Closing the audit cycle: improving short-term outcomes of oesophagectomy in a provincial hospital.

A previously published study regarding the outcomes of oesophagectomy at a provincial hospital identified issues with perioperative care (Al-Herz et al 2012). The aim of this study was to evaluate the effect of changes in management at the institution concerned. This was a cohort study which compared the outcomes of 30 patients undergoing oesophagectomy before the unit audit and 30 patients after it. Demographics, operative details, recovery parameters, and oncological data were collected retrospectively. There was a significant reduction in the use of intravenous fluid, both intraoperatively (6.6 vs 3.3L, P < 0.0001) and during the first 24 hours (9.2 vs 5.5L, P < 0.0001). Patients were extubated three days earlier (P < 0.001) after the audit, and the percentage of patients requiring tracheostomy was smaller (26.7% vs 0%, P = 0.003). The length of total hospital stay was shorter (15 vs 13 days, P = 0.035). We conclude that the publication of a unit audit changed perioperative practice and resulted in a significant improvement in the short term outcomes after oesophagectomy.

Stress-response (heat-shock) protein 90 expression in tumors of the central nervous system: an immunohistochemical study.

This retrospective study deals with the expression of stress-response (heat-shock) protein 90 (srp 90) in a series of 148 human brain tumors. Immunohistochemical procedures were employed; cells of the human breast cancer line MCF7 exposed to hyperosmolar stress served as positive controls. Deposits of reaction products were found in the cytoplasm and they displayed a granular pattern. srp 90 was detected in 14/31 meningiomas and 5/10 breast cancer metastases to the brain. The protein was also present in 6/13 glioblastomas and 7/18 astrocytomas. In addition, a positive reaction was found in 2/10 medulloblastomas, 2/14 primitive neuroectodermal tumors, 1/11 pituitary tumor, 2/21 schwannomas and 2/11 lung tumor metastases; however, oligodendrogliomas and primary malignant lymphomas were not stained. The srp 90 was detected in Western blots of meningioma tissue homogenates. No significant immunohistochemical reaction was seen with sections of normal human cerebra, brain stem, cerebella, pituitary glands and spinal cords. These results document the expression of srp 90 by a variety of primary and metastatic intracranial tumors.

Brain tumor: immunohistochemical studies on the stress-response proteins, p53 protein and proliferating cell nuclear antigen.

This retrospective immunohistochemical study compares the expression of five stress-response (heat-shock) proteins (srp's) [srp 90, srp 72, srp 27, alpha B-crystallin and ubiquitin], p53 protein and proliferating cell nuclear antigen (PCNA) in 118 primary brain tumors and 21 carcinoma metastases to the central nervous system. Serial sections of formalin-fixed, paraffin-embedded tissues were used. Most astrocytomas (9/13), ependymomas (5/5), glioblastoma multiforme (GBM) (11/12), schwannomas (19/21), meningiomas (22/23) and breast carcinoma metastases (Br-Mt) (9/10), and some medulloblastomas (5/15), primitive neuroectodermal tumors (PNETs) (5/11), pituitary adenomas (4/7) and lung carcinoma metastases (6/11), but none of 10 oligodendrogliomas had tumor cells that expressed one or more (up to five) srp's. The percentage of tumors with p53-positive cells was variable; the proportion was highest among srp-expressing GBMs (mean: 16.1%) and Br-Mts (mean: 15.3%). The mean PCNA-labeling index (LI) also varied, ranging from 1.2% in the group of pituitary adenomas to 24.5% in Br-Mts, with GBMs (20.4%) and medulloblastomas (18.4%) approaching the latter value. PCNA-LI was higher in the astrocytomas, GBMs, medulloblastomas and PNETs that expressed srp's than in those did not. A high proportion of p53-positive cells (31.3 to 59.0%) and the highest PCNA-LIs (41.0 to 49.0%) were seen in two GBMs and one Br-Mt that expressed all five srp's. We conclude that primary and metastatic tumors of the brain produce one or more stress-related proteins, and that a variable proportion of the tumor cells have immunohistochemically-detectable p53, the expression of which may depend, at least in part, on the growth potential of a given tumor.

Divergent effects of hyperosmolality on stress-response (heat shock) protein expression in cultured human tumor cells: an immunocytochemical study.

Exposing cells to adverse conditions usually elicits expression of stress-response (heat shock) proteins (srp). Here we show that hyperosmolar growth conditions do not uniformly affect srp expression in MCF-7 and HeLa S3 cells, derived from carcinoma of the breast and cervix, respectively. Thus, whereas srp 27 expression was increased in MCF-7, but not in HeLa S3, the opposite was the case with srp 72. On the other hand, hyperosmolality did not induce alpha B-crystallin or ubiquitin in either cell line. These findings show that srp expression by the human tumor cells studied is non-coordinate, suggesting that each srp is independently modulated.

Ha-ras codon 12 mutation in papillary tumors of the urinary-bladder - a retrospective study.

This report concerns the retrospective examination of low grade, low stage (Ta/T1) bladder tumors for the Ha-ras codon 12 G-->T mutation. The patients studied had a minimum of 5 years follow-up and were grouped into 3 categories: (i) patients with no recurring tumors, (ii) patients with recurring Ta/T1 tumors and (iii) patients who subsequently developed high grade, high stage tumors. A heminested, non-isotopic, allele-specific polymerase chain reaction (PCR) amplification assay was used. The codon 12 G-->T mutation was found in 10/27 specimens from patients with non-recurring Ta/T1 tumors; in 11/27 initial and 12/23 recurring Ta/T1 tumors, and in 5/8 initial Ta/T1 lesions and 8/12 subsequently developed high grade/stage tumors. Although there was no correlation between disease recurrence and mutation, these results indicate that a relatively large proportion of patients with Ta/T1 tumors of the bladder have cells with the Ha-ras codon 12 G-->T substitution.

Comparative study on the expression of stress-response protein (srp) 72, srp 27, alpha B-crystallin and ubiquitin in brain tumours. An immunohistochemical investigation.

This immunohistochemical study compares the expression of stress-response (heat-shock) protein (srp) 72, srp 27, alpha B-crystallin and ubiquitin in 86 primary human brain tumours and 21 carcinoma metastases to the central nervous system. Normal brain tissues were included for control purposes. Serial sections of formalin-fixed, paraffin-embedded tissues were used. Most meningiomas (17/23), glioblastomas (11/12) and breast carcinoma metastases (9/10) and some astrocytomas (7/13), pituitary tumours (4/9) and lung cancer metastases (5/11) had tumour cells that reacted with one or more of the antibodies used. Around 43% of the meningiomas and 25% of the glioblastomas expressed srp 72 only. Sole expression of srp 27, alpha B-crystallin or ubiquitin was seen in several tumours. Some meningiomas (3/23) and breast cancer metastases (4/10) co-expressed srp 72 and srp 27, and 1/3 of the glioblastomas co-expressed srp 27 and alpha B-crystallin. We conclude that primary and metastatic tumours of the brain produce stress-related proteins and that certain tumours concurrently express two or more srp's.

Short-term culture of exfoliated cells from the urine of patients with bladder tumors.

This report concerns the short-term culture of urothelial cells from the urine sediment of over 100 patients with bladder tumors. Primary cell outgrowth was obtained in approximately 60% of the cultures initiated. Culture outcome was not related to tumor grade, patient age, or volume of the urine sample. Around 85% of the proliferating cultures were successfully transferred into multi-compartment chamber/slides. These results suggest that the culture system may be a useful tool for the study of urothelial cells using patient material obtained by non-invasive means.

Divergent effects of butyrate on the alkaline phosphatases of SW-620 cells.

The human colon cancer cell line SW-620 produces two alkaline phosphatases (ALPs) which are not expressed by normal colon. They are the heat-stable, term-placental and the heat-labile, L-homoarginine-sensitive, liver/bone/kidney ALPs. Butyrate, an ALP inducer, has strikingly dissimilar effects on the activity of these enzymes: whereas high (2.0 mM) butyrate concentrations exclusively induce increased activity levels of liver/bone/kidney ALP, low (0.5 mM) concentrations increase the activity of both, albeit induction of term-placental ALP is less pronounced. These observations indicate that the effect of butyrate on the two ALPs is non-coordinate and suggest that their expression by SW-620 cells is independently modulated.

Expression of glial fibrillary acidic protein (GFAP) by cultured angiofibroma stroma cells from patients with tuberous sclerosis.

Large dendritic cells were cultured from facial angiofibromas of six patients with tuberous sclerosis. The cells were examined immunocytochemically for expression of selected cytoskeletal and non-structural proteins and the results compared with the staining profiles obtained with normal skin fibroblasts and normal glial cells. In similarity to normal glia, the angiofibroma stroma cells expressed glial fibrillary acidic protein (GFAP). Conversely, by analogy to fibroblasts, the abnormal stroma cells produced fibronectin and did not react with the antibody to S-100 protein. By immunogold labelling it was established that GFAP and vimentin were co-localized in intermediate filaments of the angiofibroma cells.

Concurrent mutations of coding and regulatory sequences of the Ha-ras gene in urinary bladder carcinomas.

This report concerns the study of Ha-ras gene mutations and ras p21 expression in primary tumors of the urinary bladder. Polymerase chain reaction-based techniques and computerized image analysis were used. The data obtained were related to tumor grade, DNA ploidy, and tumor invasion. A point mutation (G-->T) at Ha-ras codon 12 was found in 30 of 67 tumors. The mutation frequency was greater in grade III (65%) than in grade II (44%) tumors; no mutations were observed in grade I tumors. The mutation was observed more often in aneuploid (58%) than in diploid (28%) tumors. No other substitution at codon 12 was seen and no codon 61 mutation was detected. The tumors were also tested for the A-->G mutation at position 2719 of Ha-ras intron D. Concurrent codon 12 and intron D mutations were identified in seven high-grade aneuploid tumors; six were invasive. The levels of the ras gene product p21 were approximately 10 times higher in tumors with intron D mutation than in those without. These findings confirm on human bladder tumors the observations of the effect of synchronous exon-intron mutations reported on the bladder cancer cell line T24. Our results are the first demonstration of Ha-ras intron D alterations in human tumor tissues and suggest that concurrent mutations at codon 12 and intron D of this gene within the same tumor may contribute to the aggressive behavior of human bladder carcinomas.

Comparative immunohistochemical study on the expression of alpha B crystallin, ubiquitin and stress-response protein 27 in ballooned neurons in various disorders.

This report deals with a comparative study on the expression of alpha B crystallin, ubiquitin, stress-response protein 27 (srp 27), srp 72 and phosphorylated neurofilament protein (pNFP) by ballooned neurons in Pick's disease, Creutzfeldt-Jakob disease (CJD), amyotrophic lateral sclerosis (ALS), leptomeningeal carcinomatosis, anterior spinal artery syndrome and pellagra. Immunohistochemical techniques were used. alpha B Crystallin was expressed by the majority of ballooned neurons of Pick's disease and CJD, but not by those of the other disorders. Ubiquitin and srp 27 expression was also restricted to abnormal neurons of Pick's disease and CJD, but the proportion of stained cells was less than that expressing alpha B-crystallin. There was no evidence of ballooned neurons expressing srp 72. Except for those of pellagra patients, phosphorylated neurofilament protein (pNFP) was detected in most abnormal neurons. Our results suggest that the mechanisms involved in formation and maintenance of swollen neurons in Pick's disease and CJD may be different than those of ballooned neurons in the other entities studied.

Asymmetric distribution of oncogene products at mitosis.

Computer-assisted image analysis was used to demonstrate in exponentially proliferating human tumor cells the uneven postmitotic apportionment of several oncogene-encoded proteins (ras p21; erbB-2 p185; fos p55; myc p62). This observation may provide the explanation for the high degree of heterogeneity of postmitotic cells and the asynchrony in cell cycle traverse of cultured cells.

Ultrastructural and immunohistochemical studies on ballooned cortical neurons in Creutzfeldt-Jakob disease: expression of alpha B-crystallin, ubiquitin and stress-response protein 27.

This report concerns ultrastructural and immunohistochemical studies on ballooned neurons of ten patients with Creutzfeldt-Jakob disease (CJD). While abundant ballooned neurons and severe white matter degeneration was seen in six Japanese cases, only occasional ballooned neurons and no white matter degeneration was observed in four cases from the files of Montefiore Medical Center. Ultrastructurally, the ballooned neurons contained granule-coated fibrils of 25 to 40 nm in width and 10-nm neurofilaments. The immunohistochemical studies revealed that most ballooned neurons expressed alpha B-crystallin, with deposits of reaction products observed in the cytoplasm. A similar intracellular staining pattern was also seen with the antibody to phosphorylated neurofilament proteins (pNFP). Although the proportion of stained ballooned neurons was less, a positive reaction was also observed with antibodies against ubiquitin, stress-response protein 27 (srp 27) and synaptophysin, but not with an antibody to srp 72. Our findings suggest that expression of pNFP and synaptophysin by ballooned neurons may reflect axonal impairment and that the presence of alpha B-crystallin, srp 27 and ubiquitin may be related to the degenerative processes that neurons undergo in CJD.

Expression of stress-response (heat-shock) protein 27 in human brain tumors: an immunohistochemical study.

This report concerns the expression of the low molecular weight stress-response (heat-shock) protein 27 (srp 27) in a variety of human brain tumors. Immunohistochemical techniques were used; cells of the breast cancer line MCF7 served as positive controls. The reaction product was found exclusively in the cytoplasm. Srp 27 was detected in 5/5 breast tumor metastases to the brain and in 5/21 meningiomas. The protein was also detected in 5/11 glioblastomas and 2/5 pituitary adenomas. By comparison, positive staining was observed in only 1/15 astrocytomas and 1/7 medulloblastoma and no reaction was seen with the oligodendrogliomas, schwannomas and gangliogliomas tested. These observations demonstrate that srp 27 is expressed by certain primary intracranial tumors.

Stress-response (heat-shock) protein 72 expression in tumors of the central nervous system: an immunohistochemical investigation.

This report deals with the expression of stress-response (heat-shock) protein 72 (srp 72) in a series of 95 primary human brain tumors and 21 carcinoma metastases to the central nervous system (CNS). Immunohistochemical procedures were employed; cells of the human cervical cancer line HeLa S3 were used as positive controls. The protein was detected in 14/22 meningiomas and in 6/13 glioblastomas. Tumor cells expressing srp 72 were also found in 4/17 astrocytomas, 2/9 pituitary tumors, 2/14 primitive neuroectodermal tumors and 1/10 medulloblastomas. Whereas the majority (8/10) of the breast carcinoma metastases had tumor cells that expressed srp 72, only 2/11 lung tumor metastases were positively stained. These results document srp 72 expression by a variety of primary and metastatic tumors of the CNS.

Immunohistochemical studies on the new type of astrocytic inclusions identified in a patient with brain malformation.

Immunohistochemical studies were carried out on the new type of cerebral cortical astrocytic inclusions recently discovered in a 20-year-old patient with maldeveloped brain and micropolygyria. The inclusions appeared as eosinophilic structures (hematoxylin and eosin stain) and did not exhibit argyrophilia (modified Bielschowsky method). The inclusions were strongly stained by the antibody against S-100 protein (S 100) and to a lesser extent by the antibody to microtubule-associated protein 1B (MAP 1B). In contrast to Rosenthal fibers, the astrocytic inclusions did not react with antibodies to alpha B-crystallin, glial fibrillary acidic protein and ubiquitin. No positive reactions were obtained with antibodies against heat-shock protein 27 (HSP 27), HSP 72, actin, vimentin, desmin, cytokeratin, myelin basic protein, beta-tubulin, MAP 2, tau protein, paired helical filament, phosphorylated neurofilament protein (NFP), nonphosphorylated NFP, synaptophysin, cathepsin D, alpha 1-antichymotrypsin, alpha 1-antitrypsin and basic fibroblast growth factor. By immunoelectron microscopy, the products of the reaction with the anti-S 100 antibody appeared as heterogeneous granular deposits and with the antibody to MAP 1B they were randomly scattered throughout the astrocytic inclusions. Our results demonstrate that the immunohistochemical profile of the recently described inclusions differs from that of Rosenthal fibers. Whether the novel inclusions are involved in congenital astrocyte dysfunction and cerebral malformation remains to be established.

Effect of hyperosmolality on alkaline phosphatase and stress-response protein 27 of MCF-7 breast cancer cells.

MCF-7, a continuous cell line derived from a human breast carcinoma, exhibits very low alkaline phosphatase (ALP) activity. The enzyme is heat-stable and is inhibited by L-phenylalanine and L-phenylalanylglycylglycine, but not by L-homoarginine, 1-bromotetramisole, or levamisole. These data indicate that MCF-7 produces term-placental ALP, the oncodevelopmental enzyme form inappropriately expressed by a variety of human tumors. In contrast to human cancer cells that produce this enzyme monophenotypically, ALP activity of MCF-7 cells is not significantly increased by glucocorticoids or sodium butyrate. By comparison, exposure to hyperosmolality causes a striking increase in enzyme activity. Cycloheximide blocks this effect. The results obtained with cell-free assays were confirmed by cytochemical and immunocytochemical assays on whole cells. Because some of the agents tested in the enzyme modulation experiments affect cell proliferation, their possible effect on two stress-response proteins (srp 27 and srp 72) was also examined; specific immunocytochemical assays were used. These tests revealed that neither protein is affected by glucocorticoids; that sodium butyrate has no effect on srp 27, but alters the intracellular distribution of srp 72; and that hyperosmolality, while not significantly affecting srp 72, causes an increase in srp 27.

Protein expression in relation to the cell cycle of exponentially growing human prostatic epithelial cells.

This report concerns the study of the relationship between protein expression and the cell cycle in exponentially proliferating benign and malignant human prostate epithelial cells in short-term cultures. Multiparameter flow cytometric measurements were performed to correlate the expression of prostate-specific acid phosphatase, epithelial membrane antigen and epitectin with cell cycle progression. The expression of the three proteins was heterogeneous in G1 cells. The early post-mitotic cells exhibited the lowest levels when compared with late G1 cells, wherein the expression was many times greater. There was no further increase as the cells progressed through S and G2 + M. These findings, corroborating prior observations in other systems, suggest the possibility that the levels of the proteins studied increase during the G1 phase of the cell cycle and drop during or immediately after cytokinesis. As an alternate explanation, the heterogeneity of protein expression characteristic of G1 cells may be due, at least in part, to an asymmetric apportionment of cell constituents at mitosis.

Stimulation of human fetal astrocyte proliferation by bacterial lipopolysaccharides and lipid A.

This report concerns the effect of bacterial endotoxin [lipopolysaccharide(LPS) and lipid A] on cultured human fetal astrocytes. Exposure to 1 micrograms/ml LPS or lipid A caused a striking stimulation of the rate of proliferation of the cells. The effect was most pronounced with exponentially growing cultures. Stimulation was associated with enhance DNA synthesis as ascertained by [3H]thymidine incorporation. These findings at the cellular level may be of relevance in the elucidation of the effects of bacterial endotoxins on the developing human brain.