Annexin A5 as a New Potential Biomarker for Cisplatin-Induced Toxicity in Human Kidney Epithelial Cells

Cisplatin is a member of platinum-containing anti-cancer drugs that causes cross-linking of DNA and ultimately cancer cell apoptosis. The therapeutic function of cisplatin on various types of cancers has been widely reported but the side effects have been discovered together and nephrotoxicity has been regarded as major side effect of cisplatin. To select candidates for new sensitive nephrotoxicity biomarker, we performed proteomic analysis using 2-DE/MALDI-TOF-MS followed by cisplatin treatment in human kidney cell line, HK-2 cells, and compared the results to the gene profi le from microarray composed of genes changed in expression by cisplatin from formerly reported article. Annexin A5 has been selected to be the most potential candidate and it has been identifi ed using Western blot, RT-PCR and cell viability assay whether annexin A5 is available to be a sensitive nephrotoxic biomarker. Treatment with cisplatin on HK-2 cells caused the increase of annexin A5 expression in protein and mRNA levels. Overexpression of annexin A5 blocked HK-2 cell proliferation, indicating correlation between annexin A5 and renal cell toxicity. Taken together, these results suggest the possibility of annexin A5 as a new biomarker for cisplatin-mediated nephrotoxicity.     

Agomelatine for Depression in Schizophrenia: A Case-Series

Objectives

Agomelatine, a melatonin (MT1/MT2) receptor agonist and 5-HT2C receptor antagonist, is a new antidepressant and a potential therapeutic option for major depressive episodes and negative symptoms in persons with schizophrenia. We investigated such treatment outcomes with respect to antidepressant efficacy, safety, and tolerability.

Methods

We report a consecutive case series of seven patients with schizophrenia and comorbid major depressive symptoms who received agomelatine for a period of at least six weeks in addition to stable doses of antipsychotic agents. General psychopathology, positive, negative and depressive symptoms were assessed with standardized interviews. Relevant blood parameters were assessed.

Results

Depressive symptoms improved significantly. Positive symptoms remained stable, while negative symptoms and global psychopathology improved significantly. Agomelatine was well tolerated in most patients.

Conclusions

Our findings provide initial evidence that agomelatine is safe and efficacious in treating depressive symptoms in patients with schizophrenia. Furthermore, agomelatine seems to be effective for the treatment of negative symptoms. Randomized controlled trials are necessary to confirm these first observations.     

Quantification of the Serotonin 1A Receptor Using PET: Identification of a Potential Biomarker of Major Depression in Males

Multiple lines of research have implicated the serotonin 1A (5-HT_1A) receptor in major depressive disorder (MDD). Despite this, quantification of 5-HT_1A is yet to yield a clinically relevant MDD biomarker. One reason may be that reported sex differences in the serotonergic system confound the comparison between diagnostic groups. Therefore, this study sought to determine whether differences in 5-HT_1A binding between depressed and control subjects are affected by sex. Using positron emission tomography (PET), serotonin 1A binding was quantified in 50 patients with MDD (34 female, 16 male) and 57 healthy controls (32 female, 25 male). The subjects'' 5-HT_1A density (BP_F, equal to the product of the density of available receptors and tracer affinity), was determined by using the PET tracer [carbonyl-C-11]-WAY-100635, a selective 5-HT_1A antagonist. Results indicated that male MDD subjects had a 67.0% higher BP_F across 13 brain regions compared with male controls (df=103, p<0.0001). The greatest difference between MDD subjects and controls was in the raphe (132%, p=0.000). Furthermore, by using a threshold, male controls can be distinguished from depressed males with high sensitivity and specificity (both >80%). In females, the separation between diagnostic groups yields much lower sensitivity and specificity. This data therefore suggests a specific biosignature for MDD in males. Identification of such a biosignature could provide a deeper understanding of depression pathology, help identify those at highest risk, and aid in the development of new therapies. Further, these findings suggest that combining male and female cohorts may not be optimal for some MDD studies.     

Neurofilament Light Chain Is a Novel Biomarker for Major Depression and Related Executive Dysfunction

BackgroundEvidence suggests that major depressive disorder is related to neuroaxonal injury and that neurofilament light chain (NfL) is a biomarker of neuroaxonal injury. In addition, proinflammatory cytokines have been reported to be associated with major depression and neuroaxonal injury.MethodsForty patients with major depression and 40 age- and sex-matched healthy control participants were enrolled for the measurement of NfL and proinflammatory cytokines and assessment of executive function. General linear models were used to examine the association between NfL levels, proinflammatory cytokine levels, and executive function.ResultsPatients with major depressive disorder exhibited significantly higher NfL levels (P = .007) than the control participants. NfL levels were positively related to log-transformed levels of tumor necrosis factor-α (P = .004). Higher levels of NfL (P = .002) and tumor necrosis factor-α (P = .013) were associated with greater deficits in executive function.DiscussionNfL was a novel biomarker for major depressive disorder and related executive dysfunction. Further studies are necessary to elucidate the role of NfL in the pathophysiology of major depression and related cognitive impairment.     

Evaluation of Dextromethorphan N‐Demethylation Activity as a Biomarker for Cytochrome P450 3A Activity in Man

Abstract: The present study evaluates the usefulness of dextromethorphan N‐demethylation activity indices to reflect cytochrome P450 (CYP) 3A activity in man. Indices of dextromethorphan N‐demethylation activity were categorized as N₁=3‐methoxymorphinan/dextromethorphan, N₂=3‐hydroxymorphinan/dextrorphan, N₃=(3‐methoxymorphinan + 3‐hydroxymorphinan)/(dextromethorphan + dextrorphan). Two mg of midazolam were administered orally to 22 Japanese male volunteers, and midazolam clearance determined. Thirty mg of dextromethorphan were also orally administered to these volunteers and N₁, N₂ and N₃ indices determined by 12 hr urine collection. Results showed N₂ and N₃ were highly correlated (r>0.99, P<0.001), and significantly correlated to oral midazolam clearance (r=0.45, P<0.05); suggesting that N₂ and N₃ are more suitable than N₁ when using dextromethorphan as an index of individual CYP3A activity.     

Vitellogenin as a Biomarker of Exposure for Estrogen or Estrogen Mimics

Vitellogenin, the egg yolk precursor protein, has become a popular biomarker for measuring exposure of oviparous animals to estrogen or estrogen mimics. Vitellogenin is normally produced by females in response to normal cycles of estradiol during oogenesis. The gene for vitellogenin is also present in the livers of males but it is normally silent. Upon exposure to estrogen or to an estrogen mimic, the gene is turned on and vitellogenin is synthesized. After synthesis, it is exported into the blood where, in males, it remains until it is degraded or cleared out by the kidneys. In females, vitellogenin is taken up by the developing oocyte through receptor mediated endocytosis.There are several assays in the literature for measuring vitellogenin levels in plasma. The easiest method is through antibody based assays including ELISA (enzyme-linked immunosorbent assay) or by western blot. Competition or sandwich ELISAs are the most sensitive assays and they can detect vitellogenin in plasma in the nanogram to milligram per ml range.This chapter discusses methods for purifying vitellogenin from plasma, generating antibodies, and performing assays to measure vitellogenin.     

Evaluation of Cocoa Butter as Potential Lubricant for Coprocessing in Pharmaceutical Tablets

The lubricant activity of cocoa butter coprocessed with magnesium stearate plus talc (CMT) was compared with magnesium stearate plus talc (MT) using flow and compressional characteristics of paracetamol granules and mechanical properties of their tablets as assessment parameters. The flow of the granules quantified as Hausner's ratio, Carr's index, and angle of repose showed that CMT has a higher ability than MT to reduce densification of granules due to vibration. Compressional characteristics analyzed using density measurements and the Heckel and Kawakita plots revealed that CMT did not facilitate the increase in the densification of the granules during the filling and at low pressures, D_b. Also, CMT reduced the plastic deformation of the granules measured by the P_y-yield pressure at onset of plastic deformation and P_k-yield pressure of deformation under compression. The mechanical properties determined by the tensile strength, T, and brittle fracture index, BFI, of the tablets produced were affected by CMT. The T and BFI of tablets with CMT were lower than those of MT. The results suggest that though CMT lowered the plasticity of the granules, it improved their flow rate and assisted in producing tablets with fewer tendencies to cap or laminate. This work concluded that cocoa butter, an inexpensive and easily available lipid, is an effective and viable lubricant that can be co-processed with magnesium stearate/talc mixture for an efficient lubrication of granules and may be useful in reducing lamination and capping in formulations that are susceptible to these 2 defects of tablets.     

阿立哌唑治疗精神分裂症伴抑郁的疗效分析

目的探索精神分裂症伴抑郁的临床治疗方法,探讨阿立哌唑对治疗精神分裂症伴抑郁的临床疗效。方法随机将精神分裂症伴抑郁患者按入组顺序分为两组,观察组53例,使用阿立哌唑进行治疗;对照组52例,使用利培酮进行治疗,疗程均为5周。比较两组治疗前后的汉密顿抑郁量表(HAMD)、临床疗效的总评量表(CGI)不良反应量表(TESS)得分。结果两组治疗前后的HAMD及CGI量表评分相比均存在显著差异,观察组疗效比对照组明显,两组相比具有显著统计差异,P<0.05。而且两组显效率相比也存在显著差异,P<0.05。结论阿立哌唑对治疗精神分裂症伴抑郁疗效显著。     

Patent Evaluation: Human Amyloidin Protease as a Potential Therapeutic Agent

Summary

Novelty: The purification of human amyloidin protease, the generation of antibodies specific for the enzyme, and the isolation and in vitro expression of its cognate gene, are reported.

Biology: Methods for the purification of amyloidin protease from human tissue are provided. The enzyme has an appparent molecular weight of 80 kDa and proteolytically cleaves a Met-Asp peptide bond in the amyloid precursor protein which releases the mature Asp terminus of the β-amyloid peptide. An assay for inhibitors of the protease is described, and antibodies raised against the enzyme are provided. The isolation and characterization of the cognate gene is described together with methods for expressing the recombinant protein in vitro. Inhibitors of the enzyme may have use for the therapeutic intervention of Alzheimer's disease, and antibodies raised against it may have diagnostic use.     

Preclinical Evaluation of Reconsolidation Blockade by Clonidine as a Potential Novel Treatment for Posttraumatic Stress Disorder

Exposure to traumatic events can lead to posttraumatic stress disorder (PTSD). Current PTSD treatments typically only produce partial improvement. Hence, there is a need for preclinical research to identify new candidate drugs and to develop novel therapeutic approaches. Animal studies have indicated that fear memories can be weakened by blocking restabilization after retrieval, a process known as reconsolidation. Furthermore, evidence suggests that there are important alterations of the noradrenergic system in PTSD, and hence it may be of interest to study drugs that target this pathway. Here, we investigated the efficacy of clonidine, an α₂-adrenoreceptor agonist, to block reconsolidation in an animal model of persistent traumatic memories. Using an auditory fear conditioning paradigm in rats, we tested the efficacy of clonidine to weaken fear memory retention when administered systemically after retrieval. We evaluated dosage, number of treatments, and specificity in reconsolidation blockade. We found that postretrieval administration of clonidine disrupts fear-related memories in a dose-dependent manner and that two treatments are sufficient for maximal memory impairment. Furthermore, we determined that this effect is long lasting and specific to reconsolidation processes as shown by the selectivity to affect reactivated memories and the absence of spontaneous recovery and of postreactivation short-term memory impairment. Our results demonstrate the efficacy of systemic administration of clonidine following retrieval to persistently disrupt fear memory retention through reconsolidation blockade. This study provides important preclinical parameters for future therapeutic strategies involving clonidine to block reconsolidation as a novel treatment for PTSD symptoms.     

8-羟基脱氧鸟苷作为DNA氧化损伤标志物在疾病诊断中的应用

机体内,活性氧簇(reactive oxygen species,ROS)过度蓄积可造成DNA氧化损伤,产生8-羟基脱氧鸟苷(8-hydroxy-2’-deoxyguanosine,8-OHdG).多项研究表明,8-OHdG不仅反映整个机体平均的氧化损伤率及体内DNA氧化损伤与机体正常的修复作用,而且与糖尿病、癌症、心血管疾病的发生和病情发展有着密切的关系.本文对8-OHdG作为氧化损伤标志物在糖尿病及其并发症、癌症、心血管疾病的诊断及治疗评价中的应用进行了综述,认为8-OHdG可能成为氧化应激相关疾病的一种极有前途的生物标志物.     

Keratinocytic Vascular Endothelial Growth Factor as a Novel Biomarker for Pathological Skin Condition

Skin is an emerging target tissue in pharmaceutical and cosmetic science. Safety assessment for dermal toxicity is a critical step for development of topically applicable pharmaceutical agents and ingredients in cosmetics. Urgent needs exist to set up toxicity testing methods for dermal safety, and identification of novel biomarkers for pathological cutaneous alteration is highly required. Here we will discuss if vascular endothelial growth factor (VEGF) has a potential as a biomarker for dermal impairment. Experimental and clinical evidences for induction of keratinocytic VEGF under pathological conditions will be reviewed.     

Phospholipid Concentration in Lung Lavage Fluid as Biomarker for Pulmonary Fibrosis

Pulmonary surfactant comprised primarily of phospholipids is a phospholipid-protein complex synthesized by type II alveolar epithelial cells or Clara cells and secreted to the pulmonary alveoli. As changes have been found in phospholipid concentrations in the bronchoalveolar lavage fluid (BALF) of patients with pulmonary fibrosis, phospholipid is considered to be involved in the process of fibrois/fibrotic process. Therefore, we made a crystalline silica rat model and measured phospholipid concentrations in lung lavage fluid in order to study the relationship of phospholipid to particle-induced pulmonary fibrosis. Eight-week-old Wistar male rats (n = 35) were injected with 2 mg crystalline silica particles suspended in 0.4 ml physiological saline. Rats in the control group (n = 35) were injected with physiological saline only. There were 7 rats in each of the ten subgroups. Rats were sacrificed and dissected at 3 days, 1 wk, 1 mo, 3 mo, and 6 mo after injection. Bronchoalveolar lavage was conducted on bronchoalveoli recovered from the right lung of each rat, the lavage fluid was centrifuged, and the supernatant was used to measure phospholipid concentration. The results were compared with previously reported inflammation scores. Phospholipid concentrations in lung lavage fluid for the exposed group showed a statistically significant increase compared to the control group throughout the observation period. Moreover, when compared to histopathologically examined inflammation scores, a positive correlation was found between the two. Judging from the facts that phospholipid concentrations in lung lavage fluid increased and that this increase correlated with the severity of inflammation, this experiment indicated that phospholipids are involved in particle-induced lung disorders.     

Cyclophosphamide as a Potential Chemosterilant for Harmful Snails

Abstract: Cyclophosphamide induced changes in the ovotestis of the snail Lymnaea acuminata, the vector of the giant liverflukes Fasciola hepatica and Fasciola gigantica were studied in order to explore potential of the drug as a chemosterilant for snails. The drug caused a dose dependent reduction in the levels of DNA, RNA and proteins and the activity of the enzyme alkaline phosphatase. It increased the activity of acid phosphatase and the levels of total free amino acids in the ovotestis. While the animals showed nearly total recovery in RNA and DNA levels 7 days after termination of drug treatment, changes produced in protein, amino acid levels and phosphatase activity did not show any recovery. It appears that cyclophosphamide, while affecting its primary targets i.e. DNA and RNA, irreversibly inhibits protein synthesis through other cellular enzymes as well.