Labeling male anorectal malformations: objective evaluation of radiologic imaging before surgery.

Purpose: Prone cross-table lateral x-ray (CTLxR) and colostogram aid surgical planning for anorectal malformations (ARMs) without perineal fistulas. We suggest objective imaging tools to classify ARMs. Methods: Three observers prospectively evaluated CTLxR and colostograms of male ARM patients (2012-2022) without perineal fistulas. The level of the rectal pouch was estimated with pubococcygeal (PC) and ischiatic (I) lines. On CTLxR, we described the "pigeon sign", defined as the rectal pouch ending with a beak-like image, suspicious for a rectourinary fistula. ARM was defined as rectobulbar when the rectal pouch was below the I line, rectoprostatic when between PC and I lines, and rectovesical when above the PC line. Concordance was assessed with Fleiss' kappa. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the "pigeon sign" were calculated. Results: Thirteen patients were included in this study. The interobserver agreement on CTLxR was 69.2% (k = 0.54) on pouch ending, 84.6% (k = 0.69) on the "pigeon sign", and 76.9% (k = 0.69) on diagnosis; concordance between observers and intraoperative diagnosis was 66.6% (k = 0.56). The "pigeon sign" had 75% sensitivity, 100% specificity, 100% PPV, and 50% NPV. The interobserver agreement on colostograms was 84.6% (k = 0.77) on pouch ending and 89.7% (k = 0.86) on diagnosis; concordance between observers and intraoperative diagnosis was 92.3% (k = 0.90). Conclusion: PC and I lines and the "pigeon sign" are useful tools in examining CTLxR and colostograms. Adequate CTLxR interpretation may modify surgical strategy.

Partial protection against SIV challenge by vaccination of adenovirus and MVA vectors in rhesus monkeys.

This study explores the effect of priming rhesus monkeys with an Ad5/35 vector expressing simian immunodeficiency virus (SIV) gag and gp120, and then boosting the animals with an modified vaccinia virus Ankara (MVA) vector encoding the same antigens after a 2-month interval. The animals were intravenously challenged with 100 TCID50 of highly pathogenic SIVmac239 virus 2 months after the booster vaccination. The priming vaccination induced robust SIV-specific cell-mediated and humoral immune responses, and boosting further enhanced the cellular immunity. Vaccination reduced peak and long-term viral loads by 1-2 logs for a period of >6 months, as reflected by a reduction in both the SIV RNA and DNA levels. Of considerable interest, the immunized monkeys did not suffer from loss of CD4 T cells, particularly central memory CD4 T cells. These results demonstrate that prophylactic vaccination with Ad5/35 followed by MVA reduces viral replication and prevents CD4 T-cell loss, and that these effects may decrease the likelihood of disease progression.

DNA vaccines: capacity to induce auto-immunity and tolerance.

DNA technology has facilitated the development of plasmid-based vaccines designed to prevent viral, bacterial and parasitic infections. The rapid transition of these novel vaccines from the laboratory to the clinic raises important safety concerns. Our review examines whether DNA vaccines (i) are likely to induce systemic or organ-specific auto-immune disease and (ii) have the potential to induce tolerance rather than immunity.

[A case of adult neuroblastoma arising in the retroperitoneal space].

Neuroblastoma, common in children, rarely develops in adults. We recently treated a patient with adult neuroblastoma. A 34-year-old man complained of a swelling in right inguinal region. CT scan showed swelling of retroperitoneal and inguinal lymph nodes, and bone scintigram by 99mTc-HMDP showed an abnormal uptake in the swollen lymph nodes. Chemotherapy with CDDP (cisplatinum), VP-16 (etoposide), BLM (bleomycin), ADM (adriamycin) was not effective. Histopathological examination of a biopsy specimen revealed neuroblastoma. Another chemotherapy with CPM (cyclophosphamide), VCR (vincristine), ADM, DTIC (dacarbazine), CDDP, VP-16 was effective in decreasing the tumor size. Further high dose chemotherapy with CPM, ADM, CDDP, VP-16 combined with peripheral blood stem cell transplantation led to almost complete disappearance of the tumor and normalization of blood tumor markers (neuron specific enolase and immunosuppressive acidic protein). Retroperitoneal lymph node dissection demonstrated well-differentiated neuroblastoma in the excised tissue. Six months postoperatively, the tumor recurred in the pelvic cavity. Although chemotherapy and radiotherapy were given, he died of the disease 12 months postoperatively.

Diagnostic value of 5'-nucleotide phosphodiesterase isozyme-V for detection of liver metastasis in patients with gastro-intestinal cancer.

Serum 5'-nucleotide phosphodiesterase isozyme-V (5'-NPDase-V) in 200 patients with various gastrointestinal disorders including 178 malignant cases and 22 benign cases was measured, and the usefulness and limitations of this test for the detection of liver metastasis were studied. Of the laboratory tests performed in the 44 patients with liver metastasis, 5'-NPDase-V and carcinoembryonic antigen (CEA) (greater than 10.0 ng/ml) were positive in 68.2%. This rate was higher than that for any other laboratory test. Positive 5'-NPDase-V assays were found in 46.2% of the patients with H1 metastasis, 70% with H2 and 81% with H3. This test was considered to be useful diagnostically, when liver metastasis was of more than a certain degree. 5'-NPDase-V was positive in 85.7% of the patients who had liver metastasis and had a serum total bilirubin value of more than 2.5 mg/dl. Particular attention should be paid to the evaluation of this test when jaundice is present. In eight of the patients with liver metastasis, CEA (greater than 10.0 ng/ml) was negative, whereas 5'-NPDase-V was positive. One or both of these two markers were positive in 86.4%. A combination assay of these two markers was considered to be more useful than 5'-NPDase-V alone as a screening test for the detection of liver metastasis. In one patient who underwent hepatic lobectomy for recurrence of cancer in the liver, 5'-NPDase-V and CEA were measured before and after the operation. CEA returned to normal immediately after the operation, whereas 5'-NPDase-V became negative five months after the operation. It is presumed that the changes in 5'-NPDase-V after hepatic lobectomy were related to liver regeneration.

Detection of the ligand activity of the c-erbB-2 protein in calf serum.

We established NIH3T3 derivatives in which wild-type c-erbB-2 or activated c-erbB-2 having a point mutation in the sequence coding for the transmembrane domain was expressed. These cell lines were termed RC and A4, respectively. A4 cells but not RC or NIH3T3 cells grew even in the presence of a low concentration (0.05%) of calf serum (CS), although the rate of their proliferation was low. In media containing 0.1% CS, both A4 cells and RC cells but not NIH3T3 cells could proliferate. Furthermore, RC cells induced foci formation when cultured in media containing 0.5% and 5% CS, while growth of the parental NIH3T3 cells was contact-inhibited under these conditions. These data suggest the presence of a factor(s) which activates protein-tyrosine kinase activity of the c-erbB-2 protein. In fact, the c-erbB-2 protein prepared from RC cells showed CS-dependent protein-tyrosine kinase activity when assayed in membrane fractions.

Activity and safety of DNA plasmids encoding IL-4 and IFN gamma.

Cytokine-encoding DNA plasmids can act as 'genetic adjuvants', improving the immune response stimulated by co-administered DNA vaccines. We examined whether plasmids encoding the Th1 cytokine IFN gamma (pIFN gamma) or the Th2 cytokine IL-4 (pIL-4) have long-term effects on immune homeostasis when administered to adult mice, or alter immune maturation in neonates. Both plasmids boosted immunity against a co-administered vaccine, with pIFN gamma promoting the development of a Th1 response (characterized by the production of IgG2a antibodies), and pIL-4 preferentially stimulating a Th2 response (characterized by increased IgG1 antibody production). Both pIFN gamma and pIL-4 influenced the ratio of cells actively secreting Th1 versus Th2 cytokines, consistent with an effect on Th cell maturation. Interestingly, this effect persisted for only a few weeks and was not magnified by repeated plasmid administration. Cytokine-encoding plasmids had no long-term effect on the immune response of newborn or adult mice to subsequent antigenic stimulation, nor did they selectively induce the production of pathogenic anti-DNA autoantibodies. These results suggest cytokine-encoding plasmids may be safe as immune adjuvants.

In vivo mechanisms for the inhibition of T lymphocyte activation by long-term therapy with tacrolimus (FK-506): experience in patients with Behçet's disease.

OBJECTIVE: To examine the in vivo mechanisms of suppression of T lymphocyte function in patients with Behçet's disease (BD) undergoing long-term treatment with tacrolimus (FK-506). METHODS: Intracellular proteins were analyzed by immunoprecipitation and Western blotting. Messenger RNA expression was studied by a polymerase chain reaction-based technique. RESULTS: Interleukin-2 production was suppressed in patients treated with tacrolimus. This suppression was found to be due to inhibition of interactions between activated calcineurin (Cn) and nuclear factor of activated T cells (NF-AT), inhibition of cleavage of the autoinhibitory domain of the CnA subunit, and inhibition of heterodimer formation by CnA and CnB subunits, resulting in the absence of NF-AT in nuclei of the T cells. We found that T lymphocytes in some BD patients treated with tacrolimus had reduced amounts of FK-506 binding protein (FKBP) in their cytoplasm. CONCLUSION: Tacrolimus reduces the Cn activity of T cells in vivo by the cumulative effects of several distinct mechanisms. It is plausible that reduced amounts of FKBP may be associated with diminished clinical efficacy in some BD patients receiving prolonged treatment with tacrolimus.

Inhibition of Fas/Fas ligand-mediated apoptotic cell death of lymphocytes in vitro by circulating anti-Fas ligand autoantibodies in patients with systemic lupus erythematosus.

OBJECTIVE: The Fas/Fas ligand (FasL) system has been assigned a pivotal role in the establishment and maintenance of peripheral tolerance, and mice having defects in the Fas/FasL system are known to develop lupus-like symptoms. However, it remains unclear whether the Fas/FasL system is involved in the pathogenesis of systemic lupus erythematosus (SLE) in humans. This study examined whether there are circulating anti-FasL autoantibodies in the peripheral blood of patients with SLE that would interfere with Fas/FasL-mediated apoptosis. METHODS: Anti-FasL autoantibodies were detected by Western blot analysis using the recombinant extracellular domain of human FasL as the antigen. Apoptosis of Fas-expressing Jurkat cells, induced by recombinant soluble FasL (sFasL) in the presence of anti-FasL autoantibodies, was assessed by DNA staining with propidium iodide, followed by flow cytometric analysis. Apoptosis of Jurkat cells by cell-bound FasL was assessed by 2-color analysis, involving TUNEL staining with fluorescein isothiocyanate-dUTP and phycoerythrin-labeled anti-CD3 monoclonal antibodies. RESULTS: Among the 21 patients with SLE, 7 had IgG-isotype anti-FasL autoantibodies in their circulating blood. In addition, these autoantibodies inhibited both sFasL-mediated and cell-bound FasL-mediated apoptosis of Fas-expressing Jurkat cells. Thus, it is plausible that anti-FasL autoantibodies in patients with SLE disturb the establishment and maintenance of peripheral tolerance in vivo by inhibiting the Fas/FasL-mediated elimination of autoreactive lymphocytes. CONCLUSION: These results suggest that anti-FasL autoantibodies that inhibit Fas/FasL-mediated apoptosis are involved, at least in part, in immune abnormalities and may possibly be involved in the pathogenesis of SLE.

Liposome encapsulated doxorubicin transfer to the pelvic lymph nodes by endoscopic administration into the bladder wall: a preliminary report.

PURPOSE: Liposome encapsulated doxorubicin administered to the pelvic lymph nodes might be effective against pelvic node metastasis. We determined whether the drug can be delivered to the nodes via the bladder wall. MATERIALS AND METHODS: Ten patients underwent endoscopic administration of 10 mg. liposomal doxorubicin into the bladder wall 3 days before cystectomy. The concentration of doxorubicin in the pelvic lymph nodes obtained at operation was measured. RESULTS: No complication developed after drug administration. Almost all lymph nodes examined contained more than 100 ng./gm. doxorubicin (median 1,860). CONCLUSIONS: Endoscopic administration of liposomal doxorubicin into the bladder wall may be an alternative method to deliver doxorubicin to the pelvic lymph nodes.