Interferon gamma and regulatory T-Cells and their relation with the pathological grading of gastritis caused by helicobacter pylori

Background: Helicobacter pylori [H.pylori] is a cause of chronic-active gastritis and a majority of cases of peptic ulcer disease. It is also associated with the development of gastric adenocarcinoma, the fourth most common malignancy in the world

Aim of the study: The study aims to detect the level of interferon gamma expression and the percentage of CD4+ CD25+ T regulatory cells in patients with gastritis and their relation to the pathological grading

Methods: Gastric biopsy specimens were taken from the antrum of 30 patiens using upper GIT endoscopy. Rapid urease test [RUT], immunohistochemistry [IHC], and Hematoxylin and Eosin [H and E] stains were used to determine H.pylori status. Evaluation of the histologic features was done using H and E. IFN-gamma mRNA expression in the gastric biopsies was measured using Real-Time PCR. Flowcytometry was used for measuring the percentage of CD4+CD25+regulatory T-ells [T regs] in the peripheral blood of all the patients. The results were compared to 15 healthy control subjects

Results: H.pylori was detected in 56.7% of all patients by RUT and in 93.3% patients by H and E and in 83.3% by IHC,out of them 88% had active gastritis and all of them had chronic gastritis. There was a statistically significant positive correlation between H.pylori positive infection and the pathological grading of active gastritis [p=0.015]. However, there was no statistically significant correlation between H.pylori infection [by IHC] and the pathological grading of chronic gastritis [p=0.334]. No statistically significant difference in the levels of IFN-gamma mRNA expression between H.pylori positive and negative patients was detected. No correlation was found between the levels of IFN-gamma mRNA expression and the severity of either active or chronic gastritis. Similarly, no correlation between the percentage of CD4+ CD25+ T regulatory cells and the severity of either active or chronic gastritis was detected

Conclusion: H.pylori plays an important role in active gastritis lesion. The correlation between IFN-gamma mRNA expression and the severity of active or chronic gastritis was overshadowed by the prevalence of helminthic infection among Egyptian patients. Meanwhile, low levels of T regulatory cells among H.pylori positive and negative gastritis than controls suggests an important role of T regulatory cells in regulating the gastric mucosal inflammatory response

Correlation of synovium osteoprotegrin with magnetic resonance imaging in rheumatoid arthritis

To demonstrate the role of Osteoprote grin [OPG] expression in the synovium in the pathogenesis of rheumatoid arthritis [RA] joint damage, and correlate it with Magnetic Resonance Imaging [MRI] finding. Twenty RA patients and five controls were included. RA disease activity was assessed by disease activity score [DAS 28]. MRI examination of knee joint, including evaluation of inflammation using synovitis score and evaluation of destruction with an erosion score, were performed. Knee joint's synovial biopsy specimens were obtained, by arthroscopy, to demonstrate the degree of expression of OPG by using immunohistochemical staining with monoclonal antibodies, and to study the histopathological activity scores by histopathological examination. The OPG expression was deficient [grade 0, 1] in 70% of synovial cells and 85% of endothelial cells lining of the synovial blood vessels, and grade I and 2 in 70% of infiltrating cells in active RA synovium. There was no significant association between OPG expression score [synovial, endothelial, infiltrating cells] and disease duration, number of tender swollen joints, ESR and Larsen score. There was a significant inverse correlation between OPG expression [synovial, infiltrating] and MRI erosion score. There was a highly significant correlation between MRI synovitis score and ESR, CRP, pain score, histopathological synovium score [p<0.01] and significant negative correlation with erosion score. We concluded that decrease in OPG expression in synovium has a role in pathogenesis of joint damage in RA patients, and MRI is considered a sensitive test to detect pathological lesion in joint damage in RA patients

Clinico-pathologic assessment of the value of anti-chromatin antibodies in the diagnosis of SLE and lupus nephritis

Background: Lupus nephritis is the major cause of morbidity and mortality in systemic lupus erythematosus [SLE] patients, and renal biopsy is the most accurate method for the evaluation of the degree of renal affection. Recent studies suggest that antichromatin antibodies [anti-CHR] could help in early detection of lupus nephritis in SLE patients

Objective: To assess the prevalence of serum anti-CHR antibodies in SLE patients, to evaluate their clinical significance and their value as a marker of lupus nephritis and to correlate them with histopathological findings of lupus nephritis

Methodology: Serum anti-CHR antibodies were determined by an enzyme linked immonsorbent assay [ELISA] in 30 SLE patients, and 30 control subjects [10 cases of rheumatoid arthritis [RA], 10 cases of osteoarthritis [OA] and 10 healthy persons]. Patients and controls were also subjected to history taking, clinical examination and routine laboratory investigations including CBC, ESR, serum complement, serum creatinine, creatinine clearance; in addition to complete urine analysis, urinary protein measurement and serum albumin. Renal biopsy was performed on 20 patients of SLE with clinical and laboratory evidence of nephritis and examined by both light microscopy [LM] and electron microscopy [EM]

Results: Anti-CHR antibodies in SLE patients were positive in 20/30 [66.7%], while they were negative in all controls including cases of RA and OA, with a 66.7% diagnostic sensitivity and 100% specificity. Anti-dsDNA antibodies were detected in 13/30 [43.3%] cases of SLE only, with a 43.3% diagnostic sensitivity and 100% specificity. Positive results of anti-CHR antibodies were found in 17/20 [85%] cases of lupus nephritis [85% diagnostic sensitivity and 70% specificity]; whereas anti-dsDNA antibodies were detected in 11/20 cases [55%] only [55% diagnostic sensitivity and 80% specificity]. Significant positive correlations were found between anti-CHR antibodies and disease duration, disease activity, serum creatinine, creatinine clearance, proteinuria, ESR, ANA level and nephritic lesions seen in renal biopsy. Meanwhile, negative correlations were found with blood platelets, serum albumin, serum complement C3 level [p<0.05]. A highly significant correlation was also found between anti-CHR antibodies and disease activity score measured by ECLAM [p<0.001]. Renal biopsy findings showed that WHO class of lupus nephritis had a significant impact on anti-CHR antibody level [p<0.05], with the association of higher antibody levels with proliferative glomerular lesions and frequent electron dense deposits mainly in the subendothelial location, while no significant difference was found in case of anti-dsDNA antibodies

Conclusion: Antichromatin antibodies are more sensitive and specific than anti-dsDNA antibodies for the diagnosis of SLE and more sensitive as regards lupus nephritis. Moreover, serum level of anti-CHR antibodies can be a helpful test to expect the extent of renal affection and hence -choose patients candidate for renal biopsy

effects of coblation technology on the soft palate of rabbits. An experimental study

This study was designed to evaluate the volumetric reduction and the histologic changes of the soft palate after coblation [cold ablation] technology in experimental animals. Sixty male rabbits were included in the study divided into 3 groups: group [A], group [B] and group [C]. Each group included 10 control and 10 study animals. Exposure of the soft palate to coblation was done in study animals of the 3 groups submucosaly at 3 sites, one at midportion and 2 at lateral sides of the soft palate for one minute duration in each site. The control animals of the 3 groups were left without any interference. The animals of group A. B and C were studied 1 month, 2 months and 3 months respectively. The soft palate of each rabbit was dissected and volume was calculated. Specimens of soft palate of both control and study animals were examined by electron microscopic study. Died animals were excluded from this study. The comparison of the mean volume of soft palate in the study groups showed gradual increased reduction with time as it was more after 3 months [group C] than 2 months [group B] and 1 month [group A]. This result was confirmed by electron microscopic study as there was an increase in the amount of collagen fibers in the subepithelial layer gradually in study groups which was mild, moderate and marked in group A. B and C respectively. Cytoplasmic vacuoles were detected more in the epithelial layer of study group [A] than group [B] this means degenerative changes which is reversible as it was decreased by time and disappeared in group [C]. There is no effect on the mucosal glands of the epithelium or underlying muscles. The only histological difference between study and control groups was the increased amount of collagen fibers in the subepithelial layer. Coblation technology could be considered an effective technology for the reduction of the size of soft palate. It is safe, not painful and not invasive as it maintain the normal histology of the soft palate

Effects of methotrexate and leflunomide therapy on knee synovium of adjuvant induced arthritis as a model for rheumatoid arthritis

Leflunomide and methotrexate have proven to be efficacious in reducing joint inflammation and joint destruction in clinical models of arthritis and in rheumatoid arthritis. The objective of this study was to evaluate the effects of both drugs as well as their combination therapy on the synovium and cartilage of adjuvant arthritis as a model of rheumatoid arthritis [RA] in humans. This study was carried out on forty animals stratified into 5 groups: normal, adjuvant arthritis [AA] control, AA who received leflunomide in a dose of 20 mg/kg orally, AA who received intraperitoneal methotrexate in a dose of 0.3 mg/kg twice weekly and AA who received both leflunomide and methotrexate of the same dose given in groups 3 and 4. All animals were sacrificed after 3 weeks; the right knee was dissected and examined with light microscopy. Oxidants markers [nitric oxide [NO] and malondialdhyde [MAD]] and antioxidants markers [glutathione [GSH], erythrocyte superoxide dismutase [SOD] and ceruloplasmin [CP]] were all measured. All the treatment modalities showed variable degrees of improvement of synovial and cartilage scoring in comparison to AA [the non-treated group]. The leflunomide treated group [group 3] showed the best improvement of synovial pathology, while the combined therapy group [group 5] showed the best improvement of cartilage pathology. The oxidative stress markers showed some changes with different modalities of treatment where, nitric oxide did not change significantly between all groups. Malondialdhyde [MAD] was significantly lower in the methotrexate [MTX] treated group as compared to AA controls. Also, superoxide dismutase [SOD] was significantly lower in the leflunomide treated group, MTX treated group as well as in the group who received combined therapy as compared to AA the controls. Glutathione [GSH] level was significantly decreased with combination therapy as compared to the leflunomide treated group. Serum ceruloplasmin [CP] showed a significant decrease in its level in the MTX treated group as compared to the AA controls. MTX treatment [group 4] was the best in controlling oxidative stress markers. Further study is needed to evaluate the duration and dose effect of each drug on synovium, cartilage and oxidative markers

Sentinel node mapping in colorectal cancer: is it worth the effort?

Background: Lymphatic mapping [LM] with sentinel node [SN] biopsy is an interesting recent development in surgical oncology. This approach has the potential of accurately identifying the first lymph node [or nodes] that drain primary tumors. These nodes are the most likely to harbor metastatic or micrometastatic disease. Sentinel node mapping and the pathologic search for micrometastasis may therefore increase the accuracy of the pathologic staging, which may alter the further management and the prognosis

Aim of the Study: To evaluate the usefulness of intra-operative in-vivo and ex-vivo sentinel node mapping in colorectal cancer [CRC] resections, and its effect combined with selected pathologic focus node examination on upstaging of CRC and consequent therapeutic strategies

Patients and Methods: Twenty- nine [after exclusion of six] patients with CRC were enrolled in a study of lymph mapping [LM] with peritumoral and submucosal injection of isosulfan blue dye. In-vivo LM was undertaken intraoperatively during colon resection in 23 patients. Ex-vivo LM was done after specimen removal in 6 patients [1 rectosigmoid and 5 extraperitoneal low rectal carcinoma]. All nodes wereexamined with hematoxylin and eosin [H and E] staining; in addition, negative sentinel lymph nodes [SNs] for metastasis with H and E were multi-sectioned and examined by immunohistochemical staining with cytokeratin[CK-lHC]

Results: SNs were successfully identified in 27/29 patients [93%][at least one SN was identified]. SNs were not identified in 2 cases; one case of very low rectal cancer and the other one was a locally advanced rectosigmoid colonic cancer. LM .demonstrated primary lymphatic drainage to SNs outside the margins of conventionally surgical planned resections in 2 cases [7%] and guided multiple sections and histochemical staining that identified occult micrometastases in 3 of the SN negative patients [11%]. Upstaging was thus achieved in 5 cases [18%] using mapping and focus nodal examination. Overall, the specificity and the negative predictive value in this series were 100% and 67%, respectively, whereas the sensitivity and positive predictive value were 89% and 78% respectively. There was a significant positive correlation between the tumor T stage and lymph node metastases [P< 0.001]

Conclusion: Sentinel node mapping is easy to do intraoperatively during colorectal resections. Ex-vivo LM can be applied when in- vivo techniques are unsuccessful and could beuseful for rectal carcinoma. LM techniques appear to improve staging accuracy in CRC through detection of more node metastases as well as guiding the use of sophisticated histopathologic studies to search for occult nodal micrornetastases. It may demonstrate an unexpected pattern of lymphatic drainage requiring modification of the conventional resection