Evaluation of newer diagnostic methods for the detection and differentiation of Entamoeba histolytica in an endemic area

We evaluated the colorimetric polymerase chain reaction (PCR)-based method for the detection and differentiation of Entamoeba spp. and compared the efficacy of E. histolytica-specific antigen detection in faeces with the detection of specific antibodies to E. histolytica-specific antigen in faeces, by enzyme-linked immunosorbent assay. Faecal samples were obtained from patients attending hospital in Chandigarh, India, from March 2001 to February 2002. The PCR-based colorimetric method was found to be the most sensitive (100%) and it could differentiate between pathogenic and non-pathogenic Entamoeba spp. The present study also emphasized that the antigen detection system may prove to be a better diagnostic tool than the antibody detection system in endemic areas.     

Familial interstitial pulmonary fibrosis: A large family with atypical clinical features

A large kindred of familial pulmonary fibrosis is reported. Six members from the first two generations of this particular kindred were described more than 40 years previously; six more individuals from the third and fourth generations have also been evaluated. The proband, now 23 years of age, has mild disease; the other 11 documented affected family members all died from their disease at an average age of 37 years (range 25 to 50 years). The pathology was that of usual interstitial pneumonia, as is typical in idiopathic pulmonary fibrosis. However, the initial radiographic pattern in many of these individuals was upper lobe and nodular and, along with the young age, was atypical for idiopathic pulmonary fibrosis. Several genetic abnormalities have been associated with familial pulmonary fibrosis. The present study examined the genes coding for surfactant protein-C, ATP-binding cassette protein A3 and telomerase, and found no abnormalities.     

Active CD4+ helper T cells directly stimulate CD8+ cytotoxic T lymphocyte responses in wild-type and MHC II gene knockout C57BL/6 mice and transgenic RIP-mOVA mice expressing islet beta-cell ovalbumin antigen leading to diabetes

CD4+ helper T (Th) cells play crucial role in priming, expansion and survival of CD8+ cytotoxic T lymphocytes (CTLs). However, how CD4+ Th cell's help is delivered to CD8+ T cells in vivo is still unclear. We previously demonstrated that CD4+ Th cells can acquire ovalbumin (OVA) peptide/major histocompatibility complex (pMHC I) and costimulatory CD80 by OVA-pulsed DC (DC(OVA)) stimulation, and then stimulate OVA-specific CD8+ CTL responses in C57BL/6 mice. In this study, we further investigated CD4+ Th cell's effect on stimulation of CD8 CTL responses in major histocompatibility complex (MHC II) gene knockout (KO) mice and transgenic rat insulin promoter (RIP)-mOVA mice with moderate expression of self OVA by using CD4+ Th cells or Th cells with various gene deficiency. We demonstrated that the in vitro DC(OVA)-activated CD4+ Th cells (3 x 10(6) cells/mouse) can directly stimulate OVA-specific CD8+ T-cell responses in wild-type C57BL/6 mice and MHC II gene KO mice lacking CD4+ T cells. A large amount of CD4+ Th cells (12 x 10(6) cells/mouse) can even overcome OVA-specific immune tolerance in transgenic RIP-mOVA mice, leading to CD8+ CTL-mediated mouse pancreatic islet destruction and diabetes. The stimulatory effect of CD4+ Th cells is mediated by its IL-2 secretion and CD40L and CD80 costimulations, and is specifically delivered to OVA-specific CD8+ T cells in vivo via its acquired pMHC I complexes. Therefore, the above elucidated principles for CD4+ Th cells will have substantial implications in autoimmunity and antitumor immunity, and regulatory T-cell-dependent immune suppression.     

An Expert Support System for Breast Cancer Diagnosis using Color Wavelet Features

Breast cancer diagnosis can be done through the pathologic assessments of breast tissue samples such as core needle biopsy technique. The result of analysis on this sample by pathologist is crucial for breast cancer patient. In this paper, nucleus of tissue samples are investigated after decomposition by means of the Log-Gabor wavelet on HSV color domain and an algorithm is developed to compute the color wavelet features. These features are used for breast cancer diagnosis using Support Vector Machine (SVM) classifier algorithm. The ability of properly trained SVM is to correctly classify patterns and make them particularly suitable for use in an expert system that aids in the diagnosis of cancer tissue samples. The results are compared with other multivariate classifiers such as Na?ves Bayes classifier and Artificial Neural Network. The overall accuracy of the proposed method using SVM classifier will be further useful for automation in cancer diagnosis.     

CD4(+) T cell-released exosomes inhibit CD8(+) cytotoxic T-lymphocyte responses and antitumor immunity

T cells secrete bioactive exosomes (EXO), but the potential immunoregulatory effect of T-cell EXO is largely unknown. In this study, we generated activated ovalbumin (OVA)-specific CD4⁺ T cells in vitro via coculture of OVA-pulsed dendritic cells (DC_OVA) with naive CD4⁺ T cells derived from OVA-specific T-cell receptor (TCR) transgenic OTII mice. CD4⁺ T-cell EXO were then purified from the CD4⁺ T-cell culture supernatants by differential ultracentrifugation. CD4⁺ T-cell EXO exhibited the ‘saucer'' shape that is characteristic of EXO with a diameter between 50 and 100 nm, as assessed by electron microscopy, and contained the EXO-associated proteins LAMP-1, TCR and lymphocyte function associated antigen-1 (LFA-1), as determined by western blot. Flow cytometric analysis showed that CD4⁺ T-cell EXO expressed CD4⁺ T-cell markers (CD4, TCR, LFA-1, CD25 and Fas ligand), but to a lesser extent than CD4⁺ T cells. We demonstrated that DC_OVA took up CD4⁺ T-cell EXO via peptide/major histocompatibility complex (pMHC) II/TCR and CD54/LFA-1 interactions. OVA-specific CD4⁺ T-cell EXO from OTII mice, but not ConA-stimulated polyclonal CD4⁺ T-cell EXO from wild-type C57BL/6 mice inhibited DC_OVA-stimulated in vitro CD4⁺ T-cell proliferation and in vivo CD8⁺ cytotoxic T lymphocyte (CTL) responses and antitumor immunity against OVA-expressing B16 melanoma BL6-10_OVA cells. In addition, EXO derived from a T-cell hybridoma cell line, MF72.2D9, expressing an OVA-specific CD4⁺ TCR, had a similar inhibitory effect as OTII CD4⁺ T-cell EXO on CTL-mediated antitumor immunity. Taken together, our data indicate that antigen-specific T-cell EXO may serve as a new type of immunosuppressive reagent for use in transplant rejection and treatment of autoimmune diseases.     

Influence of environment on seed soluble carbohydrates in selected lentil cultivars

High concentrations of raffinose-family oligosaccharides (RFOs) in lentils cause stomach discomfort and reduce lentil quality for human consumption. To develop strategies for lentil quality improvement, we investigated variability, heritability and effects of environmental conditions on the content and composition of soluble carbohydrates in lentil seeds. Analyses of variance showed that cultivar and environment and their interaction had significant effects on the sugar content in lentil seeds. Glucose and sucrose contents of lentil cultivars ranged from 0.02 to 0.06 g and 1.22–1.67 g 100 g^?1 lentil meal, respectively. Total RFO content of lentil cultivars ranged from 4.5 to 5.5 mmol 100 g^?1 lentil seed meal. In all lentil cultivars, RFO content was positively correlated with glucose and sucrose contents. Analysis of RFO profiles by high performance size exclusion chromatography (HP-SEC) showed that stachyose was the major RFO in all lentil cultivars followed by raffinose and verbascose, respectively. The broad sense heritability of sucrose and RFO estimated from the analyses of variance components was 0.89 and 0.85, respectively. Lentil seed RFO content is a highly heritable trait, thus making it amenable to genetic improvement to meet consumer preferences.     

Routine pathological evaluation of tissue from inguinal hernias in children is unnecessary.

INTRODUCTION: Because unexpected disease is rare in a child's inguinal hernia sac we decided to investigate the cost of routine pathological evaluation of inguinal hernial sacs in children and the incidence of clinically significant pathological findings. METHODS: We searched the health records at the University Hospital, Saskatoon, for patients under 20 years of age who had inguinal hernia repair between 1988 and 1997. For records noting pathology findings of duct-like structures, the operative reports and histology slides were reviewed. Specimens were immunostained for muscle-specific actin. The cost of pathological evaluation was estimated using a provincial physician-billing schedule. RESULTS: During the study period, there were 488 inguinal hernia repairs in 371 patients under 20 years of age. Of these, 456 (93.4%) specimens were evaluated microscopically. There were 4 (0.88%) cases with unexpected findings diagnosed as epididymis at a cost of Can dollar 6988/case. CONCLUSION: The routine histologic evaluation of inguinal hernia sacs in children is an unnecessary expense and should be reserved for select cases at the discretion of the surgeon.     

Nonspecific interstitial pneumonia and usual interstitial pneumonia with mutation in surfactant protein C in familial pulmonary fibrosis.

Nonspecific interstitial pneumonia, a recently described form of idiopathic interstitial pneumonia, is characterized by uniform involvement of the alveolar septae with interstitial inflammation and variable amounts of fibrosis. Histological observations differentiate nonspecific interstitial pneumonia from usual interstitial pneumonia and clinically, patients with a nonspecific interstitial pneumonia pattern show better prognosis than those with usual interstitial pneumonia. We have genetically analyzed a family with a history of usual interstitial pneumonia. Most of the patients presented as adults and their biopsies showed a pattern consistent with usual interstitial pneumonia. However, three family members presented in early childhood and their biopsies revealed a nonspecific interstitial pneumonia pattern. The inheritance pattern of usual interstitial pneumonia is consistent with autosomal dominant inheritance with variable expression. DNA sequence analyses of the surfactant protein C gene in children with nonspecific interstitial pneumonia and adults with usual interstitial pneumonia exhibit a common heterozygous mutation located in exon 5. The mutation causes a Leu188 to Gln188 change in the carboxy-terminal region of prosurfactant protein C, possibly affecting peptide processing. These observations suggest that individuals with this particular mutation in surfactant protein C gene might be at increased risk of interstitial lung disease of variety of types.