Diagnosis of clinical samples spotted on FTA cards using PCR-based methods

The broad clinical presentation of Leishmaniasis makes the diagnosis of current and past cases of this disease rather difficult. Differential diagnosis is important because diseases caused by other aetiologies and a clinical spectrum similar to that of leishmaniasis [e.g. leprosy, skin cancers and tuberculosis for CL; malaria and schistosomiasis for VL] are often present in endemic areas of endemicity. Presently, a variety of methods have been developed and tested to aid the identification and diagnosis of Leishmania. The advent of the PCR technology has opened new channels for the diagnosis of leishmaniasis in a variety of clinical materials. PCR is a simple, rapid procedure that has been adapted for diagnosis of leishmaniasis. A range of tools is currently available for the diagnosis and identification of leishmaniasis and Leishmania species, respectively. However, none of these diagnostic tools are examined and tested using samples spotted on FTA cards. Three different PCR-based approaches were examined including: kDNA minicircle, Leishmania 18S rRNA gene and PCR-RFLP of Intergenic region of ribosomal protein. PCR primers were designed that sit within the coding sequences of genes [relatively well conserved] but which amplify across the intervening intergenic sequence [relatively variable]. These were used in PCR-RFLP on reference isolates of 10 of the most important Leishmania species: L. donovani, L. infantum, L. major and L. tropica. Digestion of PCR products with restriction enzymes produced species-specific restriction patterns allowed discrimination of reference isolates. The kDNA minicircle primers are highly sensitive in diagnosis of both bone marrow and skin smears from FTA cards. Leishmania 18S rRNA gene conserved region is sensitive in identification of bone marrow smear but less sensitive in diagnosing skin smears. The intergenic nested PCR-RFLP using P5 and P6 as well as PI and P2 newly designed primers showed high level of reproducibility and sensitivity. Though, it was less sensitive than kDNA minicircle primers, but easily discriminated between Leishmania species

Fournier's Gangrene

Fournier's gangrene is relatively a rare surgical disease. It is a type of necrotizing fascitis involving the genito-perineal region. Both aerobic and anaerobic pathogens lie behind the pathogenesis. In his original description J.A. Fournier thought that it is an idiopathic disease without any obvious etiology. Many controversial issues exist about the pathogenesis and treatment of the disease. To investigate the possible correlation between the clinical outcomes [hospital stay, morbidity, mortality] and the early wound closure. Retrospective analysis was done from 1997-2000, in Al-Kindy Teaching Hospital. Eleven cases were included, with variable age and sex. Several predisposing and triggering factors for the disease were found in our work. Different techniques of surgical treatment were used in wound closure after initial resuscitation and frequent mini debridements, in order to close the defect as early as possible. The outcome of our study, despite different surgical procedures used to close the wounds, was encouraging. Hospital stay, morbidity and mortality were reduced in comparison with other studies. We found that the earlier the wound was closed; a shorter hospital stay, and less morbidity and mortality could be achieved

Validity of the " modified triple test " in the first trimester screening for congenital fetal malformations in the high risk group

To assess the clinical effectiveness of the combined test using fetal nuchal translucency [NT], maternal serum free beta subunit of human chorionic gonadotrophin [beta -HCG] and pregnancy associated plasma protein -A [PAPP-A] levels during gestational weeks 10 to 14 for screening of congenital fetal malformations. Settings: Department of Obstet. and Gyn., Minia University Hospital in co-operation with Feto-Maternal Unit, Am Shams University Hospital. Study Design: One thousand pregnant women [between. 10-14 weeks' gestation] at high risk for fetal malformations were recruited. They were subjected to detailed clinical evaluation, A three-generation family history, nuchal translucency, maternal serum PAPP-A and free beta -HCG. Amniocentesis was done only for 17 cases. Ninty-four malformed fetuses were picked up out of 1000 studied high-risk pregnant women [9.4%]. The most common congenital anomalies diagnosed were the cerebrospinal anomalies [48.7%], gastrointestinal anomalies [35.4%], and cardiac anomalies [15.8%]. Anomalies detected during the first scan were 49 out of 158 [DR 31.01%, Sensitivity 41.38%, specificity 98.63%].Anomalies detected during, the second scan were 125 out of 158 [DR 86.5%, sensitivity 81.6%%, specificity 99.31%]. Amniocentesis has been done for 17 cases [DR 94.1%]. Nuchal translucency had a detection rate [DR] of 80.2% compared with 52%, 43% for free beta - HCG and PAPP-A respectively. NT + free beta -HCGhad a better DR of 86%. The sensitivity of NT in comparison with amniocentesis was 73% whereas the sensitivity of the biochemical markers [PAPP-A and free beta-hCG] was 68%, 64% respectively. When NT and the biochemical markers were combined together, the sensitivity rose up to 89.5% in relation to amniocentesis. Nuchal translucency screening for fetal malformations together with serum screening has been given grade B recommendation by the RCOG working party. As a result of advances in U/S technology; visualization of the first trimester fetus has been markedly improved. By examining fetal anatomy and NT measurement at 10-l4wk, the majority of structural [68%] and chromosomal abnormalities [79%] can be diagnosed in early pregnancy. Adding the serum markers to the age and U/S will improve detection to 90% with a 5% false positive rate

Transmissible viral proventriculitis and stunting syndrome in broiler chickens in Egypt: 1. isolation and characterization of variant infectious bursal disease virus [IBDV]

In the present study, 46 broiler chicken flocks [2-4 weeks of age in farms located in four governorates] were examined. They were affected with proventriculitis and stunting syndrome and were vaccinated with classical infectious bursal disease vaccines. The affected flocks generally showed stunting, reduced growth rate and uneven weight distribution. The virus was detected in the bursa at 72 hours PI by ELISA and electron microscope confirming the persistence of the variant IBDV in the chicks, despite the presence of IBDV classical antibodies. The present study reported, for the first time, the association of IBDV with stunting syndrome and proventriculitis in broilers in Egypt

Gullian Barre Syndrome: an electrophysiological study

The aim of this study was to confirm the presence of pure axonal degeneration in patients with GBS and to provide further underst and ing to the underlying peripheral nerve pathology. This study included 25 patients with GBS. Electrophysiological studies demonstrated the presence of 7 patients with pure myelinopathy, 17 patients with myelino-axonopathy and proved the presence of pure axonopathy in one patient. This study showed the value of electrophysiology in elucidating the diagnosis as well as lending support to the basic pathophysiologic mechanisms underlying the disease

the value of F wave in the diagnosis of acute inflammatory demyelinating polyradiculoneuropathy

This study was undertaken with the aim of assessing the role of the F wave in the detection of proximal nerve affection in patients suffering from acute inflammatory demyelinating polyradiculoneuropathy and having normal peripheral motor nerve conduction velocity. This study included 32 patients, 20 males and 12 females, their ages ranged from 16 to 70 years, fulfilling the clinical criteria of the diagnosis. It was found that, 31.2% of the patients had proximal conduction delay, in 15.6% of the patients it was distal and multisegmental in 46.8%. It was concluded that the F wave technique is essential in establishing the diagnosis in some of those patients up to 30% in whom distal conduction velocities was normal