Interstitial cells of cajal in normal human alimentary tract: an immunohistochemical study

Objectiver lnterstitial cells of Cajal [ICC] are c-kit positive immunoreactive cells which are thought to play an important role in the control of gut motility. The work aimed at studying the morphology of ICC and precisely localize their regional and transmural pattern of distribution in normal human alimentary tract. The study included 102 normal human alimentary tract specimens obtained from male patients with a mean age 37.92 +/- 8.53. All sections were stained with hematoxylin and eosin and c-kit immunohistochemical staining. Immunohistochemically stained sections were submitted for a computer aided image analytical study to detect the area percent of immunoreactive cells. The data obtained was statistically analyzed. ICC could not be demonstrated in H and E stained sections. Immunohistochemically, two morphological subtypes of ICC were recognized, a spindle bipolar and stellate multipolar forms. ICC were detected in the myenteric plexus layer of the esophagus, corpus, pylorus, small intestine, colon and rectum. Intramuscular ICC could be demonstrated in the esophagus, fundus, corpus, pylorus, colon, rectum and anal canal. ICC at the deep muscular plexus were found only in the small intestine. In the pylorus, colon and rectum, ICC were also found at the submucosal border of the circular muscle layer. The wide distribution of ICC all over the human alimentary tract is compatible with their physiological role being important mediators of gut motility

Administration of tetanus toxoid [TT] to pregnant women versus tetanus and reduced diphtheria [Td]: a randomized controlled

We conducted a clinical trial to compare the immunogenicity, reactogenicity and efficacy of Tetanus Toxoid [TT] and the combined Tetanus and reduced Diphtheria [Td] in pregnant women residing four rural communities in Egypt. The study has been designed as a randomized controlled trial. Pregnant women in each of the four villages received either TT or Td randomly. Both TT and Td vaccines are manufactured by the Egyptian Company for Biological Products and Vaccines [VACSERA] in Egypt. A total of 131 pregnant women were enrolled at the time of their antenatal care visit [at the beginning of their fifth gestational month] to one of four health units in Abu Homos district, Beheira Governorate, Egypt. Previously un-immunized women received at random 2 doses of TT or Td eight weeks apart during their pregnancy. Active outpatient follow-up for adverse events was done on the third day after each dose. Local [pain, redness and swelling] or general [fever, malaise and headache or body aches] reactions during the 3-day post-dosing interval served as the primary safety end point. Blood was collected 3 times from each subject to detect antibody level against tetanus and diphtheria by ELISA. The first sample was collected immediately before the first dose, the second before the 2[nd] dose and the third sample one week after delivery. Active surveillance home visits to all study participants were conducted twice. The first home visit was during the first week after delivery and the second one month after labor to report the health status of the mother and the baby. A total of 122 pregnant women received 2 correct doses had inter-dose intervals within the allowable range and provided 3 samples of blood, were included in per protocol analysis [62 in the TT group and 60 in the Td group]. There was no statistically significant inter group difference in the percentage of subject reporting the primary safety endpoint [fever, malaise, body ache, headache] or local reactions at the site of injection as redness and swelling, during the 3-day after each dose. There was a statistically significant greater reporting of pain at injection site in the Td group after dose I and II compared to the TT group. Home visits revealed normal mothers and babies on clinical examination in both groups. However, more babies in the TT group suffered from physiological jaundice. All women in the two groups acquired protective immunity for tetanus, determined as possession of neutralizing antibodies at titre>0.10 IU/ml after completing vaccination. However, the geometric mean titres of tetanus post dose I and II were significantly higher in vaccinees in the TT group [P<0.001]. For diphtheria, post vaccination seroprotection [titre>0.10 lU/ml] was significantly higher in the group received Id than the TT group. Geometric mean Titres of diphtheria post dose II were significantly higher in vaccinees in the Td compared to the other group [P<0.0001]. These finding demonstrared that the use o Td vaccine improves immunogenicity for both tetanus and diphtheria over the use of 'IT vaccine alone and may be recommended to replace TT in immunization of pregnant women.

Expression of human recombinase activating gene 1 and recombinase activating gene 2 [rag 1 and rag 2] in acute lymphoblastic leukemia

The process of V[D]J recombination is limited and controlled by the enzymatic activity of cytoplasmic proteins called recombinase which are the products of the activating two genes called Recombinase Activating Gene 1 and Recombinase Activating Gene 2 [RAG1 and RAG 2]. Both genes are expressed in immature B and T lymphocytes and activated V[D]J rearrangement in Ig and TCR genes which are directed in cis by recombination signals sequences [RSSs]. Also, they show variable expression in lymphoid malignancies of both B and T-cell types. This study aimed to focus on the role of the two cytoplasmic proteins [RAG1 and RAG2] in the developing of both B and T ALL, correlation to the stages of differentiation and also, their possible prognostic significance. This study included 40 newly diagnosed acute lymphoblastic leukemic patients [ALL] their age ranged from 1-10 years. Assessment of RAG1 and RAG2 expression by reverse transcriptase polymerase chain reaction [RT-PCR] was done using peripheral blood sample. RAG1 and RAG2 positive expression was higher in frequency in pediatric ALL cases compared to control group, the difference was statistically significant [P<0.01]. Among the RAG1 and RAG2 positive ALL cases 25% were pro-B-ALL and 25% CALL and 31.2% pre-B-ALL and 18.8% were early T-ALL. On the other hand RAG1 and RAG2 negative ALL cases showed higher frequency of CALL phenotype [83.3%] while pro-B ALL, pre-B ALL and early-T ALL were 4.2% and 8.3% and 4.2% respectively. The RAG1 and RAG2 initially positive ALL cases studied had poor prognosis, where 37.5% relapsed and 25%, while 37.5 were in continuous complete remission. However among the RAG1 and RAG2 negative ALL cases studied, 91.7% had good prognosis with complete remission, while only one patient [4.2%] relapsed and one died [4.2%]. A statistical significant association between RAG1 and RAG 2 positive expression and poor prognosis was noticed. RAG1 and RAG2 could be used as prognostic marker in lymphoid malignancies and its sub-classification

Serum and ascitic fluid leptin levels in cirrhotic patients: relation to other clinicopathologic parameters

Leptin is an adipocytokine peptide involved in the homeostasis of body composition. Alterations in leptin regulation have been observed in liver cirrhosis. We aimed to assess serum and ascitic fluid leptin levels in a group of patients with decompensated liver cirrhosis and to evaluate these levels in relation to tumor necrosis factor-alpha levels. We assessed both serum and ascitic fluid leptin levels by a radioimmunoassay method, in 40 patients with posthepatic cirrhosis. We calculated body mass index [BMI] as Kg/m2 in all patients and assessed other laboratory biochemical parameters of liver functions. We excluded patients with Spontaneous bacterial peritonitis and all forms of non cirrhotic ascites. Both serum and ascitic leptin levels were correlated with BMI. Ascitic fluid leptin levels [13.1 +/- 10.9 ng/ml] were twice as high as serum levels [7.0 +/- 6.4 ng/ml] and the ascitic fluid/serum ratio of leptin was >1 in all patients. We suggested that this parameter can be used as one of the characteristics of cirrhotic ascites. Serum and ascitic fluid leptin levels were positively highly correlated [r = 0.76 p<0.001] while no correlation was found between TNF-alpha levels in serum and ascites [r=0.29 p>0.05]. We concluded from this study that ascitic fluid leptin levels of cirrhotic patients with sterile ascites are on average two times higher than circulating levels of this hormone, a useful marker of characterizing cirrhotic ascites. These findings suggest that intra-abdominal production of leptin in cirrhotic ascites by peritoneal adipocytes may explain this local rise of leptin levels and this is largely regulated and interrelated to TNF-alpha. Both proinflammatory adipocytokines are related to the metabolic picture in cirrhotic patients particularly anorexia, cachexia and increased energy expenditure. This may represent a therapeutic target in the future by means of introducing anticytokine therapy or chemoprophylactic measures that may ameliorate the release of these cytokines from stimulated monocytes, T-cells or from adipocytes

Production of antigenic recombinant cholera toxin subunit B.

Cholera is a major public health problem confronting developing countries, where outbreaks occur in a regular seasonal pattern and are particularly associated with poverty and poor sanitation. Primer set was designed to flank binding region of cholera toxin subunit B [ctxB] gene that was amplified using PCR. PCR products were purified using gel purification technique. Cholera ctxB gene was cloned into cloning vector. Stop codons contained in the insert were deleted and then the insert was subcloned into pQE expression vector. Cloned vectors were subjected to DNA sequence analysis. Small-scale culture was done for preparative rCTB production and purification. Mice were immunized with purified rCTB to test ability of rCTB to elicit antibody production. Primer set showed the ability to detect and excise ctxb gene successfully form cholera genomic DNA by PCR. After deleting the stop codons, sequence analysis revealed that the insert was in open reading frame with start codon of the vector. Small bacterial cultures revealed the presence of specific band at approximately 12-125 Kda in induced culture. rCTB was able to elicit specific antibody production after animal immunization

Antigenic potential of whole cell-recombinant cholera toxin subunit B [WC-CTB] preparation

Colonisation of the small intestine by V. cholerae, a typical non-invasive pathogen, is an important early step in the pathogenesis of cholera. In the present study, trial to make cholera whole-cell vaccine with addition of recombinant B subunit of cholera toxin [rCTB] was-done. Cholera chB gene was cloned into pQE expression vector. Large-scale culture was done for preparative rCTB production and purification. Commercial CTB from V. cholerae was used for induction of polyclonal anti-CTB antibodies in mice. These polyclonal antibodies were used to test the antigenicity and identity of rCTB. Cholera whole-cell vaccine was prepared by resuspending equal volumes of dead Inaba and Ogawa vaccine strains in PBS. Orochol E Berna was used as control vaccine. Mice were divided into 4 groups [20 mice each]; group 1 [G1]: control unimmunised, group 2 [62]: rCTB immunized, group 3 [G3]: rCTB + killed Inaba and Ogawa immunised, and group 4 [G4]: Orochol immunized. Sera and faeces from all groups were collected and used in evaluation of anti-rCTB antibody levels. The spleens of animals were aseptically removed and used in lymphoproliferative assay. Small bacterial cultures revealed the presence of specific band at approximately 12-12.5 KDa in induced culture. Anti-commercial CTB antibodies were successfully prepared and used in Western blot analysis and verified the presence and antigenicity of rCTB. Large scale production and purification of rCTB resulted in 12.9 mg protein. Both serum and secretory antibody level in mice of G2 was significantly less than in mice of both G3 and G4. G3 was significantly higher than 62, while there was no significant difference between G3 and G4. G4 was significantly higher than G2, while there was no significant difference between G4 and G3. Stimulation index of splenic cells in G 1 was not significantly different from that of G2, while it was significantly lower than GB and G4. SI of G2 was significantly lower than G3 and G4. G3 was significantly higher than G1, G2, and G4. G4 was significantly higher than 01 and 02, but it was significantly lower than G3. The results in this study showed the ability of rCT B to induce immune responses in the presence of cholera bacteria more than if used alone. The inclusion of rCTB in addition to vibrios, increased systemic, intestinal, and cellular responses. Depending on previous studies, adding any new cholera strain which are present or will appear in future is possible. In addition, other vaccines [either killed bacteria or vaccine subunit] can be added to the formula indicated in this study

Concurrent weekly paclitaxel and carboplatin with thoracic radiation followed by consolidation chemotherapy for locally advanced non-small cell lung cancer [NSCLC]

To assess the feasibility and activity of chemoradiation program using paclitaxel and carboplatin-based combined-modality therapy based on the unique radiation-sensitizing properties of these agents in locally advanced unresectable stage III NSCLC. Twenty-seven patients with unresectable NSCLC were evaluable, with a median age of 54 years [range 44-69]. Eleven patients [41%] had stage IIIA and 16 [59%] had stage IIIB. Treatment consisted of paclitaxel [50 mg/m[2] i.v. infusion over 1 hour] followed by carboplatin [AUC 2.0 i.v. bolus infusion over 30 min.] administered weekly concurrently with radiation on days 1,8,15,22,29,36. Thoracic radiation was delivered in daily doses of 2 Gy to a total dose of 65 Gy over 6.5 weeks. Consolidation chemotherapy: After chemoradiation therapy, patients received an additional 4 cycles of paclitaxel 175 mg/m[2] I.V infusion over 3 hours and carboplatin at AUC 6 every 3 weeks. The overall response rate was 77.8% with CR in 7 patients [25.9%] PR in 14 patients [51.9%], SD in 5 patients [18.5%] and 1 patient [3.7%] showed progressive disease. The median overall survival was 13.5 months and 1 year and 2 year survival rates were 58% and 26% respectively. The most frequent and significantly noted toxicity in this study was esophagitis [85.2%]. The other nonhematologic toxicities included nausea and vomiting [59.2%], peripheral neuropathy [25.9%], fatigue [33.3%] and pneumonitis [11.1%]. Hematologic toxicity consisted of anemia in 12 patients [44.4%], leucopenia in 8 patients [29.6%] and thrombocytopenia in 6 patients [22.2%]. These data together with our experience suggest that, in patients with unresectable stage IIIA-IIIB NSCLC, concomitant weekly chemoradiotherapy with paclitaxel and carboplatin, followed by consolidation chemotherapy with the same regimen is feasible, generally well tolerated, and yields therapeutic results that compare favorably to those reported for other regimens. However, further randomized studies are needed to identify the optimal chemoradiotherapy regimens and schedules for treatment of unresectable stage IIIA-IIIB NSCLC patients

Concomitant boost radiation plus concurrent cisplatin for advanced head and neck carcinomas

This study was designed to assess the outcome of concurrent chemotherapy with a "concomitant boost" radiotherapy in the treatment of advanced unresectable head and neck cancer patients. Between January of 2001 and December of 2002, Thirty four patients who met the eligibility criteria were treated with combined chemoradiotherapy. Radiation consisted of 70 Gy in 40 fractions over a 6-week period [5 days / week for 4 weeks, then twice a day for 5 days for the last 2 weeks as a boost]. Concurrent chemotherapy was administered during weeks 1 and 5 of radiation therapy. Chemotherapy consisted of single agent cisplatinum. Cisplatinum was given as a daily bolus of 20 mg per square meter per day for 5 days for a total of 100 mg per square meter. Median follow-up was 22 months [6-48 months]. Complete response was recorded in 23 patients [68%] and 7 patients [21%] showed partial response. Median survival for the whole group was 30 months. The estimated 1 and 2 year rates of overall survival were 82.4% and 70.5% respectively. The median relapse free survival for complete responders was 28 months and the estimated 1 and 2 year rates of disease free survival were 80.4% and 58.4% respectively. Acute confluent mucositis [Radiation therapy Oncology Group [RTOG] grade 3] developed in 53% of patients. Long term toxicity is mainly based on xerostomia. Concurrent chemotherapy and concomitant boost radiotherapy approaches appear promising. The combination appears to be safe and effective in producing a good response, satisfactory local control and longer survival. Randomized trials comparing combined modality with the standard treatment are warranted to define the best treatment option in this group of patients with poor prognosis

Docetaxel, doxorubicin and cyclophosphamide [TAC] as first line-chemotherapy for metastatic breast cancer

This study was designed to investigate the efficacy and toxicity of docetaxel with doxorubicin and cyclophosphamide [TAC] as first-line chemotherapy for metastatic breast cancer. Forty-four patients received 3-week cycles of docetaxel 75 mg/m[2] 1 hour I. V. infusion, preceded by I. V. doxorubicin 50 mg/m[2] and cyclophosphamide 500 mg/m[2] for a maximum of 6 cycles. Complete response was achieved in 4 patients [9%]. The overall objective response rate was 70%. The objective response rates in patients with bone, lymph nodes, liver and lung involvement was 81%, 73%, 71% and 67% respectively. The median duration of response was 12 months and the median time to progression was nine months. With a median follow-up of 18 months, the median survival was 18 months whereas the 1 and 2-year survival rates were 64% and 38% respectively. The main toxicities were hematological [neutropenia in 82% of patients, anemia in 46% and thrombocytopenia in 25% of patients]. Non-hematological adverse events were infrequent and docetaxel-specific toxicities [such as fluid retention and nail changes] were mild. No anaphylaxis, cardiac toxicity or febrile neutropenia were observed in any patient. The docetaxel-doxorubicin-based TAC combination seems to be active as first-line treatment of patients with metastatic breast cancer. Although neutropenia was frequent, it was manageable. The rest of the toxicity profile seems acceptable, with no significant nonhematologic toxicities. Further randomized trials are needed to establish the role of docetaxel in the treatment strategies and natural history of breast cancer

Combined chemoradiotherapy versus radiotherapy alone in patients with esophageal cancer

This study included 61 patients with locally unresectable esophageal cancer. Thirty-one patients received radiotherapy alone [RT group], the dose of radiotherapy ranged between 60-64 Gy/6-6.5 weeks, while the remaining 30 patients received radiotherapy in a dose ranged between 45-50 Gy/4.5-5 weeks plus chemotherapy [CT-RT group]. Chemotherapy [consisting of 5-fluorouracil, leucovorin and cisplatin] was given 2-3 cycles before radiotherapy and further 2-3 cycles were given after radiotherapy. The study concluded that combined modality therapy can improve the outcome in locally unresectable esophageal carcinoma with significantly better local rate, survival, progression-free survival rates and much reduction in the incidence and timing of treatment failures

Bcl-2 and p53 expression as prognostic markers in malignant epithelial ovarian tumours [EOC]

This study aimed to investigate the prognostic and predictive relevance of Bcl-2 and P53 protein expression in epithelial ovarian carcinoma [EOC] and to evaluate the results in relation to clinicopathologic parameters, clinical outcome and response to chemotherapy. Tumor biopsy specimens from 56 consecutive and homogeneously treated patients with FIGO stages II and III epithelial ovarian cancer [EOC] were examined immunohistochemically for expression of Bcl-2 and P53 proteins. All patients received chemotherapy [CAP] after the primary surgery. A number of clinicopathologic factors [age, duration of symptoms before diagnosis, FIGO stage, histologic type, degree of differentiation, laterality, capsular invasion, peritoneal deposits, tumour size, ascites and residual disease] were studied in relation to the tissue markers Bcl-2 and P53. Both uni-and multivariate analysis of prognostic factors were performed and the relations to classical clinicopathologic parameters and response to chemotherapy were examined. The results revealed that Bcl-2 and P53 positive expression were found in 41.1% and 14.3% of the cases, respectively. Bcl-2 expression was related to age above 50 years, lower FIGO stage, absence of ascites, absence of residual disease, resistance to chemotherapy and negative P53 expression. For P53 expression, a positive relation was found with age above 50 years, duration of symptoms less than 6 months, lower FIGO stage, absence of ascites and partial response to chemotherapy. All cases with P53 positive expression were FIG0 stage II and with moderate or poor histologic differentiation. The presence of Bcl-2 expression and absence of P53 expression were associated with improved disease free survival but not overall survival. In multivariate analysis, age, FIGO stage and degree of differentiation retained independent prognostic significance with overall survival. When disease free survival was used as an end point, only FIGO stage and association groups of Bcl-2 and P53 expression were of independent significance. It was concluded that Bcl-2, P53 or an association of the two markers expression data may represent prognostic predictors in epithelial ovarian cancer [EOC]

Orchidectomy versus Orchidectomy plus cyproterone acetate in the treatment of metastatic prostatic carcinoma

This study included 42 patients all underwent orchidectomy and only 22 received cyproterone acetate in a dose of 150-200 mg PO daily. Clinical, laboratory and radiological evaluations were done during the period of the study [from January 1992 to June 1997 inclusive]. There was no high difference in the causes of death. The mean overall survival was longer in the combined treatment group [44.9 m vs. 36.9 m]. Time to disease progression was significantly longer in the combined modality group [30.7 m vs. 22.7 m]. Hot flushes were less in the combined group [31.0% vs. 60.6%] but the difference was insignificant

Primary gastrointestinal non-Hodgkin's lymphoma; clinical features, management and prognosis of 56 patients

In this study, 56 patients with primary gastrointestinal tract [GIT] non-Hodgkin's lymphoma [41 males and 15 females, mean age 41.5 years] were retrospectively evaluated. Twenty-nine patients were treated with curative surgery and adjuvant chemotherapy and 19 patients were treated with non-curative surgery plus chemotherapy and 8 patients were treated with curative surgery alone. All patients were included in the response analysis and survival. Only 33 patients were included in the disease free survivals as they had a complete response after different treatment modalities. The study confirmed the prognostic value of stage of the disease, curative surgery plus adjuvant chemotherapy in patients with primary gastrointestinal tract non- Hodgkin's lymphoma [PGITL]

Prognostic and predictive value of HER-2/Neu over expression in primary breast cancer

To investigate the presence of HER-2 over expression by immunohisto-chemistry in 81 archival breast cancer specimens and its impact on survival in relation to other prognostic factors. Paraffin embedded primary breast cancer specimens from 81 patients with stage I-III breast cancer, treated in Clinical Oncology and Nuclear Meadicine and surgical Oncology Dpartment, Mansoura University Hospital, were studied immuno-histochemically. All patients were subjected to modified radical mastectomy with axillary dissection. Locoregional postoperative radiotherapy was given to all patients with lymph node metastases or primary tumors large than 2 cm located in the medical or central area of the breast. Primenopausal node positive patients were treated with six courses of either CMF or FAC regimens. Postmenopausal node positive patients were given tamoxifen with or without CMF. HER-2neu protein over expression was detected in 28% of tumors. A significant correlation was noted between HER-2 expression, poor histologic grade and higher number of mitosis. Also, HER-2 expression was associated with shorter 5-year over all survival and disease free survival rates compared with HER-2 negative cases [52.8% V 84.8% and 44.8% V 73.9% respectively]. In a multivariate analysis, HER-2 over expression was found to be a significant predictor factor of overall and disease free survival [p=0.004 and 0.01 respectively]. HER-2/nen overxpression is an independent prognostic factor for risk of recurrence. Women with tumours without HER-2 overxpression have a good prognosic; aggressive therapy n this group in therefore difficult to justify. On the other hand, even with adjuvant chemotherapeutic treatment whose tumours exhibit HER-2 over expression have an increased risk of recurrence. Randomized trials to compare more aggressive adjuvant chemotherapy versus standard chemotherapy for women whose tumor exhibit HER-2 protein over expression are recommended

Conservative treatment versus mastectomy in the treatment of stage I and II breast cancer

Analysis of the patterns of failure for breast cancer patients who underwent modified mastectomy, or lumpectomy with axillary dissection and radiation therapy, as well as the prognostic factors that have an independent effect on treatment failures and overall survival. Sixty six patients with clinical stage I and II breast cancer were randomly assigned to undergo either modified radical mastectomy or lumpectomy, axillary, dissection and radiation therapy. All patients with positive nodes received adjuvant systemic therapy. Annual mammography was an integral component of the follow-up program. Diagnostic studies for systemic disease were performed as clinically indicated. At five years, overall survival was 79% for patients assigned to mastectomy and 91% for those assigned to lumpectomy [P=0. 97]. Disease-free survival was 74% for patients assigned to mastectomy and 72% for those assigned to lumpectomy [P=0.7]. The rate of local recurrence was 8.8% after mastectomy and 12.5% after lumpectomy [P=0.92]. Breast conservation with lumpectomy and irradiation offers results at five years that are equivalent to those obtained with mastectomy