Assessment of the osteogenic potential of alendronate on isolated adipose-derived stem cells: an ex-vivo and in-vivo study

Tissue engineering relies on the principle that mesenchymal stem cells are capable of differentiating to optimize almost all craniofacial structures. Temporary biomimetic scaffolds are necessary for accommodating cell growth and tissue genesis. The aim of this study was to evaluate the effect of alendronate on adipose-derived stem cells [ADSCs] from dogs and to compare bone regeneration in critical-sized calvarial bone defect in dogs using ADSCs in the presence and the absence of locally delivered alendronate. Seven dogs were used for the study. After isolating the adipose tissue from the inguinal pad of fat, stem cells were harvested and expanded in culture. The effect of alendronate 1 mg/ml on stem cells' osteogenic differentiation was tested for 7 days. Three critical-sized calvarial defects were created in each dog. One defect was filled with stem cells seeded on a chitosan scaffold and soaked in an osteogenic media, the second was fi lled with stem cells seeded on a chitosan scaffold and soaked with osteogenic medium, and the third one was fi lled with stem cells seeded on a chitosan scaffold. Bone formation was tested histologically after 8 weeks in each defect. Alendronate is capable of inducing osteogenic differentiation of ADSCs after 7 days of in-vitro culture. Bones such as trabeculae were deposited in alendronate and osteogenic medium defects, whereas the control group showed only fibrous tissue formation. There was no statistically significant difference in the surface area of the deposited bone trabeculae between the alendronate group and the osteogenic medium group. The surface area of individual bone trabeculae in this group was 147.99 +/- 14.803 compared with the osteogenic group. Alendronate may be used locally at a concentration of 10 mg/ml to induce osteogenic differentiation of ADSCs both in vitro and in vivo. The combination of a local, short-term alendronate treatment with ADSCs and biodegradable chitosan scaffold enhances the bone repair of a critical-sized calvarial defect in vivo

Procoagulant versus anticoagulant factors in cirrhotic patients

Liver cirrhosis leads to decreased production of clotting factors that are generally all produced in the liver except factor VIII and von Willebrand factor. However, cirrhotic patients are not protected from thrombosis. The present study aimed to assess the procoagulant and anticoagulant factors in cirrhotic patients with and without bleeding and/or thrombotic events. A total of 102 adult subjects were enroled: 51 cirrhotic patients and 51 healthy controls. After full history taking with special attention given to thromboembolic and haemorrhagic events, platelet count, serum albumin, bilirubin, international normalised ratio [INR], PT, partial thromboplastin time [PTT], hepatitis B surface antigen [HBsAg], hepatitis B core [HBc] antibodies, hepatitis C virus [HCV] antibodies, factor VIII, protein C, Protac-induced coagulation inhibition percentage [PICI%] assay and abdominal ultrasound were performed for patients and controls. Upper gastrointestinal endoscopy was conducted for the patients. Compared with control subjects, factor VIII and factor VIII/protein C were significantly higher, while protein C and PICI% were significantly lower among patients. Patients with liver cirrhosis may have a tendency for bleeding or thrombosis according to the balance of coagulant and anticoagulant status. PICI%, the assay that evaluated the functionality of the protein C anticoagulant system, was significantly lower in patients compared to control subjects. Accordingly, low PICI% and high factor VIII/protein C ratio can be taken as an index of hypercoagulability in cirrhotic patients

Potential Correlation between Carboxylic Acid Metabolites in Biomphalaria alexandrina Snails after Exposure to Schistosoma mansoni Infection

Carboxylic acids play an important role in both aerobic and anaerobic metabolic pathways of both the snail and the parasite. Monitoring the effects of infection by schistosome on Biomphalaria alexandrina carboxylic acids metabolic profiles represents a promising additional source of information about the state of metabolic system. We separated and quantified pyruvic, fumaric, malic, oxalic, and acetic acids using ion-suppression reversed-phase high performance liquid chromatography (HPLC) to detect correlations between these acids in both hemolymph and digestive gland gonad complex (DGG's) samples in a total of 300 B. alexandrina snails (150 infected and 150 controls) at different stages of infection. The results showed that the majority of metabolite pairs did not show significant correlations. However, some high correlations were found between the studied acids within the control group but not in other groups. More striking was the existence of reversed correlations between the same acids at different stages of infection. Some possible explanations of the underlying mechanisms were discussed. Ultimately, however, further data are required for resolving the responsible regulatory events. These findings highlight the potential of metabolomics as a novel approach for fundamental investigations of host-pathogen interactions as well as disease surveillance and control.

Evaluation of oligochromatography dipstick as a simplified molecular diagnostic tool for the diagnosis of cutaneous leishmaniasis

The demonstration of Leishmania parasite by microscopic examination of samples taken from skin lesions is of low sensitivity and requires expert parasitologists. On the other hand, molecular methods to detect Leishinania parasites are considered specific and sensitive. However, detection of amplicons by the classical methods is complex, expensive and needs equipments and highly trained personnel. This raises the demand for a simplified method of amplification and product detection. To evaluate the diagnostic accuracy of PCR Leishmania Oligo C-test for diagnosis of cutaneous leishmaniasis. The diagnostic accuracy of the Leishniania Oligo C-test was estimated in 30 skin scraping samples from patients presented at El Quorea specialized hospital, Taif, KSA. On the basis of parasitological detection of amastigotes by direct microscopic examination of Geimsa stained smears from skin lesions, patients were classified into confirmed cutaneous Ieishmaniasis [group A] and negative cases [group B]. Twenty patients were proved positive for Leishmania amastigotes by microscopic examination of stained smears. The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic efficacy of the Leishmania Oligo C-test were 90%, 90%, 95%, 82%, 90%, respectively. These results indicate that this simple molecular technique may be employed for rapid diagnosis of cutaneous leishmaniasis. The Leishmania Oligo C-test showed high specificity and sensitivity when compared to results obtained by microscopic examination of skin scraping samples. The assay is a promising tool for simplified molecular detection of Leishmania parasites

Carboxylic Acids as Biomarkers of Biomphalaria alexandrina Snails Infected with Schistosoma mansoni

Biomphalaria alexandrina snails play an indispensable role in transmission of schistosomiasis. Infection rates in field populations of snails are routinely determined by cercarial shedding neglecting prepatent snail infections, because of lack of a suitable method for diagnosis. The present study aimed at separation and quantification of oxalic, malic, acetic, pyruvic, and fumaric acids using ion-suppression reversed-phase high performance liquid chromatography (HPLC) to test the potentiality of these acids to be used as diagnostic and therapeutic biomarkers. The assay was done in both hemolymph and digestive gland-gonad complex (DGG) samples in a total of 300 B. alexandrina snails. All of the studied acids in both the hemolymph and tissue samples except for the fumaric acid in hemolymph appeared to be good diagnostic biomarkers as they provide not only a good discrimination between the infected snails from the control but also between the studied stages of infection from each other. The most sensitive discriminating acid was malic acid in hemolymph samples as it showed the highest F-ratio. Using the Z-score, malic acid was found to be a good potential therapeutic biomarker in the prepatency stage, oxalic acid and acetic acid in the stage of patency, and malic acid and acetic acid at 2 weeks after patency. Quantification of carboxylic acids, using HPLC strategy, was fast, easy, and accurate in prediction of infected and uninfected snails and possibly to detect the stage of infection. It seems also useful for detection of the most suitable acids to be used as drug targets.

Effect of sulphur and organic manure on thiosulphate efficiency as a nitrification inhibitor in alluvial soil

Incubation experiment was established under laboratory conditions on alluvial soil from south Giza Governorate to evaluate the effect of sulphur and organic manure on thiosulphate efficiency as a nitrification inhibitor. Ammonium sulphate and urea were applied as nitrogen sources, in addition to sodium thiosulphate and sulphur at rate of 100 mg Kg[-1] soil in mixture form with soil and organic manure at rate of 2%. Moisture was maintend at 80% of water holding capacity. Soil sample were taken every week for determination of soil acidity, ammonium, nitrate and thiosulphate concentrations. The obtained results indicated that thiosulphate alone delayed turnover of urea and Ammonium to nitrate and thus decreased soil pH. Ammonium sulphate treatment was more effective than urea one. Similar trend was noticd with a mixture of thiosulphate, sulphur and organic manure but it was more significant than thiosulphate alone. This effect can be noticed from NH[4]/NO[3] ratios which were higher in case of the mixture as compared to thiosulphate alone. The opposite relationship was observed between thiosulphate concentration in soil and nitrate release, where, it showed more significant effect by using the mixture of thiosulphate, sulphur and organic manure than as compared when thiosulphate alone was used

biochemical effects of dietary boron supplementation on immune response, steroid hormones and minerals metabolism

To investigate the effect of dietary boron [B] supplementation as sodium tetraborate on immune response, steroid hormones, and minerals metabolism in female albino rats. Eight groups each of eight female albino rats were divided as following-Group I [control group]; rats were feed on a standard basal diet. Group II; rats fed on basal diet+5 mg B/100 g diet. Group III; rats fed on basal diet+10 mg B/100 g diet. Group IV; rats fed on basal diet+15 mg B/l00 g diet. Group V; rats fed on basal diet+20 mg B/100 g diet. Group VI; rats fed on basal diet+25 mg B/100 g diet. Group VII; rats fed on basal diet+30 mg B/l00 g diet. Group VIII; rats fed on basal diet + 35 mg B/100 g diet for six weeks. The cell-mediated immune response was induced by an intradermal injection of 0.1 ml of phytohemagglutinin [PHA] [150 micro g/0.1 ml] in the flank region on both the right and left sides. Inflammatory response was measured as a change in skin thickness prior to and at 6, 12, 24 and 48h after PHA injection. In addition, serum levels of follicle stimulating hormone [FSH], luteinizing hormone [LH], progesterone, l7 beta-estradiol [E[2]], calcium, inorganic phosphorus, magnesium, total cholesterol, triacylglycerofs, and alkaline phosphatase [ALP] activity were measured. The immune response was increased by dietary boron supplementation at 6h alter injection compared to zero time. At 12h after injection the immune response had the lowest values in all rat groups fed diets containing different doses of dietary boron [5-35 mg B/100 g diet]. Serum levels of FSH and LH were not affected by boron supplementation While, serum concentration of 17 beta-estradiol increased significantly by increasing boron supplementation. In addition, serum levels of total cholesterol and tnacylglycerols were reduced in rats fed diets supplemented with 20-35 .mg B/100 g diet. In addition, serum levels of ALP activity was reduced in all treated groups compared to control group. Serum levels of calcium were increased' in rat groups fed diets containing 10-25.mg B/100 g diet, and then reduced as the supplementation of dietary boron increased [30 and 35 mg B/100 g diet]. A significant reduction was observed in serum inorganic phosphorus concentration in rats fed diets supplemented with boron at tested doses of 10-35 mg B/100 g diet. Dietary boron supplementation has regulatory effects on immune response increased estrogen levels, and maintenance of bone integrity, which may have a preventive effect on the development of osteoporosis

Detection and differentiation of entamoeba histolytica and entamoeba dispar isolates in clinical samples by PCR

A total of 140 out of 180 outpatients attended MISR University for Science and Technology Hospital complained of abdominal pain, diarrhoea and/or dysentery. Stool examination showed 47 [33.6%] had Entamoeba sp., 36 [25.7%] had cysts and 11 [7.9%] had trophozoites. Of 40 asymptomatic ones, 4 [10%] had cysts. A total of 51 positive stool samples for Entamoeba sp. [40 cysts and 11 trophozoites] were tested by Ne-sted Polymerase Chain Reaction [N-PCR] and Restriction Enzyme Digestion [RED] to clarify true E. histolytica from E. dispar. The results showed that 9/51 [17.6%] had E. dispar, while 31 [60.8%] had E. histolytica and 11 [21.6%] had dual infection with both E. histolytica and E. dispar. All E. histolytica PCR proved cases were from the symptomatic group, 11 had trophozoites and 34 had cysts. Thus, the result showed the potential use of molecular tools in detection of E. histolytica and E. dispar, and is a promising tool for epidemiology, particularly to differentiate pathogenic and non pathogenic Entamoeba sp

[Studies on staphylococcus aureus in milk and yoghurt treated by bifidobacteria]

Milk samples and lab-made yoghurt were used to investigate the antibacterial effect of bifidobacte-ria bifidum on Staphylococcus aureus pathogen. All samples were analyzed microbiologically for presence of S. aureus when fresh and after 2 and 5 days of storage at refrigeration temperature lor milk and yoghurt respectively proven to be contaminated with S. aureus from infected, non treated dairy cows and expermintally milk inoculated with S. aureus [5x 10[4] cfu/ml]. The obtained results indicated that there were adequate difference in S.aureus count in the examined samples treated with Bifidobacteria bifidum from zero time till the 2[nd] day of storage in bifudus milk. The reduction of S.aureus were 100% at 2[nd], 3[rd], 4[th] and 5[th] days of storage in bifudus yoghurt manufactured from inoculated milk with S.cinmis. The current study indicates that Bifido-bacteria may restrict and /or inhibit S. aureus in milk and yoghurt?

Co-infection with cryptosporidium parvum and cyclospora cayetanensis in ummunocompromised patients

Forty nine stool specimens collected from severe diarrheic patients. Eight were suffering from Hodgkin's lymphoma, and the rest were suffering from acute lymph plastic leukaemia. All were examined microscopically for protozoan parasites mainly, Cryptosporidium parvum and Cyclospora cayetanensis. Of the patients, 34 [69.4%] were positive and 15 [30.6%] were negative by both microscopy and nested PCR. An additional 12 [24.5%] who were negative by microscopy were positive by nested PCR. Stool examination revealed 16 cases with C. parvum, and 6 with C. cayetanensis, and 3 cases showed mixed infection. The results were compared with the established nested PCR assay to detect DNA directly from stool specimens. The patients <3 years old more affected by Cryptosporidium infection, unlike Cyclospora sp. Infection was in older age groups, which reflected the modes of parasite' transmission. Diarrheal illness was stronger for Cyclospora than for Cryptosporidium. After the extraction of DNA from stool, a 402-bp fragment of C. parvum, and 602 bp fragment of C. cayetanensis was amplified. The amplified products, 194-bp DNA fragment for C. parvum, and 306 bp DNA fragment of C. cayetanensis were used for the second run. This study indicated that primers were specific for DNA of C. parvum and C. cayetanensis. PCR detected a total of 22 [44.9%] positives for C. parvtim infection [6 negative by AF stool examination], and 12[24.5%] positives for C. cayetanensis. Infection [6 negative by AF stool examination], 7[14.3%] showed mixed infection [4 negative by AF stool examination], all microscopic negative specimens were positive by successive stool examination. Microscopy exhibited sensitivity of 72.7% for C. parvwn, 50% for C. cayetanensis and 100% specificity for both parasites compared to 100% sensitivity and specificity with PCR. So, PCR is more sensitive and easier to interpret but required more hands-on time to perform and is more expensive. However, PCR batch analysis reduces the cost considerably

Alphafetoprotein in screening for congenital hypothyroidism

This study was conducted on 500 full-term neonates and 25 older patients with congenital hypothyroidism [CH], newly or previously diagnosed. Alphafetoprotein [AFP] was elevated in two neonates. In one, persistent elevation of AFP and thyroid stimulating hormone [TSH] with low thyroxine [T4] were found [congenital hypothyroidism]. In the other, AFP, TSH and T4 levels normalized [transient hypothyroidism]. The mean AFP level in new CH patients was significantly higher than in previously diagnosed patients, and was higher in CH patients than in controls. Significant relationships were found between AFP and T4, AFP and TSH, and AFP and age. AFP is a sensitive indicator of thyroid status and can be used as a screening test for hypothyroidism from the first day of life and in follow-up of CH patients

Evaluation of cytology and colposcopy in single station management of cervical intraepithelial neoplasia using large loop excision of the transformation zone

In this study, one hundred and twelve patients had an excisional treatment for their cervical intraepithelial lesions. It can be concluded that the colposcopic assessment of the cervical intraepithelial lesions provides a safe reliable guidance to the single step management 'select and treat' of patients referred to the colposcopy clinic with abnormal cervical cytology. The cytological grading of the cervical intraepithelial cervical neoplasia is not a substitute for colposcopic evaluation and histological diagnosis. The referral of patients with low grade SIL or atypical cytology for colposcopic evaluation is mandatory, especially if it is recurrent or persistent

probability distribution for the permanent teeth loss

The main purpose of this study is to determine the probability distribution for the permanent teeth. Design: The study analysis based upon common facts in this area. Sample: Patients from Saddam Hospital were chosen for the study purposes. Among [354] patients have been [49%] males and [51%] females. Intervention: The permanent teeth have been divided in terms of anterior as group A, canines and premolars as group B, and molars as group C. Main outcome measures: X2 test was applied to evaluate the goodness of fit in the sample. The study presents a significant correlation in terms of teeth losses and age factors. In terms of groups, we find [for male]: the negative exponential distribution for group A, normal distribution for both groups B and C while the normal distribution for group A and Weible distribution for both groups B and C are the best distributions for fitting

Systemic prostacyclin in cirrhotic patients

A study of urinary excretion of 6-ketoprostaglandin F1oc in cirrhotic patients as a reliable index of systemic prostacyclin was done in order to understand the mechanism responsible for renal vascular homeostasis in cirrhotic patients. The study was done on 20 patients with cirrhosis and 20 control normal children. Patients without ascites and ascitic patients without renal failure showed increased urinary excretion of immunoreactive 6-ketoprostaglandin F1oc that was believed to reflect renal prostaglandin production and was found to be significantly increased in patients without ascites and in ascetic patients with preserved renal function. Increased renal production of vasodilator prostaglandin in cirrhosis was commonly interpreted as a defence against vasoconstrictor influences on renal hemodynamics. Enhanced renal prostaglandin synthesis in cirrhosis occurs either independently of elevated vasoconstrictor hormones or the activated vasoconstrictor systems enhance prostaglandin synthesis as a compensatory mechanism

Role of exchange transfusion in the treatment of neonatal septicemia

This study included 30 neonates [20 males and 10 females] with advanced neonatal sepsis whose gestational age ranged between 30-42 weeks [mean 35.16 +/- 0.12 weeks]. All the cases were subjected to full history taking, thorough clinical examination and investigations which included complete blood picture, C reactive protein and blood culture. All the cases underwent either a single or double volume exchange transfusion with fresh whole blood. The phagocytic power was estimated twice before and after exchange transfusion. All the cases were thrombocytopenic and had variable total leucocytic counts. Mortality rate was significantly increased in those neonates with leucopenia. C reactive protein was positive in 83.3% of cases. Blood culture was positive in all cases, Klebsiella was the commonest organism isolated [90% of cases]. Both phagocytic index and lytic index [tests for phagocytic power] increased significantly after exchange transfusion. The overall mortality was 53.3% in spite of the antimicrobial treatment and the exchange transfusion and it was highest among prematures less than 34 weeks. There was clinical improvement in 66.6% of cases after exchange: complete blood picture revealed improvement after the exchange especially for the percent stab cells. So, the judicious use of exchange transfusion is recommended especially in neonates with significant bleeding tendency or significant neutropenia.:AUTM:

Frequency of collagen disorders in Egyptian children - A one year study

172 patients with definite diagnosis of collagen diseases were submitted to this retrospective study. It aimed at studying the general frequency of collagenosis, the frequency of the different types, the predominant age, the sex distribution and the common presentations in Egyptian children [1-15 years]. It was found that the overall frequency of collagenosis was 4.4%. The frequency of JRA was 0.15%. SLE, DM and HSP was 0.03% for each, MCTD 0.08% and RHD 4.3%. All connective tissue diseases showed female predominance, except HSP where the only case reported was a boy and JRA that showed equal sex distribution. The age distribution, the commonest presentation and the laboratory aspects varied from one disease to another

Chromatographic studies on experimental candida keratitis

Chromatographic analysis and soluble protein contents of aqueous humour and crystalline lens were done in induced ocular infection of rabbit's eyes by Candida albicans in both treated and non-treated inflamed eyes as well as their normal controls. The results showed a significant decrease in the soluble protein contents and molecular weights in aqueous humour and crystalline lens of non-treated eyes. The topical treatment showed an important response which made the treated eyes to be slightly affected by the Candida albicans infections. We conclude from this study that early treatment prevents severe ocular complications and column chromatography is a sensitive technique for protein analysis of crystalline lens and aqueous humour