La ecografía carotídea es útil para predecir enfermedad coronaria y mortalidad en pacientes en hemodiálisis / Prognostic value of carotid intima media thickness and wall plaques in hemodialysis patients

Introducción: La enfermedad cardiovascular es la principal causa de muerte en los pacientes urémicos en hemodiálisis (HD). La ecografía carotídea es una herramienta sencilla y no invasiva para conocer el estado aterosclerótico de los pacientes. Objetivo: Conocer las asociaciones clínicas del grosor íntima-media carotídeo (GIM) y de la placa carotí- dea y su valor predictivo sobre el riesgo de enfermedad co- ronaria y la mortalidad. Metodología: Estudio prospectivo en el que se incluyeron 60 pacientes estables en HD (68 ± 13 años, 48% hombres, 50% diabéticos, tiempo en HD de 32 ± 11 meses) y 274 controles, semejantes en edad y sexo. El período de seguimiento fue de 66 ± 13 meses. Determi- naciones: Datos demográficos y clínicos, analítica general y niveles séricos de homocisteína y folato. Se midió el GIM mediante ecocardiografía 2D. Resultados: El GIM fue mayor en los pacientes en HD que en el grupo control (0,947 ± 0,308 frente a 0,619 ± 0,176 mm; p <0,001). El GIM se correlacionó con la edad (r = 0,268; p = 0,038), con la condición de diabético (r = 0,650; p <0,001) y la de hiper- tenso (r = 0,333; p = 0,012), pero no con colesterol total, HDL, LDL, triglicéridos, homocisteína o folato. Los pacien- tes con enfermedad coronaria, enfermedad vascular peri- férica o ictus tenían un GIM mayor que los que no presen- taban dichas afecciones (1,156 ± 0,371 frente a 0,875 ± 0,285 mm; p <0,001; 1,205 ± 0,374 frente a 0,911 ± 0,231 mm; p = 0,007; 1,195 ± 0,264 frente a 0,844 ± 0,251; p <0,001, respectivamente). Se encontraron datos similares respecto a la presencia de placas en la pared carotídea. Durante el período de seguimiento fallecieron 36 pacientes, 24 de los cuales (67%) por causa cardiovascular, cuyo GIM fue mayor (1,020 ± 0,264 frente a 0,858 ± 0,334 mm; p = 0,044). La supervivencia a la finalización del período de es- tudio fue significativamente mejor en el cuartil inferior de GIM (72%) que en el superior (20%). La presencia de pla- cas carotídeas fue predictor independiente de mortalidad cardiovascular. Conclusiones: El GIM y las presencia de pla- cas carotídeas se relacionan con algunos de los factores clá- sicos de riesgo cardiovascular como la edad, la diabetes o la hipertensión en pacientes urémicos. Su medición es útil para predecir la enfermedad coronaria y la mortalidad a largo plazo en los paciente urémicos (AU)

ntroduction: Cardiovascular disease and other complica- tions of atherosclerosis are the most common cause of death in patients with chronic renal failure in maintenance hemodialysis (MHD). Carotid ultrasonography is a simple no invasive tool to investigate the vascular system, by means of intima media thickness (IMT) measurement and carotid wall calcifications. Objective: To determine IMT and the presence of plaques, and their possible clinical re- lationships; finally we tried to investigate whether they would predict cardiovascular morbidity and mortality in patients in MHD. Methods: We studied 60 MHD patients (age 68 ± 13 years, 48% male, 50% diabetics, tiem on MHD 32 ± 11 months) and a control group of 274 people matched for age and sex. Follow-up period was 66 ± 13 months. Measurements: Demographic and clinical data, serum levels of homocysteine (tHcy), folic acid (FA) and B 6 and B 12 vitamins. IMT was measured by high-resolution B- mode ultrasonography. Results: IMT was higher in MHD patients than in those in the control group (0.947 ± 0.308 vs 0.619 ± 0.176 mm; P <0.001). IMT was related with age (r = 0.268; P = 0.038), diabetic (r = 0.650; P <0.001) and hy- pertensive condition (r = 0.333; P = 0.012), but not wih lipids, tHcy or FA. Similar findings were found with the presence or not of carotid plaques but serum LDL-choles- terol levels were also related (r= –0.280; P = 0.031). Patients who suffered from coronary artery disease, peripheral ar- tery disease or stroke had higher IMT than those without those events (1.156 ± 0.371 vs 0.875 ± 0.285 mm; P <0.001; 1.205 ± 0.374 vs 0.911 ± 0.231 mm; P = 0.007; 1.195 ± 0.264 vs 0.844 ± 0.251; P <0.001 respectively). Something similar ocurred with the presence of plaques. During the follow- up period 36 patients (60%) died, 67% of them due to car- diovascular causes. IMT was higher in patients who expired than those who survived (1.020 ± 0.264 vs 0.858 ± 0.334 mm; P = 0.044). The survival rate during the observation was significantly lower in the final IMT fourth (20%) than in the first (72%) (P = 0.014). The presence of carotid plaques was an independent predictor of cardiovascular mortality. Conclusions: These findings suggests that meas- urement of carotid IMT and the presence of wall plaques are useful tools to predict cardiovascular events and mor- tality in patients in MHD (AU)

[Carotid ultrasound: prevention of heart disease and mortality on haemodialysis]. / La ecografía carotídea es útil para predecir enfermedad coronaria y mortalidad en pacientes en hemodiálisis.

INTRODUCTION: Cardiovascular disease and other complications of atherosclerosis are the most common cause of death in patients with chronic renal failure in maintenance hemodialysis (MHD). Carotid ultrasonography is a simple non-invasive tool to investigate the vascular system, by means of intima media thickness (IMT) measurement and carotid wall calcifications. OBJECTIVE: To determine IMT and the presence of plaques, and their possible clinical relationships; finally we tried to investigate whether they would predict cardiovascular morbidity and mortality in patients in MHD. METHODS: We studied 60 MHD patients (age 68 +/- 13 years, 48% male, 50% diabetics, time on MHD 32 +/- 11 months) and a control group of 274 people matched for age and sex. Follow-up period was 66 +/- 13 months. MEASUREMENTS: Demographic and clinical data, serum levels of homocysteine (tHcy), folic acid (FA) and B6 and B12 vitamins. IMT was measured by high-resolution B-mode ultrasonography. RESULTS: IMT was higher in MHD patients than in those in the control group (0.947 +/- 0.308 vs 0.619 +/- 0.176 mm; P < 0.001). IMT was related with age (r = 0.268; P = 0.038), diabetic (r = 0.650; P < 0.001) and hypertensive condition (r = 0.333; P = 0.012), but not wih lipids, tHcy or FA. Patients who suffered from coronary artery disease, peripheral artery disease or stroke had higher IMT than those without those events (1.156 +/- 0.371 vs 0.875 +/- 0.285 mm; P < 0.001; 1.205 +/- 0.374 vs 0.911 +/- 0.231 mm; P = 0.007; 1.195 +/- 0.264 vs 0.844 +/- 0.251; P < 0.001 respectively). Something similar occurred with the presence of plaques. During the follow-up period 36 patients died (60%), 67% of them due to cardiovascular causes. IMT was higher in patients who died than those who survived (1.020 +/- 0.264 vs 0.858 +/- 0.334 mm; P = 0.044). The survival rate during the observation period was significantly lower in the final IMT fourth (20%) than in the first (72%) (P = 0.014). The presence of carotid plaques was an independent predictor of cardiovascular mortality. CONCLUSIONS: These findings suggests that measurement of carotid IMT and the presence of wall plaques are useful tools to predict cardiovascular events and mortality in patients in MHD.

Biological and prognostic role of acid cysteine proteinase inhibitor (ACPI, cystatin A) in non-small-cell lung cancer.

BACKGROUND: Acid cysteine protease inhibitor (ACPI) is an intracellular protein, often linked to neoplastic changes in epithelium and thought to have an inhibitory role in malignant transformation. AIM: To analyse the expression and prognostic role of ACPI in non-small-cell lung cancer (NSCLC). METHOD: Histological samples from 199 patients with resected NSCLC were stained immunohistochemically for the expression of ACPI in normal and preneoplastic bronchial epithelium, and in various types of lung carcinomas. RESULTS: A normal bronchial epithelium showed positive staining for ACPI in the basal cells, whereas the upper two-thirds of the dysplastic epithelium was ACPI positive. High staining for ACPI was found in 74% (91/123) of squamous-cell carcinomas, whereas 16% (8/49) of adenocarcinomas and 30% of (8/27) large-cell carcinomas showed the high expression of ACPI (p<0.001). Among squamous-cell carcinomas, low expression of ACPI was correlated with poor tumour differentiation (p=0.032). In the whole tissue, reduced expression of ACPI was associated with tumour recurrence (p=0.024). In overall survival (OS) and disease-free survival (DFS) analyses, the histological type of the tumour (both p<0.001) and stage of the tumour (p=0.001, p=0.013, respectively) were related to patient outcome. Low expression of ACPI in tumour cells was associated with poor OS and DFS (p<0.041, p=0.004, respectively). In multivariate analysis, ACPI did not retain its prognostic value, whereas the traditional factors were the most important prognostic factors. CONCLUSIONS: ACPI expression is linked with the malignant transformation of the bronchial epithelium and predicts a risk of tumour recurrence as well as poor rate of survival for the patients. However, ACPI does not have any independent prognostic value in NSCLC.

Role of apolipoprotein E epsilon 4 allele on chronic allograft nephropathy after renal transplantation.

Lipid abnormalities may contribute to chronic allograft nephropathy (CAN). Apolipoprotein E (ApoE) gene polymorphism regulates lipoprotein metabolism, but little is known about an association between CAN and this polymorphism. The ApoE gene (E3/E4) polymorphism was typed by PCR assay (99 E3/E3, 28 E3/E4, 1 E4/E4) on 128 consecutive renal transplant patients with functioning grafts for more than 3 years (6.7 +/- 2.8 years). Twenty-eight patients with histological CAN were compared with 100 patients who had no clinical evidence of chronic rejection (no proteinuria and sCr < 2.5 mg%). As expected, univariate analysis revealed that patients with CAN experienced a greater acute rejection rate (78% vs 21%; P=.001), a higher serum creatinine (3.6 +/- 1.7 vs 1.4 +/- 0.5 mg%; P=.0001), and an older organ donor (43 +/- 20 vs 29 +/- 13 years; P=.0001). The lipid profiles (total cholesterol and triglycerides levels) were similar in both groups with 60% in each group receiving anti-lipemic drugs. Interestingly, the ApoE epsilon 4 allele was overrepresented in the group with CAN (39% vs 17%, P=.019). Logistic regression analysis showed that the epsilon 4 allele was an independent predictor of CAN (OR: 3.4; CI 95%: 1.07 to 11; P=.040) as were donor age and acute rejection episodes. In conclusion, an interaction between risk factors and genetic factors may determine CAN in this population. This finding may help to target prophylactic interventions in these recipients.

Atlantic salmon (Salmo salar L.) skin contains a novel kininogen and another cysteine proteinase inhibitor.

We describe the purification and characterization of two novel cysteine proteinase inhibitors found in Atlantic salmon skin. One of these, salmon kininogen, has a molecular mass of 52 kDa as determined by matrix-assisted laser desorption/ionization time-of-flight MS, is multiply charged with pI values of 4.0, 4.2 and 4.6 and shows homology to kininogens including the bradykinin motif. The other, salarin, has a molecular weight of 43 kDa, a pI of 5.1 and shows weak homology to cysteine proteinases. Both proteins are N- and O-glycosylated and inhibit papain and ficin but not trypsin.

Rhabdovirus-induced apoptosis in a fish cell line is inhibited by a human endogenous acid cysteine proteinase inhibitor.

To determine the mechanisms of cell death in rhabdovirus-infected cells, we studied the infection of the epithelial papilloma of carp cell line with spring viremia of carp virus. Studies using electron microscopy, confocal microscopy, and agarose gel electrophoresis revealed changes in cell morphology and DNA fragmentation indicative of apoptosis. The virus-induced apoptosis was inhibited in cells treated with a human endogenous acid cysteine proteinase inhibitor.

Concentrations of lysosomal cysteine proteases are decreased in renal cell carcinoma compared with normal kidney.

Renal cell carcinoma contains significantly lower concentrations of the lysosomal cysteine proteases, cathepsins B, C, H, L and S, than does normal kidney, as shown by several methods, such as activity determination, enzyme-linked immunosorbent assay, immunoblotting and immunohistochemistry. The same low levels of enzyme activity and concentration have been determined in renal cell carcinoma metastases in the lung. Our results on the decreased concentration of cysteine peptidases at the protein level would seem to conflict with earlier results on an increased concentration of the cathepsin L mRNA in renal cell carcinoma.

The possible place of cathepsins and cystatins in the puzzle of Alzheimer disease: a review.

Lysosomal proteinases (cathepsins) and their endogenous inhibitors (cystatins) have been found to be closely associated with senile plaques, cerebrovascular amyloid deposits, and neurofibrillary tangles in Alzheimer disease (AD). Further, profound changes in the lysosomal system seem to be an early event in "at-risk" neurons of AD brains. There is an ongoing controversy as to whether lysosome-associated proteolytic mechanisms are causally related to the development and/or further progression of the disease. The present article deals with some arguments "pro" and "contra" an involvement of the endosomal/lysosomal pathway in amyloidogenesis as a cardinal process in AD. Other putative targets of acidic proteinases and their natural inhibitors in the pathogenesis of AD (such as formation of neurofibrillary tangles and regulation of apolipoprotein E) are also discussed.

Accessory cells in physiological lymphoid tissue from the intestine: an immunohistochemical study.

We report a study of the organization of accessory cell populations, in normal mucosal lymphoid tissue from small intestine (8 cases), large intestine (6) and appendix (9) using a panel of monoclonal antibodies and polyclonal antisera in paraffin-embedded tissue. Two populations were identified in dome areas, one positive for acid cysteine proteinase inhibitor and HLA class II (WR18) only and the second positive for S-100 protein, CD68, and WR18 and negative for acid cysteine proteinase inhibitor and factor XIIIa. Superficial colonic mucosal and small intestinal villous tip macrophages stained positively with CD68 and WR18 only, while deeper cryptal and submucosal populations exhibited additional positivity for factor XIIIa, but both populations were negative for acid cysteine proteinase inhibitor and S-100 protein. Germinal centre macrophages were positive for CD68, WR18 and acid cysteine proteinase inhibitor and negative for factor XIIIa, and S-100 protein. T zone dendritic cells included a population which stained positively for S-100 protien, WR18 and were negative for factor XIIIa, CD68 and acid cysteine proteinase inhibitor, an immunophenotype typical of interdigitating dendritic reticulum cells. This distribution of phenotypically identifiable accessory cell subpopulations was apparent at all three sites examined. We suggest that the specialized subpopulations of dendritic cells staining for S-100 protein and for acid cysteine proteinase inhibitor which are restricted to the dome areas, may have a potential role in the transfer of antigen across the epithelium to the germinal centres, while factor XIIIa appears to identify a tissue macrophage population with a potential role in stromal modulation distant from direct antigen challenge.

Expression of acid cysteine proteinase inhibitor (ACPI) in the normal human prostate, benign prostatic hyperplasia and adenocarcinoma.

Acid cysteine proteinase inhibitor (ACPI or cystatin A) is a protein (12 kDa) which inhibits the action of several cysteine proteinases, e.g. cathepsins B, H, L and S. In this study the cellular location of ACPI has been immunohistochemically investigated in the normal human prostate, in benign prostatic hyperplasia (BPH) and in adenocarcinoma. ACPI was found in the basal epithelial cells of the normal prostate. The secretory epithelial cells did not express ACPI. In the hyperplastic prostate, the expression of ACPI was decreased and it was also expressed more focally in the basal cells. Hyperplastic basal cells also expressed ACPI. In prostatic adenocarcinoma, no ACPI expression was found. The absence of ACPI expression was obvious and if the sections contained both benign and malignant cells, only the benign glandular structures always expressed ACPI. The results suggest that expression of ACPI might be related to prostatic epithelial cell proliferation and differentiation. Possibly the detection of ACPI in tissue sections might be helpful in identifying prostatic adenocarcinoma, especially in cases with small carcinomatous foci.

Immunolocalization of cystatin A in condylomatous and dysplastic lesions of the human uterine cervix: correlation with the presence and type of human papillomavirus infection.

Cystatin A is the major cysteine proteinase inhibitor in human squamous epithelia. We investigated the occurrence of cystatin A in normal, condylomatous, and dysplastic lesions of the cervix with or without human papillomavirus (HPV) infection. Cystatin A was detected by immunohistochemistry and HPV infection by DNA hybridization techniques. In the normal uterine cervix, cystatin A was seen throughout the epithelium, except in the basal and parabasal cell layers. In condylomatous lesions, the staining intensity was similar to that in normal epithelium. In low-grade cervical intraepithelial neoplasias (CINs), reduced staining was seen in the lower third of the epithelium; in high-grade CINs, a reduction in staining intensity was also seen in the middle and upper thirds. Cystatin A staining in epithelia and nuclei was negative in highly cellular and poorly differentiated CIN III. The cytoplasmic staining of cystatin A did not correlate with presence or type of HPV DNA. In the high-grade CINs infected with HPV types 16 and 18, however, cystatin A staining was more often confined to the nuclear compartment.

Acid cysteine proteinase inhibitor in cutaneous lymphocytic infiltrates.

Acid cysteine proteinase inhibitor (ACPI, cystatin A) is normally present in squamous epithelium and dendritic cells of lymphoid follicles. Its expression is altered both in proliferative and malignant squamous epithelium and in neoplastic lymphoid follicles. The expression of ACPI in the lymphoid infiltrates of cutaneous psuedolymphomas and B-cell lymphomas was studied. Eighteen pseudolymphomas from 15 patients were divided into three groups according to the proportion of B and T lymphocytes. The B-cell-type lesions with well-developed follicles and germinal centers showed a pronounced ACPI expression in dendritic cells. Varying amounts of ACPI-positive cells were present in the mixed B- and T-cell-type and also in the T-cell-type lesions. The labeled cell population was distinct from the factor XIIIa-positive dermal dendrocytes, S-100-positive histiocytes, and HAM 56-positive histiocytes. Malignant lymphomas contained a few haphazardly arranged ACPI-positive cells with short dendrites and granular cytoplasm. It was concluded that follicular dendritic cells can be reliably labeled with ACPI antiserum in cutaneous pseudolymphomas. The structure and distribution of ACPI-containing cells in malignant cutaneous B-cell lymphomas is altered when compared with pseudolymphomas.

Identification of acid cysteine proteinase inhibitor (cystatin A) in the human thymus.

BACKGROUND: Acid cysteine proteinase inhibitor (ACPI, also called cystatin A) is a protein that is present in the epithelial cells of the skin and in the dendritic reticulum cells of lymphoid tissues. In this study the presence and cellular localization of ACPI in the thymus was investigated. METHODS: The cellular and topographical location of ACPI was immunohistochemically demonstrated in the normal thymus of man. RESULTS: ACPI was found in the cells of the Hassall's corpuscles and in many medullary cells. Most of these cells were epithelial cells, as shown by the results of immunohistochemical cytokeratin and epithelial membrane antigen stainings. Also, some individual cytokeratin negative but S-100 positive medullary reticular dendritic cells were stained with ACPI. CONCLUSIONS: The finding that ACPI is constantly present in the thymus at restricted and specific cellular locations leads to the suggestion that protease inhibitors may play a role in specific thymic functions.

Cystatin A-like immunoreactivity is widely distributed in human brain and accumulates in neuritic plaques of Alzheimer disease subjects.

The cellular localization of cystatin A, an endogenously occurring inhibitor of lysosomal thiol proteases (cathepsins B, H, L and S), was studied immunohistochemically in human postmortem brain using the peroxidase-antiperoxidase method. Both polyclonal and monoclonal antibodies to cystatin A were employed. Western blot analysis revealed one molecular form of the inhibitor in human brain extracts. Its molecular weight was about 13,000. Immunostaining appeared in a sizeable population of neurons and a few cells surrounding cerebral blood vessels (pericytes). In Alzheimer disease subjects cystatin A was found in many neuritic plaques. Possible functional consequences with regard to a role of cystatin A in the inhibition of the Alzheimer amyloid precursor protein (APP)-clipping enzyme, cathepsin B, are discussed.

Lysosomal proteinases as putative diagnostic tools in human neuropathology: Alzheimer disease (AD) and schizophrenia.

The cathepsin B, D and L were studied by immunohistochemical techniques in the human postmortem brain. The enzyme were primarily localized in neurons. Makroglial cells were seldom immunostained. It is shown that cathepsins B and D frequently occur in neuritic plaques of Alzheimer victims, thereby raising the question, whether or not cathepsin immunohistochemistry is a useful tool in the diagnosis of this disease. Furthermore, we identified certain glial cells to be immunoreactive for cathepsins in schizophrenics.

Immunolocalization of cystatin A in neoplastic, virus and inflammatory lesions of the uterine cervix.

Cystatin A was immunohistochemically demonstrated in the normal squamous epithelium of the uterine cervix, particularly in the parabasal and superficial cell layers whereas it was absent or scanty in the basal cells and in areas with parakeratosis. Cystatin A was also found in neoplastic lesions (dysplasia, carcinoma in situ and squamous cell carcinoma), but less abundant than in normal squamous epithelium. The immunoreaction in intraepithelial neoplasia was closely related to the degree of morphological maturation of the squamous cells with more abundant cystatin A in low grade dysplasia and less in high grade dysplasia and carcinoma in situ. In squamous cell carcinoma, cystatin A was often abundant in highly differentiated areas and almost absent in poorly differentiated ones. Cystatin A was found in the squamous epithelium in herpes and in condylomatous lesions. It was also found in the cytoplasm of neutrophils, but not in lymphocytes and plasma cells. In unspecific cervicitis, cystatin A was found extracytoplasmatically as small vesicles in the epithelial-stromal junction. The implications of cystatin A in neoplastic, virus, and inflammatory processes are discussed.

Primary structure of bovine cathepsin S. Comparison to cathepsins L, H, B and papain.

The primary structure of bovine cathepsin S was determined by combining results of protein and peptide sequencing with the sequence deduced from nucleic acid sequencing. Using polymerase chain reaction (PCR) technology, cDNA clones commencing at amino acid 22 of the mature enzyme and continuing through the 3' untranslated region of bovine cathepsin S mRNA were isolated and sequenced. The open reading frame in these overlapping clones correctly predicts the determined amino acid sequence of 13 tryptic peptides derived from purified bovine spleen cathepsin S. The deduced amino acid sequence shows that mature bovine cathepsin S consists of 217 amino acids corresponding to a molecular weight of 23.7 kDa. Cathepsin S belongs to the papain superfamily of lysosomal cysteine proteinases and shares 41% identity with papain. Amino acid sequence identities of bovine cathepsin S to human cathepsins L, H, and B are 56%, 47% and 31% respectively.

Antigenic expression of cathepsin B in aged human brain.

The lysosomal thiol proteinase, cathepsin B, has been localized in different regions of aged human brain by use of the peroxidase-antiperoxidase technique. Cathepsin B-immunoreactive material was detected in multiple neurons of human hippocampus, neocortical area A 10, prefrontal gyrus and nuc. basalis of Meynert as well as in single white matter astrocytes. In brains of Alzheimer disease-affected subjects cathepsin B was revealed in neuritic plaques too. Possible functional consequences with regard to normal aging, neuropeptide metabolism and pathological changes are discussed.