Assessment of Cognitive Flexibility in Jordanian Diabetic Patients by Wisconsin Card Sorting and Trail Making Tests: Implications with Demographic, Anthropometric and Therapeutic Variables.

Purpose: Cognitive flexibility is a mental ability that aids in smoothly alternating between them tasks in the brain. Diabetes Mellitus (DM) is a, common disorder that has been associated with impairments in cognitive functions. This research is a retrospective case-control study aimed at establishing a clear relationship between cognitive flexibility and diabetes among Jordanians, considering demographic, anthropometric, and therapeutic variables. Patients and Methods: The Wisconsin Card Sorting Test (WCST)-64 item and the Trail Making Test (TMT) assessed cognitive flexibility in 268 people with diabetes and healthy control. Demographic, therapeutic data were collected. We also measured waist-to-hip ratio (WHR) and body mass index (BMI). As the variables were non-normally distributed, non-parametric statistical tests were used to examine differences (Kruskal-Wallis) and correlation (Spearman) between variables. Results: The patient group did worse on the WCST In contrast to the control group, patients exhibited more significant delays for both Part A and Part B of the TMT (p<0.05). Males had higher WCST conceptual level responses than females. In addition, participants with professional jobs showed less delay in TMT Part A (p<0.05). Age was positively correlated with WCST's total errors and TMT's Parts A and B (p<0.05). BMI was negatively correlated with the WCST's conceptual level of responses and positively correlated with TMT's Part B (p<0.05). In addition, urea and albumin levels were positively correlated with TMT's Part A (p<0.05). Furthermore, creatinine was positively correlated with WCST's total errors and TMT's Part A (p<0.05). Conclusion: Some measures of cognitive flexibility are associated with DM status in the studied sample of Jordanians and other variables (educational levels, occupation, lifestyle, average duration of illness, and age).

Dental pulp stem cells ameliorate D-galactose-induced cardiac ageing in rats.

Background: Ageing is a key risk factor for cardiovascular disease and is linked to several alterations in cardiac structure and function, including left ventricular hypertrophy and increased cardiomyocyte volume, as well as a decline in the number of cardiomyocytes and ventricular dysfunction, emphasizing the pathological impacts of cardiomyocyte ageing. Dental pulp stem cells (DPSCs) are promising as a cellular therapeutic source due to their minimally invasive surgical approach and remarkable proliferative ability. Aim: This study is the first to investigate the outcomes of the systemic transplantation of DPSCs in a D-galactose (D-gal)-induced rat model of cardiac ageing. Methods. Thirty 9-week-old Sprague-Dawley male rats were randomly assigned into three groups: control, ageing (D-gal), and transplanted groups (D-gal + DPSCs). D-gal (300 mg/kg/day) was administered intraperitoneally daily for 8 weeks. The rats in the transplantation group were intravenously injected with DPSCs at a dose of 1 × 106 once every 2 weeks. Results: The transplanted cells migrated to the heart, differentiated into cardiomyocytes, improved cardiac function, upregulated Sirt1 expression, exerted antioxidative effects, modulated connexin-43 expression, attenuated cardiac histopathological alterations, and had anti-senescent and anti-apoptotic effects. Conclusion: Our results reveal the beneficial effects of DPSC transplantation in a cardiac ageing rat model, suggesting their potential as a viable cell therapy for ageing hearts.

Enhanced autophagy and phagocytosis of apoptotic lymphocytes in splenic macrophages of acute ethanol-treated rats: Light and electron microscopic studies.

Autophagy is a prosurvival mechanism for the clearance of damaged cellular components, specifically upon exposure to various stressors. In lymphoid organs, excessive ethanol consumption increases lymphocyte apoptosis, resulting in immunosuppression. However, ethanol-induced autophagy and related phagocytosis of apoptotic lymphocytes in the spleen have not been studied yet. Adult male Wistar rats were injected intraperitoneally either with 5 g/kg ethanol or phosphate-buffered saline (as a control group) and then sacrificed 0, 3, 6, and 24 hours after injection. Light and transmission electron microscopy (TEM) findings indicated enhanced T cell apoptosis in the white pulps of ethanol-treated rats (ETRs) compared with the control group, which peaked at 6 h and was associated with the accumulation of tingible body macrophages (TBMs). These macrophages exhibited an upregulated autophagic response, as evidenced by enhanced LC3-II (a specific marker of autophagosomes) expression, which peaked at 24h. In addition, double labeling immunofluorescence of LC3-II with lysosomal markers revealed the enhanced formation of autolysosomes in TBMs of ETRs, which was associated with suppression of p62 immunostaining, indicating the enhanced autophagic flux. Interestingly, this elevated autophagic response in ETR TBMs was accompanied by evidence of LC3-associated phagocytosis (LAP) of apoptotic splenocytes. This is based on TUNEL/LC3-II double labeling and TEM observations of phagosomes containing apoptotic bodies, enclosed within phagosomal membranes adjacent to the autophagic vacuoles. It can be concluded that enhanced prosurvival autophagy in splenic TBMs of ETRs and clearing of apoptotic lymphocytes via LAP may contribute to preventing secondary necrosis and autoimmune diseases.

Evaluating cognitive and affective abilities among medical students: behavioural and medicolegal perspectives.

Medical students attending university for the first time experience a new environment, full of significant social, cultural, and intellectual challenges. Moreover, drug abuse and bullying among university students are major global concerns. The aim of the current study was to assess the impact of medicolegal issues on undergraduate and postgraduate students. It is a cross-sectional survey-based study, with each set of questions investigating cognitive functions, aggression, personality, and exposure to medicolegal issues. Males and those with a chronic disease have been significantly exposed to medicolegal issues; exposed students were significantly older than nonexposed ones. The scores of aggression were significantly higher among exposed and male students. The cognitive scores were higher for the students from rural areas than in urban areas, and females were more neurotic than males. The current study recommends conducting campaigns to educate university students on the importance of formally disclosing unethical behaviors and listening to the victims to facilitate overcoming their negative feelings. As many victims feel comfortable disclosing victimization to their friends, we recommend conducting peer educational programs to help friends support their colleagues regarding unethical misconduct.

Prevalence of eating disorders among Saudi female university students during the COVID-19 outbreak.

Eating disorders are multifaceted problems with various risk factors, including the sociocultural context, social media, society's beauty standards, personality, and genetics. The coronavirus disease 2019 (COVID-19) pandemic has been a cause of stress among university students, as well as inducing changes in their physical activity and eating habits. Objective: The objectives of this study were to evaluate the changes in body mass index and risk of developing eating disorders among university students during the COVID 19 pandemic. Methods: This was a cross-sectional study of 1004 female students recruited from a university in Riyadh, Saudi Arabia. Data were collected from December 2020 to March 2021 through a self-administered questionnaire comprising three parts: sociodemographic items, the Eating Attitudes Test, and an evaluation of behavioral changes during the COVID-19 pandemic. Results: Most participants were aged 18-24 years, single, lived with their parents, and had a moderate to high family income. There was a significant relationship between the risk of developing eating disorders and marital status (p < 0.001). College type (p < 0.003), fast food consumption (p = 0.010), and engaging in exercise (p < 0.001) were also significant factors. Based on categorizations of risk levels derived from the literature, about 31.5% of the participants had a high risk of developing eating disorders. Conclusion: According to our results, eating disorders are relatively common among Saudi female undergraduate students. Thus, educational programs that aim to increase this population's awareness concerning appropriate nutrition and body weight are needed.

Human dental pulp stem cells attenuate streptozotocin-induced parotid gland injury in rats.

OBJECTIVE: Diabetes mellitus causes deterioration in the body, including serious damage of the oral cavity related to salivary gland dysfunction, characterised by hyposalivation and xerostomia. Human dental pulp stem cells (hDPSCs) represent a promising therapy source, due to the easy, minimally invasive surgical access to these cells and their high proliferative capacity. It was previously reported that the trophic support mediated by these cells can rescue the functional and structural alterations of damaged salivary glands. However, potential differentiation and paracrine effects of hDPSCs in diabetic-induced parotid gland damage have not been investigated. Our study aimed to investigate the therapeutic effects of intravenous transplantation of hDPSCs on parotid gland injury in a rat model of streptozotocin (STZ)-induced type 1 diabetes. METHODS: Thirty Sprague-Dawley male rats were randomly categorised into three groups: control, diabetic (STZ), and transplanted (STZ + hDPSCs). The hDPSCs or the vehicles were injected into the rats' tail veins, 7 days after STZ injection. Fasting blood glucose levels were monitored weekly. A glucose tolerance test was performed, and the parotid gland weight, salivary flow rate, oxidative stress indices, parotid gland histology, and caspase-3, vascular endothelial growth factor, proliferating cell nuclear antigen, neuronal nitric oxide synthase, endothelial nitric oxide synthase, and tetrahydrobiopterin biosynthetic enzyme expression levels in parotid tissues were assessed 28 days post-transplantation. RESULTS: Transplantation of hDPSCs decreased blood glucose, improved parotid gland weight and salivary flow rate, and reduced oxidative stress. The cells migrated to the STZ-injured parotid gland and differentiated into acinar, ductal, and myoepithelial cells. Moreover, hDPSCs downregulated the expression of caspase-3 and upregulated the expression of vascular endothelial growth factor and proliferating cell nuclear antigen, likely exerting pro-angiogenic and anti-apoptotic effects and promoting endogenous regeneration. In addition, the transplanted cells enhanced the parotid nitric oxide-tetrahydrobiopterin pathway. CONCLUSIONS: Our results showed that hDPSCs migrated to and survived within the STZ-injured parotid gland, where functional and morphological damage was prevented due to the restoration of normal glucose levels, differentiation into parotid cell populations, and stimulation of paracrine-mediated regeneration. Thus, hDPSCs may have potential in the treatment of diabetes-induced parotid gland injury.

Quercetin prevents myocardial infarction adverse remodeling in rats by attenuating TGF-ß1/Smad3 signaling: Different mechanisms of action.

This study investigated the anti-remodeling and anti-fibrotic and effect of quercetin (QUR) in the remote non-infarcted of rats after myocardial infarction (MI). Rats were divided as control, control + QUR, MI, and MI + QUR. MI was introduced to the rats by ligating the eft anterior descending (LAD) coronary artery. All treatments were given for 30 days, daily. QUR persevered the LV hemodynamic parameters and prevented remote myocardium damage and fibrosis. Also, QUR supressed the generation of ROS, increased the nuclear levels of Nrf2, and enhanced SOD and GSH levels in the LVs of the control and MI model rats. It also reduced angiotensin II, nuclear level/activity of the nuclear factor NF-κß p65, and protein expression of TGF-ß1, α-SMA, and total/phospho-smad3 in the LVs of both groups. Concomitantly, QUR upregulated LV smad7 and BMP7. In conclusion, QUR prevents MI-induced LV remodeling by antioxidant, anti-inflammatory, and anti-fibroticα effects mediated by ROS scavenging, suppressing NF-κß, and stimulating Nrf-2, Smad7, and BMP7.

Osteoporosis knowledge, attitudes, and practices among female Princess Nourah University students in Riyadh, Saudi Arabia.

BACKGROUND AND OBJECTIVE: Osteoporosis is characterized by decreased normal bone density. More than 8.9 million fractures worldwide annually are caused by osteoporosis; these fractures are a significant cause of morbidity and mortality. Evidence suggests that the modification of several lifestyle habits could assist in lowering the incidence of osteoporosis. However, limited studies have been conducted in Saudi Arabia to assess the knowledge, attitudes, and lifestyles associated with osteoporosis among college-age females. This study aimed to provide evidence to assist in the development of effective strategies against osteoporosis. MATERIALS AND METHODS: This cross-sectional study was conducted at Princess Nourah Bint Abdul Rahman University (PNU), in February 2018; a self-administered questionnaire was used. The different components of the questionnaire assessed knowledge, attitudes, and lifestyles with regard to osteoporosis. The participants were divided into groups on the basis of their age as follows: juniors, 17-20 years of age; seniors, 21-25 years of age. RESULTS: Of the 250 included participants, 122 (49%) and 128 (51%) were seniors and juniors respectively. Only 16% of all participants achieved a good score on the knowledge questionnaire; in particular, knowledge regarding osteoporosis risk factors was inadequate. Media was the only source of information of the included participants. Only 49% of participants believed that osteoporosis is a serious disease. Overall, only 32% and 27% of juniors and seniors are consumed sufficient dairy products, and 13% and 11% of juniors and seniors engaged in physical exercise, respectively. CONCLUSION: Osteoporosis misconceptions were extremely prevalent among PNU students, as was poor knowledge and lifestyle habits regarding osteoporosis. Information regarding osteoporosis presented through the media needs to be revised and simplified. Concerned institutions should combine their efforts eventually practice. Information about osteoporosis presented through media need to be revised, simplified, and implement a national program to improve osteoporosis awareness and prevention.

Transplantation of human dental pulp stem cells in streptozotocin-induced diabetic rats.

Type 1 diabetes mellitus (T1DM) is a chronic metabolic disease caused by the destruction of pancreatic ß-cells. Human dental pulp stem cells represent a promising source for cell-based therapies, owing to their easy, minimally invasive surgical access, and high proliferative capacity. It was reported that human dental pulp stem cells can differentiate into a pancreatic cell lineage in vitro; however, few studies have investigated their effects on diabetes. Our study aimed to investigate the therapeutic potential of intravenous and intrapancreatic transplantation of human dental pulp stem cells in a rat model of streptozotocin-induced type 1 diabetes. Forty Sprague Dawley male rats were randomly categorized into four groups: control, diabetic (STZ), intravenous treatment group (IV), and intrapancreatic treatment group (IP). Human dental pulp stem cells (1 × 106 cells) or vehicle were injected into the pancreas or tail vein 7 days after streptozotocin injection. Fasting blood glucose levels were monitored weekly. Glucose tolerance test, rat and human serum insulin and C-peptide, pancreas histology, and caspase-3, vascular endothelial growth factor, and Ki67 expression in pancreatic tissues were assessed 28 days post-transplantation. We found that both IV and IP transplantation of human dental pulp stem cells reduced blood glucose and increased levels of rat and human serum insulin and C-peptide. The cells engrafted and survived in the streptozotocin-injured pancreas. Islet-like clusters and scattered human dental pulp stem cells expressing insulin were observed in the pancreas of diabetic rats with some difference in the distribution pattern between the two injection routes. RT-PCR analyses revealed the expression of the human-specific pancreatic ß-cell genes neurogenin 3 (NGN3), paired box 4 (PAX4), glucose transporter 2 (GLUT2), and insulin in the pancreatic tissues of both the IP and IV groups. In addition, the transplanted cells downregulated the expression of caspase-3 and upregulated the expression of vascular endothelial growth factor and Ki67, suggesting that the injected cells exerted pro-angiogenetic and antiapoptotic effects, and promoted endogenous ß-cell replication. Our study is the first to show that human dental pulp stem cells can migrate and survive within streptozotocin-injured pancreas, and induce antidiabetic effects through the differentiation and replacement of lost ß-cells and paracrine-mediated pancreatic regeneration. Thus, human dental pulp stem cells may have therapeutic potential to treat patients with long term T1DM.

Zeaxanthin exerts protective effects on acetic acid-induced colitis in rats via modulation of pro-inflammatory cytokines and oxidative stress.

Ulcerative colitis is a common intestinal inflammatory disease characterized by upregulation of pro-inflammatory cytokines and oxidative stress. Zeaxanthin is a nutritional carotenoid that belongs to the xanthophyll family of pigments. It exerts potent anti-inflammatory and antioxidative effects. The present study aimed to evaluate the effect of zeaxanthin on acetic acid-induced ulcerative colitis in rats. Rats were randomly categorized into five groups: control, zeaxanthin, acetic acid, acetic acid + zeaxanthin, and acetic acid + prednisolone groups. Zeaxanthin (50 mg/kg/day) or prednisolone (5 mg/kg/day) was orally administered for 14 days before induction of ulcerative colitis. On the 15th day, colitis was induced by transrectal administration of 3% acetic acid. The rats were sacrificed 24 h after rectal instillation and their colon tissues were examined. Pretreatment with zeaxanthin significantly reduced disease activity index, wet colon weight, ulcer area, macroscopic scores, and histological changes. Zeaxanthin also effectively downregulated the levels of myeloperoxidase and malondialdehyde, upregulated the enzymatic activity of superoxide dismutase and catalase, and raised glutathione levels. With regard to anti-inflammatory mechanisms, zeaxanthin suppressed tumor necrosis factor-alpha, interferon-gamma, interleukin-6, interleukin-1 beta, and nuclear transcription factor kappa B levels, and inhibited nitric oxide synthase and cyclooxygenase-2 protein expression. Our results indicate that oral administration of zeaxanthin ameliorates acetic acid-induced colitis in rats via antioxidative effects and modulation of pro-inflammatory cytokine and mediator activity. Therefore, zeaxanthin may be an effective therapeutic candidate for the treatment of ulcerative colitis.

Avocado soybean unsaponifiables ameliorates cartilage and subchondral bone degeneration in mono-iodoacetate-induced knee osteoarthritis in rats.

Osteoarthritis (OA), the most common type of arthritis, is a disabling progressive disease mainly affecting the elderly and is becoming a major public health problem. Current therapies for OA provide only palliative pain relief and therapeutic candidates that are able to slow the progression of structural deterioration is a major unmet need for this disorder. Avocado soybean unsaponifiables (ASU) has proven its safety and effectiveness in clinical studies of knee osteoarthritis (OA); however, whether ASU exerts structure-modifying effects is still to be elucidated. There are limited studies that have explored the underlying mechanisms of ASU's beneficial effects in animal models of OA. To this end, this study is the first to evaluate the effects of ASU in a rat model of mono-iodoacetate (MIA)-induced knee OA. OA was induced in rats by knee intra-patellar injection of MIA. Oral administration of ASU (27.5 mg/kg per day for 3 weeks) was initiated 3 weeks after MIA injection. We analysed the knee samples using light and electron microscopy. In addition, we used immunohistochemistry to investigate the expression of inducible nitric oxide synthase (iNOS), tumour necrosis factor-α (TNF-α), and matrix metalloproteinase-13 (MMP-13) in OA cartilage and subchondral bone. ASU significantly attenuated the synovium, cartilage, and subchondral degeneration. In addition, it reduced the expression of TNF-α and MMP-13 in OA cartilage and the expression of iNOS in both OA cartilage and subchondral bone. These results provide evidence of the structure-modifying abilities of ASU via its anti-oxidative and anti-inflammatory properties, in addition to its ability to modulate MMP-13 activity. This work suggests that ASU can be used as a potential disease-modifying treatment for OA.

Is Aquaporin-3 a Determinant Factor of Intrinsic and Extrinsic Aging? An Immunohistochemical and Morphometric Study.

Aquaporin-3 (AQP3) is an aquaglyceroporin that plays a role in skin hydration, cell proliferation, and migration. The aim of this work was to investigate the expression of AQP3 in sun-exposed and sun-protected human skin from different age groups to understand the relationship between AQP3 and skin aging. Using standard immunohistochemical techniques, sun-exposed and sun-protected skin biopsies were taken from 60 normal individuals. AQP3 was expressed in the basal and the suprabasal layers, sparing the stratum corneum, in all specimens. Dermal expression was detected in fibroblasts, endothelial cells, and adnexa. Sun-protected skin showed a significantly higher epidermal H-score and percentage of expression (P=0.002 and <0.001, respectively) compared with sun-exposed skin. The AQP3 expression intensity showed a gradual decrease from the 20 to 35-year-old group to the 35 to 50-year-old group, with the least immunoreactivity in the above 50-year-old group. A significant difference was detected in the H-score in favor of the 20 to 35-year-old group in sun-exposed and sun-protected skin (P<0.001 for both). A significant negative correlation was noted between the AQP3 expression percentage and the age in sun-exposed (r=-0.64, P<0.001) and sun-protected skin (r=-0.53, P<0.001). In conclusion, the skin dryness observed in intrinsic and extrinsic aged skin may be explained, at least in part, by AQP3 downregulation. This may open new avenues sufficient to control skin texture and beauty. Its interaction in skin protein organization and gene polymorphism can also be tackled in future research. In addition, clinical trials using AQP3 topical applications should be carried out to evaluate its effectiveness in the reversal of age-related skin changes.

Protective role of garlic oil against oxidative damage induced by furan exposure from weaning through adulthood in adult rat testis.

Furan is produced in a wide variety of heat-treated foods via thermal degradation. Furan contamination is found to be relatively high in processed baby foods, cereal products, fruits juices, and canned vegetables. Several studies have demonstrated that furan is a potent hepatotoxin and hepatocarcinogen in rodents. However, few studies have investigated the toxic effects of furan in the testis. In addition, the exact mechanism(s) by which furan exerts toxicity in the testis has not been fully elucidated. In this study, we investigated the potential of furan exposure from weaning through adulthood to induce oxidative stress in adult rat testis, as well as the potential of garlic oil (GO) to ameliorate the induced toxicity. Our results reveal that furan administration significantly reduced serum testosterone levels and increased the levels of malondialdehyde (MDA); furthermore, furan administration decreased significantly the enzymatic activity of testicular antioxidants, including glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) and induced histopathological alterations in the testis. GO co-administration ameliorated the reduction in testosterone levels and dramatically attenuated the furan-induced oxidative and histopathological changes. In addition, Go significantly down-regulated the increased caspase-3 and cytochrome P450 2E1 (CYP2E1) expression in the furan-treated testis. To the best of our knowledge, this study is the first to demonstrate the furan-induced oxidative changes in the adult rat testis and the protective role of GO to ameliorate these changes through its antioxidant effects and its ability to inhibit CYP2E1 production.

Mobilization of endogenous bone marrow-derived stem cells in a thioacetamide-induced mouse model of liver fibrosis.

The clinical significance of enhancing endogenous circulating haematopoietic stem cells is becoming increasingly recognized, and the augmentation of circulating stem cells using granulocyte-colony stimulating factor (G-CSF) has led to promising preclinical and clinical results for several liver fibrotic conditions. However, this approach is largely limited by cost and the infeasibility of maintaining long-term administration. Preclinical studies have reported that StemEnhance, a mild haematopoietic stem cell mobilizer, promotes cardiac muscle regeneration and remedies the manifestation of diabetes. However, the effectiveness of StemEnhance in ameliorating liver cirrhosis has not been studied. This study is the first to evaluate the beneficial effect of StemEnhance administration in a thioacetamide-induced mouse model of liver fibrosis. StemEnhance augmented the number of peripheral CD34-positive cells, reduced hepatic fibrosis, improved histopathological changes, and induced endogenous liver proliferation. In addition, VEGF expression was up-regulated, while TNF-α expression was down-regulated in thioacetamide-induced fibrotic livers after StemEnhance intake. These data suggest that StemEnhance may be useful as a potential therapeutic candidate for liver fibrosis by inducing reparative effects via mobilization of haematopoietic stem cells.

Beneficial effects of bone marrow-derived mesenchymal stem cell transplantation in a non-immune model of demyelination.

Multiple sclerosis (MS) is an autoimmune disease characterized by demyelination and axonal loss throughout the central nervous system. Most of the previous studies that have been conducted to evaluate the efficacy of mesenchymal stem cells (MSCs) have utilized immune models such as experimental autoimmune encephalomyelitis (EAE). However, with this experimental setting, it is not clear whether the MSCs exert the functional improvement via an indirect consequence of MSC-mediated immunomodulation or via a direct replacement of the lost cells, paracrine actions, and/or an enhancement of endogenous repair. This study is the first to demonstrate the capability of intravenously injected bone marrow-derived MSCs (BM-MSCs) to migrate, engraft, and improve the demyelination in the non-immune cuprizone model of MS. The ultrastructural analysis conducted in this study revealed that the observed histological improvement was due to both reduced demyelination and enhanced remyelination. However, the detected remyelination was not graft-derived as no differentiation of the transplanted cells towards the oligodendroglial phenotype was detected. In addition, the transplanted cells modulated the glial response and reduced apoptosis. These results suggest that the therapeutic potential of BM-MSCs for MS is not only dependent on their immunosuppressive and immunomodulatory nature but also on their ability to enhance endogenous repair and induce oligo/neuroprotection. Proving the efficacy of BM-MSCs in a non-immune model of MS and evaluating the underlying mechanisms should enrich our knowledge of how these cells exert their beneficial effects and may eventually help us to enhance and maintain an efficacious and sustainable cell therapy for MS.

Neuroprotective effect of ginger in the brain of streptozotocin-induced diabetic rats.

Diabetes mellitus results in neuronal damage caused by increased intracellular glucose leading to oxidative stress. Recent evidence revealed the potential of ginger for reducing diabetes-induced oxidative stress markers. The aim of this study is to investigate, for the first time, whether the antioxidant properties of ginger has beneficial effects on the structural brain damage associated with diabetes. We investigated the observable neurodegenerative changes in the frontal cortex, dentate gyrus, and cerebellum after 4, 6, and 8 weeks of streptozotocin (STZ)-induced diabetes in rats and the effect(s) of ginger (500 mg/kg/day). Sections of frontal cortex, dentate gyrus, and cerebellum were stained with hematoxylin and eosin and examined using light microscopy. In addition, quantitative immunohistochemical assessments of the expression of inducible NO synthase (iNOS), tumor necrosis factor (TNF)-α, caspase-3, glial fibrillary acidic protein (GFAP), acetylcholinesterase (AChE), and Ki67 were performed. Our results revealed a protective role of ginger on the diabetic brain via reducing oxidative stress, apoptosis, and inflammation. In addition, this study revealed that the beneficial effect of ginger was also mediated by modulating the astroglial response to the injury, reducing AChE expression, and improving neurogenesis. These results represent a new insight into the beneficial effects of ginger on the structural alterations of diabetic brain and suggest that ginger might be a potential therapeutic strategy for the treatment of diabetic-induced damage in brain.

Profiling metabolite changes in the neuronal differentiation of human striatal neural stem cells using 1H-magnetic resonance spectroscopy.

OBJECTIVE: Neural stem cells (NSCs) have been found to play an increasing clinical role in stroke. However, at present, it is not yet possible to noninvasively monitor their differentiation once implanted into the brain. METHODS: Here, we describe the use of high-resolution H-magnetic resonance spectroscopy (MRS) to define a metabolite profile of undifferentiated human striatal NSCs from the STROC05 cell line and their differentiation after 3-weeks of treatment with purmorphamine. RESULTS: The undifferentiated conditions were characterized by ~95% of cells expressing nestin and ~77% being Ki67(+)ve, indicating that these were still proliferating. Phosphophocholine+glycerophosphocholine (PC+GPC) as well as myo-Inositol (mI) were increased in these cells. PC+GPC and mI were markedly reduced upon differentiation, potentially serving as markers of the NSC state. Upon differentiation (~45% neurons, ~30% astrocytes, ~13% oligodendrocytes), the concentration of many metabolites decreased in absolute value. The decreasing trend of the N-acetyl-aspartate level was observed in differentiated cells when compared with NSCs. An increase in plasmalogen (enriched in myelin sheets) could potentially serve as a marker of oligodendrocytes. CONCLUSION: These metabolite characteristics of undifferentiated and differentiated NSCs provide a basis for exploration of their possible use as markers of differentiation after cell transplantation.

Implantation of undifferentiated and pre-differentiated human neural stem cells in the R6/2 transgenic mouse model of Huntington's disease.

BACKGROUND: Cell therapy is a potential therapeutic approach for several neurodegenetative disease, including Huntington Disease (HD). To evaluate the putative efficacy of cell therapy in HD, most studies have used excitotoxic animal models with only a few studies having been conducted in genetic animal models. Genetically modified animals should provide a more accurate representation of human HD, as they emulate the genetic basis of its etiology. RESULTS: In this study, we aimed to assess the therapeutic potential of a human striatal neural stem cell line (STROC05) implanted in the R6/2 transgenic mouse model of HD. As DARPP-32 GABAergic output neurons are predominately lost in HD, STROC05 cells were also pre-differentiated using purmorphamine, a hedgehog agonist, to yield a greater number of DARPP-32 cells. A bilateral injection of 4.5x105 cells of either undifferentiated or pre-differentiated DARPP-32 cells, however, did not affect outcome compared to a vehicle control injection. Both survival and neuronal differentiation remained poor with a mean of only 161 and 81 cells surviving in the undifferentiated and differentiated conditions respectively. Only a few cells expressed the neuronal marker Fox3. CONCLUSIONS: Although the rapid brain atrophy and short life-span of the R6/2 model constitute adverse conditions to detect potentially delayed treatment effects, significant technical hurdles, such as poor cell survival and differentiation, were also sub-optimal. Further consideration of these aspects is therefore needed in more enduring transgenic HD models to provide a definite assessment of this cell line's therapeutic relevance. However, a combination of treatments is likely needed to affect outcome in transgenic models of HD.

Purmorphamine increases DARPP-32 differentiation in human striatal neural stem cells through the Hedgehog pathway.

Transplantation of neural stem cells (NSCs) is a promising therapeutic approach for Huntington's disease (HD). HD is characterized by a progressive loss of medium-sized spiny neurons (MSNs) in the striatum. DARPP-32 (dopamine and cyclic AMP-regulated phosphoprotein, 32 kDa) is expressed in 98% of these MSNs. To establish an effective cell therapy for HD, the differentiation of human NSCs into MSNs is essential. Enhancing differentiation of NSCs is therefore an important aspect to optimize transplant efficacy. A comparison of 5 differentiation protocols indicated that the Hedgehog agonist purmorphamine (1 µM) most significantly increased the neuronal differentiation of a human striatal NSC line (STROC05). This 3-fold increase in neurons was associated with a dramatic reduction in proliferation as well as a decrease in astrocytic differentiation. A synergistic effect between purmorphamine and cell density even further increased neuronal differentiation from 20% to 30% within 7 days. Upon long-term differentiation (21 days), this combined differentiation protocol tripled the number of DARPP-32 cells (7%) and almost doubled the proportion of calbindin cells. However, there was no effect on calretinin cells. Differential expression of positional specification markers (DLX2, MASH1, MEIS2, GSH2, and NKX2.1) further confirmed the striatal identity of these differentiated cells. Purmorphamine resulted in a significant upregulation of the Hedgehog (Hh) signaling pathway (GLI1 expression). Cyclopamine, an Hh inhibitor, blocked this effect, indicating that purmorphamine specifically acts through this pathway to increase neuronal differentiation. These results demonstrate that small synthetic molecules can play a pivotal role in directing the differentiation of NSCs to optimize their therapeutic potential in HD.