In vitro cytotoxicity of folate-silica-gold nanorods on mouse acute lymphoblastic leukemia and spermatogonial cells

Objective: The purpose of this study was to evaluate in vitro cytotoxicity of gold nanorods [GNRs] on the viability of spermatogonial cells [SSCs] and mouse acute lymphoblastic leukemia cells [EL4s]

Materials and Methods: In this experimental study, SSCs were isolated from the neonate mice, following enzymatic digestion and differential plating. GNRs were synthesized, then modified by silica and finally conjugated with folic acid to form F-Si-GNRs. Different doses of F-Si-GNRs [25, 50, 75, 100, 125 and 140 microM] were used on SSCs and EL4s. MTT [3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide] proliferation assay was performed to examine the GNRs toxicity. Flow cytometry was used to confirm the identity of the EL4s and SSCs. Also, the identity and functionality of SSCs were determined by the expression of specific spermatogonial genes and transplantation into recipient testes. Apoptosis was determined by flow cytometry using an annexin V/propidium iodide [PI] kit

Results: Flow cytometry showed that SSCs and EL4s were positive for Plzf and H-2kb, respectively. The viability percentage of SSCs and EL4s that were treated with 25, 50, 75, 100, 125 and 140 microM of F-Si-GNRs was 65.33 +/- 3.51%, 60 +/- 3.6%, 51.33 +/- 3.51%, 49 +/- 3%, 30.66 +/- 2.08% and 16.33 +/- 2.51% for SSCs and 57.66 +/- 0.57%, 54.66 +/- 1.5%, 39.66 +/- 1.52%, 12.33 +/- 2.51%, 10 +/- 1% and 5.66 +/- 1.15% for EL4s respectively. The results of the MTT assay indicated that 100 microM is the optimal dose to reach the highest and lowest level of cell death in EL4s and in SSCs, respectively

Conclusion: Cell death increased with increasing concentrations of F-Si-GNRs. Following utilization of F-Si-GNRs, there was a significant difference in the extent of apoptosis between cancer cells and SSCs

Impaired memory and evidence of histopathology in CA1 pyramidal neurons through Injection of Abeta1-42 peptides into the frontal cortices of rat

Introduction: Alzheimer's disease [AD] is one of the most common neurodegenerative disorders, which has much benefited from animal models to find the basics of its pathophysiology. In our previous work [Haghani, Shabani, Javan, Motamedi, and Janahmadi, 2012], a non-transgenic rat model of AD was used in electrophysiological studies. However, we did not investigate the histological aspects in the mentioned study

Methods: An AD model was developed through bilateral injection of amyloid-beta peptides [Abeta] into the frontal cortices. Behavioral and histological methods were used to assess alterations in the memory and [ultra]structures. Furthermore, melatonin has been administered to assess its efficacy on this AD model

Results: Passive avoidance showed a progressive decline in the memory following Abeta injection. Furthermore, Nissl staining showed that Abeta neurotoxicity caused shrinkage of the CA1 pyramidal neurons. Neurodegeneration was clearly evident from Fluoro-jade labeled neurons in Abeta treated rats. Moreover, higher NF-kappaB immunoreactive CA1 pyramidal neurons were remarkably observed in Abeta treated rats. Ultrastructural analysis using electron microscopy also showed the evidence of subcellular abnormalities. Melatonin treatment in this model of AD prevented Abeta- induced increased NF-kappaB from immunoreaction and neurodegeneration

Discussion: This study suggests that injection of Abeta into the frontal cortices results in the memory decline and histochemical disturbances in CA1 pyramidal neurons. Furthermore, melatonin can prevent several histological changes induced by Abeta

Chronic ritalin administration during adulthood increases serotonin pool in rat medial frontal cortex

Ritalin has high tendency to be abused. It has been the main indication to control attention deficit hyperactivity disorder. The college students may seek for it to improve their memory, decrease the need for sleep [especially during exams], which at least partially, can be related to serotonergic system. Therefore, it seems worthy to evaluate the effect of Ritalin intake on mature brain. There are many studies on Ritalin effect on developing brain, but only few studies on adults are available. This study was undertaken to find Ritalin effect on serotonin transporter [SERT] density in medial frontal cortex [MFC] of mature rat. Thirty male Wistar rats were used in the study. Rats were assigned into five groups [n = 6 per group]: one control, two Ritalin and two vehicle groups. Twelve rats received Ritalin [20 mg/kg/twice a day] orally for eleven continuous days. After one week of withdrawal and another two weeks of rest, in order to evaluate short-term effects of Ritalin, six rats were sacrificed. Another six rats were studied to detect the long-term effects of Ritalin; therefore, they were sacrificed 12 weeks after the previous group. The immunohistochemistry was performed to evaluate the results. Immunohistochemistry studies showed a higher density of SERT in both 2 and 12 weeks after withdrawal from Ritalin intake in MFC of rat and there was no significant difference between these two groups. Our findings demonstrated both short- and long-term effects of Ritalin on frontal serotonergic system after withdrawal period

Do clinical and demographic features of patients with upper-gastrointestinal cancer affect their health-related quality of life?

Oesophagogastric [OG] cancer as a globally common and deadly malignancy, which is widely spread in Northeast Iran, has an extensive impact on health-related quality of life [HRQL]. Demographic and histopathologic changes have been apparent in oesophagogastric cancer, therefore. HRQL could be used, as an outcome, to assess and determine the efficacy and impact of cancer care. A consecutive sample of upper-gastrointestinal cancer patients admitted to the main oncology/ radiotherapy departments in the North-East of Iran were recruited into the study. All participants completed the European Organisation for Research and Treatment of Cancer [EORTC] QLQ-C30 and QLQ-OG25questionnaires in a face to face interview. Of the total 275 patients participated in the study, 54% had oesophageal, 34% stomach and 12% OG junction cancers. About 73.1% had TNM [tumour, node, metastasis] staging; of which 69% were in stage III and IV. The most common type of cancer in oesophagus was Squamous Cell Carcinoma [SCC] [95.3%] in lower third, Adenocarcinoma in stomach [97.8%] and in the OG junction [93.8%]. Patients with stomach or OG junction tended more to present in higher stages [P < 0.001]. Unlike QLQ-C30, the EORTC QLQ-OG25 was able to differ patients significantly in anxiety scale [P = 0.01], body image, chocking and weight loss [P < 0.05]. Those who had self care ability had better quality of life scores [P < 0.001] in more scales and items. SCC is predominant type of upper GI cancer in Khorasan provinces similar to the high risk area in Northern Iran. The specific health-related quality of life tool [EORTC QLQ-OG25] was able to distinguish most of the symptoms in patients with upper GI cancer

[ immunohistochemical assessment of ALDH1 activity in breast cancer and it's correlation with pathologic features]

Aldehyde dehydrogenase 1 [ALDH1] is a marker of normal and malignant human mammary stem cells that has been reported to be associated with poor prognosis. Studies on the detection of ALDH1+cells can help the treatment of patients with breast cancer. The aim of this study was to determine the activity of ALDH1 in breast cancer and its relationship with the pathological features of the tumors. ALDH1 activity was studied by immunohistochemistry in 121 paraffin-embedded histological samples of breast cancer patients from Department of Pathology of Milad Hospital, Tehran, Iran during 2006-2007. The relationship of ALDH1 with the pathological features of the tumors [size, grade, lymph node metastasis and vascular invasion] was also investigated. Eighty-five percent of breast cancer samples expressed ALDH1 in their cytoplasm with a wide range of intensity [weak, moderate and strong], while 18 samples [14.9%] were completely negative. The majority of cases [97.1%] showed ALDH1 positivity in the stroma of tumors which varied from weak [2.9%] to strong [73.5%]. ALDH1 H-score [ALDH1% x intensity] of tumor cells varied from 0 to 240 [mean=80]. ALDH1 H-score was 80 in 59 [48.8%] samples. There was no statistically significant relationship between ALDH1 H-score and age [P=0.358], tumor size [P=0.375], tumor grade [P=0.207], lymph node metastasis [P=0.125] or vascular invasion [P=0.190]. ALDH1 activity was demonstrated in 85.1% of breast cancer samples although its level of expression was not correlated with the pathologic features of breast tumors

Neuroprotective effect of exogenous melatonin on dopaminergic neurons of the substantia nigra in ovariectomized rats

Melatonin has receptors in substantia nigra pars compacta [SNc] and regulates development of dopaminergic [DA] neurons. This study was undertaken to determine ability of melatonin to protect SNc dopaminergic neuron loss induced by estrogen deficiency in ovariectomized rats. Female rats were randomized into four groups of seven each: control, ethanol sham, ovariectomy [ovx] and ovx with melatonin [ovx + m]. In ovx, ovaries were removed. Ovx + m group was intraperitoneally injected with melatonin for 10 days, while the ethanol sham group received only ethanol. All rats were perfused with 4% paraformaldehyde, midbrains removed, fixed and paraffin embedded, then processed for Nissl and tyrosine hydroxylase staining [IHC]. Ten sections of SNc in Nissl and IHC staining were analyzed in each animal, Nissl stained and tyrosine hydroxylase [TH] immunoreactive cells were counted in five experimental groups randomly. Data was analyzed using SPSS by ANOVA and /-test. Differences were considered significant for P<0.05. There was less cell number in ovx compared to control and ethanol sham groups significantly [P<0.001]. The ovx + m group had more cells than the ovx group in the SNc significantly [P<0.001]. Furthermore, there was significant decrease of TH positive cell number in the ovx group compared to control and ethanol sham groups [P<0.05]. The number of TH immunoreactive cells-was higher in ovx + m compared to the ovx group [P<0.05]. These findings can be compared with human and used in clinical application for prevention of DA neuron death of SNc after ovariectomy

Immuno-histochemical localization of endothelial nitric oxide synthase in testicular cells of men with non-obstructive azoospermia

Non obstructive azoospermia [NOA] is one of the causes of male infertility in which spermatogenesis process is disturbed. Recent studies suggested the possible role of endothelial nitric oxide synthase [eNOS] in spermatogenesis process. The aim of the present study is to evaluate the expression of eNOS in human testicular tissue in men with NOA and men with normal spermatogenesis by using immunocytochemistry. In this case-control study, testicular biopsies were obtained from 10 men with NOA and 7 men with normospermia who were attended to infertility center for diagnosis or infertility treatment. Immunohistochemistry was used to localize the isoform of eNOS in these tissues and the intensity of staining was semi quantitively assessed. In addition, the histopathological evaluation was examined in both groups. The isoform of eNOS enzyme activity was detected in the cytoplasm of sertoli and leydig cells in both groups. There was, however, a considerable variability in the intensity of staining between two groups. The expression of eNOS in Leydig cells in control group was significantly [p<0.05] higher than those in the NOA group. In contrast, expression of eNOS in Sertoli cells in NOA was more than those in the control group. eNO Simmune staining was absent in the normal germ cells but was intense in the abnormal germ cells with piknotic neucleous. The most histopathological finding were hypospermatogenesis [27.2%], Sertoli cell only syndrome [18.1%] and tubular fibrotic [13.6%]. These results suggested that increase level of eNOS may play an important role in the apoptosis process in the abnormal germ cells and disturbance of spermatogenesis process

BRCA1 protein expression level and CD44[+] phenotype in breast cancer patients

CD44[+]/CD24[-/low] breast cancer cells have tumour-initiating properties with stem cell-like features. Breast cancer gene 1 [BRCA1] is a tumour suppressor gene that plays a crucial role in DNA repair and maintenance of chromosome stability. The clinicopathological features of breast cancer in BRCA1 mutation carriers suggest that BRCA1 may function as a stem-cell regulator. In the present experimental study we examined the expression and localization of the BRCA1 protein and investigated the prognostic value as well as its relationship with the putative cancer stem cell [CSC] marker [CD44] in 156 tumour samples from a well-characterized series of unselected breast carcinomas using immunohistochemistry. Statistical analysis of the data was performed using SPSS software version 16 [Chicago, IL, USA]. In breast tumours, the loss of nuclear expression was detected in 23 cases [15%], whereas cytoplasmic expression of BRCA1 was observed in 133 breast carcinomas [85%]. Altered BRCA1 expression was significantly associated with high grade and poor prognosis breast tumours [p=0.006]. We further established an inverse significant correlation between BRCA1 expression levels and CD44[+] cancer cell phenotype [p=0.02] Loss of BRCA1 expression is a marker of tumour aggressiveness and correlates with CD44[+] tumour cell phenotype. Taken together, the present study supports the idea that the loss of BRCA1 results in persistent errors in DNA replication in breast stem cells and provides targets for additional carcinogenic events