Hexosaminidase as a new potential marker for larynx cancer

ObjectivesLarynx squamous cell carcinoma is one of the most common forms of cancer in the area of the neck. The aim of our study was to investigate the activities of N-acetyl-β-d-hexosaminidase (HEX) in larynx cancer compared with the specimens from the healthy space of the tumor that served as controls.Design and methodsLarynx cancer (n = 15) and normal healthy tissue around the tumor (n = 15) were collected from the patients during total laryngectomy. Specimens were immediately frozen in ? 80 °C. To assess hexosaminidase activity, release of p-nitrophenol from p-nitrophenol derivatives was used.ResultsWe observed a significantly higher activity of the investigated enzyme in all laryngeal cancer specimens compared with that in healthy tissue homogenates. The differences were statistically significant.ConclusionsIt could be assumed that HEX may release particular sugars from the ends of oligosaccharide chains of glycocalyx proteins, changing adhesive forces binding together cells, and the communication between cells and elements of extracellular matrix.     

Hyperhomocysteinemia is a risk factor for cancer and a new potential tumor marker

Plasma homocysteine (Hcy), a well-known independent risk factor for coronary heart disease, is also a risk factor for cancer. Results from our studies indicate that Hcy could be used as a tumor marker. We found elevated circulating total homocysteine (tHcy) in cancer patients even though they were not treated with anti-folate drugs. In serial specimens from cancer patients undergoing treatment, the change of tHcy coincided with the concentration of tumor markers. The rapid proliferation of tumor cells contributed to the much higher concentrations of circulating tHcy. Both concentrations of tHcy and tumor marker would increase in parallel during the growth of tumor cell, but only the Hcy concentration would decline in response to tumor cell death. Several biochemical changes, including folate deficiency, oxidative stress, aberrant DNA methylation, and production of homocysteine thiolactone have been identified in association with hyperhomocysteinemia, which explained why elevated homocysteine eventually led to carcinogenesis. Conceivably, tHcy may be used as a more accurate tumor marker for monitoring cancer patients during treatment, and hyperhomocysteinemia as a risk factor for carcinogenesis.     

Procollagen I amino-terminal propeptide as a potential marker for multiple myeloma

ObjectivesInvestigating relationship between bone markers, cytokines and conventional prognostic parameters in patients with multiple myeloma (MM) and to assess the clinical application of bone turnover markers.Design and methodsSixty-four patients with MM were examined before treatment and followed for survival for over 7 years. Serum concentrations of bone markers and cytokines were determined by the Roche and R&D kits, respectively. Standard deviation scores (SDS) were employed to normalize values.ResultsCollagen fragments (β-CTX) were elevated in 47%, procollagen I amino-terminal propeptide (PINP)—in 28%, and osteocalcin (OC) in 11% of patients. The values of the SDS of PINP and OC, but not β-CTX significantly decreased with MM stage. β-CTX inversely correlated with vascular endothelial growth factor (VEGF) and albumin, and directly correlated with serum macrophage colony-stimulating factor (M-CSF). OC values correlated with albumin and β2-microglobulin. PINP inversely correlated with LDH. The SDS values of PINP were significantly lower in MM patients with advanced bone disease.ConclusionsCirculating PINP concentration may be a useful marker for monitoring of treatment of multiple myeloma patients with bone lytic lesions, in particular, of patients treated with preoteasome inhibitors.     

DNA integrity as a potential marker for stool-based detection of colorectal cancer

BACKGROUND: Molecular genetic analysis of DNA in patient stools has been proposed for screening of colorectal cancer (CRC). Because nonapoptotic cells shed from tumors may contain DNA that is less degraded than DNA fragments from healthy colonic mucosa, our aim was to show that DNA fragments isolated from stools of patients with CRC had higher integrity than DNA isolated from stools of patients with healthy colonic mucosa. METHODS: We purified DNA from the stools of a colonoscopy-negative control group and patients with CRC and examined the relationship between long DNA fragments and clinical status by determining stool DNA integrity, using oligonucleotide-based hybrid captures with specific target sequences in increasingly long PCR reactions (200 bp, 400 bp, 800 bp, 1.3 kb, 1.8 kb, 24 kb). DNA fragments obtained from CRC patients were compared with fragments obtained from colonoscopy-negative individuals for length and/or integrity. RESULTS: DNA fragments isolated from CRC patients were of higher molecular weight (>18 bands detected of a total of 24 possible bands) than fragments isolated from fecal DNA of the colonoscopy-negative control group. CONCLUSIONS: The presence of long DNA fragments in stool is associated with CRC and may be related to disease-associated differences in the regulation of proliferation and apoptosis. An assay of fecal DNA integrity may be a useful biomarker for the detection of CRC.     

Serum LINE-1 hypomethylation as a potential prognostic marker for hepatocellular carcinoma

BACKGROUND: We investigated the clinical implications of global hypomethylation, one of the most consistent epigenetic changes in cancer, in the sera of patients with hepatocellular carcinoma (HCC). METHODS: Combined bisulfite restriction analysis PCR was used to assess the methylation status of LINE-1 repetitive sequences in genomic DNA derived from sera of 85 patients with HCC, 73 patients with cirrhosis, 20 healthy carriers of hepatitis B virus (HBV) and 30 healthy controls. RESULTS: Serum genome hypomethylation, the percentage of unmethylated LINE-1, was significantly increased in patients with HCC (P<0.001). The levels of serum LINE-1 hypomethylation at initial presentation correlated significantly with the presence of HBsAg, large tumor sizes, and advanced tumor stages classified by the CLIP score. Multivariate analyses showed that serum LINE-1 hypomethylation was a significant and independent prognostic factor of overall survival. CONCLUSION: Serum LINE-1 hypomethylation may serve as a prognostic marker for patients with HCC.     

The potential use of urine cell free DNA as a marker for cancer

Introduction: Although the role of circulating cell free DNA in cancer has been widely demonstrated, less is known about the role of urine cell free DNA (UcfDNA). UcfDNA can serve as a ‘liquid biopsy’ for urological and non-urological tumors, as it carries information on DNA from cells exfoliated in urine and from circulation.

Areas covered: We review the studies on UcfDNA as a source of biomarkers for cancer, focusing on the new techniques and the differences between urological and non-urological tumors. We searched Pubmed for articles published between 1998 and 2016 with the following key words and phrases: ‘urine’ and ‘cell free DNA’ or ‘liquid biopsy’ or ‘cancer’.

Expert commentary: Despite the few papers published on this topic, UcfDNA is an important component of ‘liquid biopsy’, a useful and non-invasive tool for cancer diagnosis, prognosis and treatment monitoring, containing a wide range of genetic information.     

Serum antiguanosine antibodies as a marker for SLE disease activity and pathogen potential

BACKGROUND: This article reviews research conducted on the immunogenicity of the nucleosides of DNA, especially guanosine, the most immunologically active nucleoside. Discussed is the relationship between circulating antibodies to guanosine, their potential role in SLE disease activity, the binding properties of monoclonal antiguanosine antibody (4H2) compared to polyclonal antiguanosine antibodies in humans with SLE, cell membrane penetration by these antibodies and their interference with signal transduction possibly related to their binding to mitochondria and their apparent GTPase activity. METHODS: Enzyme-linked immunosorbent assay methodology was used to show clinical relationships between antiguanosine antibody levels and disease activity in SLE. These results are discussed along with methods of detecting cell penetration by this antibody using special staining techniques, laser-scanning microscope detection of mitochondrial localization, and interference of cAMP and pKA production/activation. Additionally, there is some discussion regarding the assay used to detect enzymatic activity of antiguanosine antibodies. RESULTS: Enhanced circulating levels of antiguanosine antibodies in patients with SLE correlate closely with SLE disease activity. Other factors are discussed that support the pathogenic potential of these antibodies, including their ability to penetrate lymphocytes, bind to mitochondria, inactivate mitochondrial function, interfere with signal transduction, and their potential enzymatically activity. CONCLUSIONS: Antiguanosine antibodies correlate with SLE disease activity and may be pathogenically important in SLE by interfering with signal transduction, inactivating mitochondrial and cell function in patients with SLE.     

Procalcitonin as a diagnostic marker and IL-6 as a prognostic marker for sepsis

The diagnosis and prognosis of sepsis after antimicrobial therapy among systemic inflammatory response syndrome (SIRS) patients were evaluated with the biomarkers procalcitonin (PCT), interleukin-6 (IL-6), C-reactive protein (CRP), erythrocyte sedimentation rate, and white blood cell counts.     

一种新型前列腺癌标记物AMACR(P504S)的研究进展

前列腺癌是男性最常见的真皮外恶性肿瘤。一种新近确认的前列腺癌的特异性分子标记物α-甲酰-辅酶A-消旋酶(AMACR,又称P504S)应用于临床,促进了前列腺癌的诊断及鉴别诊断。现对该分子标记物的研究现状作一综述。     

Recombinant VirB5 protein as a potential serological marker for the diagnosis of bovine brucellosis

The molecular tag vaccine against Brucella abortus and serological testing are the main methods of prevention of brucellosis used currently. They can discriminate vaccinated animals and humans from those naturally infected. In this study, we constructed a gene deletion mutant strain, B. abortus S19 virB5 with a molecular tag. Recombinant VirB5 was expressed and purified for evaluation as a diagnostic reagent for bovine brucellosis. In total, 400 sera samples were tested using a VirB5 antigen-based enzyme-linked immunosorbent assay (ELISA) and the results were compared with those of the standard tube agglutination test (SAT). This showed that the sensitivity was 88.2%, specificity was 97.8% and accuracy was 94.8%. Recombinant VirB5 could also be used to discriminate B. abortus-infected mice from mice infected with the B. abortus S19 virB5 mutant strain. It was concluded that recombinant VirB5 could be used as a potential antigen and serological marker for the diagnosis of bovine brucellosis.     

A novel assay for cobalt-albumin binding and its potential as a marker for myocardial ischemia-a preliminary report

We initially observed a phenomenon of reduced in vitro binding of exogenous cobalt [Co(II)] to the N-terminus of human serum albumin (HSA) in emergency chest pain patients with early onset unstable angina and myocardial infarction. We then developed a colorimetric assay to measure cobalt-HSA binding and record the results in absorbance units (ABSU). In a preliminary clinical study of 139 emergency patients with acute chest pain, 99 patients with evidence of myocardial ischemia (Group 1) had elevated assay levels (mean ABSU ± SD; 0.519 ± 0.086) compared to 40 patients (Group 2) with no evidence of ischemia (0.316 ± 0.092) (p < 0.00001). In Group 1, 95 of 99 (96.0%) patients had levels higher than a decision threshold of 0.400 ABSU and in Group 2, 37 of 40 (92.5%) samples had higher cobalt binding capacity (ABSU ≤ 0.400). Further studies are warranted to determine if an assay measuring altered cobalt-HSA binding is a clinically useful diagnostic test to rule out myocardial ischemia.     

Serum serotonin as unexpected potential marker for staging of experimental hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the primary cancer of the liver. The present study aimed to assess the potential role of the endogenous regulators of angiogenesis like neurotransmitters, as possible HCC biomarkers. Five groups of rats were used in this study (8 rats per each): control healthy group (I), four intoxicated groups (II, III, IV, and V) used for induction of HCC with a single IP dose of diethylnitrosamine (DENA), 200 mg/kg. Groups II, III, IV, and V were sacrificed after 8, 16, 24, and 32 weeks of DENA injection respectively. Serum levels of epinephrine, nor-epinephrine, serotonin, and dopamine of all animals were estimated using high performance liquid chromatography technique coupled with fluorescence detector (HPLC-FLD). Development of HCC was confirmed histopathologically. Our results showed a significant increase in 3 neurotransmitters (epinephrine, nor-epinephrine, and serotonin) in DENA intoxicated HCC rat model. Only serotonin exhibited a significant increase in early histological stage HCC development (16 weeks post DENA injection) in comparison to alpha-fetoprotein (AFP), (24 weeks post DENA injection). These results suggest that neurotransmitters (Epinephrine and Norepinephrine) may have a role as a biomarker for late histological stage HCC. Like AFP, while serotonin may be used for early stage HCC.     

Plasmepsins as potential targets for new antimalarial therapy

Malaria is one of the major diseases in the world. Due to the rapid spread of parasite resistance to available antimalarial drugs there is an urgent need for new antimalarials with novel mechanisms of action. Several promising targets for drug intervention have been revealed in recent years. This review addresses the parasitic aspartic proteases termed plasmepsins (Plms) that are involved in the hemoglobin catabolism that occurs during the erythrocytic stage of the malarial parasite life cycle. Four Plasmodium species are responsible for human malaria; P. vivax, P. ovale, P. malariae, and P. falciparum. This review focuses on inhibitors of the haemoglobin-degrading plasmepsins of the most lethal species, P. falciparum; Plm I, Plm II, Plm IV, and histo-aspartic protease (HAP). Previously, Plm II has attracted the most attention. With the identification and characterization of new plasmepsins and the results from recent plasmepsin knockout studies, it now seems clear that in order to achieve high-antiparasitic activities in P. falciparum-infected erythrocytes it is necessary to inhibit several of the haemoglobin-degrading plasmepsins. Herein we summarize the structure-activity relationships of the Plm I, II, IV, and HAP inhibitors. These inhibitors represent all classes which, to the best of our knowledge, have been disclosed in journal articles to date. The 3D structures of inhibitor/plasmepsin II complexes available in the protein data bank are briefly discussed and compared.     

胃癌患者血清巨噬细胞移动抑制因子水平的临床意义

目的：检测胃癌患者血清巨噬细胞移动抑制因子（MIF）的浓度水平,并评估其在胃癌诊断中的应用价值。方法用ELISA方法检测2007年2月至2010年4月在我院手术胃癌患者106例,其中男71例,女35例,平均年龄56.2岁（22～78岁）。所有病例均经病理学确认为原发性胃癌）、48例胃炎患者（接受胃镜检查诊断为慢性胃炎）及50例健康（健康体检者,排除一切肿瘤以及炎症性相关疾病）对照者血清MIF水平,同时测定癌胚抗原CEA。通过多因素Logistic回归分析,利用受试者工作特征（ROC）曲线分析MIF、CEA以及联合诊断对胃癌的诊断价值。结果胃癌患者血清中MIF平均浓度为4586 pg/ml,均高于单纯胃炎患者（1602.4 pg/ml）和健康对照（1105.6 pg/ml）,差异均有显著统计学意义（P均＜0.05）。胃癌患者血清MIF的水平与患者性别、年龄、病理类型不相关。但胃癌患者血清MIF水平随着肿瘤分期的升高而升高,差异有显著统计学意义（P＜0.05）。伴有淋巴结转移胃癌患者MIF水平显著高于未转移者（P＜0.05）。根据ROC曲线分析,MIF检测胃癌的临界值为3018 pg/ml时诊断效能最大,敏感度为79.4%,特异度为85.1%。MIF与CEA联合检测诊断价值高于单独诊断（P＜0.05）。结论胃癌患者血清中存在高水平的MIF,MIF可能为胃癌潜在的肿瘤标志物,MIF联合CEA可提高胃癌的诊断率,值得临床应用。     

A new minimally invasive method for the transtubal,microendoscopic application of fluids to the middle ear

The Eustachian (or auditory) tube is of central importance for the regulation of ambient air pressure changes within the middle ear spaces. Dysfunction of the Eustachian tube usually leads to chronic inflammatory changes of the middle ear.

The aim of the present feasibility study was to investigate an alternative, minimally invasive approach for the application of fluids to the middle ear via the Eustachian tube. This so‐called transtubal application (TTA) was conducted in a prospective, non‐randomized study with a total of ten subjects. The TTA approach consisted of placing a microendoscope within the Eustachian tube under local anaesthesia via its epipharnygeal opening. Subsequently, fluids were applied through an additional working channel after microendoscopic evaluation. Therefore the subjects were positioned supine‐laterally and had to swallow actively. The successful fluid application into the middle ear was evidenced by microendoscopy of the tympanic membrane (visualization of the fluid level). In all cases, a successful application could be evidenced. Side effects (e.g. pain, mucosal injuries, microbleedings) were not observed.

This new technique (TTA) offers the opportunity of a minimally invasive approach to treat tubal dysfunction and possibly other middle ear diseases by local fluid and/or drug application.     

Serum progranulin as a predictive marker for high activity of antineutrophil cytoplasmic antibody‐associated vasculitis

BackgroundThis study investigated whether serum progranulin could act as a predictive marker for high disease activity of antineutrophil cytoplasmic antibody (ANCA)‐associated vasculitis (AAV).MethodsFifty‐eight AAV patients were included in this study. Clinical and laboratory data were obtained at blood collection. The Short‐Form 36‐Item Health Survey Physical and Mental Component Summaries (SF‐36 PCS and SF‐36 MCS), Birmingham Vasculitis activity score (BVAS), Five‐Factor Score (FFS), and Vasculitis Damage Index (VDI) were assessed as AAV‐specific indices. Whole blood was collected and serum samples were isolated and stored at −80°C. Serum progranulin concentration was quantified by ELISA kits.ResultsThe median age of patients was 63.0 years (19 men). The median BVAS was 11.0, and the median serum progranulin level was 49.0 ng/ml. Serum progranulin was significantly correlated with BVAS, FFS, erythrocyte sedimentation rate, C‐reactive protein level, SF‐36 PCS, haemoglobin, and serum albumin. Severe AAV was arbitrarily defined as the highest tertile of BVAS (BVAS ≥16). When the cut‐offs of serum progranulin were set as 55.16 ng/ml and 43.01 ng/ml for severe AAV, AAV patients with serum progranulin ≥55.16 and 43.01 ng/ml had significantly higher risks of severe AAV than those without (relative risk (RR) 4.167 and 4.524, respectively).ConclusionsProgranulin might play an anti‐inflammatory role in AAV pathogenesis and serum progranulin could be used as a predictive marker for high activity of AAV.     

Troponin-T as a serum marker for myocardial infarction

We report here our experience with serum troponin T(TnT), measured with the sandwich immunoassay introduced by Boehringer Mannheim as a marker for myocardial infarction. We assayed TnT in serial serum samples from 30 patients with time courses of serum CK, CK-MB, AST, and LD that we consider typical of acute myocardial infarction (MI). In every patient but one, TnT rose in parallel with both CK-MB and AST, but remained elevated significantly longer. The ratios of the elevations of the different markers varied from patient to patient with marked variation in the ratio of TnT to CK-MB. There appeared to be a significant association between the magnitude of that ratio with the level of ALT. J. Clin. Lab. Anal. 11:125–128, 1997. © 1997 Wiley-Liss, Inc.     

Telomerase as a potential marker for inflammation and cancer detection in bronchial washing: A prospective study

ObjectivesThe diagnosis of lung cancer remains difficult especially in peripheral tumors, given the absence of relevant markers and of sensitive imaging techniques. Telomerase is a ribonucleotide enzyme responsible for the immortalization of cancerous cells and seems to increase in bronchial aspirates of lung cancer patients. The purpose of our study is to further investigate the value of telomerase measurement in bronchial aspirates as a diagnostic tool for lung cancer.Design and methodsRandom 82 bronchial aspirates were obtained from patients undergoing bronchoscopy to diagnose any lung illness including inflammation and cancer. Cytology examination, quantification of proteins by Bradford method, and telomerase activity measurement by quantitative Real-time PCR were performed. Out of 82 specimens, 11 were excluded because of hemolysis, absence of elements or lack of final diagnosis. ROC curve analysis was done.ResultsA significant difference in telomerase activity average was noted between normal patients and those with inflammation and cancer. Discriminatory capacity of telomerase activity was: for cancer vs. non cancer, AUC = 0.74 (95% CI: 0.62–0.84), sensitivity = 78%, specificity = 72%, Negative Predictive Value = 87%, at cut-off > 0.46 atmol/mg protein/20 min; for cancer vs. normal, AUC = 0.87 (95% CI: 0.72–0.96), se = 78%, sp = 92%, NPV = 71%, at cut-off > 0.46; for cancer vs. inflammation, AUC = 0.69 (95% CI: 0.55–0.80), se = 74%, sp = 70%, NPV = 79%, at cut-off > 1.03, and for inflammation vs. normal, AUC = 0.76 (95% CI: 0.62–0.88), se = 79%, sp = 77%, NPV = 59%, at cut-off > 0.ConclusionTelomerase activity in bronchial aspirates is a promising diagnostic marker for lung cancer and inflammation detection.