Aging and stem cell therapy: ampk as an applicable pharmacological target for rejuvenation of aged stem cells and achieving higher efficacy in stem cell therapy

In recent years, tissue regeneration has become a promising field for developing stem cellbased transplantation therapies for human patients. Adult stem cells are affected by the same aging mechanisms that involve somatic cells. One of the mechanisms involved in cellular aging is hyperactivation of mechanistic target of rapamycin complex 1 [mTORC1] and disruption of 50 adenosine monophosphate-activated protein kinase [AMPK]. Aging of stem cells results in their

impaired regenerative capacity and depletion of stem cell pools in adult tissue, which results in lower efficacy of stem cell therapy. By utilizing an effective therapeutic intervention for aged stem cells, stem cell therapy can become more promising for future application. mTORC1 inhibition is a practical approach to preserve the stem cell pool. In this article, we review the dynamic interaction between sirtuin [silent mating type information regulation 2 homolog] 1, AMPK, and mTORC1. We propose that using AMPK activators such as 5-aminoimidazole-4- carboxamide ribonucleotide, A769662, metformin, and oxidized nicotinamide adenine dinucleotide [NAD+] are practical ways to be employed for achieving better optimized results in stem cell-based transplantation therapies

Overexpression of indoleamine 2,3-dioxygenase correlates with regulatory T cell phenotype in acute myeloid leukemia patients with normal karyotype

BACKGROUND: Production of immunosuppressive enzymes such as indoleamine 2,3-dioxygenase (IDO) is one of the strategies employed by hematologic malignancies, including acute myeloid leukemia (AML), to circumvent immune surveillance. Moreover, IDO has the ability to convert CD4+CD25− conventional T cells into regulatory T cells (Tregs). In this study, we evaluated the expression of IDO in cytogenetically normal acute myeloid leukemia (CN-AML) patients and its correlation with the Treg marker, FOXP3, as well as clinical and laboratory parameters. METHODS: Thirty-seven newly diagnosed CN-AML patients were enrolled in our study along with 22 healthy individuals. The expression of the IDO and FOXP3 genes was analyzed by SYBR Green real-time PCR. RESULTS: Both IDO and FOXP3 were highly upregulated in CN-AML patients compared to control groups (P=0.004 and P=0.031, respectively). A positive correlation was observed between IDO and FOXP3 expression among AML patients (r=0.512, P=0.001). Expression of IDO and FOXP3 showed no significant correlation with laboratory parameters such as white blood cell and platelet counts, hemoglobin levels, bone marrow blast percentage, gender, and FLT3 mutation status (P>0.05). CONCLUSION: Higher IDO expression in CN-AML patients may be associated with an increased Treg phenotype which may promote disease progression and lead to poor prognosis of CN-AML patients.

Additional cytogenetic aberrations in chronic myeloid leukemia: a single-center experience in the Middle East

BACKGROUND: Additional cytogenetic aberrations are associated with disease progression in chronic myeloid leukemia (CML). This study was conducted to determine the type and frequency of these aberrations and their relationship with hematologic and molecular findings in the Middle East. METHODS: In this retrospective study, 134 well-established cases of CML were selected from 2010 to 2016. Their hematologic phase and type of fusion gene were determined. Finally, their karyotypes were analyzed and reported according to ISCN 2013. RESULTS: Patients had a mean age of 44 years. Twenty-two patients (16.4%) showed additional cytogenetic aberrations. Nine patients (6.7%) harbored a variant Philadelphia chromosome, and most were in the chronic phase. Seventeen patients (12.7%) had major and minor route abnormalities. There was a significant relationship between additional cytogenetic aberrations and major molecular response (P=0.032). Patient survival in the group with additional cytogenetic aberrations was significantly lower (49.7±11.1 mo) than that in the group without additional cytogenetic aberrations (77.3±3.1 mo) (P=0.031). CONCLUSION: This study revealed the same frequency of additional cytogenetic aberrations in CML as found in previous studies. Additional chromosomal aberrations led to shorter survival and lower rates of achievement of a major molecular response.

PAX-5 positive anaplastic large cell lymphoma presenting by dysphagia; a case report

Anaplastic large cell lymphoma [ALCL] is a distinct pathologic entity with characteristic morphologic, im-munophenotypic and cytogenetic features. Obstructive symptoms are rare presentation of ALCL. We report a 16-year-old boy who initially presented with dysphagia. Upper gastrointestinal endoscopy revealed severe stenosis with an infiltrative process starting from 24 cm of incisors in lower esophagus Esophageal mucosal biopsy demonstrated lymphomatous involvement that ancillary tests confirmed the diagnosis of ALCL, ALK [kinase-positive], and PAX5 positive. The patient responded to CHOP-based chemotherapy. This case illustrated an unusual presentation of primary Non Hodgkin lymphoma of esophagus

[Huge intraabdominal Kaposiform hemangioendothelioma without Kasabach-Merritt syndrome in a 7 months year old male]

Kaposiform hemangioendothelioma [KHE] is an intermediate vascular tumor which occurs in children and commonly associated with Kasabach-Merritt syndrome [KMS]. It commonly occurs in skin, trunk, extremities, and retroperitoneum. We report a 7 months year old boy with a huge intaabdominal KHE without KMS

Molecular genotyping of toxoplasma gondii in human spontaneous aborted fetuses in Shiraz, southern Iran

Congenital toxoplasmosis is associated with variable complications including encephalitis, microcephaly, hydrocephaly, hepatitis, lymphadenopathy and even intrauterine death. Presence of Toxoplasma gondii in human placenta may induce congenital infection. The aim of this study was to determine the genotypes of Toxoplasma gondii infection in human spontaneous aborted fetuses in Shiraz, south of Iran. Five hundred and forty two paraffin-embedded blocks of aborted placenta were collected, from two university-affiliated hospitals in Shiraz. Occurrence of spontaneous abortion was confirmed by examine of the slides. After re-cutting of the blocks and dewaxing, semi-nested PCR assay was used to detect the fragments of T. gondii B1 gene in the samples. Also direct molecular genotyping was performed on positive samples with Restriction Fragment Length Polymorphism-PCR analysis on the SAG2 gene. Among the 542 tissue samples, the B1 gene was amplified from 78 [14.4%] of cases with the semi nested PCR and typed by RFLP. The genotype of Toxoplasma strains of 65 [out of 78] PCR-positive samples were evaluated and 54 out of 65 [83.1%] were found to be type II and 11 out of 65 [16.9%] were type I. Considering the high level of Toxoplasma infection in aborted fetuses in this study, Toxoplasma might largely contribute to spontaneous abortion in this area of Iran

Colony forming unit endothelial cells do not exhibit telomerase alternative splicing variants and activity

Endothelial progenitor colony forming unit-endothelial cells [CFU-EC] were first believed to be the progenitors of endothelial cells, named endothelial progenitor cells. Further studies revealed that they are monocytes regulating vasculogenesis. The main hindrance of these cells for therapeutic purposes is their low frequency and limited replicative potentials. This study was undertaken to determine telomerase activity and alternative splicing variants in CFU-EC as a potential cause of limited replicative capacity in these cells. CFU-EC were isolated from peripheral blood using a standard cell culture assay. Colonies were detached mechanically and alternative splicing variant mRNA were evaluated using real-time PCR. Telomerase enzyme activity was assessed using telomerase repeat amplification protocol. The same procedures were done on the cancer cell line Calu6 as the positive control. The cultured peripheral blood mononuclear cells formed colonies with spindle-shaped monocytic cells sprouted from the clusters. These morphological characteristics fulfill the definition of CFU-EC. Telomere length amplification protocol assay revealed no telomerase activity and real-time PCR showed no expression of telomerase enzyme mRNA in CFU-EC. Both parameters were significantly higher in the cancer cell line Calu6 taken as the positive control. The absence of telomerase activity in the CFU-EC is a result of pre-transcriptional regulation of gene expression rather than other mechanisms for controlling telomerase activity such as post-transcriptional modifications. This finding can explain the limited proliferative activity of CFU-EC cells. We propose that absence of telomerase activity in CFU-EC can be attributable to their more mature monocytic nature that needs further investigations

Expression of UBE2Q2, a putative member of the ubiquitin-conjugating enzyme family in pediatric acute lymphoblastic leukemia

Acute lymphoblastic leukemia [ALL] is a cancer of the white blood cells most commonly found in childhood with a peak incidence at 2-5 years of age. The ubiquitin degradation pathway facilitates degradation of damaged proteins and regulates the growth and stress response. This pathway is activated in various cancers, including ALL. It has been previously reported that the newly characterized human gene UBE2Q2, a putative member of the ubiquitin-conjugating enzyme family, is over-expressed in the tumor mass and invasive epithelium in head and neck squamous cell carcinoma and breast cancer. Here, we have used quantitative reverse transcriptase polymerase chain reaction [RT-PCR] to assess expression of the UBE2Q2 gene in bone marrow samples of 20 children with ALL. Whole blood samples of 20 normal children were used as control specimens. RT-PCR revealed the expression of UBE2Q2 mRNA in 80% of the bone marrow samples from ALL patients as well as in 85% of leukemic normal peripheral blood cells. According to the results of quantitative RT-PCR, the levels of UBE2Q2 mRNA expression in the bone marrow cells of 11 out of the 20 children with ALL [55%] were significantly higher [> 2-47 fold] than those in blood cells of normal children. Our data suggest that the newly characterized human gene, UBE2Q2, may have implications for the pathogenesis of ALL and could be used for molecular diagnosis purposes in the future

prevalence of human papilloma virus in esophageal squamous cell carcinoma

Carcinomas of esophagus, mostly squamous cell carcinomas, occur throughout the world. There are a number of suspected genetic or environmental etiologies. Human papilloma virus [HPV] is said to be a major etiology in areas with high incidence of esophageal carcinoma, while it is hardly detectable in low incidence regions. This study was designed to evaluate the prevalence of HPV in esophageal squamous cell carcinoma [ESCC] cases diagnosed in Pathology Department, Medical School, Shiraz University of Medical Sciences. DNA material for PCR amplification of HPV genome was extracted from formalin-fixed paraffin-embedded tissue blocks of 92 cases of ESCC, diagnosed during 20 years from 1982 to 2002. Polymerase chain reaction was performed for amplification and detection of common HPV and type specific HPV-16 and HPV-18 genomic sequences in the presence of positive control [HPV-18 and HPV positive biopsies of uterine exocervix] and additional internal controls i.e. beta-globin and cytotoxic T lymphocyte antigen 4 [CTLA4]. Good amplification of positive control and internal controls was observed. However, no amplification of HPV genome was observed. There is no association between HPV infection and the development of esophageal squamous cell carcinoma in the cases evaluated

Serum HER2 level in epithelial ovarian cancer

The proto-oncogene HER2 plays a key role in the control of cellular proliferation. Its overexpression has been reported to be associated with a poor prognosis in cancer, particularly in breast cancer. In the present study, serum HER2 levels were investigated in patients diagnosed with epithelial ovarian cancer. Serum HER2 levels were detected by an ELISA commercial kit in 51 patients and 33 healthy individuals. The mean serum HER2 level was found to be significantly higher in patients than healthy controls [P=0.005]. In 29% of patients, serum HER2 levels were higher than the cut-off value. HER2 serum level was not associated with tumor stage at diagnosis. Elevation of HER2 in a high proportion of patients with epithelial ovarian cancer further strengthens the importance of this molecule in the pathogenesis of ovarian cancer

Hydatid cyst of ovary: a case report

Echinococcus granulosus is considered the major cause of human hydatid cysts. Usually the duration of cyst formation is 10-20 years. This period shortens significantly upon rupture of a primary cyst. The literature describes low incidence of primary involvement of ovary as a site of hydatid cyst formation. Our case is the first report on ovarian hydatid cyst in Iran. A 60-year-old woman was presented with abdominal pain in the left lower quadrant area. Paraclinical data were suggestive of neoplasia and preoperative diagnosis was ovarian tumor. During laparotomy, multiple cysts resembling hydatid cysts were observed in the left ovary. Pathological examination confirmed the diagnosis of hydatid cyst. Although there is a small possibility of secondary ovarian echinococcal disease, it is more probable for this case to be primary infection, as the patient had developed ovarian hydatid cysts 15 years after hepatic involvement and recurrence after 30 months is very uncommon

Polymerase chain reaction; detection of human papilloma virus [HPV] type - 16 and 18 infection in laryngeal and oral squamous cell carcinoma in southern Iran

One fifth of cancers world wide are associated with viral infection. Epidemiologic and biomolecular evidence suggested that Human Papilloma Virus [HPV] infection may be associated with the development of head and neck cancer. [1] To clarify the role of HPV infection in head and neck cancers. [2] To evaluate the presence of HPV DNA in laryngeal and oral squamous cell carcinoma in southern Iran and comparison of results with studies in other regions Department of Otolaryngology-Head and Neck Surgery, Khallili Hospital, Shiraz Medical University Iran From 2003 to 2006. Eighty three [83] patients with Squamous Cell Carcinoma [SCC] of the larynx, 40 patients with benign mucosal lesion of the larynx [control], 47 patients with SCC of oral cavity and 10 patients with benign oral lesion were studied for the presence, of HPV DNA by Polymerase Chain Reaction [PCR]. None of the laryngeal SCCs or control group was positive for HPV DNA. Only 3/47 specimens from oral SCC were positive for HPV DNA. Oral control group was negative for HPV DNA. The present work suggests that HPV infection has not important role in carcinogenesis of laryngeal or oral SCC in southern Iran. However a multi center case-control study is needed to clarify this association

Agreement rate of skin prick test with tissue eosinophil count in patients with nasal polyps

The pathogenetic mechanism of nasal polyps remains unknown, although allergy has been cited as an important factor in the etiology of nasal polyposis. Currently there is no definite histological criterion for differentiation of allergic from inflammatory nasal polyp. However, in a few studies, tissue eosinophil count has been used for this. This study aimed to find out the agreement rate of skin prick test and tissue eosinophil count in patients with nasal polyposis. Twenty five patients [18 males, 7 females] with nasal polyp were enrolled in this study. For each patient tissue sample from polyp material was taken for histopathological investigation. Moreover, skin prick test was performed for each patient using eleven common aeroallergens. Skin prick test was positive in 48% of the patients. Tissue eosinophil count of more than 50% was found in 75% of skin prick positive and in 69.2% of skin prick negative patients. Also tissue eosinophil count of more than 50% was found in 69.2% of patients with typical allergic symptoms as well as 75% of patients without allergic symptoms. No agreement was found between skin prick tests and tissue eosinophil counts in patients with nasal polyp. Also no difference was found between the tissue eosinophil counts in allergic and non allergic patients. Considering these results, it can be concluded that having a high tissue eosinophil count in patients with nasal polyp does not indicate that the polyp is allergic

T-cell tolerance following bacterial glutamic acid decarboxylase [GAD] feeding in streptozotocin-induced diabetes

Autoimmune type 1 diabetes mellitus is caused by T-cell mediated immune destruction of the insulin-producing a-cell in pancreatic islets of Langerhans. Specificity of the auto-antibodies and of the auto-reactive T-cells has been investigated, in which several auto-antigens were proposed. To determine whether glutamic acid decarboxylase [GAD] feeding would induce oral tolerance of either T-cell or B-cell compartment in streptozotocin [STZ] diabetic rats. Rats in the experimental group were fed 2 mg/kg of GAD [extracted from Escherichia coli] 14 days before intra-peritoneal injections of streptozotocin [30 mg/kg body weight for 5 consecutive days]. Two control groups were considered: diabetic control group, which underwent STZ injections without receiving GAD, and normal control group. Systemic response was compared between the three groups. T-cells response was assessed by a proliferation assay of spleen cells and those of the B-cells by enzyme-linked immunosorbent assay [ELISA] for anti-GAD specific antibodies in serum. Compared with the diabetic control group, a significant reduction was observed only in the proliferative response of spleen cells, but not in the level of anti-GAD antibody. GAD feeding induces systemic T-cell tolerance in STZ-induced diabetes

p53 protein expression and its relation to the apoptotic index in prostate adenocarcinoma

Bakgorund: Prostate cancer is one of the most commonly diagnosed cancers in males. Tumor suppressor gene p53 plays an important role in causing cell cycle arrest and allowing apoptosis to proceed

Objective: To investigate the expression of p53 protein and its relation to apoptosis and prostate cancer traditional prognostic indicators

Methods: In this study expression of p53 was examined in paraffin-embedded tissues from 50 cases of prostate carcinoma by immunohistochemistry and evaluated using an index of staining. Correlation between p53 expression and apoptosis was detected by TUNEL method. Pathological grade, Gleason score and stage of carcinoma were also determined

Results: P53 expression was observed in 48 of 50 cases [26-100% of tumor cells] with mean staining index of 141 +/- 65. A significant association between p53 expression and pathologic grade [r=0.37, p=0.004] and Gleason score [r= 0.4, p=0.009] of patients was observed. Apoptosis was detected in only 6 patients

Conclusions: p53 expression showed no correlation with apoptotic index. No correlation between p53 expression and stage or apoptosis and clinicopathological characteristics of patients was found. p53 expression showed a significant correlation with differentiation status of the prostate carcinoma and can be helpful as a prognostic marker. Decreased level of apoptosis observed in our cases was not correlated with p53 expression indicating the possible role of other regulatory molecules involved in the apoptosis