Efficacy of Human Platelet Rich Fibrin Exudate vs Fetal Bovine Serum on Proliferation and Differentiation of Dental Pulp Stem Cells

BACKGROUND AND OBJECTIVES: The imperative role of dental pulp stem cells (DPSCs) in regenerative therapy demands an in-vitro expansion which must deal with the safety and ethical problems associated with fetal bovine serum (FBS). The primary aim of this study was to compare the effects of human platelet rich fibrin (hPRF) exudate Vs FBS on proliferation and osteodifferentiation of human dental pulp stem cells (hDPSCs). The secondary one was to determine the optimum concentration of hPRF exudate inducing hDPSCs proliferation and osteodifferentiation. METHODS: The direct method was used to prepare hPRF exudate. hDPSCs were isolated from impacted mandibular third molars of twelve donors by the outgrowth method. For cell viability and proliferation rate testing, 96 well plates were used and the assay was done in duplicate and the trial repeated four times under the same conditions. Six wells were used to contain 10% FBS, serum free media, 1%, 5%, 10% and 20% concentrations of hPRF exudates, respectively. The proliferation assay was carried out by MTS tetrazolium cell proliferation assay kit and Elisa reader. The study design for osteodifferentiation protocol was exactly as the proliferation one and instead the assay was carried out by alizarin red with Elisa reader. RESULTS: Compared to 10% FBS, 10% hPRF exudate was the optimum concentration for hDPSCs proliferation, while 1% hPRF exudate was the optimum concentration for osteodifferentiation of hDPSCs. CONCLUSIONS: Avoiding the risk of zoonosis which may be occurred with FBS, it is recommended to use 10% hPRF exudate for proliferation and 1% for osteodifferentiation.

The Effect of Different Routes of Injection of Bone Marrow Mesenchymal Stem Cells on Parotid Glands of Rats Receiving Cisplatin: A Comparative Study

BACKGROUND AND OBJECTIVES: Cisplatin is a powerful antitumor chemotherapeutic agent that is widely used in the treatment of many cancers but it has many side effects on many organs including salivary glands. Bone marrow is considered to be a rich environment that comprises many types of stem cells of which BMSCs (Bone marrow mesenchymal stem cells) are the most studied with potentiality to differentiate into many cell types. This study was conducted to evaluate the effect of different routes of injection of BMSCs on parotid glands of rats receiving cisplatin. METHODS AND RESULTS: Sprague-Dawley rats were divided into 3 groups: a negative control group receiving phosphate buffered saline, a positive control group receiving cisplatin, and an experimental group where rats received cisplatin and then received iron oxide-labeled BMSCs by either intravenous or intraparotid routes or both. Animals were sacrificed at periods of 3,6,10 and 15 days after cisplatin injection, then histological, ultrastructural and immunohistochemical studies were done. The experimental stem cell treated group showed better histological features and increased PCNA proliferation index when compared to the control. The systemic and combination groups showed better results than the local group. Iron oxide-labeled cells were detected with Prussian blue stain. CONCLUSIONS: This study proved that BMSCs can improve cisplatin induced cytotoxicity in parotid glands. Systemic administration showed to have a better effect than local intraparotid administration and comparable effect to combined administration.

Study of cardiac abnormalities in children with Gaucher disease

Gaucher disease is the most prevalent lysosomal storage disease which results from inherited deficiency in the glucocerebrosidase enzyme. Three main clinical forms have been described, type I non-neurophatic, type II acute neuropathic and type III subacute neuropathic [1]. In this study, we present specific characteristics, as well as our experience in diagnosing the cardiac abnormalities in a group of Egyptian patients with this disease. The study included 22 patients with Gaucher disease attending Children's Hospital, Alexandria University. The recombinant enzyme imiglucerase [cerezyme] was given in a dose of 60 IU/kg/2 weeks [2]. Hemoglobin, plasma chitotriosidase and abdominal ultrasound were assessed before starting therapy and every 6 months. Molecular analysis was done to 17 patients. At presentation, the mean age was 7.94 +/- 6.26 years. 6 patients [27.2%] had type I, 16 patients had type III Gaucher disease [72.7%]. The commonest genotype was homozygous L444P which was present in 11 patients [50%] followed by homozygous D409H found in three patients [13.6%]. Gaucher's disease leads to deposition of glucocerebrosides in various organs. Recently, type IIIC Gaucher's disease, homozygous for the D409H mutation, has been identified; this is an ultra-rare cardiac variant with progressive calcification of aortic and/or mitral heart valves[3]. In this study the cardiac evaluation through clinical examination and investigations of the cases revealed that [50%] had positive cardiac findings

Telomerase activity, cytokeratin 20 and cytokeratin 19 in urine cells of bladder cancer patients

This work aims to search for markers suitable for the screening of bladder cancer, which should be specific, sensitive, reproducible, non-invasive and at acceptable cost. The study included 50 patients diagnosed as bladder cancer [35 TCC, 15 SCC] of different stages and grades, 30 patients with various urothelial diseases, besides 20 apparently healthy subjects of matched age and sex to the malignant group. A random midstream urine sample was collected in a sterile container for the determination of telomerase by RT-PCR, keratin 19 by ELSA CYFRA 21-i IRMA kit, keratin 20 by RT-PCR and immunohistochemical staining, and urine cytology. For all parameters [telomerase, Ki9, K20 and cytology] the malignant group was significantly different from both the benign and the control groups. None of the four studied parameters was correlated to the stage of the disease, and when it comes to grade, only Ki9 showed a significant positive correlation with grade both in TCC and SCC. When ROC curves for all parameters were compared, Ki9 had the largest area under the curve, and then comes K20. K 19 may be used as a biological marker for the diagnosis of bladder cancer Ki9 could not be used for differential diagnosis of different types of bladder cancer, meanwhile it could be a marker for differentiation that decreases in less differentiated tumors. As a tumor marker, K20 reflects inability to differentiate tumor type or grade in TCC, while in SCC of the bladder it is correlated with the grade. As a method, RT-PCR is superior to immunostaining for the detection of bladder cancer, meanwhile K20 immunohistochemistry [IHC] results were much better than urine cytology as a bladder cancer screening test. Haematuria and inflammation reduced the specificity of telomerase assay, which reduced its validity as a tumor marker of bladder cancer. Ki9 and K20 are the best candidates as screening tests for the diagnosis of bladder cancer, representing the highest sensitivity and specificity, beside the radiological and histopathology. Meanwhile, telomerase, although it was a sensitive enough marker, it reflected a high false positive rate

Long- term cardiac sequelae in patients with initial isolated rheumatic arthritis: a study of 100 patients in Alexandria, Egypt

The study was undertaken to assess the risk of development of rheumatic heart disease [RHD] in patients with initial isolated rheumatic arthritis [RA] and to identify the risk factors behind it. Clinical data of 100 patients with initial RA, who met the revised Jones criteria for acute rheumatic fever [ARF], diagnosed at the Alexandria University Children's Hospital, and their diagnoses dated back at least one year were retrospectively reviewed. Age at diagnosis was 7.9 +/- 3.29 years [mean +/- SD, range 3-16 years]. Echocardiographic findings at the initial episode were obtained from the hospital charts. The average duration of follow up of our patients was 7.5 +/- 4.42 years [range 1-21 years], and age at the time of enrollment was 15.4 +/- 5.09 years [mean +/- SD, range 7-31 years]. Complete clinical and echocardiographic examinations were done for all patients to detect the presence and severity of cardiac involvement [if any]. Our results showed that 23 patients had got one or more recurrences during the period of follow-up with a total of 39 episodes of recurrent rheumatic activity. Migratory polyarthritis constituted 66.7% of these episodes, carditis 17.9%, and 15.4% were in the form of chorea. Twelve of the 23 patients had definite valve lesions, although 6 of them did not experience carditis during recurrences. Seventeen patients had no clinical evidence of recurrent rheumatic activity, still developed RHD. So, a total of 29 of our patients [29%] developed RHD at the end of follow- up period. About 60% of them developed RHD within the first 5 years of the acute episode. Mitral regurgitation was the commonest valve lesion. Echo proved to be of great value in detecting cases with abnormal valve regurgitation in the absence of clinically audible murmur [silent murmur]. Patients with initial isolated RA should be carefully monitored, undergone frequent echo examinations for early detection of valve lesion which will influence the prognosis of these patients. Development of RHD in these patients should be considered while deciding the duration of secondary prophylaxis in these patients

Profile of secondary prophylaxis among children with rheumatic heart disease in Alexandria, Egypt

A cross-sectional study was conducted in specialist children's hospitals in Alexandria, which aimed to evaluate the current regimen of secondary prophylaxis for children suffering from rheumatic heart disease. Two-thirds of the patients had complied with their prophylactic regimen. Prophylactic failure occurred in one-third of the patients, raising doubts about the efficacy of the brands of penicillin prescribed. Recurrence of rheumatic fever was recorded in 37.3% of the patients, with semiurban or rural residence and non-compliance with secondary prophylaxis the significant risk factors. These unsatisfactory findings suggest the need for a more effective strategy of primary and secondary prophylaxis for controlling rheumatic fever in our community

Anthropometric measurments of the Egyptian ear auricle

Anthropometric measurements of the human ear auricle have been done focusing on the length, width, insertion and inclination of the auricle. Four hundred subjects of both genders aged from one month to 55 years old were included in this study. The averages of length, width, insertion and inclination were measured in each age group, recorded and analyzed. It was found that the rate of growth in length, insertion, width and inclination was 45, 30, 24 and 24% successively in the age period from newborn up to less than nine years age. In the next age periods from nine years to less than 15 years, the rate of growth was 28, 12, 8 and 2.5% as regards the width, length, insertion and inclination. After the age of 15 years, the growth of the auricle was minimal

Evidence of valve regurgitation by Doppler echocardiography in patients with rheumatic fever without clinical carditis

Because of the observation that patients with acute rheumatic fever without clinical carditis develop later rheumatic heart disease, and the postulation that such patients may have a subclinical [silent] carditis in the initial attack, we studied the heart in a group of such patients using Doppler echocardiography. Twenty five patients with recent isolated rheumatic arthritis and 25 patients with recent isolated rheumatic chorea in their initial attack were the subject of the study. They all had no past history of rheumatic fever or rheumatic heart disease. Doppler studies were done one to 28 days from the onset of symptoms. It was found that 50% of the patients had Doppler evidence of valve regurgitation, most commonly of the mitral and aortic valves. The severity of regurgitation was mild in most cases [75%] and moderate in the rest. 28% of the sample had two valves affected simultaneously. The results explain the finding of rheumatic heart disease in patients initially free of carditis several years after the first attack. They demonstrate that the use of Doppler echocardiography in the diagnosis of carditis in initial rheumatic fever attacks is essential They also demonstrate the importance of more prolonged bed rest, treatment with corticosteroids, and possibly life time prophylaxis in rheumatic fever patients who have Doppler evidence of valve regurgitation in the absence of acardiac murmur

Cholesterol gall stone dissolution using MTBE: study of efficacy, safety and prevention of recurrence

Twenty-three patients, 4 males and 19 females [mean age 38.6 years], with symptomatic noncomplicated cholesterol gall stone[s] were treated by percutaneous transhepatic direct contact MTBE dissolution. All patients were evaluated clinically and ultrasonographically before and after dissolution. Also, CT scan evaluation was done before dissolution to detect any calcification and to assure good gall bladder bed. Patients were classified into two groups: Group 1 [12 patients including 2 patients with residual stones], received ursodeoxycholic acid for three months after dissolution and group 2 [11 patients] did not receive ursodeoxycholic acid therapy. In 21 patients, first time gallbladder puncture was successful [91%], and in 2 obese patients second punctures [5 and 2 months later], were successful using stiffer catheter. Average dissolution time was 8.47 hours. Gall stone dissolution rate ranged from 0.98 to 3.4 cm/hour. The rate of dissolution was increased with the increase in stone number and stone burden. Complete stone dissolution was achieved in 21 patients [91.3%], residual gall bladder sediment not casting posterior shadow on ultrasound examination was detected in 3 patients [13.04%]. Residual stone fragments escaped dissolution in 2 patients [8.69%]. Side effects of the procedure were minimal and no major complications occurred. Results of 3 years follow up showed only one genuine recurrence [10%] out of 10 patients who received prophylactic post dissolution therapy in group 1, while 5 out of the 11 patients in group 2 showed recurrence [45.45%]. Direct contact MTBE cholesterol gall stone dissolution is safe and effective alternative for surgery. Post dissolution ursodeoxycholic acid therapy for 3 months is highly effective in preventing recurrence of stones

effects of the gut flora and portal bacteraemia on schistosomal hepatic fibrosis

Very high percentage of gastrointestinal colonization occurs in bilharzial patients. Portal bacteremia of gastrointestinal origin could be demonstrated in a good percentage of bilharzial patients. Bacterial colonization is enhanced by gastrointestinal stasis and hypoacidity of gastric juice. Liver function tests are disturbed and returned to normal following antibiotic therapy. Bacterial overgrowth in the GIT plays a major role in the production of hypoalbuminemia. Immunoglubulines [G, M and A] are increased in bilharzial patients in response to both schistosomal and bacterial antigens. The continuous antigenic stimulation and increased antigenic load by bacterial antigens from the GIT are factors sharing in hepatic affection after cessation of bilharzial infestation

Synthesis of some peptides containing o-and p-amino methyl benzoate, aminobenzamide, phthalamide and therephthalamide residues

It is well established that several substituted o-and p-aminobenzoic acid and phthalamide derivatives have played an important role as nitrogen donor ligands in the synthesis of a wide variety of metal complexes. They also possess bacteriostatic, tuberculostatic and fungicidal activities. Recently, we have reported the synthesis of several N-tosylpeptide derivatives and some of which were found to be biologically active. As a continuation of this work the synthesis of some N-tosyldi-and tri-glycyl-o-[or-p-] amino methylbenzoate and the corresponding N, N'-bis derivatives of c-and p-aminobenzamide, phthalamide and terephtalamide derivatives [VII-XXII] and the study of their biological activities and copper [II] complexes are described in this note

HLA antigens in pulmonary tuberculosis

A group of 15 patients with pulmonary tuberculosis was examined. Typing for HLA, A and B antigens was performed. The frequencies of HLA antigens in this group were compared with those of 100 controls

Detection of hepatitis B surface antigenaemia in patients with schistosomal hepatic fibrosis

The incidence of hepatitis B surface antigen [HB[s]Ag] among 80 patients with schistosomal hepatic fibrosis was determined using the new ELIZA technique. The correlations between the detectable antigen and disturbed liver functions as well as any accompanying clinical manifestations were discussed

association between HLA antigens and diabetes mellitus

HLA antigens were determined in 25 patients with diabetes mellitus. When the data are compared with other studies, there is a definite positive association of juvenile onset diabetes with HLA-B[8]. Analysis of the data indicated that HLA-B[8] possessed a primary association with the diabetogenic gene[s] among studied Egyptian population sample

Folate status in schistosomal hepatic fibrosis patients

The present study was carried on SHF cases to detect any evidence of folate deficiency as has been reported in cases of alcoholic cirrhosis. This work was carried out on 60 male cases of SHF cases of different stages [20 hepatomegalic cases, 15 hepatosplenomegalic cases and 25 cases with shrunken liver and splenomegaly]. and 15 matching normal controls. Estimation of serum folate, red cell folate and hepatic folate although showed variable degree of significant reduction in SHF cases when compared to normal controls, but the results of folate parameters did not reach the level of folate deficiency. Study of M.C.V., average neutrophilic lobe index and myelogram did not reflect any evidence of folate deficiency. Absence of folate deficiency in the studied groups of SHF may be explained by the high folate content of our average national Egyptian diet