Accompanying a semi-nested PCR assay to support histopathology findings of fungal keratitis in formalin-fixed paraffin-embedded corneal samples.

BACKGROUND: Fungal species are responsible for 40%-50% of all microbial keratitis cases. Due to the low amount of extracted DNA in ocular Formalin-fixed Paraffin-embedded (FFPE) samples, selecting a reliable molecular method is a substantial issue in this field. METHODS: Sixty-six samples were collected via the penetrating keratoplasty (PK) technique. Histopathology assays were performed using hematoxylin-eosin (H&E) and periodic acid Schiff (PAS) staining methods. The ITS1/ITS4 and ITS1/ITS2 primer pairs were used in a semi-nested polymerase chain reaction (PCR) to target the universal internal transcribed spacer (ITS) region. Some PCR results were validated through sequencing. RESULTS: Fungal DNA was detected in 44 of 66 samples (66.7%), and histopathology was positive for 41 of 66 samples (62.1%). Of 41 histopathologically proven fungal-positive cases, 39 were PCR-positive (95%). Moreover, of 44 PCR-positive samples, 39 (88.6%) were histopathology-positive, and 5 (11.3%) were histopathology-negative. Totally in 39 cases (59%), both histopathology and PCR yielded positive results. The Kappa agreement rate between the two diagnostic methods, including histopathology and PCR, was 0.77. Sensitivity, specificity, positive predictive value, and false predictive value were reported as 88.64%, 90.9%, 95.12%, and 80%, respectively. CONCLUSION: As we reached the acceptable Kappa agreement rate, we concluded that applying the semi-nested PCR assay is a promising method for supporting the evidence by histopathology. Finally, we suggest targeting more specific gene regions using primer pairs that amplify smaller amplicon sizes and surveying novel molecular methods such as NGS to achieve higher sensitivity and Kappa agreement rates.

Investigating the Effect of Hydroalcoholic Extract of Licorice Root to Prevent Ovariectomy-Mediated Complications.

Introduction: The importance of women's health and the quality of life after menopause is a critical issue. To prevent disability and menopause complications as well as avoid the side effects of hormone replacement therapy (HRT), in this study, licorice hydroalcoholic extract (Glycyrrhiza uralensis roots) was evaluated as a natural remedy. Methods: Seventy-two female Sprague-Dawley rats were divided into six groups: control group, Sham-operated group, Glycyrrhiza (Gly) 30% group, and ovariectomized group as well as two ovariectomized groups treated with Gly 10% and Gly 30%. Normal saline and different treatments were administered orally for 8 weeks. At the end of the study, calcium, alkaline phosphatase, estrogen, and progesterone levels in the ovariectomized rats were determined. Moreover, the stereological and histopathological changes in uterine tissue in all groups were determined. Phytochemical analyses were also performed to determine the total phenolic content and antioxidant potential of the extract. Result: The hydroalcoholic extract of licorice root exhibited considerable effect to improve calcium, estrogen, and progesterone levels in the ovariectomized rats. Also, hydroalcoholic extract of licorice root successfully decreases the amount of alkaline phosphatase (ALP) level. The stereological and histopathological findings confirmed the therapeutic potential of this extract. The considerable effects of hydroalcoholic extract of licorice root could be due to high amounts of phytoestrogens with similar estrogen-like structures. Considerable total phenolic content and antioxidant activity were also seen in licorice root extract. Conclusion: Hydroalcoholic extract of licorice root due to containing high amounts of phytoestrogens with similar chemical structures to estradiol notably improves biochemical parameters as well as stereological and histopathological markers of uterine tissues in ovariectomy rats, so it could be a potential agent for prevention and/or treatment as hormone replacement therapy in healthy middle-aged and/or older women.

Prognostic significance of immunoscore related markers in bladder cancer.

BACKGROUND: The significance of total and specific subpopulations of tumor-infiltrating lymphocytes (TILs) in cancer is now well-documented. In the present study, we investigated the relevance of CD3+, CD8 +, CD45RO +, and FOXP3 + TILs to the prognosis and survival of patients with bladder cancer and the disease's clinical-pathological parameters. METHODS: Infiltration of each subset was immunohistochemically evaluated in both stromal and intratumoral regions of tumor tissues from 85 patients with urothelial cell carcinoma of the bladder, with known survival. RESULTS: Our results indicated that intratumoral CD45RO+ lymphocytes were significantly higher in high-grade tumors than in low-grade ones (P = 0.028). The frequencies of intratumoral CD3+ (P = 0.002), CD8 + (P = 0.008), intratumoral (P = 0.002), and stromal (P = 0.017) CD45RO+ lymphocytes were also higher in patients with muscular invasion than those without invasion. The frequencies of intratumoral CD3+ (P = 0.043), CD8+ (P = 0.003), CD45RO+ (P = 0.023), and total CD45RO+ (P = 0.015), showed variation in patients with different T-stage, as well; mostly increased in T2 versus Ta and T1. Comparing patients in different stages revealed an increase in the frequencies of total CD3+ (P = 0.011), intratumoral CD3+ (P = 0.006), total CD8+ (P = 0.012), intratumoral CD8+ (P = 0.009) and stromal CD8+ (P = 0.034), as well as total and stromal CD45RO+ lymphocytes (P = 0.01 and P = 0.034, respectively) in stage II comparing to stage I, while the frequencies of stromal CD3+ (P = 0.077) and CD8+ (P = 0.053) cells tended to be decreased in stage III compared to stage II. CONCLUSIONS: We collectively observed that the frequency of immune cells, especially CD45RO+, CD3+, and CD8+ lymphocytes, were significantly higher in early-progressed tumors. This observation could be explained by continuous and prolonged stimulation of immune cells with tumor antigens during tumor progression or an increase in the recruiting factors, especially in the early stages, to eliminate tumor cells. However, with tumor progression to the late stages, the inhibitory microenvironment provided by tumor cells suppresses or changes the functionality of the effector and memory immune cells to help tumor growth. However, more functional studies with larger sample sizes are needed to reveal the real status of the immune system in patients with bladder cancer.

COVID-19-associated facial cutaneous mucormycosis superinfection: A potentially life-threatening disease.

A 49-year-old male was involved in an accident and an abdominal computer tomographic examination revealed papillary renal cell carcinoma of the right kidney. During hospitalization, the patient was infected with COVID-19. In the following COVID-19 treatment, a black dot developed on the right side of the head and face. Antifungal therapy and surgical debridement were initiated and gradual improvement was observed.

And the Oscar Goes to Peripheral Blood Film for the Detection of Lead Poisoning in a Complicated Toxic Patient: A Case Report with a Review of Laboratory Clues.

BACKGROUND: Peripheral blood smear examination is an invaluable laboratory test, which provides the complete hematologic and/or nonhematologic picture of a case. In addition to verifying the results of automated cell counters, it has the potential to identify some pathologic and morphologic changes that remain hidden using the cell counters alone. Case Presentation. A 40-year-old man with a three-year history of alcohol intake and marijuana abuse presented with severe lower extremities of the bone and abdominal pain. Physical examination showed high blood pressure, high pulse rate, and abdominal tenderness. He underwent extensive laboratory and imaging tests, and cholecystectomy and bone marrow studies were associated with no definite diagnosis. Right after all these invasive, expensive, and time-consuming investigations during a month, finding coarse basophilic stippling in the red blood cells in the peripheral blood smear by an expert led to the final diagnosis. Elevated blood lead level and the presence of ring sideroblasts in the bone marrow study confirmed the diagnosis of lead poisoning, and the patient responded well to chelator therapy in a short period. CONCLUSION: This case clearly showed the value of peripheral blood smear review and its impact on patient care. In order not to lose the cases, laboratories are recommended to design their own policy for peripheral blood smear review. The peripheral blood smear is the fastest, simplest, and most available screening test, which can prevent many misdiagnoses and malpractices. It provides rich morphological information, among which basophilic stippling is highly suggestive of lead poisoning.

Evaluating a semi-nested PCR to support histopathology reports of fungal rhinosinusitis in formalin-fixed paraffin-embedded tissue samples.

BACKGROUND: Fungal rhinosinusitis (FRS) encompasses a various spectrum of diseases. Histopathology is the "reference method" for diagnosing FRS, but it cannot determine the genus and species. Moreover, in more than 50% of the histopathologically proven cases, the culture elicited no reliable results. This study was an attempt to evaluate the diagnostic efficiency of semi-nested polymerase chain reaction (PCR) from formalin-fixed paraffin-embedded (FFPE) functional endoscopic sinus surgery (FESS) in FRS patients. METHODS: One hundred ten specimens were subjected to DNA extraction and histopathology examination. The amplification of the ß-globin gene by conventional PCR was used to confirm the quality of extracted DNA. The semi-nested PCR was performed using ITS1, ITS2, and ITS4 primers during two steps. Sequencing the internal transcribed spacer region (ITS1-5.8S-ITS2) to identify causative agents was performed on PCR products. RESULTS: Sixty-four out of 110 samples were positive by histopathology evidence, of which 56 samples (87.5%) were positive by PCR. Out of 46 negative samples by histopathological methods, five samples (10.9%) yielded positive results by PCR. Sensitivity, specificity, positive predictive value, and negative predictive value of the semi-nested PCR method were reported 87.5%, 89.2%, 92.7%, and 85.2%, respectively. The kappa factor between PCR and histopathological methods was 0.76, indicating substantial agreements between these two tests. CONCLUSION: Due to the acceptable sensitivity and specificity of the present method, it might be used to diagnose fungal sinusitis infections along with microscopic techniques. This method is recommended to confirm the diagnose of suspected fungal sinusitis with negative histopathology results.

Myofibroblastoma of the Breast: A Morphologic and Immunohistochemical Study of Three Cases.

Myofibroblastoma (MFB) of the breast is an uncommon entity of benign spindle neoplasms of the breast. This tumour possesses a broad spectrum of histomorphological patterns. Distinguishing of myofibroblastoma variants from malignant mimics of this benign neoplasm is essential for pathologists to avoid further invasive surgical procedures. In this article, we report the clinical, morphological, and immunohistochemical features of three cases, including two females and one male patient with mammary myofibroblastoma with emphasis on the histomorphological findings. As there is not yet enough information about MFB, more reports of MFB are still required to more clarify the pathogenesis and potential predisposing factors of this rare type of breast tumours.

Thiopurine S-methyltransferase and Pemphigus Vulgaris: A Phenotype-Genotype Study.

BACKGROUND & OBJECTIVE: Thiopurine drugs are considered as a treatment modality in various autoimmune disorders including pemphigus vulgaris (PV). These drugs are metabolized by an enzyme "Thiopurine S-methyl transferase" (TPMT). Various variants of this enzyme may have decreased activity leading to serious drug side effects. To investigate the phenotype and genotype of TPMT in PV patients receiving thiopurine drugs. METHODS: A total of 50 patients (29 women and 21 men) with pemphigus vulgaris treating with standard dose of Thiopurine drugs were selected. Sex, age, result of liver function test and complete blood count were recorded. Genotyping of two common non-functional allele (TPMT*2 and TPMT*3C) by Allele-specific and RFLP-PCR was performed. TPMT enzymatic level was determined by an ELISA based method. RESULTS: Of patients, 36 (72%) were found to have normal TPMT level; and 12, (24%) had higher level of enzyme and 2, 4% had low TPMT enzyme, but none of the patients showed mutant TPMT*2 and TPMT*3C alleles. None of the patients showed hepatotoxicity and bone marrow suppression. CONCLUSION: The phenotypic assay based on ELISA method may have false positive and misleading results but genotyping using PCR-RFLP and allele specific PCR is accurate, simple and cost-effective and can be used in patients decided to undergo thiopurine treatment.

A novel frame-shift deletion in FANCF gene causing autosomal recessive Fanconi anemia: a case report.

BACKGROUND: Fanconi anemia (FA) is a heterogeneous genetic disorder characterized by congenital anomalies, early-onset bone marrow failure, and a high predisposition to cancers. Up to know, different genes involved in the DNA repair pathway, mainly FANCA genes, have been identified to be affected in patients with FA. CASE PRESENTATION: Here, we report clinical, laboratory and genetic findings in a 3.5-year-old Iranian female patient, a product of a consanguineous marriage, who was suspicious of FA, observed with short stature, microcephaly, skin hyperpigmentation, anemia, thrombocytopenia and hypo cellular bone marrow. Therefore, Next Generation Sequencing was performed to identify the genetic cause of the disease in this patient. Results revealed a novel, private, homozygous frameshift mutation in the FANCF gene (NM_022725: c. 534delG, p. G178 fs) which was confirmed by Sanger sequencing in the proband. CONCLUSION: Such studies may help uncover the exact pathomechanisms of this disorder and establish the genotype-phenotype correlations by identification of more mutations in this gene. It is the first report of a mutation in the FANCF gene in Iranian patients with Fanconi anemia. This new mutation correlates with a hematological problem (pancytopenia), short stature, and microcephaly and skin hyperpigmentation. Until now, no evidence of malignancy was detected.

Sebaceous Lymphadenoma of Parotid: Imaging, Cytological, and Histological Findings in Detail.

Sebaceous lymphadenomas are rare and account for less than 1% of primary salivary gland tumors. These rare tumors are mostly found in men older than 50 years. The clinicopathological features of these tumors are poorly understood and no definite causative factor has been reported for them till now. They are not often diagnosed prior to surgery, which could be due to their rarity and lack of enough preop radiological and cytological findings. Few case reports have been published in literature about their pathogenesis and accompanying malignant lesions. The cytological and imaging findings have been dealt with in some articles. Here we describe the histologic, cytologic, and radiologic findings of sebaceous lymphadenomas of parotid gland, all together, and discuss their differential diagnoses in various diagnostic methods.

Effect of Methanolic Extract of Corn Silk on Cisplatin-Induced Nephrotoxicity in Rats.

BACKGROUND: Cisplatin is a cytotoxic agent in cancer therapy. Nephrotoxicity is considered as a side effect of cisplatin usage. Using rate models, we studied the possible protective impact of corn-silk (CS) extract against cisplatin-induced nephrotoxicity. MATERIALS AND METHODS: Thirty-five experimental rats were divided into five groups (n=7 per each group) as follow: C1: Control received distilled water only; C2: received one dose of cisplatin, and CS: received 300 mg/kg/day of CS. Both CS1 and CS2 received 200 and 300 mg/kg/day of the CS extract orally, individually, for eight consecutive days. CS1 and CS2 received a single dose of cisplatin on the first day only. The specific biochemical markers and histopathological alterations were evaluated. RESULT: According to our results, cisplatin administration could have induced severe degeneration in all parts of the nephron tubules and liver. Pre-treatment with CS exhibited a significant decrease in the malondialdehyde (MDA) levels as compared to the values obtained after treatment with cisplatin alone (P<0.01). Moreover, the CS extract with 200 mg dose showed significant (P<0.01) protection against the cisplatin-induced elevation of blood urea nitrogen. Further, the serum levels of alanine transaminase and aspartate transaminase were higher in the cisplatin-treated groups, when compared to the control group (P<0.05). Furthermore, the hepatic function was also improved in cisplatin-treated animals, which were pre-treated with CS. CONCLUSION: CS has the potential to attenuate nephrotoxicity and lipid peroxidation induced by cisplatin in rats.

Fine-Needle Aspiration Biopsies of Ovarian Masses: A Reliable Technique.

OBJECTIVE: In gynecology, fine-needle aspiration (FNA) has an overall accuracy of 94.5% in differentiation between benign and malignant tumors. The purpose of this study was to determine reliable cytological criteria for categorizing ovarian masses into benign and malignant categories, their subtypes, and also to evaluate FNA accuracy in the diagnosis of ovarian tumors in relation to histopathological findings. STUDY DESIGN: A prospective study was performed on all patients with a preoperative diagnosis of ovarian tumor who were referred to our hospital between August 2013 and August 2015. During surgery, FNA was performed using an 18-gauge needle by a pathologist. Aspirated material was spread on clean glass slides and stained with Papanicolaou and Wright-Giemsa stains. The cytological findings and results were compared with the histological diagnosis. RESULTS: Of the 81 cases in this study, there was a discrepancy between the cytological and histological diagnosis in 9 cases. The overall cytological diagnostic accuracy in our study was 88.9% with a sensitivity and specificity of 78.1 and 95.5%, respectively. CONCLUSION: FNA of an ovarian mass is a minimally invasive procedure with acceptable diagnostic accuracy, especially when differentiating between benign and malignant lesions, and can be considered as a useful diagnostic modality for choosing an appropriate management course.

Prevalence of Endometriosis in Malignant Epithelial Ovarian Tumor.

BACKGROUND: The present study aims to assess the prevalence and histological characteristics of endometriosis in different types of ovarian surface epithelial tumors. METHODS: Microscopic slides of 110 ovarian tumors (89 malignant and 21 borderlines) were reviewed from 2008 to 2013 in two major gynecological centers affiliated with the Shiraz University of Medical Sciences, Shiraz, Iran. The presence or absence of endometriosis and transitions from atypical endometriosis to carcinoma were also histologically evaluated. Chi-square and t-test were used to compare the study groups. RESULTS: The mean age of the patients was 49.93 ± 9.36 years in the Endometriosis-Associated Ovarian Carcinomas (EAOC) group and 50.18 ± 12.8 years in the non-EAOC group. Among the 110 patients, 28 (25.4%) had endometriosis. According to ovarian cancer subtype 67% (4/6) of clear cell adenocarcinoma, 65% (11/17) of endometrioid adenocarcinoma, 28% (7/25) of low grade serous adenocarcinoma, 4% (1/25) of high grade serous adenocarcinoma, 30% (4/13) of borderline serous tumor, and 25% (1/4) of mixed carcinoma had endometriosis. None of the mucinous borderline tumors and mucinous adenocarcinoma cases had endometriosis. Moreover, 23 cases had typical endometriosis, while 14 had atypical endometriosis. On the other hand, 19 cases had both typical and atypical endometriosis. Furthermore, transition from atypical endometriosis to carcinoma was seen in 11 cases. CONCLUSION: Clear cell and endometrioid carcinoma are the most common types of EACO. Atypical endometriosis was more commonly seen in endometrioid and clear cell carcinomas which are included in type I ovarian cancer. Thus, it can be concluded that atypical endometriosis is a precursor for type I ovarian cancer.

Infantile fibrosarcoma of ethmoid sinus, misdiagnosed as an adenoid in a 5-year-old child.

Infantile fibrosarcoma of head and neck is rare and the presence of this tumor in ethmoid sinus is even more uncommon. To the best of our knowledge, <5 cases have been reported in the last 20 years in the English literature, so far, only one of which has been infantile type in a 15 months old girl. In this case report, we will explain our experience with a rare case of infantile fibrosarcoma originating from ethmoid sinus in a 5-year-old boy who presented with dyspnea and epistaxis. After biopsy, it was diagnosed as fibrosarcoma of sinus origin.

Gastrointestinal Basidiobolomycosis, a Rare and Under-diagnosed Fungal Infection in Immunocompetent Hosts: A Review Article.

Gastrointestinal Basidiobolomycosis (GIB) is an unusual, rare, but emerging fungal infection in the stomach, small intestine, colon, and liver. It has been rarely reported in the English literature and most of the reported cases have been from US, Saudi Arabia, Kuwait, and Iran. In the last five years, 17 cases have been reported from one or two provinces in Iran, and it seems that it has been undiagnosed or probably unnoticed in other parts of the country. In this review, we explored the English literature from 1964 through 2013 via PubMed, Google, and Google scholar using the following search keywords: BasidiobolomycosisBasidiobolus ranarumGastrointestinal Basidiobolomycosis In this review, we attempted to collect all clinical, pathological, and radiological findings of the presenting patients; complemented with previous experiences regarding the treatment and prognosis of the GIB. Since 1964, only 71 cases have been reported, which will be fully described in terms of clinical presentations, methods of diagnosis and treatment as well as prognosis and follow up.

Cardiac capillary hemangioma in a patient with atypical chest pain and normal ECG.

Cardiac hemangiomas account for less than 2% of primary cardiac tumors. They can present with palpitation, arrhythmia, right ventricular outflow tract obstruction, pseudoangina, and in more extreme cases sudden death. Our case report presents a 41-year-old man with atypical chest pain, who had a normal heart examination, electrocardiogram, and an unremarkable family and medical history. His echocardiography demonstrated a round semimobile mass in the right ventricle. The diagnosis was confirmed by both transesophageal echocardiography and cardiac magnetic resonance imaging. The tumor was surgically excised and sent for pathology, which was reported as capillary hemangioma. .