Histological and Physiological Studies of the Effect of Bone Marrow-Derived Mesenchymal Stem Cells on Bleomycin Induced Lung Fibrosis in Adult Albino Rats

BACKGROUND@#Lung fibrosis is considered as an end stage for many lung diseases including lung inflammatory disease, autoimmune diseases and malignancy. There are limited therapeutic options with bad prognostic outcome. The aim of this study was to explore the effect of mesenchymal stem cells (MSCs) derived from bone marrow on Bleomycin (BLM) induced lung fibrosis in albino rats. @*METHODS@#30 adult female albino rats were distributed randomly into 4 groups; negative control group, Bleomycin induced lung fibrosis group, lung fibrosis treated with bone marrow-MSCs (BM-MSCs) and lung fibrosis treated with cell free media. Lung fibrosis was induced with a single dose of intratracheal instillation of BLM. BM-MSCs or cell free media were injected intravenously 28 days after induction and rats were sacrificed after another 28 days for assessment. Minute respiratory volume (MRV), forced vital capacity (FVC) and forced expiratory volume 1 (FEV1) were recorded using spirometer (Power lab data acquisition system). Histological assessment was performed by light microscopic examination of H&E, and Masson’s trichrome stained sections and was further supported by morphometric studies. In addition, electron microscopic examination to assess ultra-structural changes was done. Confocal Laser microscopy and PCR were used as tools to ensure MSCs homing in the lung. @*RESULTS@#Induction of lung fibrosis was confirmed by histological examination, which revealed disorganized lung architecture, thickened inter-alveolar septa due excessive collagen deposition together with inflammatory cellular infiltration. Moreover, pneumocytes depicted variable degenerative changes. Reduction in MRV, FVC and FEV1 were recorded. BM-MSCs treatment showed marked structural improvement with minimal cellular infiltration and collagen deposition and hence restored lung architecture, together with lung functions. @*CONCLUSION@#MSCs are promising potential therapy for lung fibrosis that could restore the normal structure and function of BLM induced lung fibrosis.

Histological and Physiological Studies of the Effect of Bone Marrow-Derived Mesenchymal Stem Cells on Bleomycin Induced Lung Fibrosis in Adult Albino Rats

BACKGROUND@#Lung fibrosis is considered as an end stage for many lung diseases including lung inflammatory disease, autoimmune diseases and malignancy. There are limited therapeutic options with bad prognostic outcome. The aim of this study was to explore the effect of mesenchymal stem cells (MSCs) derived from bone marrow on Bleomycin (BLM) induced lung fibrosis in albino rats. @*METHODS@#30 adult female albino rats were distributed randomly into 4 groups; negative control group, Bleomycin induced lung fibrosis group, lung fibrosis treated with bone marrow-MSCs (BM-MSCs) and lung fibrosis treated with cell free media. Lung fibrosis was induced with a single dose of intratracheal instillation of BLM. BM-MSCs or cell free media were injected intravenously 28 days after induction and rats were sacrificed after another 28 days for assessment. Minute respiratory volume (MRV), forced vital capacity (FVC) and forced expiratory volume 1 (FEV1) were recorded using spirometer (Power lab data acquisition system). Histological assessment was performed by light microscopic examination of H&E, and Masson’s trichrome stained sections and was further supported by morphometric studies. In addition, electron microscopic examination to assess ultra-structural changes was done. Confocal Laser microscopy and PCR were used as tools to ensure MSCs homing in the lung. @*RESULTS@#Induction of lung fibrosis was confirmed by histological examination, which revealed disorganized lung architecture, thickened inter-alveolar septa due excessive collagen deposition together with inflammatory cellular infiltration. Moreover, pneumocytes depicted variable degenerative changes. Reduction in MRV, FVC and FEV1 were recorded. BM-MSCs treatment showed marked structural improvement with minimal cellular infiltration and collagen deposition and hence restored lung architecture, together with lung functions. @*CONCLUSION@#MSCs are promising potential therapy for lung fibrosis that could restore the normal structure and function of BLM induced lung fibrosis.

Assessment of circulating MCP-1 level and 2518A>G gene polymorphism in systemic lupus erythematosus / Avaliação do nível de MCP-1 circulante e polimorfismo do gene 2518A> G no lúpus eritematoso sistêmico

Lupus nephritis (LN) is a major contributor to morbidity and mortality in patients with Systemic Lupus Erythematosus (SLE). This study aims to investigate the possible role of a functional polymorphism in the regulatory region of the monocyte chemo-attractant protein-1 (MCP-1) gene and MCP-1 blood level in the diagnosis of LN and in correlating the MCP-1 blood levels with disease activity. The study included 56 SLE patients and 56 controls. All the SLE patients suffered from LN. An analysis of MCP-1 gene polymorphism by polymerase chain reaction was performed followed by restriction fragment length polymorphism (PCR-RFLP) analysis and MCP-1 blood level was determined using the ELISA technique. Calculation of Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) was performed. Serologic tests included the determination of antinuclear antibody (ANA) and anti-double-stranded (ds) DNA antibodies, Complement C3 and C4 levels. A significant increase in the frequency of genotype A/G and a decrease in the frequency of genotype A/A were found among patients with active LN compared to inactive LN. There was a statistically significant difference in the blood level of MCP-1 between LN patients and controls. Also, MCP-1 blood levels were significantly higher in active LN patients than inactive LN. A significant positive linear correlation was detected between MCP-1 blood level and SLEDAI, creatinine, and 24 hours protein in LN patients. These results suggest that an A/G genotype together with the measurement of the blood level of MCP-1 can be a useful tool for detection and follow up of active LN.

A nefrite do lúpus (LN) é um dos principais contribuintes para a morbidade e mortalidade em pacientes com o Lúpus Eritematoso Sistémico (LES). Este estudo tem como objetivo investigar o possível papel de um polimorfismo funcional na região reguladora do gene da proteína quimioatraente de monócitos-1 (MCP-1) e do nível sanguíneo de MCP-1 no diagnóstico de LN e na correlação do sangue de MCP-1 níveis com atividade da doença. O estudo incluiu 56 pacientes com LES e 56 controles. Todos os pacientes com LES sofriam de LN. Uma análise do polimorfismo do gene MCP-1 por reação em cadeia da polimerase foi realizada seguida pela análise do polimorfismo do comprimento do fragmento de restrição (PCR-RFLP) e o nível sanguíneo do MCP-1 foi determinado pela técnica ELISA. O cálculo do índice de atividade da doença sistêmica do lúpus eritematoso (SLEDAI) foi realizado. Os testes sorológicos incluíram a determinação de anticorpos antinucleares (ANA) e anticorpos anti-DNA de fita dupla (ds), níveis de Complemento C3 e C4. Um aumento significativo na frequência do genótipo A/G e uma diminuição na frequência do genótipo A/A foram encontrados entre os pacientes com LN ativo em comparação com o LN inativo. Houve uma diferença estatisticamente significante no nível sanguíneo de MCP-1 entre pacientes com LN e controles. Além disso, os níveis sanguíneos de MCP-1 foram significativamente mais altos em pacientes com LN ativo do que com LN inativo. Uma correlação linear positiva significativa foi detectada entre o nível sanguíneo de MCP-1 e SLEDAI, creatinina e proteína de 24 horas em pacientes com LN. Esses resultados sugerem que um genótipo A/G, juntamente com a medição do nível sanguíneo de MCP-1, pode ser uma ferramenta útil para a detecção e acompanhamento do LN ativo

Electron microscopic study of the left caput epididymal epithelium of adult albino rats in an experimental left varicocele model

Varicocele is a vascular lesion characterized by dilatation and tortuosity of the pampiniform plexus of the spermatic cord. It is one of the most common causes of male infertility. Most studies have focused on testicular function. Meanwhile, there are limited data in the literature on its impact on the epididymal structure. The aim of this study was to assess the effect of an experimental left varicocele on the histological structure of the left caput epididymis of adult albino rats and the cauda epididymal sperm count. Forty adult male albino rats [220-250 g] were randomly allocated to four equal groups. In group I [control group], rats did not undergo any surgical intervention. In group II [sham-operated group], animals underwent sham operation. The rats of groups III and IV underwent partial ligation of the left renal vein to create a varicocele and were sacrificed after 4 weeks and 8 weeks, respectively. Left caput epididymal strips of five rats of each group were harvested and prepared for light and electron microscopic examinations. Sperm samples were obtained from the left cauda epididymides of the remaining animals of each group for sperm count, and the results were statistically analyzed. There was a positive correlation between the sperm count and the duration of varicocele. Group III rats revealed a significantly lower sperm count as compared with the control and sham-operated groups. A significantly greater reduction in the sperm count was found in group IV rats as compared with group III rats. Histologically, varicocele-induced left caput epididymal alterations mainly involved the principal cells. They included cytoplasmic vacuolation and widening of the intercellular spaces in semithin sections of group III rats. Ultrastructurally, increased vesicular and vacuolar contents of the apical cytoplasm along with dilatation of Golgi saccules and rough endoplasmic reticulum were observed. The tubular lumina depicted several retained defective spermatids and spermatozoa with retained cytoplasmic droplets. With an increase in varicocele duration [in group IV], apparent thinning out of the lining epithelium with few stereocilia was observed in semithin sections. Ultrastructurally, the principal cells revealed a relatively dense cytoplasm and rather heterochromatic nuclei. The lumina were bordered with short, sparse microvilli and showed several defective spermatozoa. Experimental left varicocele induces evident histological alterations of the ipsilateral caput epididymis of adult rats and also a reduction in the caudal sperm count, both of which are duration dependent

BRCA1 [l85 del AG] mutation among Egyptian breast cancer female patients

This study was conducted to estimate the frequency of BRCA1 [1 85delAG] mutation among Egyptian female patients with breast cancer. Forty selected female patients with breast cancer, 80 of their female relatives and 10 healthy females as a control group were included in this study. Result: The age of onset of breast cancer was below 40 years in 25 [62.5%] patients and above 40 years in 15 [37.5%] patients.There were significant differences among the patients regarding the age at menarche before 13 years [P=0.011, P<0.05], onset of breast cancer [P=0.000, P<0.001], parity [P=0.000, P<0.001], first delivery before 30 years of age [P=0.04, P<0.05], breast feeding [P=0.002, P<0.05], and positive family history [P=0.000, P<0.001]. The frequency of BRCA1 [1 85delAG] mutation was found among 10% of the patients group .Eight percent of patients with early onset below 40 years and 13.5% of patients with onset after 40 years were heterozygotes for the mutation. Three percent of patients with unilateral breast cancer, 40% of patients with bilateral breast cancer and 50% of patients with breast ovarian cancer were carrying the mutation. Our results indicated that breast ovarian cancer and bilateral breast cancer patients were likely to have BRCA1 [l85delAG] mutation than in unilateral breast cancer

relation between the duration of inhalation abuse of a volatile substance and its effect on the testis. a histological and biochemical study in adult male albino rats

Volatile substance abuse in general, and toluene inhalation in particular, for their neuropsychological effects, represents a significant problem in many developed and developing countries. The present work was designed to investigate the histopathological changes in the testis of adult male albino rats, induced by toluene vapour inhalation over different periods. The present study was carried out on forty adult male albino rats with body weights ranging from 60-100g. The animals were categorized into two groups: Group I: [Control Group] included ten rats received no treatment, Group II: [Toluene inhalants] included thirty adult rats exposed to toluene vapour inhalation. A clean dry piece of cotton was soaked with toluene liquid and placed in the covered cages three times daily, each for about thirty minutes for six days per week. These animals were subdivided into three equal subgroups according to the exposure period; Subgroup [A]: ten rats exposed to toluene vapour for two weeks, Subgroup [B]: ten rats exposed to toluene vapour for eight weeks, Subgroup [C]: ten rats exposed to toluene vapour for twelve weeks. At the end of each duration of the experiment, animals were scarificed by decapitation using light ether anesthesia after taking blood samples. I- Histological examination: Specimens were taken from the testis of all rats and processed for examination by light microscope using haematoxylin and eosin stain and ultrastructural study using the transmission electron microscope. II. Hormonal assay: The concentration of testosterone level, luteinizing hormone [LH] and follicle - stimulating hormone [FSH] were estimated by radio immunoassay. III. GAS chromatography: Concentration of toluene vapour in the blood was measured by High performance liquid chromatography. IV. Statistical analysis: The one way ANOVA test was applied to estimate the significant values of the hormonal assay for serum testosterone, LH and FSH and the 5% level of significance was chosen. The histopathological changes observed in the testis of rats exposed to toluene inhalation demonstrated its potentials to induce cytotoxic effects on the spermatogenic cells, Sertoli cells and the interstitial cells of Leydig. The severity of the toluene damaging potentials appeared to be dependent on and directly proportionate to the duration of toluene inhalation. So, the histological changes were mild and scattered in the testis specimens of group A [2 weeks inhalation] and was more severe in both eight and twelve weeks groups. The correlation between high performance liquid chromatography for toluene gas in blood, the biochemical gonadal and gonadotrophin hormonal assay and the histological assessment, explored the various mechanisms that were incorporated in the establishment of the toluene induced testicular injury. The present study proved the undoubting evidences for the damage potentials of toluene on the testis as the major reproductive organ in the male. Furthermore, the study showed the direct proportionality between the toxic effects of toluene vapor and the length of the exposure duration. Yet, the observed histological alterations were highly suggestive for a probable impaired reproduction in experimental animals which needs further study

Study of autonomic function in liver cirrhosis and hepatocellular carcinoma

Background and Aim: cardiovascular autonomic neuropathy is a known, but often unrecognized complication of liver cirrhosis and it can lead to many adverse effects including increased risk of mortality. However, few published studies are available about autonomic dysfunction in non alcoholic liver disease. Considering the adverse prognostic implications of autonomic neuropathy, the aim of the present study was to assess cardiovascular autonomic function in patients with liver cirrhosis and patients with hepatocellular carcinoma

Methods: The study included 60 cirrhotic patients [13 females and 47 males with mean age 53.5+/-7.6 years], 40 patients with hepatocellular carcinoma [8 females and 32 males with mean age 54.3+7.95], and 20 age and sex matched healthy controls. Clinical examination beside laboratory and radiological investigations necessary for diagnosis were done. Cardiovascular autonomic function using the standard tests was examined in patients and controls. We studied the presence and extent of autonomic dysfunction in the patients in relation to clinical and laboratory characteristics

Results: Compared to control subjects, both cirrhotics and hepatoma cases had impaired autonomic function tests, prolonged QTc [P<0.001] and higher autonomic function scores [P<0.001]. HR response to deep breathing was impaired more in hepatoma group than in cirrhotics [P<0.001]. 54 of cirrhotics [90%] and 37 of hepatoma patients [92.5%], had abnormal results of one or more autonomic function tests. No significant difference was found between cirrhosis and hepatoma groups as regard the distribution of autonomic dysfunction [P=0.245]. Parasympathetic dysfunction was more prevalent than sympathetic one in cirrhosis group [E:11.7% and D:41.7%, VS 36.7%] and also in hepatoma group [E:27.5% and D:32.5%, VS 32.5%]. Cirrhotics with autonomic neuropathy had significantly higher rate of CV autonomic neuropathy symptoms [P=0.002], higher rate of ascites [P<0.001], lower BMI [P<0.001], lower serum albumin [P<0.001] and higher INR [P=0.008] than those without neuropathy. In patient groups, combined AD increased in frequency according to child class [P<0.001 and <0.05 respectively], also the abnormal autonomic tests were significantly related to child class, serum albumin and INR. We concluded that AD, mainly parasympathetic is present with comparable frequency in liver cirrhosis and hepatoma patients and is related to the severity of liver failure. So, the standard autonomic function tests should be used during evaluation of such cases, also HR response to DB may help in screening cirrhotics for hepatoma

Abbreviation: AD: Autonomic dysfunction: AFP: alpha fetoprotein: AN: Autonomic neuropathy: BMI; body mass index; CV: cardiovascular: D: definite; DB deep breathing: DBP: diastolic blood pressure: E: early: FBS: fasting blood sugar HCC: hepatocellutar Carcinoma: HR: heart rate; HR LS: heart rate response to standing: HRV: heart rate variability; M+SD: mean + standard deviation; PH: postural hypotension; PPS: postprandial sugar: QTc: Corrected QT interval; SBP: systolic blood pressure: VR: Valsalva ratio