Amelioration of exosome and mesenchymal stem cells in rats infected with diabetic nephropathy by attenuating early markers and aquaporin-1 expression.

Diabetic nephropathy (DN) is a prevalent diabetic microvascular condition. It is the leading cause of kidney disease in the advanced stages. There is no currently effective treatment available. This research aimed to investigate the curative potentials of exosomes isolated from mesenchymal stem cells affecting DN. This study was performed on 70 male adult albino rats. Adult rats were randomized into seven groups: Group I: Negative control group, Group II: DN group, Group III: Balanites treated group, Group IV: MSCs treated group, Group V: Exosome treated group, Group VI: Balanites + MSCs treated group and Group VII: Balanites + exosome treated group. Following the trial period, blood and renal tissues were subjected to biochemical, gene expression analyses, and histopathological examinations. Results showed that MDA was substantially increased, whereas TAC was significantly decreased in the kidney in the DN group compared to normal health rats. Undesired elevated values of MDA levels and a decrease in TAC were substantially ameliorated in groups co-administered Balanites aegyptiacae with MSCs or exosomes compared to the DN group. A substantial elevation in TNF-α and substantially diminished concentration of IGF-1 were noticed in DN rats compared to normal health rats. Compared to the DN group, the co-administration of Balanites aegyptiacae with MSCs or exosomes substantially improved the undesirable elevated values of TNF-α and IGF-1. Furthermore, in the DN group, the mRNA expression of Vanin-1, Nephrin, and collagen IV was significantly higher than in normal healthy rats. Compared with DN rats, Vanin-1, Nephrin, and collagen IV Upregulation were substantially reduced in groups co-administered Balanites aegyptiacae with MSCs or exosomes. In DN rats, AQP1 expression was significantly lower than in normal healthy rats. Furthermore, the groups co-administered Balanites aegyptiacae with MSCs or exosomes demonstrated a substantial increase in AQP1 mRNA expression compared to DN rats.

Diagnostic performance of kidney injury molecule-1 for detection of abnormal urinary albumin-to-creatinine ratio in type 2 diabetes mellitus.

In this study, we aimed to investigate whether serum and urinary levels of kidney injury molecule 1 (KIM-1) are increased in type 2 diabetes mellitus (DM) patients with albuminuria. While the correlation of urinary KIM-1 with renal impairment has been well established, the association with serum KIM-1 has not yet been so well documented. The present pilot study included Egyptian patients with type 2 diabetes (T2D): A total of 84 patients with T2D (age 49-64 years; 20 men) were included in the analysis of the present study. They were divided into two groups according to Alb/Cr ratio: the first group included 32 patients (38.1%) with abnormal Alb/Cr ratio (38.6 mg/g·Cr), and the second group included 52 patients with normal Alb/Cr ratio (17.9 mg/g·Cr). Serum and urinary KIM-1 were measured using enzyme-linked immunoassays. There was a significant difference between both serum and urine KIM-1 and Alb/Cr ratio. Patients with abnormal Alb/Cr ratio had significantly higher serum and urinary KIM-1. These results yielded sensitivity (75.0%) and specificity (96.2%) regarding serum KIM-1 with a cut-off point of 37.5 pg/mL, and sensitivity (93.8%) and specificity (88.5%) regarding urinary KIM1 with a cut-off point of 32.00 ng/g.Cr. Serum KIM-1 and urinary KIM-1 were significantly correlated with eGFR and Alb/Cr ratio. In this pilot study, we found that urinary, serum, and urinary levels of KIM-1 are associated with significant impairments in renal function among diabetic patients. Our results also showed that serum and urine KIM-1 can be used as potential biomarkers for diabetic kidney disease.

DNA integrity in diagnosis of premalignant lesions.

BACKGROUND: Carcinogenesis is a dynamic process which traditional biopsying can not keep up with. Saliva as fluid in the vicinity of the tumor can offer better insights to this process. This study aimed to identify the accuracy of salivary DNA integrity index in differentiating between oral premalignant lesions and oral cancer. MATERIAL AND METHODS: This phase II diagnostic test accuracy study included 93 patients divided into three groups: 30 oral cancer patients, 33 patients with oral premalignant lesions divided into 21 oral lichen planus patients and 12 patients with leukoplakia and 30 normal individuals who acted as controls. Oral rinse was collected from all participants and they all underwent conventional visual and tactile examination, and patients with oral lesions had the diagnosis confirmed by histopathological examination of tissue biopsy. DNA integrity index was determined as the ratio between ALU247 and ALU115 measured by qPCR. RESULTS: There was no statistically significant difference regarding ALU115, ALU247 and DNA integrity index between the three study groups. The index was significantly higher in the oral cancer group than the oral lichen planus patients, while no significant difference was found between the oral cancer and the leukoplakia cases. The DNA integrity index sensitivity, specificity, positive and negative predictive values were 73%, 45%, 55% and 65% respectively. CONCLUSIONS: Salivary DNA integrity index showed poor diagnostic abilities in differentiating between the oral cancer and premalignant lesions.

Toll-like receptor 7 mRNA is reduced in hepatitis C-based liver cirrhosis and hepatocellular carcinoma, out-performs alpha-fetoprotein levels, and with age and serum aspartate aminotransferase is a new diagnostic index.

BACKGROUND: Hepatitis B and C viruses are leading causes of liver cirrhosis and hepatocellular carcinoma (HCC). Toll-like receptor 7 (TLR-7) has been implicated in the pathogenesis of HCC linked to hepatitis B. We hypothesised a role of leukocyte TLR-7 mRNA in hepatitis C related liver cirrhosis and HCC, using alpha-fetoprotein (AFP) and liver function tests as comparators. METHODS: We recruited 102 patients with HCV-related HCC, 97 with HCV-related liver cirrhosis and 60 healthy controls. Quantification of TLR-7 mRNA was performed using real-time PCR, AFP and routine LFTs by standard techniques. RESULTS: TLR-7 mRNA levels were significantly lower in HCC patients compared to cirrhotic patients and lower again in healthy controls (p < 0.001 for trend). In multivariate analysis, age, aspartate transaminase (AST), AFP, and TLR-7 mRNA were significant predictors of HCC. The ROCC/AUC for age, AST and TLR-7 mRNA were all between 0.64 and 0.78 (all P < 0.01), but for AFP was 0.57 (95% CI 0.48-0.65, P = 0.09). We derived an index score using age, AST and TLR-7 mRNA for the diagnosis of HCC. The ROCC/AUC for the index was superior to all three root indices in the prediction of HCC. The index linked significantly with the Tokyo and Vienna liver cancer staging systems, but not with those of the CLIP and Okuda systems, in distinguishing HCC from liver cirrhosis. CONCLUSION: The combination of TLR-7 mRNA levels with age and AST improves the performance of TLR-7 in HCC diagnosis, out-performs alpha-fetoprotein and predicts early HCC.

A novel serum metabolome score for breast cancer diagnosis.

INTRODUCTION: Early detection of breast cancer is important in diagnosis and treatment, and so in enhancing patient survival and reducing death rates. Because of the low diagnostic sensitivity and specificity of widely used breast cancer biomarkers such as CA15-3, we hypothesised that a panel of new metabolic markers would provide superior sensitivity and specificity for this disease. MATERIAL & METHODS: We recruited 120 women with malignant breast cancer, 47 with benign breast disease and 55 females as a healthy control group. Metabolites 8-hydroxy-2'-deoxyguanosine, 1-methylguanosine, and 1-methyl adenosine were detected and quantified by gas chromatography-mass spectrometry, CA15.3 by ELISA. Cut-off values of individual and combined metabolome with CA15-3 were analysed using the receiver operating characteristics curve (ROCC) to test the efficiency of the candidate metabolome in identifying breast cancer. RESULTS: The overall linear trend of biomarkers across the groups was significant with highest levels in breast cancer (all p < 0.05). Using cut-off values of CA15-3, 8-hydroxy-2'-deoxyguanosine, 1-methylguanosine and 1-methyl adenosine of 30.5 U/l, 15.0 µg/l, 18.5 µg/l and 22.0 µg/l, respectively, diagnostic performance analyses of combined metabolome with CA15-3 gave a ROCC area under the curve of 0.94 (95% CI: 0.91-0.98)(p < 0.01) with good sensitivity (88.8%), specificity (86.8%) and efficiency (90.6%). Unlike CA15.3, the highest levels of each of the metabolite were in the early stage of breast cancer. CONCLUSION: The diagnostic combination test of candidate metabolome with CA15.3 may be a useful tool for the early detection of breast cancer and used as a metabolomics signature in this disease.

Persistent left superior vena cava in patient with right atrial myxoma undergoing open heart surgery. A case report and review of literature.

Persistent left superior vena cava (PLSVC) is a rare and asymptomatic congenital cardiovascular anomaly. Being asymptomatic, PLSVC was usually discovered while performing interventions (such as insertion of central lines, Swan-Ganz catheters, or placing pacemakers) through the left internal jugular vein or left subclavian veins. Commonly, PLSVC is detected not only as an isolated congenital anomaly, but also it can be associated with many other cardiac anomalies. Also, presence of a dilated coronary sinus on echocardiography should raise the suspicion of PLSVC. The diagnosis should be confirmed by contrast venography or computed tomography angiography. The present case is a female patient, 29 year old, who was undergoing elective excision of a right atrial mass, with closure of patent foramen ovale, and she had end-stage renal failure on regular hemodialysis three times weekly through a permicath inserted in the right subclavian vein.

Salivary microRNAs in oral cancer.

OBJECTIVE: This study investigated the use of three salivary microRNAs (miRNA-21, miRNA-184, and miRNA-145) as possible markers for malignant transformation in oral mucosal lesions. MATERIALS AND METHODS: Salivary whole unstimulated samples were collected from a study group of 100 subjects, consisting of 20 clinically healthy controls, 40 patients with oral potentially malignant disorders (PMDs) [20 with dysplastic lesions and 20 without dysplasia], 20 with biopsy-confirmed oral squamous cell carcinoma (OSCC), and 20 with recurrent aphthous stomatitis (RAS) as disease controls. Total RNA was isolated and purified from saliva samples using the microRNA Isolation Kit (Qiagen, UL). miRNA expression analysis was performed using qRT-PCR (Applied Biosystems). RESULTS: There was a highly significant increase in salivary miRNA-21 and miRNA-184 in OSCC and PMD (with and without dysplasia) when compared to healthy and disease controls (P < 0.001). Conversely, miRNA-145 levels showed a highly significant decrease in OSCC and PMD overall (P < 0.001). RAS cases showed no significant difference from normal controls in any measured miRNA (P > 0.05). The only microRNA to discriminate between OSCC and PMD with dysplasia was miRNA-184. When receiver operating characteristic curves were designed for the three miRNAs, cutoff points delineating the occurrence of malignant change were a fourfold increase in miRNA-21 with specificity 65% and sensitivity 65%, a 0.6 decrease in miRNA-145, with specificity 70% and sensitivity 60%, and a threefold increase of miRNA-184, with specificity 75% and sensitivity 80%. Calculating the area under the curve revealed that miRNA-184 was the only one among the studied miRNAs that provided good diagnostic value. CONCLUSION: Salivary determination of the miRNAs tested might furnish a noninvasive, rapid adjunctive aid for revealing malignant transformation in oral mucosal lesions, particularly miRNA-184.

Identification of Mediterranean mutation in Egyptian favism patients.

OBJECTIVES: Identify and screen the G6PD Mediterranean mutation in favism patients by applying a Amplification Refractory Mutation System Polymerase Chain Reaction (ARMS-PCR). PATIENTS AND METHODS: A total of 114 unrelated Egyptians patients were included in the present study; their ages ranged between (2-9) years with male to female ratio 4.5:1. G6PD activity was determined qualitatively from red cell hemolysate during attack. The G6PD Mediterranean mutation in patients has been identified by ARMS-PCR. RESULTS: G6PD deficiency was detected in 87.7%, (n=100). The frequency of G6PD Mediterranean mutation was (94.7%), (n=108). The association between G6PD deficiency and Mediterranean mutation was a highly significant. CONCLUSIONS: Glucose-6-phosphate dehydrogenase Mediterranean mutation is one of the most common mutations causing G6PD deficiency among Egyptian children with favism.

Oxidative stress and antioxidant defense in Egyptian favism patients.

BACKGROUND: Favism occurs as the result of intolerance to the ingesting of fava beans or to the inhalation of pollen from the Vicia faba plant. Patients with favism are always Glucose-6-phosphate dehydrogenase (G6PD)-deficient, but not all G6PD-deficient individuals develop hemolysis as a result of fava beans consumption. PATIENTS AND METHODS: Blood samples were collected from children with favism (n = 55) between age (2-12 years) on EDTA tubes divided into 3 groups: group 1 control group (n = 15), group 2 before blood transfusion (during hemolytic action) (n = 20) and group 3 after blood transfusion (treated) (n = 20). RESULTS: It was found that in group 2 GSH level was significantly low; (1.11 ± 0.39, p < 0.001) compared to controls (26.31 ± 5.26, p < 0.001). In group 3 after blood transfusion Level of GSH rose but remained lower than normal level (5.88 ± 2.33, p < 0.001) compared to controls. As for oxidative stress parameters, both levels of H2O2 and MDA were highly significant in group 2; (213.49 ± 57.56, p < 0.001), (98.05 ± 22.34, p < 0.001) compared to controls (3.75 ± 1.164, p < 0.001), (7.38 ± 2.07, p < 0.001), respectively. Moreover, in group 3 after blood transfusion, levels of H2O2 and MDA were decreased but remained high compared to controls (66.55 ± 22.49, p < 0.001), (47.18 ± 9.62, p < 0.001) sequentially. Also, there was a negative correlation between GSH that acts as antioxidant defense enzyme and each one of oxidative stress parameters MDA & H2O2. However, there was a positive correlation between H2O2 and MDA. CONCLUSIONS: From this study, it could be concluded that the favic patients have high oxidative stress (H2O2 and MDA) more than normal individuals and less antioxidant defense (GSH). With the passage of time these individuals, cells would be more vulnerable for H2O2-induced senescence.  

Interleukin 10 receptor alpha subunit (IL-10RA) gene polymorphism and IL-10 serum levels in Egyptian atopic patients.

OBJECTIVES: To assess the value of serum interleukin (IL) 10 levels as an immunological marker in atopy and to determine the role of an IL-10RA gene single nucleotide polymorphism (SNP) (serine 138-to-glycine exchange [S138G]) in the pathogenesis of atopic diseases. METHODS: Seventy-five patients with atopic disorders were compared with 25 age-matched healthy volunteers. Serum total immunoglobulin (Ig) E and IL-10 levels were measured by enzyme-linked immunosorbent assay and the IL-10RA gene S138G variant was screened by multiplex allele-specific polymerase chain reaction. RESULTS: There was a significant association between G allele frequencies of the S138G variant (62%, 60% and 68% for atopic asthma, atopic dermatitis, and allergic rhinitis, respectively) in atopic patients compared to in controls. There were significant differences in mean IgE levels but not mean serum IL-10 levels between the allelic variants in atopy groups. CONCLUSION: The IL-10RA gene SNP S138G may contribute to susceptibility to atopic diseases but serum IL-10 level is not a sensitive indicator in atopy.

Papillon-Lefèvre syndrome: neutrophil function in 15 cases fron 4 families in Egypt.

OBJECTIVE: The primary purpose of this study was to investigate the periodontal pathologic cause of Papillon-Lefèvre syndrome by comparing, with respect to neutrophil function, probands with Papillon-Lefèvre syndrome from 4 families in Egypt, unaffected siblings of the probands, and age-matched and gender-matched control subjects. STUDY DESIGN: Family histories and clinical dermal and oral manifestations of Papillon-Lefèvre syndrome were evaluated for 15 affected members of 4 families with the syndrome, 10 siblings of the probands, and 7 age-matched and gender-matched controls. Phagocytic and intracellular killing (lytic activity) of polymorphonuclear neutrophils was evaluated for all subjects according to a modification of the method of Wilkinson; opsonization was evaluated according to a modification of the methods of Cutler et al. Data were analyzed by means of analysis of variance. RESULTS: Family pedigrees were plotted, and consanguinity was noted in 3 of the 4 families with Papillon-Lefèvre syndrome. The means and SDs for phagocytic killing, lytic activity, and opsonization indices were as follows: probands, 4.76+/-1.99, 0.42+/-0.20, and 0.84+/-0.07; unaffected siblings, 10.4+/-1.3, 3.3+/-0.3, and 0.84+/-0.07; controls, 10.8+/-0.8, 3.5+/-0.6, and 0.85+/-0.05. The phagocytic killing and lytic activity indices demonstrated significance between the probands and both siblings and controls (P<.0005), whereas the opsonization index did not demonstrate significance between groups. CONCLUSIONS: Significantly decreased neutrophil function in probands with Papillon-Lefèvre syndrome was demonstrated with respect to neutrophil phagocytotic and lytic activity but not with respect to opsonization. Therefore, specific neutrophil dysfunction appears to be etiologically involved in this disorder.

A new concept for the pathogenesis and treatment strategies for oral lichen planus.

Mounting evidence points out to the immunologic basis of lichen planus (LP). Tumour necrosis factor (TNF-alpha) is a cytokine with multiple biologic activities, some of which may be relevant to the pathogenesis of LP. The present study was thus conducted to assess its production by peripheral blood leucocytes in patients with active lesions of oral lichen planus (OLP) compared to age matched healthy control volunteers. For the assessment, the quantitative immunometric sandwich technique was employed. Significantly greater amounts of TNF-alpha were found in serum samples of OLP patients compared to healthy individuals, suggesting a possible association between this cytokine and the pathogenesis of OLP and pointing out to the primary initiative role played by altered keratinocytes. This pointed out the beneficial effect of anti-TNF drugs in the management of OLP.

Bacteriological and clinical assessment of tetracycline as root conditioning in adjunct to periodontal surgery.

The present study was conducted on 14 quadrants from seven solicited patients. After initial phase of plaque control, six different clinical parameters were recorded and scaling and root planning performed. In each of the seven patients two Modified Widman flaps were done. One served as a control and the other as experimental in which irrigation by Tetracycline-Hcl solution 100 mg/ml for 5 minutes was performed. On comparing the two treatment modalities the tetracycline-Hcl root conditioning showed a better improvement in all the clinical parameters tested. Tetracycline irrigation gave less bacterial counts than the control group right after irrigation, however after two weeks the bacterial counts increased again and were insignificantly different in the two groups.

A bacteriologic aid in the differential diagnosis of periapical and periodontal abscesses.

The aim of the present investigation was to determine whether spirochaetal count could help differentiate between periodontal and periapical abscesses. 23 draining abscesses were included, where 15 were diagnosed as periodontal and 8 as periapical, based on clinical and radiographic data. Exudate samples were collected aseptically and examined by both darkfield microscopy and Gram-stained smear. The results revealed a statistically high increase in spirochaetal count in periodontal abscesses compared to periapical ones. The obvious difference appeared adequate to differentiate both lesions and establish a rapid chairside diagnostic aid.

Prepubertal periodontitis: a genetic analysis.

Genetic factors were recognized as possible predisposing factors in the etiopathogenesis of prepubertal periodontitis (PP). Consequently, the present work was carried out to determine the possible mode of inheritance and if there were any associated chromosomal aberrations in cases with PP. The study included 3 families with probands manifesting PP (the families included 18 individuals). Pedigree analysis was carried out as well as chromosomal analysis. The results pointed out to the possibility of new mutations arising due to various environmental factors (including the use of pesticides and their handling), in addition to a possible autosomal recessive inheritance. Chromosomal analysis showed no association between a certain chromosomal aberration and PP. An interesting finding was that a girl proband proved to be Xo female, i.e. a case of Turner's syndrome. This could be considered the first report of Turner's syndrome with PP.

Effects of sheesha and cigarette smoking on pulmonary function of Saudi males and females.

The effects of sheesha or cigarette smoking on pulmonary function values were examined in 441 and 154 male and female smokers respectively. Cigarette smokers showed a marked decline in pulmonary function values with increasing age that was more pronounced than in non-smokers. VC, FEV1 and FVC mean values of male sheesha smokers were significantly lower than those of corresponding non-smokers with all pulmonary function measurements demonstrating a decline with age in sheesha smokers, particularly in the age group 20-49. The extent of the decline in pulmonary function measurements in young and middle aged female sheesha smokers (20-45 years) was not as pronounced as that of older female smokers. It is concluded that sheesha smoking as well as cigarette smoking, will produce similar harmful effects on the function of ventilatory capacity of both male and female subjects and increase the risk of developing obstructive airway disease, with sheesha smokers being at a greater risk.

A study of carboxyhaemoglobin levels of cigarette and sheesha smokers in Saudi Arabia.

A single carboxyhaemoglobin (COHb) estimation of late evening blood sample among non-smokers, cigarette smokers, and sheesha smokers was evaluated among Saudis. The COHb level in smokers of 15 to 40 cigarettes a day ranged between 0.7 and 10.3 with a mean value of 6.1 +/- 2.58 COHb. Values among sheesha smokers ranged between 6.5 and 13.9 with a mean value of 8.8 +/- 1.83, significantly higher than those of cigarette smokers (P less than 0.001) for a given degree of exposure to tobacco smoke.