comparative study on some immunodiagnostic techniques currently used for the diagnosis of lymphatic filariasis in Egypt

Since 2002, the Egyptian Ministry of Health and Population in collaboration with the WHO implemented a lymphatic filariasis elimination campaign in Egypt. Yet, surveillance and follow-up measures detected asymptomatic microfilaraemic cases among the at risk population. The present work aimed to evaluate and compare tile diagnostic value of filarial immunoassays currently used in clinical as well as research institutions in Egypt regarding their sensitivity, specificity and reliability. Accordingly, Dirofilaria immitis adult crude antigen was evaluated as a capture molecule for anti-human filarial antibodies in serum of patients with occult as well as overt infections. A comparative study was done on a total of 82 subjects. Serum samples from all subjects were tested for the presence of anti-fliarial antibodies, utilizing reagents currently used for the diagnosis of lymphatic filariasis in Egypt. Tile commercially available Dirofilaria immitis adult crude antigen was used to detect serum IgG by indirect ELISA, lgG4 ELISA, Dot-ELISA and dipstick ELISA techniques. Among 18 microfilaraemic cases, indirect ELISA, IgG4 ELISA, Dot-ELISA and dipstick ELISA techniques were positive in 61.1%, 100%. 77.8% and 77, respectively. Corresponding figures among 20 symptomatic amicrofilaraemic cases were 95%, 0%. 85% and 80% with 75%, 95.5%, 88.6% and 88.6% specificity, respectively. Although IgG4 ELISA was more sensitive and specific in detection of microfilaraemic asymptomatic group than other ELISA techniques, Dot ELISA was found to be relatively more sensitive than other techniques among all groups of patients examined. The use of Dirofilaria immitis adult worm crude antigen in ELISA tests revealed an acceptable degree of sensitivity and specificity, in addition to its commercial availability. It is a reliable reagent with high immunodiagnostic potential

Nuclear accumulation of beta-catenin is a common and early event during neoplastic progression of Barrett esophagus

The majority of the adenocarcinomas arising in Barrett esophagus manifest clinically at an advanced stage and have a poor prognosis. As a result of this, much attention has been directed toward the exploration of markers for neoplastic progression in Barrett esophagus. The objective of the present study was to evaluate the role of beta-catenin expression in the progression of Barrett esophagus to adenocarcinoma by immunohistochemical analysis in 25 cases with adenocarcinoma and premalignant lesions and to examine its relationship to various prognostic factors. In our study, abnormal beta-catenin expression consisting of the loss of membranous staining and the appearance of the nuclear staining was found in 23 cases of esophageal adenocarcinoma [92%]. Of patients with the 23 tumors showing abnormal beta-catenin expression. 16 cases [64%] nuclear staining was present. This nuclear reaction was associated significantly with progression from metaplasia to low-grade dysplasia [P = 0.01]. In addition, in glands with clear histological transition from metaplasia to low-grade dysplasia. focal nuclear accumulation of beta-catenin was found only in the low-grade dysplastic areas. Also, we proved that the prevalence of reduced expression of beta-catenin on the membrane with or without nuclear staining increased significantly from low-grade to high-grade dysplasia to adenocarcinoma [P > 0.05]. According to clinicopathological correlation, abnormal beta-catenin expression was highly expressed in high grade tumors [G[3]] and tumors with lymph node metastasis with equal incidence 100%. Also, we found no gross difference in beta-catenin expression between Tis-T[1] and T[2]-T[3] tumors. 88.9% and 93.75%. respectively. These results indicate that abnormal expression of beta-catenin is a common and early event during neoplastic progression in Barrett esophagus and could be a valuable prognostic marker. Also it could be used to identify patients with Barrett esophagus who run a high risk of disease progression

Genes regulating apoptosis: an immunohistochemical study of BCL-2 and P53 expression in colorectal adenomas and carcinomas

Evidence is accumulating for the failure of apoptosis as an important factor in the evolution of colorectal cancer and its poor response to adjuvant therapy. The proto-oncogene Bcl-2 is a known inhibitor of apoptosis that may allow the accumulation and propagation of cells containing genetic alterations. Inactivation of P[53] the tumour suppressor gene are common in human colorectal cancer.This study aimed at evaluating the expression of Bcl-2 and P[53] proteins in colorectal adenomas versus carcinomas for better understanding of their role in the process of colorectal carcinogenesis and correlate their expression with the clinicopathologic features of these tumours. So we analyzed the expression of these markers in 11 colorectal adenomas and 14 carcinomas with the adjacent normal mucosa. Staining patterns were assessed semiquantitatively and correlated with each other and with age, sex, tumour size, site, tumour differentiation and Dukes' classification. A higher proportion of adenomas [54.5%] than carcinomas [36.4%] expressed BCL-2, Bcl-2 expression was more frequent in well differentiated carcinomas than moderately and poorly differentiated carcinomas. A lower proportion of adenomas [35.7%] than carcinomas [64.3%] expressed P[53] A total of [18.2%] of adenomas and [13.3%] of carcinomas expressed both markers. We suggested that Bcl-2 provides a survival advantage in the proliferative compartment of normal crypts of colorectal neoplasms, however, its expression is lost during the evolution from adenomas to carcinomas, whereas P[53] expression is increased. Although P[53] expression doesn't delineate aggressive behaviour and tumour progression in colorectal cancer, Bcl-2 provides a favorable marker for good differentiation in these tumours

Evaluation of circulating immune complexes in filariasis

Seventy five individuals were selected from El Korein city [endemic area for filariasis in Sharkia Governorate]. They were classified into 4 groups: Group I [20 asymptomatic microfilaraemic]. Group II [20 symptomatic microfilaraemic including 7 patients with fever and retrograde lymphangitis, 5 patients with lymphadinitis and 8 patients with pitting oedema]. Group III [25 symptomatic amicrofilaraemic with chronic filarial presentation: 15 cases with elephantiasis, 5 cases with hydrocoele and 5 cases with chylurea]. Group IV [10 endemic control].Additional fifth group of healthy control from zagazig city which is non endemic for filariasis [l0 non endemic control]. Each group was subjected to complete history, clinical examination, urine and stool analysis, Thick blood film stained by Giemsa stain. Sera separated and tested for detection of circulating antigens and circulating immune complexes using polyclonal antibodies by sandwich ELISA technique, circulating antigens were detected in 85% and 75% of symptomatic microfilaraemic and asymptomatic microfilaraemic groups respectively with high significant difference [P < 0.001] compared to non endemic control. Antigenaemia was detected in 32% of chronic filariasis and in 20% of endemic control but with non significant difference [P> 0.05]. Circulating immune complexes were detected in 100% in both symptomatic microfilaraenic and chronic cases, but in 50% of asymptomatic microfilaraemic and in 20% of endemic control.O.D. reading showed high significant difference with control group [P < 0.001]. Patients with chylurea showed highest level of circulating immune complexes among all clinical presentation while patients with fever and retrograde lymphangitis showed highest level among acute cases. From these results we can conclude that presence of circulating immune complexes in sera of endemic control put this group in risk of developing clinical disease by time

Humoral immune response before and after treatment of hydatid cyst

Twenty five patients infected with hydatid cyst were chosen from the inpatient clinics of Tropical Medicine Department of Zagazig University Hospital and were divided into three groups. The first group, included 8 patients, were given medical treatment with mebendazole in a dose of 10 mg/kg body weight/day each in six cycles of six weeks with free intervals of 2 weeks, the second group include 12 patients were undergoing surgical intervention by complete excision of the cyst and the third group include 5 nontreated patients considered as +ve control group. The fourth group was additional 5 healthy matched individuals taken as -ve control group. Each individual was subjected to physical and clinical examination, ultrasonography blood picture for differential leucocytic count and detection of Ig[G], Ig[M] and haemagglutinating antibodies by ELISA and IHAT respectively. All parameters were tested, before and every six months up to two years posttreatment. Before treatment: the level of immunoglobulins Ig[G], Ig[M] and haemagglitinating antibodies [HA], were high in infected cases. Ig[G] level was 1.286 +/- 0.582 and 1.338 +/- 0.449 in medical and surgical groups respectively versus 0.064 +/- 0.0362 in -ve control group with statistical high significant difference [P>0.001]. Ig[M] level was 0.5005 +/- 0.373 and 0.624 +/- 0.246 in medical and surgical treated groups respectively versus 0.057 +/- 0.333 in -ve control group with statistical significant difference [P>0.05]. HA titre was 1280.875 +/- 721.311 and 1365.33 +/- 630.22 in medical and, surgical groups, respectively versus 76.8 +/- 53.54 in -ve control group with statistical high significant difference [P>0.001]. After medical treatment: the level of immunoglobulins Ig[G], Ig[M] and HA was decreased in infected cases to reach normal level. Ig[G] level was gradually decreased to reach normal level in 12.5 and 25% of cases at 18 and 24 months respectively and it still elevated in 62.5% of cases for more than 2 years. Ig[M] level was gradually decreased to reach normal level in 37.5%, 62.5% and 12.5 of cases at 12, 18 and 24 months respectively. HA titre was gradually decreased to reach normal level in 37.5%, 37.5% and 12.5% of cases at 12, 18 and 24 months respectively and it still elevated in 12.5% of cases for more than 2 years postoperative. The level of Ig[G], Ig[M] and HA was decreased to become normal. Ig[G] level was decreased to reach normal in 16.67% and 58.33% of cases at 18 and 24 months respectively and it still elevated in the remaining 25% of cases for more than 2 years. IgM level was decreased to reach normal level in 56%, 33.3% and 8.33% of cases at 12, 18 and 24 months respectively and it still elevated in the remaining 8.33% of cases for more than 2 years. HA titre was decreased to reach normal level in 41.6%, 25% and 16.67% of cases respectively and it still elevated in remaining 16.67% of cases for more than 2 years. Eosinophilia was found in 30% of cases. There was marked improvement in the symptomatology after surgical intervention while after medical treatment there was mild improvement. Ultrasonography showed decrease in size of cyst after medical treatment through 1 1/2 year