Prostasin, a potential tumor marker in ovarian cancer: a pilot study

INTRODUCTION: Ovarian cancer is generally diagnosed at advanced stages of the disease; therefore, poor prognoses are typical. The development of tumor markers is thus of utmost importance. Prostasin is a protease that in normal tissues is highly expressed only in the prostate gland and seminal fluid. A previous study showed that prostasin is highly overexpressed in ovarian cancer cell lines. This study sought to evaluate the expression of prostasin in ovarian cancer. METHODS: Fresh tumor samples of ovarian epithelial cancer (n: 12) were analyzed for expression of prostasin mRNA (messenger ribonucleic acid) by conventional and real time quantitative PCR (polymerase chain reaction). As a standard control, a normal prostate sample was analyzed. RESULTS: Using conventional PCR, prostasin was detected in all but one sample. Using quantitative PCR, prostasin was over-expressed in all but one of the samples as compared to the control (prostate). CONCLUSIONS: These findings indicate that prostasin is overexpressed in many epithelial ovarian cancers. Further studies of prostasin as a potential biomarker for this disease are warranted.

Hepatocyte growth factor [HGF], HGF activator [HGFA] and c-met expression in rheumatoid synovial tissues in relation to disease activity

We examined the localization and mRNA expression of HGF, HGFA and c-Met in synovial tissues [ST] in rheumatoid arthritis [RA] in relation to disease activity to characterize its biologic function in that disease. Immunohistochemical staining and RT-PCRfor HGF, HGFA and c-Met were performed on ST specimens from 34 RA-patients and 20 osteoarthritis [OA] controls. Synovial fluid [SF] samples were taken from all RA and OA for measuring HGF with ELISA technique. Immunohistochemical staining of RAST revealed that HGFA and c-Met were strongly expressed infibroblasts, macrophages, endothelial cells and less so on synovial lining cells. But HGF was expressed faintly in macrophages andfibroblasts. While, in OAST, HGFA and c-Met were detected in the same cells as RAST but in a different distribution. HGF was localized in vascular endothelial cells. RT-PCR showed HGF, HGFA and c-Met mRNA in all RAST and all OAST. HGF levels in SF samples were higher in RA patients [range 5.6-39.2 ng/ml and mean 26.3 +/- 1.2 ng/ml] than OA controls [range 4.2-37.5 ng/ml and mean 11.2+2.4 ng/ml]. The differences were statistically significant [p<0.001]. A non-significant correlation was found between HGF-SF levels and disease activity score [DAS] [p>0.5]. HGFA, HGF and c-Met mRNA are expressed in ST in RA and OA. Lack of correlation between HGF-SF levels and DAS indicated that HGF played a regulatory role in the immunopathogenesis of RA

Frecuencia de la variante 23968A>G del gen PCSK9 en individuos chilenos con enfermedad coronaria confirmada por angiografía y controles / Frequency of 23, 968A>G polymorphism at the PCSK9 gene in Chilean subjects with coronary artery disease confirmed by angiography and controls

Introducción: Variaciones comunes en el gen PCSK9 han sido asociadas a elevadas concentraciones de colesterol plasmático. Entre ellas, el polimorfismo 23968A>G ha sido relacionado con hipercolesterolemia poligénica en individuos europeos y japoneses. Objetivos: Determinar la frecuencia de la variante 23968A>G del gen PCSK9 en pacientes con enfermedad arterial coronaria y controles, y evaluar su posible contribución al desarrollo de esta patología en individuos chilenos. Métodos: Se efectuó genotipificación de la variante 23968A>G del gen PCSK9 a 218 individuos no relacionados de la Región de La Araucanía: 110 con enfermedad coronaria confirmada por angiografía (estenosis >70 por ciento) y 108 sujetos controles, mediante la técnica de reacción en cadena de la polimerasa seguida de restricción enzimática (PCR-RFLP). Resultados: La distribución de genotipos para el polimorfismo 23968A>G del gen PCSK9 en los pacientes con enfermedad coronaria (AA: 95.5 por ciento, AG: 4.5 por ciento, GG: 0 por ciento) y controles (AA: 95.4 por ciento, AG: 4.6 por ciento, GG: 0 por ciento) fue similar (p= 0.769). La frecuencia del alelo mutado G fue también semejante entre casos y controles (0.0227 vs. 0.0231, p=0.771). La odds ratio (OR) relacionada al alelo mutado G fue 0.98 (I.C. 95 por ciento: 0.28 - 3.44) confirmando la ausencia de asociación. Conclusiones: El polimorfismo 23968A>G del gen PCSK9 no contribuye para el desarrollo de enfermedad arterial coronaria en la población estudiada.

Background: Common variants of the PCSK9 gene are associated to elevation of plasma cholesterol levels. Among them, the 23968A>G variant has been related to polygenic hipercholesterolemia in European and in Japanese subjects. Aim: to determine the frequency of the 23968A>G variant of the PCSK9 gene in patients with coronary artery disease and controls and to estimate its significance in the development of CAD in Chilean subjects. Methods: Genotypes of the 23968A>G variant of the PCSK9 gene were determined in 218 unrelated subjectsin the Region de La Araucania. One- hundred ten had angiography proven significant coronary artery disease(>70 percent stenosis) and 108 were controls. PCR followed by enzymatic restriction was used. Results: The genotype distribution for 23,968A>G variant in CAD patients (AA: 95.5 percent, AG: 4.5 percent, GG: 0 percent) and controls (AA: 95.4 percent, AG: 4.6 percent, GG: 0 percent) was similar (p=0.769). The relative frequency of mutated 23,968G allele in CAD and controls was not significantly different (0.0227 vs. 0.0231, p=0.771). Moreover, the odds ratio (OR) shown that the G allele was not associated with CAD (OR: 0.98; 95 percent C.I. = 0.28 - 3.44, P = NS). Conclusion: The 23968A>G polymorphism of the PCSK9 gene is not related to the presence of CAD in this population.

Study of leukocytic hydrolytic enzymes in patients with acute stage of coronary heart disease.

BACKGROUND: Coronary heart disease (CHD) is a major killer worldwide. Atherosclerosis, which is the basis of CHD, is believed to be an inflammatory disorder. Though various aspects of atherosclerosis are extensively studied, leukocytic hydrolytic enzymes are not studied very well with respect to CHD. AIM: This study was planned to assess changes associated with leukocytic hydrolases in CHD patients. SETTING AND DESIGN: A tertiary care hospital; case-control study. MATERIALS AND METHODS: 106 patients with acute myocardial infarction, 60 patients with unstable angina and 45 healthy controls were included in the study. Acid phosphatase, lysozyme, adenosine deaminase (ADA) and cathepsin-G levels were estimated from leukocytes. Reduced glutathione (GSH) and malondialdehyde (MDA) levels were measured. STATISTICAL ANALYSIS: Statistical comparison of data was done using student's t-test (unpaired). Correlation difference was calculated by using Pearson correlation coefficient. RESULTS: Significantly higher levels of acid phosphatase, lysozyme, ADA with lower levels of cathepsin G in leukocytes were observed in CHD group. We also found significantly higher levels of serum MDA with lower concentrations of blood GSH in CHD group. In diabetic CHD group, significantly higher levels of leukocytic acid phosphatase, lysozyme, ADA and serum MDA with lower levels of cathepsin G and blood GSH were observed. CONCLUSIONS: Our study indicates that leukocyte hydrolytic enzymes, mainly acid phosphatase, lysozyme and ADA were more active in CHD patients and may contribute to inflammation related with CHD. Its also indicates that leukocyte cathepsin-G may have antiinflammatory role.

ANCA: serology in Wegener's granulomatosis.

BACKGROUND AND OBJECTIVES: Wegener's granulomatosis (WG) is being increasingly diagnosed in India, which exists in two forms, the 'limited Wegener's granulomatosis' (LWG) having upper respiratory tract (URT) and lower respiratory tract (LRT) involvement and the 'classical Wegener's granulomatosis' (CWG), with the triad of URT, LRT involvement along with kidney involvement. Cytoplasmic ANCA (C-ANCA) or anti-Proteinase3 (anti-PR3), which is highly diagnostic for WG, rarely perinuclear ANCA (P-ANCA) may exist. AIMS: To detect anti-neutrophil cytoplasmic antibodies (ANCA) and correlate it with serological, hematological parameters, and the Birmingham Vasculitis Activity Score (BVAS). SETTINGS AND DESIGN: Twenty-three clinically and histopathologically proven WG (16 CWG, 7 LWG) were studied. MATERIAL AND METHODS: C-ANCA and P-ANCA patterns were identified by immunofluorescence and specificities were confirmed by 'alpha granule' enzyme linked immunosorbent assay (ELISA), anti-PR3, anti-MPO (myeloperoxidase) and anti-Lactoferrin (anti-LF) by ELISA. RESULTS: LRT involvement was seen in 91.3%, URT in 78.3%, and renal manifestations in 69.6% cases. The BVAS in CWG was significantly higher than BVAS in the LWG. Decreased hemoglobin, increased WBC counts, ESR, CRP and Creatinine were seen in CWG as compared to LWG. The C-ANCA was present in 65.2% patients and P-ANCA in 13% cases. Anti-PR3 was seen in 69.6% patients and anti-LF in 17.4% cases. Severity of disease and ANCA was higher in CWG than in LWG. CONCLUSIONS: Vasculitis syndromes are known to overlap and many go undetected; therefore ANCA testing, along with the clinical and histopathological observations may be helpful in early detection and management of WG cases.

Serine proteases in female serum, new markers in polycystic ovary syndrome

Prostatic Specific Antigen [PSA] is present in very low concentration in female sera., but it can be measured with high sensitivity assays. Polycystic ovary syndrome [PCOS] is a disorder characterized by hyperandrogenism and chronic anovulation. Serum Testosterone is increased in PCOS patients. Women with higher levels of androgen may have higher levels of PSA compared with women with normal levels of androgen. Hirsutism represents a state of androgen excess in women. Women with PCOS usually suffer from hyperandrogenism and were selected to test the hypothesis in this study. Twenty five PCOS females were investigated for the level of PSA in their sera, using chemillurninescent technique. PSA level was found to be higher than normal, and correlated with testosterone levels. Our data suggest that measurement of this serine protease in serum may aid in the diagnosis of PCOS patients

Mucosal mast cell-derived chondroitin sulphate levels in and worm expulsion from FcRgamma-knockout mice following oral challenge with Strongyloides venezuelensis

Mucosal mast cell-derived chondroitin sulphates (sulphated proteoglycans) were assayed in gut washings and homogenate of FcRgamma-knockout (KO) and wild-type (WT) C57BL/6 mice challenged with Strongyloides venezuelensis in order to assess their possible role in secondary immunity against enteric nematodes. Groups of immune KO and WT mice were challenged by oral gavage with 300 infective larvae (L3). Establishment of infection was assessed by daily faecal analysis to determine the number of eggs per gram of faeces (EPG) and by adult worm recovery on days 5 and 13 post challenge. Mucosal mast cell (MMC) counts were done on days 5 and 13 post challenge while MMC-derived chondroitin sulphates in gut washings (days 1 and 5) and homogenate (day 8) were assayed by high performance liquid chromatography (HPLC). Results showed that patent infection occurred in challenged KO but not WT mice despite significantly higher mastocytosis in jejunal sections of KO than WT mice (p<0.001). Similarly but against prediction, significantly higher concentration of MMC-derived chondroitin sulphates was observed in gut homogenate of KO than WT mice (p<0.05). In contrast, significantly higher concentration of chondroitin sulphates was observed in gut washings of WT than KO mice (p<0.05). These results suggest that MMC in KO mice failed to release sufficient amount of sulphated proteoglycans into the gut lumen as did the WT mice, which may have been part of the hostile environment that prevented the establishment in and eventual expulsion of adult S. venezuelensis from the gut of WT mice following challenge.

Tryptase and fatal anaphylaxic reaction / 法医学杂志

OBJECTIVE@#To investigate the relationship between tryptase in serum and anaphylaxis.@*METHODS@#The concentrations of tryptase in the sera of heart blood in three persons died from anaphylaxis shock were detected by ELISA. The first sample was obtained from a man, aged 38, died of injecting Amikacin. The second sample was obtained from a man, aged 42, died of injecting Cephradine. The third sample was from a woman, aged 39, died of injecting Lincomycin. All samples were stored in -20 degrees C.@*RESULTS@#The concentrations of tryptase in sera were 52 ng/ml, 121 ng/ml and 0.73 ng/ml. It was unknown why the concentration of tryptase in the third sample was normal.@*CONCLUSION@#In fetal anaphylaxia reaction tryptase measurement is a useful indicator, but the diagnosis is not to be based on the test alone.