hematoxylin and eosin (H&E), methenamine silver nitrate (Gomori-Grocott), and picrosirius to qualitative and quantitative analyses of the granulomas and the presence of fungal lesions. The numbers of CFUs/g counted in the lungs were 189.8 +/- 20.64, 353.6 +/- 46.21, 547.2 +/- 108.1, and 295.2 +/- 89.17 in the first, second, fourth, and eighth weeks, respectively. One week after infection, inflammatory cells and reticular and collagens fibers, the latest typical of fibrosis, were detected. After 2 and 4 weeks, a progressive intensification of the infection and fibrosis was observed, but in week 8 a more organized granuloma was evident, with macrophages, epithelioid cells, and yeasts in the central portion, and intense peripheral basophilia. Pycnotic structures typical of apoptotic bodies were observed in weeks 1 and 8. The different histologic staining used acted as a fundamental tool for the study of the morphologic organization of granuloma formation.

Pneumocystis jiroveci Pneumonia Mimicking Miliary Tuberculosis in a Kidney Transplanted Patient

Bilateral interstitial infiltration in chest radiography, which may be fine granular, reticular or of ground glass opacity, is the typical radiographic findings of Pneumocystis jiroveci pneumonia. Recently, atypical radiographic features, including cystic lung disease, spontaneous pneumothorax or nodu lar opacity, have been reported intermittently in patients with P. jiroveci pneumonia. We report the case of a 29‐year‐old woman with a transplanted kidney whose simple chest radiography and HRCT scan showed numerous mi liary nodules in both lungs, mimicking miliary tuberculosis (TB). Under the presumptive diagnosis of m iliary TB, empirical anti‐TB medication was started. However, Grocott methenamine silver nitrate staining of a transbronchial lung biopsy tissue revealed P. jiroveci infection without evidence of TB. These findings suggest that e ven in TB‐endemic area other etiology such as P. jiroveci as well as M. tuberculosis should be considered as an etiology of miliary lung nodules in immuno-compromised patients.Key Words: Pneumocystis carinii (Pneumocystis jiroveci), Kidney transplantation, Bronchoscopy, Tuberculosis, Miliary

Outbreak of severe disseminated aspergillosis in a flock of ostrich (Struthio camelus)

This study was undertaken to describe clinical, mycological and histopathological findings in black neck ostriches affected with severe aspergillosis in a flock including 80 birds, near Tehran, Iran. The signs included anorexia, depression, notable weight loss, diarrhoea, severe respiratory distress and death. Grossly, the lungs showed numerous white to yellow caseous nodules and the walls of the thoracic and abdominal air sacs were thickened with inflammatory exudates containing cellular debris, necrotic masses and green mold colonies. Multiple nodules were observed in the liver, spleen and gastrointestinal tract as well. Histopathologically, there were conidial heads and fungal hyphae in the air sacs and multifocal necrotic and granulomatous lesions with septated and dichotomously branched hyphae in various tissues, which were stained with haematoxylin and eosin and Grocott's methenamine silver nitrate. Aspergillus fumigatus was isolated in various tissues taken from affected ostriches.

Paecilomyces lilacinus Scleritis With Secondary Keratitis

To report a case of Paecilomyces lilacinus scleritis with secondary keratitis after initially diagnosed immune-based scleritis. An 82-year-old man was diagnosed with immune-based scleritis and treated with oral cyclophosphamide 50 mg twice daily. However, multiple scleral abscesses and a fibrinoid aqueous reaction developed 3 months later. Infectious scleritis was suspected. The culture from the necrotic sclera grew P. lilacinus. Despite treatment with antimicrobials, the infection progressed to the cornea. The medication included topical natamycin suspension 5% and fluconazole 2 mg/mL hourly, as well as oral itraconazole 100 mg daily. Debridement of the necrotic tissue and intracameral injection with amphotericin-B were performed. The infection resolved 4 months later, although its resolution was accompanied by development of phthisis bulbi, and the visual acuity remained light perception. Infection by P. lilacinus can occur in a patient with scleritis. Infectious scleritis is a particular risk in cases involving diabetes mellitus, prior scleritis, previous surgery, and immunosuppression. Early detection of the microorganism and aggressive treatment are necessary to eradicate such an infection; however, the prognosis for vision recovery can remain poor despite treatment.