Piriformis pyomyositis, an unusual presentation of leg pain post partum--case report and review of literature.

Piriformis pyomyositis is defined as a subacute infection of skeletal muscles associated with systemic infectious symptoms. In the literature it rarely occurs postpartum. We report a case of piriformis pyomyositis involving a parturient and review the published cases available in the literature.

Potential predictors of chemotherapy response in ovarian cancer--how do we define chemosensitivity?

OBJECTIVE: The aim of this study was to assess whether microvessel density (measured by CD31), vascular endothelial growth factor (VEGF) or multidrug resistance (MDR1) could determine the response to chemotherapy or act as prognostic factors in ovarian cancer. METHODS: Seventy-nine ovarian specimens were immunostained. Pearson correlation, 1-way ANOVA and chi-square were used for univariate analysis. Kaplan Meier survival curves were used, log-rank was used for univariate analysis and a Cox proportional hazards regression model was used for multivariate evaluation. Response to chemotherapy was assessed after 6 months and again after 1 year. RESULT: Quantifying VEGF proved to be a valuable independent prognostic indicator in progression-free survival (PFS) (p<0.05) and overall survival (OS) (p<0.0001). VEGF correlated with response to chemotherapy at the 6-month interval (r=0.446, p<0.001) but failed to correlate at the 1-year interval. Increased staining with CD31 was associated with decreased PFS (p<0.01) and OS (p<0.01) in univariate but not multivariate analysis. MDR1 failed to act as a prognostic marker or as a predictor of response to chemotherapy. CONCLUSION: VEGF correlates with response to chemotherapy at the 6-month but not the 12-month interval. What should our criteria be for determining sensitivity to chemotherapy? CD31, VEGF and MDR1 do play a role in some ovarian malignancies but other factors are likely to be involved and perhaps molecular profiling will determine which factors will be important for determining the response to chemotherapy.

p16INK4A, CDC6, and MCM5: predictive biomarkers in cervical preinvasive neoplasia and cervical cancer.

AIM: To analyse and compare expression patterns of three potential biomarkers-p16(INK4A), CDC6, and MCM5-and evaluate their use as predictive biomarkers in squamous and glandular cervical preinvasive neoplasia. METHODS: Immunocytochemical analysis of p16(INK4A), MCM5, and CDC6 expression was performed on 20 normal, 38 cervical intraepithelial neoplasia 1 (CIN1), 33 CIN2, 46 CIN3, 10 squamous cell carcinoma, 19 cervical glandular intraepithelial neoplasia (cGIN), and 10 adenocarcinoma samples. Staining intensity was assessed using a 0-3 scoring system. p16(INK4A), MCM5, and CDC6 expression was also examined in ThinPrep slides exhibiting mild, moderate, and severe dyskaryosis. Human papillomavirus (HPV) was detected using a modified SYBR green assay. Fluorogenic polymerase chain reaction (PCR) and solution phase PCR were used for specific HPV typing. RESULTS: All three markers showed a linear correlation between expression and grade of dysplasia. p16(INK4A) and MCM5 protein expression was upregulated in all grades of squamous and glandular dysplasia. CDC6 protein was preferentially expressed in high grade lesions and in invasive squamous cell carcinoma. CONCLUSION: p16(INK4A) expression was closely associated with high risk HPV infection-all grades of squamous and glandular cervical lesions were immunohistochemically positive. MCM5 staining intensity was independent of high risk HPV infection, highlighting its potential as a biomarker in both HPV dependent and independent cervical dysplasia. CDC6 may be a biomarker of high grade and invasive lesions of the cervix, with limited use in low grade dysplasia. p16(INK4A) was the most reliable marker of cervical dysplasia. Combinations of dysplastic biomarkers may be useful in difficult diagnostic cases.

p16INK4A positivity in benign, premalignant and malignant cervical glandular lesions: a potential diagnostic problem.

A wide array of immunohistochemical markers have been evaluated with respect to their specificity in staining dysplastic cervical cells in cervical biopsies and cervical cytological smears. However, there is still a significant demand for better biomarkers to identify neoplastic cervical glandular and squamous epithelial cells precisely. The CDKN2A gene, located on chromosome 9p21, encodes the tumour suppressor protein, p16INK4A, which decelerates the cell cycle by inactivating CDK4 and CDK6. The aim of this study was to compare and contrast the expression pattern of p16INK4A in benign and neoplastic glandular lesions and tubo-endometrioid metaplasia. All cases in each category displayed some p16INK4A expression. Adenocarcinoma and in situ cases showed a combination of intense nuclear and cytoplasmic staining. It was observed that all cases of tubo-endometrioid metaplasia showed occasional nuclear positivity and definite cytoplasmic staining. These findings may have important implications for the potential utility of p16INK4A as a biomarker for glandular dysplastic lesions. While p16INK4A has been demonstrated to be an excellent marker of cervical dysplasia in squamous neoplastic lesions of the cervix, it has potential pitfalls in cervical glandular lesions that may limit the utility of this biomarker in resolving the nature of suspicious glandular lesions, particularly in cytopathology.

p16INK4A as a marker for cervical dyskaryosis: CIN and cGIN in cervical biopsies and ThinPrep smears.

AIM: To examine the potential of p16(INK4A) as a biomarker for dysplastic squamous and glandular cells of the cervix in tissue sections and ThinPrep smears. METHODS: Immunocytochemical analysis of p16(INK4A) expression was performed on 22 normal cervical tissue samples, five cervical glandular intraepithelial neoplasia (cGIN), 38 cervical intraepithelial neoplasia 1 (CIN1), 33 CIN2, 46 CIN3, and 10 invasive cancer cases (eight squamous and two adenocarcinomas). All samples were formalin fixed and paraffin wax embedded, and immunohistochemical analysis was carried out using a mouse monoclonal anti-p16(INK4A) antibody after antigen unmasking. The staining intensity was assessed using a 0 to 3 scoring system. In addition, the expression status of p16(INK4A) was examined in 12 normal ThinPrep smears, one smear exhibiting cGIN, and a total of 20 smears exhibiting mild, moderate, and severe dyskaryosis. Human papillomavirus (HPV) detection was carried out using a modified SYBR green assay system. Fluorogenic polymerase chain reaction (PCR) and solution phase PCR were used for specific HPV typing. RESULTS: p16(INK4A) immunoreactivity was absent in all normal cervical tissues examined. Dysplastic squamous and glandular cells were positive for p16(INK4A) expression in all cases included in this study, except for one CIN3 case. p16(INK4A) expression was mainly nuclear in CIN1 cases, and both nuclear and cytoplasmic in CIN2, CIN3, cGIN, and invasive cases. All cases positive for HPV expressed the p16(INK4A) protein, although not all cases found positive for p16(INK4A) were HPV positive. In general, the p16(INK4A) staining intensity was lower in cases negative for HPV or those containing a low risk HPV type. CONCLUSION: This pattern of overexpression demonstrates the potential use of p16(INK4A) as a diagnostic marker for cervical squamous and also glandular neoplastic lesions. In addition, the technique can be used to identify individual dyskaryotic cells in ThinPrep smears. Thus, p16(INK4A) is a useful marker of cervical dyskaryosis.

Serous papillary adenocarcinomas of the ovary display heterogeneity in their response to chemotherapy.

The response of ovarian serous papillary adenocarcinomas to various cytotoxic drugs was examined using the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) (MTS) cytotoxicity assay. Thirty tumors were collected and organized into four groups according to histologic grade and FIGO stage: stage III, grade 2; stage III, grade 3; stage IV, grade 2; and stage IV, grade 3. The MTS chemosensitivity assay was performed on each tumor to examine the response to cisplatin, paclitaxel, hycamtin and the combination of cisplatin and paclitaxel. Ovarian adenocarcinomas of similar stage and grade displayed varying responses to the same drug. A lower concentration of the drug was often as effective as the peak plasma concentration. For some specimens combination therapy was more effective for inhibiting tumor growth, and for others single-agent therapy gave a better response. A chemosensitivity/resistance profile is recommended before deciding on appropriate chemotherapy.

Robotic enzyme amplification: a comparison of some kinetic properties of bovine liver, Candida utilis and Proteus sp. glutamic dehydrogenases.

NADP(H)-specific Bakers yeast glucose 6-phosphate dehydrogenase (BYG6PDH) was paired, in turn, with each of three different source glutamate dehydrogenases (GDHs): NAD(P)-specific bovine liver (BLGDH), NADP-specific Candida utilis (CUGDH) and NADP-specific Proteus sp. (PSGDH) to constitute three enzyme cycling systems; (i) BYG6PDH/BLGDH; (ii) BYG6PDH/CUGDH; and (iii) BYG6PDH/PSGDH. When incorporated into an enzymatic cycling/amplification system for NAD kinase and run on a centrifugal fast analyzer (CFA), the microbial source enzyme CUGDH gave rise to a seven-fold greater amplification rate [21.5 x 10(3) cycles-1 (cph)] relative to that realized (3 x 10(3) cph) using the BYG6PDH/BLGDH cycling pair previously reported. Either of these cycling systems can be used as a flexible and general-purpose module for robotic amplification and data collection of NADP(H) linked enzymes as a user's requirements dictate. Although the BYG6PDH/PSGDH cycling pair produced a respectable cycling rate (14.4 x 10(3) cph), for reasons discussed the PSGDH enzyme was not considered a suitable replacement for BLGDH in an NADP(H) cycling system.

Topographical localization of lipofuscin pigment in the brain of the aged fat-tailed dwarf lemur (Cheirogaleus medius) and grey lesser mouse lemur (Microcebus murinus): comparison to iron localization.

The present study was undertaken to explore the distribution of lipofuscin in the brain of cheirogaleids by autofluorescence and compare it to other studies of iron distribution. Aged dwarf (Cheirogaleus medius) and mouse (Microcebus murinus) lemurs provide a reliable model for the study of normal and pathological cerebral aging. Accumulation of lipofuscin, an age pigment derived by lipid peroxidation, constitutes the most reliable cytological change correlated with neuronal aging. Brain sections of four aged (8-15 year old) and 3 young (2-3 year old) animals were examined. Lipofuscin accumulation was observed in the aged animals but not in the young ones. Affected regions include the hippocampus (granular and pyramidal cells), where no iron accumulation was observed, the olfactory nucleus and the olfactory bulb (mitral cells), the basal forebrain, the hypothalamus, the cerebellum (Purkinje cells), the neocortex (essentially in the pyramidal cells), and the brainstem. Even though iron is known to catalyse lipid oxidation, our data indicate that iron deposits and lipofuscin accumulation are not coincident. Different biochemical and morphological cellular compartments might be involved in iron and lipofuscin deposition. The nonuniform distribution of lipofuscin indicates that brain structures are not equally sensitive to the factors causing lipofuscin accumulation. The small size, the rapid maturity, and the relatively short life expectancy of the cheirogaleids make them a good model system in which to investigate the mechanisms of lipofuscinogenesis in primates.

Plasminogen activators and inhibitors in ovarian adenocarcinomas.

The fibrinolytic process was examined in cultures from malignant epithelial ovarian tumors and compared to normal ovarian tissue. The fibrinolytic enzymes, urokinase plasminogen activator (uPA) antigen and activity, tissue plasminogen activator (tPA) antigen and activity, plasminogen activator inhibitor 1 (PAI-1) and plasminogen activator inhibitor 2 (PAI-2) antigen were measured in short-term primary cultures at weekly intervals over a 3-week period. The cultures comprised of 15 ovarian adenocarcinomas and 12 normal ovarian tissue samples. The concentration of uPA antigen (p < 0.01) and activity (p < 0.01) and (p < 0.05) levels were higher in malignant specimens on all 3 count days. Activity levels of tPA were elevated significantly in malignant specimens on days 7 and 21 (p < 0.05). No significant difference was found between PAI-1 levels. PAI-2 antigen levels were significantly higher in the tumor specimens on days 14 and 21 (P < 0.01). These data indicate that uPA may have a significant role in the biology of ovarian cancer and may be an important factor in early tumor spread. Further work is required on the effects of intervention in this biological process.

Ocular lens NAD kinase: partial purification and metabolic implications.

The ocular lens displays a significant amount of NADP(H) dependent metabolic traffic, but the origin of this cofactor has not been established. Size exclusion chromatography of bovine lens crude extract on a Sephacryl S300-HR column fitted with an eluate concentrator revealed two bands with NAD kinase activity, based on enzymatic cycling with signal amplification of the column fractions using a Cobas-Fara II centrifugal fast analyzer. Ve/Vo ratios from the chromatographic runs suggest that the relative molecular weight values lie within the ranges 8.91-3.98 x 10(5) and 2.04-1.26 x 10(5), respectively, for these two bands. An approximately 10-fold enhancement of enzyme activity over the crude fraction is realized from the chromatography step. Results point to NAD kinase as the source generator of this anchoring and linking cofactor for the oxidative stress and pentose phosphate enzyme systems, respectively.

Laminar organization of acetylcholinesterase and cytochrome oxidase in the lateral geniculate nucleus of prosimians.

Hess and Rockland [Hess and Rockland (1983) Brain Res. 289, 322-325] proposed that the distribution of acetylcholinesterase within the lateral geniculate nucleus might correlate with the daily activity patterns shown by primates. In diurnal primates, the magnocellular laminae show a greater acetylcholinesterase reaction product. In nocturnal primates, the parvocellular laminae are more heavily stained. We have examined the laminar distribution of acetylcholinesterase and cytochrome oxidase in the lateral geniculate nucleus of a series of rare prosimian primates. In all prosimians examined, the most dense acetylcholinesterase reaction product is seen in the parvocellular layers of the lateral geniculate nucleus. Heavy cytochrome oxidase activity is seen in both the magnocellular and parvocellular layers, but not the koniocellular layers of the prosimian lateral geniculate nucleus. We have also employed a polyclonal antibody to choline acetyltransferase to examine the laminar organization or cholinergic activity in the Galago (Bushbaby) lateral geniculate nucleus. We report that choline acetyltransferase immunoreactivity does not correlate with acetylcholinesterase activity in the prosimian lateral geniculate nucleus. Although the lateral geniculate nucleus is more immunoreactive than most other thalamic structures and although the intercalated koniocellular laminae demonstrate somewhat lighter choline acetyltransferase immunoreactivity, no great difference in staining intensity is seen between the parvocellular and magnocellular laminae. In addition, we examined the phenotype of known inputs to assess the laminar specificity of cholinergic projections to the bushbaby lateral geniculate nucleus. Layer VI of primary visual cortex, which is known to be a source of acetylcholinesterase in the parvocellular layers, does not contain cholinergic cells, nor does the pretectal nucleus, which projects mainly to the parvocellular layers. The parabigeminal nucleus is cholinergic; however, this nucleus is known to project to the koniocellular layers, along with the non-cholinergic superior colliculus. Finally, the pedunculopontine tegmental nucleus, which provides a strong input to many regions of the thalamus, including the lateral geniculate nucleus, is cholinergic. The laminar organization of its input to the lateral geniculate nucleus is not known. Increased acetylcholinesterase reaction product within the parvocellular layers of the lateral geniculate nucleus is common to all strepsirhine primates. The pattern is also seen in the only two nocturnal haplorhine primates, Tarsius and Aotus (owl monkey). The relation of this increased acetylcholinesterase activity to cholinergic function remains unclear.(ABSTRACT TRUNCATED AT 400 WORDS)

Plasma exchange in the management of severe rhesus haemolytic disease.

Over a ten year period from 1977 to 1987, 30 mothers with severe rhesus haemolytic disease and expected fetal loss were treated by plasma exchange using a blood cell separator. All patients had at least on previous stillborn or neonatal death due to haemolytic disease of the newborn. Of the 30 patients, 19 pregnant women were successfully treated. Overall, 53% (16 babies) survived intact. Three of the deaths were due to causes other than erythroblastosis foetalis.

Adaptation of an enzymatic cycling assay for NADP(H) measurement to the COBAS-FARA centrifugal analyzer.

NADP(H) measurements by enzymatic amplification are described in which the interface step between cycling (glucose-6-phosphate and glutamic dehydrogenases) and indicator (6-phosphogluconic dehydrogenase) enzymes has been reconfigured, permitting the entire operation to run as a continuous assay on a centrifugal fast analyzer. This is accomplished by using the sequential load feature of the analyzer and incorporating either sodium dodecyl sulfate (SDS) or SDS and hydrogen peroxide as kill reagents to replace the thermal step (destruction of cycle enzymes by boiling). The ability of SDS to render a cycle inoperative during the run time of the indicator enzyme depends on the inherent resistivity and absolute amount of its enzyme proteins to this surfactant. Criteria used to judge the efficacy of a potential kill reagent are based on the sample blank time-response curve and the cycle product recovery by the indicator enzyme. Various other enzyme cycling systems which can be fitted to the centrifugal fast analyzer are highlighted.

Fertility following perforated appendicitis in girls.

A study was undertaken to determine fertility status in a group of adult females who as children had been operated on for perforated appendicitis between 1957 to 1975. The 389 girls operated on for perforated appendicitis were reviewed. Their ages ranged from 10 months to 13 years at the time of appendicectomy. Of these girls, 276 were now 20 to 43 years old, and they were contacted by means of a mail questionnaire, and personal interview wherever necessary. It proved possible to contact 181 women; 102 of them were married and 79 were unmarried. Eight-four of the married women (82%) had one or more children. Nine unmarried women also had one or more children. Eighteen married women who have no children were studied in detail. Five women were on contraceptives, two desired pregnancy but had not conceived, and one patient was separated from her husband. Two patients had conceived and aborted, and two were married to infertile men. Of the remaining six patients who had been investigated for infertility, no demonstrable cause of infertility was found in three. Of the other three patients, one showed evidence of bilateral tubal occlusion secondary to pelvic inflammatory disease, one has had a right ectopic pregnancy followed by two abortions, and the third patient was found to have a pituitary adenoma. Our data show that perforated appendicitis before puberty has little if any role in the aetiology of tubal infertility.

Polyol dehydrogenase: purification. Evidence for multiple forms and some properties of the dominant variant of the horse liver enzyme.

Purification of horse-liver polyol dehydrogenase (PDH) on DE52 anion-exchange cellulose reveals the presence of three fractions with enzyme activity. These appear in the breakthrough volume (PDH-3) and the salt gradient (PDH-1, -2) respectively. The major band of activity (greater than approximately 90%) is found in the PDH-2 fraction. A reexamination of sheep-liver polyol dehydrogenase also reveals the presence of three bands of activity, with the dominant fraction (PDH-3) corresponding to the preparation described by Smith (Biochem. J., 83, 135-144, (1962)). The interaction between horse-liver (and sheep-liver) PDH and Blue Sepharose CL-6B is found to be endothermic. This property is utilized in the final purification step. Horse-liver PDH-2 has a molecular/subunit weight of approximately 85,000/approximately 28,000, a Stokes' radius of 3.8 nm, and an isoelectric point of 7.4.

Fate of pancreatic nodules induced by raw soya flour in rats.

We have previously shown that rats fed raw soya flour (RSF) for more than four months develop hyperplastic foci of pancreatic acinar cells, which undergo malignant change if feeding RSF is continued throughout the life of the animals. The tendency to undergo malignant change is augmented by the additional use of a genotoxic carcinogen such as azaserine. The present study has sought to examine the reversibility of the focal neoplastic change in the pancreas. Rats fed RSF for 24 weeks and then given a diet not containing soya flour (NSC) had a normal pancreas when killed after 60 weeks of study. When RSF was fed for only 36 weeks, however, some of the rats developed pancreatic cancer even though the diet had been switched to NSC. Similarly, while azaserine in the dose used in the present study does not produce pancreatic cancer in our strain of Wistar rats, coincident administration of RSF for 12 weeks (but not for six weeks) resulted in progression to pancreatic adenoma. Although change from RSF to NSC after 30 weeks resulted in rapid reduction in pancreatic weight and content of RNA, neoplastic foci persisted and became frankly malignant. We conclude that phenotypic reversion to normal of the RSF diet- and azaserine-treated rat pancreas is only possible if RSF alone is fed continuously for not more than about 24 weeks or six weeks if the rats have been exposed to a pancreatic initiating carcinogen.