Mohammad Vasei

Diagnosis of cytomegalovirus (CMV) in biopsies relies on detecting classic viral cytopathic effects (CPE) in tissue. These effects are not always apparent, and confirmatory tests are necessary. This study aimed to compare the results of the different diagnostic tests for CMV detection in colitis, including PCR on fresh and formalin-fixed paraffin-embedded (FFPE) tissue, immunohistochemistry (IHC), histology, and plasma PCR, in association with the clinical course. In this prospective study, CMV-PCR was performed on fresh tissue (FT) and FFPE tissue, and IHC was conducted on colon biopsies from 153 children with colitis referred to Children Medical Center Hospital (Tehran, Iran) from 2015 to 2019. The results of different diagnostic methods were evaluated in association with the clinical and histopathological findings. Fifty out of 153 (32%) cases had positive FT-CMV PCR. Forty of these fifty positive samples and 21 of 103 negative ones had concomitant FFPE biopsy tissue. FFPE-PCR and IHC were positive in 17 (42.5 %) and 2 (5%) out of 40 FT-PCR-positive cases, respectively. The two IHC-positive cases had positive FFPE-PCR and high CMV-DNA plasma levels and showed histologically active colitis and CPE. Remarkably, 14 (35%) cases were identified with positive FT-PCR without any evidence of colitis in histopathology. During follow-up, FT-PCR-positive inflammatory bowel disease (IBD) cases treated with antiviral drugs showed a poorer outcome than the non-treated cases (P=0.03). A high positive rate was observed for both FT- and FFPE-CMV PCR, with a poor association with histology. IHC positivity was associated with high plasma CMV DNA levels and the presence of CPE. The efficacy of anti-CMV treatment in colitis cases should be verified through randomized controlled clinical trials. A preprint version of this article is available online at doi: 10.22541/au.168052684.43457923/v1.

Background: Gastric cancer is the fourth leading cause of cancer-related deaths in the world. The identification of gastric cancer subtypes related to recognizable microbial agents may play a pivotal role in the targeted prevention and treatment of this cancer. The current study is conducted to define the frequency of Epstein-Barr virus (EBV) infection in gastric cancers of four major provinces, with different incidence rates of gastric cancers, in Iran. Methods: Paraffin blocks of 682 cases of various types of gastric cancer from Tehran, South and North areas of Iran were collected. Twelve tissue microarray (TMA) blocks were constructed from these blocks. Localization of EBV in tumors was assessed by in situ hybridization (ISH) for EBV-encoded RNA (EBER). Chi-squared test was used to evaluate the statistical significance between EBV-associated gastric cancer (EBVaGC) and clinicopathologic tumor characteristics. Results: Fourteen out of 682 cases (2.1%) of gastric adenocarcinoma were EBER-positive. EBER was positive in 8 out of 22 (36.4%) of medullary carcinomas and 6 out of 660 (0.9%) of non-medullary type, which was a statistically significant difference (P<0.001). The EBVaGCs were more frequent in younger age (P=0.009) and also showed a trend toward the lower stage of the tumor (P=0.075). Conclusion: EBV-associated gastric adenocarcinoma has a low prevalence in Iran. This finding can be due to epidemiologic differences in risk factors and exposures, and the low number of gastric medullary carcinomas in the population. It may also be related to gastric tumor heterogeneity not detected with the TMA technique.

Introduction: The discovery of gene editing techniques has opened a new era within the field of biology and enabled scientists to manipulate nucleic acid molecules. CRISPR-Cas9 genome engineering has revolutionized this achievement by successful targeting the DNA molecule and editing its sequence. Since genomic changes are the basis of the birth and growth of many tumors, CRISPR-Cas9 method has been successfully applied to identify and manipulate the genes which are involved in initiating and driving some neoplastic processes. Methods: By review of the existing literature on application of CRISPR-Cas9 in cancer, different databases, such as PubMed and Google Scholar, we started data collection for "CRISPR-Cas9", "Genome Editing", "Cancer", "Solid tumors", "Hematologic malignancy" "Immunotherapy", "Diagnosis", "Drug resistance" phrases. Clinicaltrials.gov, a resource that provides access to information on clinical trials, was also searched in this review. Results: We have defined the basics of this technology and then mentioned some clinical and preclinical studies using this technology in the treatment of a variety of solid tumors as well as hematologic neoplasms. Finally, we described the progress made by this technology in boosting immune-mediated cell therapy in oncology, such as CAR-T cells, CAR-NK cells, and CAR-M cells. Conclusion: CRISPR-Cas9 system revolutionized the therapeutic strategies in some solid malignant tumors and leukemia through targeting the key genes involved in the pathogenesis of these cancers.

Background: Gastric cancer is one of the most common cancers worldwide. Human bocavirus (HBoV), a recently isolated virus, has been investigated for its role in many respiratory and enteric diseases. Few studies have reported its presence in solid tumors, such as lung and colon cancers. The aim of this study was to detect the presence of the HBoV1 genome in gastric adenocarcinoma, which has not yet been evaluated. Methods: Formalin-fixed paraffin-embedded (FFPE) blocks of 189 gastric tumors and 50 blocks of non-tumor gastric tissue products from elective weight reduction operations were collected. DNA extraction and real-time polymerase chain reaction (PCR) were performed for HBoV1 detection. DNA sequencing was performed using ABI Genetic Analyzer series 3500. Results: The mean age of the patients was 60±13.33 years. Tumors were more common in males than females (2.5/1). HBoV1 PCR was positive in 34 (18%) cases of GC and 10 (20%) cases of chronic gastritis (P>0.05). There was no association between age, sex, stage, and histologic subtype of the tumor and HBoV1 positivity (P>0.05) in tumor samples. The rate of intestinal metaplasia and presence of lymphoid stroma were also not more frequent in HBoV1-positive tumors (P>0.05). Conclusion: The HBoV1 can be detected in a relatively high proportion of Iranian patients with gastric cancer (18%) with no predilection for specific subtypes and no association with the degree of lymphocytic infiltration. HBoV1 can also be observed in approximately 20% of chronic gastritis cases. Further comprehensive studies are needed to elucidate the role of HBoV1 in gastric cancer development.

For diagnosis of CMV infection in colon biopsies, immunohistochemistry (IHC) and polymerase chain reaction (PCR) can be helpful besides pathologic examination. We performed CMV IHC and PCR on formalin–fixed paraffin-embedded (FFPE) and fresh tissue (FT) in clinically suspicious children for colitis and compared the results with the patients’ clinical course. In a prospective cohort study, among 153 candidate children for the colonoscopy due to colitis, 50 showed positive CMV PCR on FT. PCR on FFPE tissue was positive in 17/40 (42.5 %) cases, of which 2/40 (5%) had positive IHC. Low titer plasma CMV DNA level was seen in 7/17 FFPE-PCR positive cases. Two patients had a high plasma CMV DNA level, who were the two IHC-positive cases. Fecal calprotectin level showed an association with disease histologic activity, but not with CMV positivity on either of the methods of detection. IHC positivity was more likely to be associated with a high plasma CMV DNA level. There was a high rate of false positivity by CMV PCR test on fresh tissue that was neither associated with histopathology nor with the CMV DNA plasma level. Antiviral treatment was correlated with a poorer outcome in patients with inflammatory bowel disease (P=0.035). Conclusion: The PCR result of FT is correlated neither to histology nor to the serum CMV-DNA level. High fecal calprotectin level is associated with active lesions in histopathology. Large randomized clinical trials are recommended to assess the efficacy of antiviral treatment in acute colitis cases that have a high copy number of CMV in the blood or positive results on IHC.

Background: Triple-negative breast cancer (TNBC) accounts for 15% of all breast cancer (BC) cases and is a severe type of BC. Since medicinal herbs containing biocompatible substances that are accepted by patient more than chemical therapeutics, they can be considered a safe option for treating BC. Objective: This study evaluated the effect of Sambucus Ebulus (S. ebulus) extract on a model of TNBC. Methods: S. ebulus extract was prepared using petroleum ether, ethyl acetate, and methanol. The petroleum ether extract was fractionated and analyzed using vacuum liquid chromatography and GC-MS, respectively. MDAMB- 231 and MCF-10A were used as TNBC and normal breast cells, respectively. Flowcytometry and MTT assays were performed to evaluate cell cycle, apoptosis, and viability of the cells. Gene expression analysis was performed using RT-qPCR. Nude mouse allograft tumor models were used, and pathological sections were evaluated. Results: The findings indicated that S. ebulus extract remarkably decreased cell proliferation and viability. The extract had no toxicity to the normal breast cells but efficiently killed the cancer cells. Cell cycle- and apoptosisrelated gene expression showed that fraction 4 of S. ebulus extract significantly increased the expression of Bax, Bak, P53, and c-MYC. Conclusion: This study showed satisfactory results of the effect of S. ebulus extract on clearing BC cells both in vitro and in vivo. Thus, S. ebulus extract may be a safe herbal compound for eliminating BC cells without toxicity to host cells.

Moeinadin Safavi, M.D. and Mohammad Vasei, M.D.. Ann Lab Med 2021;41:240-2. https://doi.org/10.3343/alm.2021.41.2.240

Background: Acute respiratory distress syndrome (ARDS) is a fatal complication of coronavirus disease 2019 (COVID-19). There are a few reports of allogeneic human mesenchymal stem cells (MSCs) as a potential treatment for ARDS. In this phase 1 clinical trial, we present the safety, feasibility, and tolerability of the multiple infusions of high dose MSCs, which originated from the placenta and umbilical cord, in critically ill COVID-19-induced ARDS patients.Methods: A total of 11 patients diagnosed with COVID-19-induced ARDS who were admitted to the intensive care units (ICUs) of two hospitals enrolled in this study. The patients were critically ill with severe hypoxemia and required mechanical ventilation. The patients received three intravenous infusions (200 × 10⁶cells) every other day for a total of 600 × 10⁶human umbilical cord MSCs (UC-MSCs; 6 cases) or placental MSCs (PL-MSCs; 5 cases).Findings: There were eight men and three women who were 42 to 66 years of age. Of these, six (55%) patients had comorbidities of diabetes, hypertension, chronic lymphocytic leukemia (CLL), and cardiomyopathy (CMP). There were no serious adverse events reported 24–48 h after the cell infusions. We observed reduced dyspnea and increased SpO2 within 48–96 h after the first infusion in seven patients. Of these seven patients, five were discharged from the ICU within 2–7 days (average: 4 days), one patient who had signs of acute renal and hepatic failure was discharged from the ICU on day 18, and the last patient suddenly developed cardiac arrest on day 7 of the cell infusion. Significant reductions in serum levels of tumor necrosis factor-alpha (TNF-α;P < 0.01), IL-8 (P < 0.05), and C-reactive protein (CRP) (P < 0.01) were seen in all six survivors. IL-6 levels decreased in five (P = 0.06) patients and interferon gamma (IFN-γ) levels decreased in four (P = 0.14) patients. Four patients who had signs of multi-organ failure or sepsis died in 5–19 days (average: 10 days) after the first MSC infusion. A low percentage of lymphocytes (< 10%) and leukocytosis were associated with poor outcome (P = 0.02). All six survivors were well with no complaints of dyspnea on day 60 post-infusion. Radiological parameters of the lung computed tomography (CT) scans showed remarkable signs of recovery.Interpretation: We suggest that multiple infusions of high dose allogeneic prenatal MSCs are safe and can rapidly improve respiratory distress and reduce inflammatory biomarkers in some critically ill COVID-19-induced ARDS cases. Patients that develop sepsis or multi-organ failure may not be good candidates for stem cell therapy. Large randomized multicenter clinical trials are needed to discern the exact therapeutic potentials of MSC in COVID-19-induced ARDS.