Immunohistochemical localization of actinomycin D in human melanoma tumor cells

Human melanoma tumor colonies and derived cell lines (81-46a) were exposed to Actinomycin D (Act D) in vitro in order to study subcellular sites of drug-cell interaction. Light microscopy of 13 day-old colonies previously treated for 24 hours with 0.01 microgram/ml, 0.1 microgram/ml, and 1 microgram/ml Act D revealed a decrease in colony size and organization, an increase in pyknotic nuclei, and an accumulation of cell debris concomitant with an increase in Act D concentration. The 81-46a cells were treated with 0.1 microgram/ml Act D for 15, 30, and 60 minute intervals, followed by Act D antibodies. Immunofluorescence microscopy revealed both cytoplasmic, vesicular, filamentous, and nuclear variations in drug distribution with increased exposure. Cell viability studies indicated a decrease in viable cells as exposure time to Act D increased.     

Pseudohypoparathyroidism with paradoxical increase in hypocalcaemic seizures due to long-term anticonvulsant therapy

A case of pseudohypoparathyroidism is described. Unusual features included the apparent absence of a familial history and the long delay in clinical diagnosis. Dental evidence is presented which dates the metabolic abnormality back to at least the age of 2, yet symptoms did not appear until the age of 12 and the correct diagnosis was not made until she was 32. The role of long-term anticonvulsant therapy in the paradoxical exacerbation of the patient''s hypocalcaemic seizures is discussed.     

Mesangiocapillary glomerulonephritis: A long-term study of 40 cases

Forty cases of mesangiocapillary glomerulonephritis are reviewed for whom both light and electron microscopy and full clinical data were available. Immunofluorescence microscopy (IF) was performed on 23 cases and complement screening (CH50, C4 and C3) on 25 cases, with follow-up period of 5–22 y. The results of EM revealed 17 cases (42 per cent.) of Type I and 23 cases (58 per cent.) of Type II MCGN but only 52 per cent. of Type II cases were correctly identified by light microscopy. Epimembranous deposits were seen as frequently in Type II as in Type I (26 per cent. and 30 per cent.) and fragmentation of glomerular capillary basement membranes (GBM) was seen in 27 per cent. of Type I cases. Overall patient survival was 49 per cent. at 10 y and that of patients who presented with nephrotic syndrome was poor (39 per cent. at 10 y). Persistent hypo-complementaemia with C3 Nephritic Factor was present in 40 per cent.; the survival of these patients was less than those with normal complement levels (70 per cent. vs 100 per cent. at 5 y) and they were also more likely to develop renal failure. Renal failure was more likely to develop in those with a creatinine clearance of < 100 ml/min at presentation and where the biopsy showed substantial crescents in >20 per cent. of glomeruli. Mean CH50, C3 and C4 was lower in the hypocomplementaemic as compared to normocomplementaemic patients, and there were no differences between Type I and Type II. IF showed immuno-globulins and fibrin as well as C3 in both Type I and Type II cases. Our results support the concept of an immune-complex mediated phase in both types of MCGN, and we further suggest that (a) epimembranous deposits are common in both Type I and Type II and (b) cases with fragmentation of the GBM should be designated Type Ia.     

Leukaemic liver infiltration in AKR mice prematurely induced by immunization with human or allogeneic liver protein.

An attempt was made to produce a model of autoimmune hepatitis in AKR mice by immunization with allogeneic liver extract or a purified (human) liver-specific lipoprotein (LSP). Although a mononuclear infiltrate was found in the liver in recipients of purified liver antigen and to a lesser degree in recipients of allogeneic extract, this was associated with the development of an acute thymus-derived leukaemia. This mouse strain is normally susceptible to leukaemia after a latent period of approximately 6 to 9 months but the immunization schedule markedly reduced the age of onset of this disease.     

Changes in plasma amino acid patterns in chronic alcoholic patients during ethanol withdrawal syndrome: their clinical implications

Changes or imbalances in plasma amino acid patterns during withdrawal from ethanol were recorded in six randomly selected male chronic alcoholic patients (age range 23-47 years). Duration of drinking ranged from 4-15 years and their average daily amount of ethanol intake was more than 100G. Plasma amino acids (taurine, threonine, serine, glutamate, glutamine, proline, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, tryptophan, ornithine, lysine and arginine) were estimated by autoanalyzer in all patients on admission before starting conventional detoxification therapy for ethanol withdrawal syndrome, and during therapy on day 3 and day 6. On admission, there was a statistically significant rise in the plasma levels of almost all aminoacids, particularly glutamate, glutamine, phenylalanine, proline, glycine, methionine, cysteine, lysine, tyrosine, valine, isoleucine, leucine, serine, threonine, alanine and arginine (in comparison to those of normal controls) in five out of six patients. During the following six days of treatment and total abstinence, the pattern of plasma aminoacid levels did not change significantly despite considerable clinical improvement. Plasma tryptophan levels were undetectable in all patients on admission, day 3 and also on day 6 except in one patient with lesser amount and shorter duration of drinking, the levels just returned to within normal range only on day 6. Plasma levels of histidine and taurine were found to be slightly lower than normal.