Covid-19-Associated Mucormycosis: Histopathology of the Deadly Fungal Infection

Abstract Introduction Many patients suffered from rhino-orbital-cerebral mucormycosis during the coronavirus disease 2019 (COVID-19) pandemic in India. Diabetes is a known risk factor of COVID-19 infection and mucormycosis. Objective The present study was done to describe the clinical spectrum and histopathological findings of mucormycosis in COVID-19 patients and their outcomes. Methods A cross-sectional study was done over a period of two and half months. The biopsy samples or scrapings from sinonasal or periorbital tissue of 38 patients were analyzed. Hematoxylin & Eosin (H&E stain) slides were evaluated along with Grocott-Gomori methenamine-silver and Periodic acid-Schiff stains to highlight the fungal elements. Results The male to female ratio was 2.5:1, and the mean age of the subjects was 53 years old. A total of 68.4% (n = 26/38) of the patients had diabetes as a comorbidity, 84.2% (n = 32/38) had a history of steroid intake, and 55.3% (n = 21/38) were given supplemental oxygen during their treatment. The common presentations were nasal blockage, discharge, eye pain, headache, and altered mentation. The sites of biopsy were: nasal cavity 76.3% (n = 29/38), periorbital fat/orbit 21.1% (n = 8/38), maxillary sinus 15.8% (n = 6/38) and ethmoid sinus 13.2% (n = 5/38). In 76.3% (n = 29/38) cases, broad, irregular, nonseptate, and right-angle branching hyphae were seen on H&E-stained tissue sections. Conclusion COVID-19 led to various complications in individuals affected by it. Mucormycosis was one such lethal complication. An early diagnosis and prompt treatment is crucial to control the progression of the disease and improve outcomes.

Ovarian hydatid cyst: an uncommon site of presentation

ABSTRACT Hydatid cyst is a parasitic infestation caused by Echinococcus larvae. Hydatid cyst of the ovary is a highly unusual presentation. Herein, we present a case of a young woman who complained of episodic lower abdominal pain. Ultrasound of the abdomen revealed a multi-cystic left adnexal mass measuring 86 mm x 67 mm. A possibility of ovarian cystic neoplasm was suggested. Unilateral salpingo-oophorectomy was performed. On histopathological examination, a cyst measuring 8.0 x 5.5 x 4.5 cm was found, replacing the entire ovary. The cyst cavity was filled with serous fluid and multiple pearly white membranous structures, giving a multiloculated appearance. Microscopic examination showed a cyst lined by a lamellar membrane containing protoscolices and hooklets. Hydatid disease is a zoonotic ailment caused by tapeworms (Echinococcus granulosus or, less commonly, Echinococcus multilocularis). The definitive hosts are carnivores. Humans are the accidental intermediate hosts. The hydatid cyst commonly affects the liver and the lungs. The primary hydatid cyst of the ovary is quite rare, with few case reports in the literature. In most cases, symptoms are vague, and the lesion is misdiagnosed as benign or malignant ovarian cystic neoplasm on clinical and radiological examination. Ovarian hydatid cyst is treated by surgery with ovarian cystectomy as the gold standard. The possibility of a hydatid cyst should be kept under differential diagnoses while evaluating the cystic diseases of the ovary.

Anorectal balloon cell melanoma: a rare variant

ABSTRACT Balloon cell melanoma is a rare presentation of malignant melanoma, usually on the skin, with less than 100 cases reported. Mucosal BCM is even rarer, with only one case of anorectal BCM reported in English literature. The diagnosis is based on the histopathologic findings of a tumor composed of large, foamy melanocytes, with or without pigmentation, and confirmed by immunohistochemical studies showing expression for melanocytic markers. The foam cell appearance of the tumor cells and the lack of melanin pigment lead to a diagnostic dilemma, mostly when presented at an unusual location. Herein, we report a case of balloon cell melanoma at the anorectal junction in a 73-year-old male patient complaining of constipation and bleeding per rectum. Surgical resection was performed with no evidence of recurrence after three years of close follow-up. We believe this case will raise awareness among the medical community to consider this tumor a differential diagnosis in rectal masses.

A Study of the Clinical, Pathological and Radiological Profile of Lung Cancer in Non-Smokers

Aim and Objective: To study the clinical, pathological andradiological profile of lung cancer in non-smokers in a tertiarycare center.Methods: 53 non-smokers diagnosed with lung cancerattending the Department of Respiratory Medicine, KingGeorge’s Medical College from September 2015 to August2017 were enrolled. Record of all diagnostic investigations andprocedures performed namely transthoracic fine needleaspiration cytology (FNAC) and biopsy, bronchoscopy,thoracoscopy, closed pleural biopsy, lymph-node FNAC andbiopsy, routine blood and sputum examinations and a detailedhistory were obtained. Data was analysed retrospectively.Results: The mean age of presentation was 53.8±11.6 years.Majority were females (60.4%). Most common presentingsymptom was cough (84.9%). Mean duration of symptoms was6.9 months. Pallor was the most common clinical examinationfinding (41.5%). Mass with effusion was the most commonradiological lesion (45.3%). 22.6% masses were centrallylocated. Transthoracic biopsy could diagnose 32 (60.4%)cases. Adenocarcinoma was the most common type in bothmales (76.2%) and females (78.1%). Epidermal growth factorreceptor (EGFR) mutation was positive in 46.3% ofadenocarcinoma. Exon 19 deletion was the more commonmutation.Conclusion: Lung cancer among never smokers is a distinctclass with risk factors and genetic features discrete from thoseassociated with tobacco smoke. Indoor air pollutants as well asETS are definitely implicated risk factors. Targetable mutationsare commoner in non-smokers and hence mutation testingshould always be done in such patients. It is important toconduct studies about the diverse characteristics of this entityto consolidate our knowledge of this growing group of cancer.

Prognostic Value of CA19-9 and CA242 in Gallbladder Cancer - An Exploratory Study

Introduction: There are, at present, no biomarkers topredict to prognosis of Gallbladder cancer. We conducted aprospective exploratory study to evaluate the role to serum CA19-9 and CA 242 as prognostic markers.Material and Methods: We enrolled consecutive patients forthis study and CA 19-9 and 242 were estimated from venoussamples. Association of these markers with clinical variablesand median overall survival (OS) difference between patientswho has raised versus normal levels of these markers wasdetermined.Results: Sixty-two patients were enrolled for this study.Forty-four (71%) patients had elevated CA19-9. Thirty-nine(62.9%) patients had an raised CA242 levels. CA 19-9 wasfound to be significantly associated with the presence ofjaundice (p=0.038) and advanced stage (p=0.009). MedianOS of patients who had elevated CA 19-9 was 5.73 monthscompared to 8.33 months in patients who had normal CA19-9. The difference was not statistically different (p= 0.15).Median OS for patients who had elevated CA 242 was 5.53months, which was inferior to those who had normal levels(9.1 months). This difference approached, but was notstatistically significant (p=0.055).Conclusion: This is the first study to show associationbetween CA 19-9 and stage of disease in GBC. At present,CA 19-9 and CA 242 cannot be recommended as prognosticmarkers. However, role of CA 242 needs to be examined ina larger cohort of patients of GBC to establish its usefulness.

Do phosphatase of regenerating liver‑3, matrix metalloproteinases‑2, matrix metalloproteinases‑9, and epidermal growth factor receptor‑1 predict response to therapy and survival in glioblastoma multiforme.

Context: Poor survival of the glioblastoma multiforme (GBM) has been attributed in part to the invasive nature of the lesion making complete surgical removal near impossible. Phosphatase of regenerating liver‑3 (PRL‑3), matrix metalloproteinases‑2 and ‑9 (MMP‑2 and MMP‑9), and epidermal growth factor receptor (EGFR‑1) play a role in invasive nature of tumor cells. Aims: This study was conducted to evaluate PRL‑3, MMP‑2, MMP‑9, and EGFR‑1 (markers) expression in cases to GBM and to correlate their expression with therapy response and survival. Settings and Design: GBM cases (n = 62) underwent surgery followed by radiation (n = 34) and chemoradiation (n = 28). Using WHO Response Evaluation Criteria in Solid Tumors criteria response to therapy was assessed at 3 months and cases followed up for survival. Subjects and Methods: Expression of markers was assessed by immunohistochemistry as a percentage of positive tumor cells in hot spots. Statistical Analysis Used: Kaplan–Meier, ANOVA, Chi‑square test, univariate, and multivariate Cox‑regression analysis was done. Results: Response to therapy was evident in 54.8% cases of responders with the mean survival of 494.03 ± 201.13 days and 45.2% cases of non responders (278.32 ± 121.66 days) with P = 0.001. Mean survival for the patient’s opted chemoradiation was 457.43 ± 222.48 days which was approximately 3 months greater than those who opted radiation alone (P = 0.029). We found PRL‑3 overexpression was an independent, significant, poor prognostic factor for survival by multivariate analysis (P = 0.044). Cases negative for MMP’s and EGFR showed increased survival, but the difference was insignificant. Conclusion: PRL‑3 expression appears to be related to an adverse disease outcome.

Revisiting epidermal growth factor receptor in glioblastoma multiforme: Does it play a role in response to therapy.

Background and Aim: Glioblastoma multiforme (GBM) are the most aggressive class of cancer of central nervous system with hallmark characteristics that include rampant proliferation, necrosis, and endothelial proliferation. Epidermal growth factor receptor (EGFR) has been implicated as the primary contributor to glioblastoma initiation and succession. The present study was designed to evaluate EGFR protein expression in GBM as predictor of response to therapy and survival. Materials and Methods: Epidermal growth factor receptor was assessed by immunohistochemistry as a percentage of positive tumor cells in hot spots (10 high-power fields). The study group comprised of 35 cases of GBM. All cases underwent surgical resection and subsequently underwent radiotherapy (n = 17) or radiotherapy with adjuvant temozolomide chemotherapy (n = 18). Immediate response to therapy was assessed at 3 months using World Health Organization response evaluation criteria in solid tumors criteria and cases followed up for survival. Results: Twenty-four cases (68.6%) expressed EGFR while 11/35 (31.4%) cases were negative. Response to therapy was evident in 21/35 cases (60.0%) and 14/35 were (40.0%) nonresponders. Mean EGFR protein expression in responders was 37.23 ± 33.70 and in nonresponders was 59.5 ± 39.46 (P = 0.542). The percentage of responders which were EGFR negative was 72.7% and while response in EGFR positive cases was observed in 54.2%. Mean survival in EGFR positive and negative GBM was 394.37 ± 189.11 and 420.54 ± 191.23 days, respectively. Conclusion: The EGFR negative cases appear to respond better to therapy, however, the difference is not statistically significant (P = 0.298). Further, EGFR protein expression does not play a definitive role in predicting survival. This is an original study evaluating EGFR in terms of therapeutic response.

Increased expression of ADAM33 protein in asthmatic patients as compared to non-asthmatic controls.

Background & objectives: ADAM33 is a member of a family of genes that encode membrane-anchored proteins with a disintegrin and a metalloprotease domain, primarily expressed in lung fibroblasts and bronchial smooth muscle cells. ADAM33 has been identified as a risk factor for asthma and is known as a gene associated with airway remodelling. The present study was conducted with the aims to investigate the expression of ADAM33 protein in patients of asthma and non-asthmatic controls, and to assess if the expression of ADAM33 protein relates with severity of asthma. Methods: A total of 35 subjects, including 27 patients with asthma and eight non-asthmatic controls were included using Global Initiative for Asthma guidelines 2005. Bronchial biopsy tissues were collected and paraffin sections were made to store all study samples. Immunohistochemistry was performed using standardized protocol. Results: An increase in expression of ADAM33 protein was observed in the epithelium, smooth muscle and mesenchymal cells of asthma cases when compared to controls but there was no relationship with severity of asthma. Interpretation & conclusions: A higher expression of ADAM 33 protein was seen in asthma patients compared to controls. Large prospective studies need to be done with adequate study design to confirm these preliminary finding.

Epstein-Barr virus DNA load and its association with Helicobacter pylori infection in gastroduodenal diseases

Helicobacter pylori and Epstein-Barr virus (EBV) infections are common worldwide. Although H. pylori infection is a major factor in gastroduodenal diseases, its role in association with EBV infection is unknown. Objective: To study the association of H. pylori infection and EBV DNA load in patients with gastroduodenal diseases. Methods: Biopsy samples were collected from 200 adult patients [non-ulcer dyspepsia (NUD) 100, peptic ulcer disease (PUD) 50, gastric carcinoma (GC) 50] undergoing upper gastrointestinal endoscopy. H. pylori infection was diagnosed by rapid urease test, culture, histopathology, PCR and Q-PCR. EBV DNA was detected by non-polymorphic Epstein-Barr nuclear antigen-1 (EBNA-1) gene based Q-PCR. Results: In patients with GC and PUD, EBV DNA was detected more often than NUD (GC versus NUD = 90 percent versus 37 percent, p < 0.001; PUD versus NUD = 70 percent versus 37 percent, p < 0.001). The dual prevalence of H. pylori infection and EBV DNA was significantly higher in patients with GC and PUD than in those with NUD. Median copy number of EBV DNA was considerably higher in GC and PUD than NUD (p < 0.01). The copy number of EBV DNA was significantly higher in H. pylori infected patients (p = 0.015). The number of ureA gene copies was also found to be significantly higher in PUD and NUD with presence of EBV DNA. However, in GC no significant difference was seen between EBV positive and negative status. Conclusion: There was a trend for higher EBV DNA load in H. pylori positive individuals suggesting a probable role of H. pylori in modulating the conversion of EBV to its lytic phase.

A study to evaluate asbestos fiber burden in lung and pleural malignancies.

Background: There is scarcity of data on asbestos fiber burden in lung and pleural malignancies. Aim: To evaluate asbestos fiber burden in biopsy samples of suspected lung and pleural malignancies. Study Design: This was a single-centre, observational study. Study Period: From August 2010 to July 2010. Setting: Department of Pulmonary Medicine, CSMMU, UP, Lucknow, a tertiary care hospital in India. Study Population: Suspected cases of lung and pleural malignancy. Materials and Methods: Biopsy tissues taken by computed tomography (CT)-guided biopsy, bronchoscopic biopsy, and pleural biopsy by Cope's needle were analyzed for histopathology and asbestos burden by Haq et al.'s method. Results: 20 patients were studied. Mean fiber burden was 9.25 × 10 4 fibers/g. Average burden in lung malignancies (11 patients) was 9.178 × 10 4 fibers/g and in pleural tissue (9 patients) was 9.332 × 10 4fibers/g. Among the different cell types, mean fiber burden in squamous cell carcinoma was 8.99 × 10 4 fibers/g, in adenocarcinoma was 9.71 × 10 4 fibers/g, and in small cell carcinoma was 7.54 × 10 4 fibers/g. Mean fiber burden in bronchoscopic endobronchial biopsy tissue was 10.69 × 10 4 fibers/g, while in CT-guided biopsy was 8.60× 10 4fibers/g. Conclusion: Maximum number of fibers was found in adenocarcinoma.

Proliferating cell nuclear antigen, p53 and micro vessel density: Grade II vs. Grade III astrocytoma.

Histological classification and grading are prime procedures in the management of patients with astrocytoma, providing vital data for therapeutic decision making and prognostication. However, it has limitations in assessing biological tumor behavior. This can be overcome by using newer immunohistochemical techniques. This study was carried out to compare proliferative indices using proliferating cell nuclear antigen (PCNA), extent of p53 expression and micro vessel morphometric parameters in patients with low grade and anaplastic astrocytoma. Twenty-five patients, each of grade II and grade III astrocytoma were evaluated using monoclonal antibodies to PCNA, p53 protein and factor VIII related antigen. PCNA, p53-labeling indices were calculated along with micro vessel morphometric analysis using Biovis Image plus Software. Patients with grade III astrocytoma had higher PCNA and p53 labeling indices as compared with grade II astrocytoma (29.14 plus/minus 9.87% vs. 16.84 plus/minus 6.57%, p 0.001; 18.18 plus/minus 6.14% vs. 6.14 plus/minus 7.23%, p 0.001, respectively). Micro vessel percentage area of patients with grade III astrocytoma was also (4.26 plus/minus 3.70 vs. 1.05 plus/minus 0.56, p 0.001), higher along with other micro vessel morphometric parameters. Discordance between histology and one or more IHC parameters was seen in 5/25 (20%) of patients with grade III astrocytoma and 9/25 (36%) of patients with grade II disease. PCNA and p53 labeling indices were positively correlated with Pearson's correlation, p less than 0.001 for both). Increased proliferative fraction, genetic alterations and neovascularization mark biological aggressiveness in astrocytoma. Immunohistochemical evaluation scores over meet the challenge of accurate prognostication of this potentially fatal malignancy.

Utility of cysticercus fasciolaris antigen in Dot ELISA for the diagnosis of neurocysticercosis.

Background: Clinical diagnosis of neurocysticercosis (NC) is established by CT scan and MRI. However, absolute diagnosis is not possible in a fair number of cases, and serological assays are used as adjunct. Besides, CT scan and MR imaging are resource-intensive tests and not practical for screening in endemic areas. Aim: To provide a low-cost, efficient, and reproducible assay for the detection of antibodies against cysticerci. Hence we have attempted to standardize and evaluate the diagnostic utility of the cysticercus fasciolaris antigen in a Dot ELISA assay for diagnosis of NC. Setting and Design: Tertiary hospital-based, case-control series. Materials and Methods: Confirmed cases of NC diagnosed by presence of ring lesions in CT scan or MR imaging with presence of scolex were taken as positive controls (n = 50). Negative controls (n = 50) included subjects with normal CT scan studies (n = 30) and diseased controls with ring lesions in CT scan confirmed to be neurotuberculosis (n = 20). Dot ELISA was standardized and validated with commercially available ELISA (UBI, USA) using sera from the study groups. Statistical Analysis: Chi-square test was used to compare the immunodiagnostic performance of the two tests. P value less than .05 (P < 0.05) was considered significant. Results: The Dot ELISA had a sensitivity of 88% and specificity of 74% with a positive predictive value of 77.19% and negative predictive value of 81.06%. Likelihood ratios for a positive and a negative test were 3.4 and 0.2. The sensitivity and specificity of commercial ELISA were 92% and 84% respectively. Difference between the performances of the two tests was not significant statistically. Conclusions: Dot ELISA has sensitivity and specificity comparable to ELISA for the diagnosis of NC. The test is simpler, not requiring expertise and instrumentation. Further validation of the test as a screening tool is required.