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T-cell receptor Vbeta gene usage by lymphocytes infiltrating human renal allografts.

T cell lines have been derived from human kidney allograft biopsies using mitogenic stimulation. Southern blotting using a T-cell receptor (TCR) Cbeta probe revealed an oligoclonal pattern of rearranged bands in all 12 samples analysed. In some cases, differences in band patterns were noted between independent cultures from the same biopsy. Most T-cell clones derived from 2 biopsies showed different patterns of rearranged bands. The polymerase chain reaction (PCR) was used to study TCR Vbeta gene usage in allograft-derived T-cell cultures. This was more sensitive and more informative than Southern blotting and revealed that most TCR Vbeta genes were expressed in T cells from biopsies showing cellular rejection. The potential usefulness of this technique to quantify TCR V gene usage in allospecific T-cell populations is discussed.

DNA↗

Nerve fibre proliferation in interstitial cystitis.

The aetiology of pain in interstitial cystitis is not understood, although it has been reported to be due to release of mediators from mast cell granules. Cystolysis and intravesical instillation of dimethyl sulphoxide have been shown to relieve pain in this condition. We have studied the nerve population within the bladder wall using immunohistochemical stains for protein gene product 9.5. A group of 18 cases of chronic interstitial cystitis and 12 controls; neuropathic bladder (n = 1), chronic bacterial cystitis (n = 3), systemic lupus erythematosus cystitis (n = 2) and normals (n = 6), were investigated. There were significantly more nerve fibres within the sub-urothelial and detrusor muscle layers in chronic interstitial cystitis than there were in normals. Patients with chronic cystitis of other aetiology did not have a significant increase in nerve fibre density within the bladder wall suggesting a specific association between nerve fibre proliferation and interstitial cystitis. Cystolysis is shown to deplete selectively the submucosal nerve plexuses without altering the nerve density within detrusor muscle. This finding explains the desensitisation of the bladder without impairment of detrusor function after this procedure.

Cell Division↗

Does the mast cell have an intrinsic role in the pathogenesis of interstitial cystitis?

In order to examine the role of mast cells in the inflammatory bladder disease interstitial cystitis, mast cells isolated from the human bladder of normal and diseased tissue were challenged with a range of secretagogues. Calcium ionophore A23187 and anti-IgE caused histamine release from all bladder mast cells in a dose-related manner. Mast cells from the diseased tissue were far more responsive than those from the normal tissue. Mast cells from the muscle of normal bladder were responsive towards substance P and compound 48/80. However, mast cells from interstitial cystitis bladder did not release significant amounts of histamine with these two secretagogues.

Calcimycin↗

Both soluble and membrane-bound forms of Flt3 ligand enhance tumor immunity following "suicide" gene therapy in a murine colon carcinoma model.

In prodrug-activated ("suicide") gene therapy, tumor cells are transfected with the gene for an enzyme that converts an inactive prodrug, such as ganciclovir (GCV), to a toxic compound. Transfected cells are killed on administration of GCV, as also are untransfected "bystander" cells. The ability of the dendritic cell stimulatory cytokine Flt3 ligand (Flt3-L) to modulate prodrug-activated gene therapy has been investigated. Transfectants of the murine colon carcinoma MC26 were generated expressing soluble (FLS) and membrane-bound forms of Flt3-L. They were inoculated together with wild-type MC26 cells and cells expressing herpes simplex virus-1 (HSV1) thymidine kinase into BALB/c mice, which were then administered GCV. Expression of Flt3-L or FLS prevented regrowth of tumor in most mice, which was comparable to the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF), while tumors recurred in all mice receiving "suicide" gene therapy alone. Recurring tumor cells were resistant to direct killing by GCV but sensitive to "bystander" killing in vitro. Mice without tumor recurrence were rechallenged with unmodified MC26 cells. Of those mice given transfectants expressing GM-CSF, Flt3-L, or FLS, approximately 50% were immune to rechallenge. These mice also showed cytotoxic and proliferative responses to MC26 cells. These experiments show that both soluble and membrane-bound forms of Flt3-L were able to induce a protective immune response to colon carcinoma cells in a fashion similar to GM-CSF.

Animals↗

Collecting duct carcinoma of the renal medulla presenting with paraplegia.

We report an interesting case of a patient with collecting duct carcinoma arising from the left kidney who presented with paraplegia secondary to metastases. The diagnosis was based on CT and histology. To our knowledge this is the first case of collecting duct carcinoma to present with paraplegia. The literature review also highlights the rarity of this disease with less than a hundred cases reported to date and the aggressive nature and poor prognosis despite prompt interventions.

Adult↗

Primary uretero-iliac fistula: the unusual source of haematuria.

Uretero-iliac fistula is a rare cause of frank haematuria. The aetiology of such fistula is commonly iatrogenic. We present a unique case of a primary aorto-iliac fistula in the absence of an aneurysm or arteriovenous malformation. The diagnosis was demonstrated by ureteroscopy and real-time retrograde ureterogram. Multiple arterial embolisation of the fistula had failed, and the patient underwent a successful ureterolysis and ligation of fistula. We demonstrate the diagnostic difficulties and treatment dilemma of such rare cause of haematuria.

Aged↗

Pharmacokinetics of azidothymidine during late pregnancy in Long-Evans rats.

Azidothymidine (Zidovudine) is used to treat symptomatic human immunodeficiency virus infection. It has the ability to reverse seropositivity as well as decrease serum viral titers, and perhaps decrease potential for transmission of the virus. An animal model (pregnant Long-Evans rats) was used to investigate the potential for placental transmission of azidothymidine. The drug crosses the placenta to reach concentrations in the placenta and fetus that are comparable to 75% and 58%, respectively, of those in the maternal serum by 2 hours after administration. By 4 to 6 hours after administration azidothymidine concentrations in the placenta and fetal liver significantly exceed maternal concentrations.

Animals↗

Placental transfer of cefazolin and piperacillin in pregnancies remote from term complicated by Rh isoimmunization.

Although the administration of prophylactic antibodies for intrauterine transfusion is controversial, little information is available regarding placental transfer of antibiotics administered to the mother, or whether the presence of hydrops affects this placental transfer. Sixteen intravascular intrauterine transfusions were performed in 10 patients. Seven (10 procedures) patients were given 2 gm of cefazolin before the procedure and samples were obtained by fetal vascular access. Three patients (six procedures) were given 4 gm of piperacillin and samples were similarly obtained. Specimens were obtained for fetal serum, maternal serum, and amniotic fluid antibiotic concentration. The mean serum cefazolin concentration in hydropic fetuses was 18.04 +/- 3.37 micrograms/ml, and in nonhydropic fetuses the concentration was 21.02 +/- 17.8 micrograms/ml (p = 0.72). The mean fetal serum concentration of piperacillin was 22 +/- 12 micrograms/ml. The placental transfer of both drugs was similar. We conclude that the transplacental passage of these antibiotics is prompt and that the presence of hydrops does not significantly impair the passage of cefazolin.

Blood Transfusion, Intrauterine↗

Changes in T-lymphocyte subpopulations during pregnancy complicated by human immunodeficiency virus infection.

OBJECTIVE: We describe changes in T-lymphocyte subpopulations in pregnancies complicated by human immunodeficiency virus infection. STUDY DESIGN: T-lymphocyte counts were performed every trimester and post partum on all patients with human immunodeficiency virus infection. Patients with a CD4 count < 500 cells/mm3 were offered zidovudine after the first trimester. We performed a regression analysis of repeated measures on the total lymphocyte, CD4, and CD8 counts. RESULTS: Twenty-three patients were studied, 10 (43%) of whom took zidovudine. CD4, CD8, and total lymphocyte counts decreased in patients not taking zidovudine but remained stable in patients receiving zidovudine. The differences were not statistically significant. CONCLUSIONS: CD4 and CD8 counts may decrease during pregnancies complicated by human immunodeficiency virus infection, primarily because of decreases in the total lymphocyte count. The use of zidovudine may prevent this decline. The clinical use of absolute CD4 counts during pregnancy requires further study.

Acquired Immunodeficiency Syndrome↗

T cell receptor gamma gene status of human alpha/beta+ and gamma/delta+ T cell clones: absence of V9JP rearrangements in alpha/beta+ clones is not a result of a lack of rearrangements involving more 5' J gamma segments.

T cell receptor (TCR) gamma gene rearrangements were examined in panels of human T cell clones expressing TCR alpha/beta or gamma/delta heterodimers. Over half of the alpha/beta+ clones had both chromosomes rearranged to C gamma 2 but this was the case for only 20% of the gamma/delta+ clones. While more than half of the gamma/delta+ clones showed a V9JP rearrangement, this configuration was absent from all 49 alpha/beta+ clones analysed. However, this was not a result of all rearrangements being to the more 3' J gamma genes as 11 alpha/beta+ clones had rearrangement(s) to JP1, the most 5' J gamma gene segment. Both alpha/beta+ and gamma/delta+ clones showed a similar pattern of V gamma gene usage in rearrangements to J gamma 1 or J gamma 2 with a lower proportion of the more 3' genes being rearranged to J gamma 2 than for the more 5' genes. Several alpha/beta+ and several gamma/delta+ clones had noncoordinate patterns of rearrangement involving both C gamma 1 and C gamma 2. Eleven out of fourteen CD8+ clones tested had both chromosomes rearranged to C gamma 2 whereas all clones derived from CD4-8- cells and having unconventional phenotypes (CD4-8- or CD4+8+) had at least one C gamma 1 rearrangement. Twelve out of twenty-seven CD4+ clones also had this pattern, suggesting that CD4-8+ clones had a tendency to utilize more 3' J gamma gene segments than CD4+ clones. There was some evidence for interdonor variation in the proportions of TCR gamma rearrangements to C gamma 1 or C gamma 2 in alpha/beta+ clones as well as gamma/delta+ clones. The results illustrate the unique nature of the V9JP rearrangement in gamma/delta+ clones and the possible use of a sequential mechanism of TCR gamma gene rearrangements during T cell differentiation is discussed.

Antigens, Differentiation, T-Lymphocyte↗

M&B 28,767: a potent anti-secretory and anti-ulcer PG analogue. A comparative study with 16, 16' dimethyl PGE2 methylester.

M&B 28,767 [(+/-)11-deoxy-16-phenoxy-omega-tetranor PGE1] and 16, 16'-dimethyl PGE2 methylester (DMPG) were compared for their effects on gastric acid secretion (GAS) and gastric ulceration (GU), employing various laboratory models. In anaesthetised rats, GAS was stimulated by a continuous i.v. infusion of pentagastrin (30 micrograms/kg/h), and PG analogues were perfused through the stomach for 1 h. M&B 28,767 (3-15 micrograms/kg/h) and DMPG (3-60 micrograms/kg/h) reduced GAS in a dose-related manner, the ED50 values being 4 and 15 micrograms/kg/h respectively. In conscious rats possessing indwelling gastric cannulae, oral doses of M&B 28,767 (0.025-0.1 microgram/kg) and DMPG (0.50-1.0 microgram/kg) caused a prolonged inhibition of pentagastrin-stimulated GAS. M&B 28,767 was 17 times more potent than DMPG; the respective ED50 values were 0.036 and 0.6 microgram/kg. Indomethacin-induced ulceration in rats, was reduced by both M&B 28,767 and DMPG; the respective ED50 values being 3.0 and 0.8 micrograms/kg. Both compounds given orally increased gastrointestinal motility in mice; M&B 28,767 (1-3 mg/kg) and DMPG (0.1-0.3 mg/kg) caused diarrhoea, the former being about 0.1 times as potent as the latter. In another test, M&B 28,767 (0.5-5.0 mg/kg) and DMPG (10-40 micrograms/kg) overcame morphine-induced constipation in a dose-related manner, the respective ED50s being 0.9-1.4 mg/kg and 20-40 micrograms/kg. Thus, M&B 28,767 had a better profile of activity than DMPG as an antisecretory and antiulcer agent.

16,16-Dimethylprostaglandin E2↗