[The patient has died now what? : Step by step-what happens with patients after their death?] / Patient tot was nun? : Schritt für Schritt ­ was passiert mit dem Patienten nach seinem Tod?

The determination of a person's death, the subsequent post-mortem examination and the creation of the death certificate are part of regular medical work. The post-mortem examination, which is exclusively a medical duty, must be carried out immediately after the determination of death and includes in particular the definition of the cause of death and the type of death, which, in the case of a non-natural or unexplained death, entails further investigations by the police or public prosecutor and sometimes also forensic examinations. This article aims to shed more light on the possible processes after the death of a patient.

An advanced method for the immunohistochemical detection of nitric oxide synthase (NOS) in the female genital tract.

The expression of nitric oxide synthase (NOS) in male and female urogenital tissues has been investigated by using conventional light microscopical immunoperoxidase staining. We present an improved immunohistochemical method for the specific and simultaneous detection of endothelial and neuronal NOS (eNOS/nNOS) in vaginal tissue. Specific antibodies have been used in combination with the tyramide signal amplification method. We found a subepithelial meshwork of varicose nerve fibers. A subpopulation of fibers presented immunoreactivity specific for nNOS. Epithelial cells also showed cytoplasmatic labeling for nNOS. Arteries presenting signals for eNOS in their endothelial layer were found in close proximity to nNOS-positive nerve fibers.

Age-, tumor-, and metastatic tissue-associated DNA hypermethylation of a T-box brain 1 locus in human kidney tissue.

BACKGROUND: While a considerable number of tumor-specific hypermethylated loci have been identified in renal cell cancer (RCC), DNA methylation of loci showing successive increases in normal, tumoral, and metastatic tissues could point to genes with high relevance both for the process of tumor development and progression. Here, we report that DNA methylation of a locus in a genomic region corresponding to the 3'UTR of the transcription factor T-box brain 1 (TBR1) mRNA accumulates in normal renal tissues with age and possibly increased body mass index. Moreover, a further tissue-specific increase of methylation was observed for tumor and metastatic tissue samples. RESULTS: Biometric analyses of the TCGA KIRC methylation data revealed candidate loci for age-dependent and tumor-specific DNA methylation within the last exon and in a genomic region corresponding to the 3'UTR TBR1 mRNA. To evaluate whether methylation of TBR1 shows association with RCC carcinogenesis, we measured 15 tumor cell lines and 907 renal tissue samples including 355 normal tissues, 175 tissue pairs of normal tumor adjacent and corresponding tumor tissue as well 202 metastatic tissues samples of lung, bone, and brain metastases by the use of pyrosequencing. Statistical evaluation demonstrated age-dependent methylation in normal tissue (R = 0.72, p < 2 × 10-16), association with adiposity (P = 0.019) and tumor-specific hypermethylation (P = 6.1 × 10-19) for RCC tissues. Comparison of tumor and metastatic tissues revealed higher methylation in renal cancer metastases (P = 2.65 × 10-6). CONCLUSIONS: Our analyses provide statistical evidence of association between methylation of TBR1 and RCC development and disease progression.

A switch from epithelial to mesenchymal properties correlates with lymphovascular invasion in squamous cell carcinoma of the penis.

The purpose of the study was to assess the incidence and prognostic role of epithelial-to-mesenchymal-transition (EMT) in squamous cell carcinoma of the penis (SCCP). Sixty tumor specimens of surgically treated SCCP patients characterized by a central histopathologic review were stained with antibodies against E-cadherin, N-cadherin, ß-catenin, and vimentin. Staining profiles were scored by two independent raters, and correlated with pertinent clinical and pathological parameters and cancer-specific mortality (CSM; median follow-up: 34 months, interquartile range: 6-60 months). Correlation statistics proved good interobserver agreement in staining evaluation (K-values between 0.62 and 1.00, all p<0.001). Based on consensus decision between both raters, 36 study cases (60%) showed a switch from E-cadherin to N-cadherin (as a hallmark of EMT), which correlated with the presence of lymphovascular invasion (ρ=0.287, p=0.026) while failing to interfere with CSM. Although cadherin switch was correlated with a loss of membranous ß-catenin expression (ρ=0.629, p<0.001), none of the study cases showed nuclear ß-catenin expression, and only three EMT cases (8.3%) had tumor buds revealing vimentin expression. Our data suggest that roughly half of surgically treated SCCP cases exhibit EMT, a parameter correlating with lymphovascular invasion. However, further studies are clearly needed to validate the so far unresolved possible role of cadherin switch in terms of predicting nodal spread in patients with SCCP. Moreover, the apparently complex mechanisms driving EMT in SCCP should be explored by future studies, as knowledge about these might provide a so far unexploited basis for the development of novel targeted therapies against SCCP with metastatic dissemination.

Melanocortin receptor agonists in the treatment of male and female sexual dysfunctions: results from basic research and clinical studies.

INTRODUCTION: Over the last 20 years, basic and clinical research activities studying the male and female sexual responses have led to several pharmacological options to treat male erectile dysfunction (ED) and female arousal and orgasmic disorders. While some strategies exclusively focus on peripheral mechanisms--such as nitric oxide/cyclic GMP signaling, which is known to play a role in the control of genital vascular and nonvascular smooth muscle--others have considered the central pathways involved in mediating arousal and orgasmic functions in females as well as the induction of penile erection in males. Aside from dopaminergic agonists, drugs known to target the central melanocortin system have also been assumed to have a promising potential in the treatment of female and male sexual dysfunctions. AREAS COVERED: The present review summarizes the achievements that have been made in the clinical development of melanocortin receptor (MCR) agonists (melanotan I, melanotan II, bremelanotide) for the treatment of symptoms of sexual arousal and orgasmic disorders in adult females and ED in males. EXPERT OPINION: The data available at present have facilitated our understanding of how the melanocortin pathway regulates both the male and female sexual functions. Indeed the data warrant further investigation to demonstrate the impact of the activation of MCRs by specific agonists on penile erection and female arousal and orgasm function.

Diagnostic and prognostic impact of peritumoral stromal remodeling in patients with surgically treated invasive penile squamous cell cancer.

Stromal remodeling (SR), characterized by focal loss of CD34(+) fibrocytes paralleled by a gain of α-smooth muscle actin (α-SMA)-positive myofibroblasts, has been reported in several cancer types. However, the role of SR in invasive penile squamous cell cancer (PSC) has not been investigated so far. We compared 90 surgically treated PSCs (study group) and 55 control specimens (33 foreskins and 22 differentiated penile intraepithelial neoplasias) for the presence of stromal CD34(+) fibrocytes and α-SMA-positive myofibroblasts scored by independent raters. Multivariate proportional hazards regression analysis was used to assess the impact of staining profiles on cancer-specific mortality of the 90 PSCs (median follow-up, 32 months; interquartile range, 6-64). The incidence of SR differed significantly between study and control group specimens (51.1% versus 9.1%; P < .001). Five years postsurgically, 24% and 46% of the study patients without and with SR had succumbed to their PSC (P = .010). After adjusting for the age at the time of surgery, type of surgery, tumor size, Broders' grade, pT stage, and nodal status, study patients with SR showed 3.76-fold increased cancer-specific mortality (95% confidence interval, 1.3-10.5; P = .012). Our findings suggest that SR might have prognostic as well as some limited differential diagnostic value in terms of delineating invasive PSC from preinvasive lesions. However, our preliminary data clearly need to be validated by larger advanced studies in the future.

Phosphodiesterase type 1, calcitonin gene-related peptide and vasoactive intestinal polypeptide are involved in the control of human vaginal arterial vessels.

OBJECTIVE: The vagina makes a major contribution to the normal female sexual response cycle. An increase in vaginal blood flow is considered a key event in the mechanism of sexual arousal. Recent research has focused mainly on the cyclic GMP pathway and phosphodiesterase type 5 (PDE5, cyclic GMP specific PDE) in the control of vaginal vascular smooth muscle, whereas only little is known on the role of other key proteins and mediators of cyclic nucleotide mediated signaling in this process. The aim of the present study was to evaluate in the human vagina, by means of immunohistochemistry, the expression and distribution of phosphodiesterase type 1 (PDE1, known to hydrolize both cyclic AMP and cyclic GMP) in relation to calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and protein gene product 9.5 (PGP 9.5). STUDY DESIGN: Sections of human vagina (full wall specimens) were incubated with antibodies directed against PDE1, CGRP, VIP, PGP 9.5 and alpha-actin, followed by exposure to fluorochrome-labelled secondary antibodies. Visualization was commenced by means of laser fluorescence microscopy. RESULTS: Microscopic examination revealed a dense meshwork of PGP 9.5-positive nerve fibers innervating the sections of vaginal wall. Small vessels interspersing the tissue presented dense staining for PDE1 in their smooth musculature. Blood vessels were seen surrounded by PDE1-immunoreactive longitudinal smooth muscle fibers. The vessels were also found innervated by PGP-positive varicose nerve fibers characterized by the expression of CGRP. Some fibers presented immunosignals specific for VIP. CONCLUSION: Key mediators of the cyclic AMP and cyclic GMP pathways are co-localized in nerves seen in close proximity to vascular smooth muscle expressing PDE1. These findings suggest that both signaling cascades are involved in the control of vaginal blood flow.

Avanafil for the treatment of erectile dysfunction: initial data and clinical key properties.

Orally active, selective inhibitors of phosphodiesterase type 5 (PDE 5, cyclic GMP PDE), such as sildenafil, tadalafil and vardenafil, are currently the first-choice treatment options for the clinical management of erectile dysfunction (ED) of various etiologies and severities. However, a significant number of patients remain dissatisfied with the available therapies due a lack of efficacy or discomfort arising from adverse events. Several new PDE5 inhibitors, among which are avanafil (TA-1790), lodenafil, mirodenafil, udenafil, SLX-2101, JNJ-10280205 and JNJ-10287069, have recently been approved and introduced into the market or are in the final stages of their clinical development. Avanafil (marketed in the US under the brand name STENDRA(™)) has been developed by VIVUS Inc. (Mountain View, CA, USA) and has recently received approval from the US Food and Drug Administration (FDA) for use in the treatment of male ED. The drug has demonstrated improved selectivity for PDE5, is rapidly absorbed after oral administration with a fast onset of action and a plasma half-life that is comparable to sildenfil and vardenafil. In phase II and phase III clinical trials that included a large number of patients, avanafil has been shown to be effective and well tolerated. Owing to its favorable pharmacodynamic and pharmacokinetic profile, avanafil is considered as a promising new option in the treatment of ED. The present article summarizes the initial data and clinical key properties of avanafil.

[Post-mortem examination prior to cremation--an instrument to verify the quality of medical post-mortems and uncover non-natural deaths?]. / Die Krematoriumsleichenschau--Instrument zur Qualitätskontrolle der ärztlichen Leichenschau und Aufdeckung nichtnatürlicher Todesfälle?

The external post-mortem examination, its deficient quality and possible causes have been the subject of numerous political and professional discussions. The external post-mortem examination is the basis for the decision whether further criminal investigations are required to clarify the cause of death. It is thus an essential instrument to ensure legal certainty. Before cremation, a second external post-mortem examination is performed by a public medical officer to make sure that errors of the first post-mortem are corrected. In the present study, cases were retrospectively analyzed in which a forensic autopsy had been ordered on the basis of the results of the post-mortem examination performed before cremation. The entries on the death certificate regarding the manner and cause of death were compared with the autopsy results. Between 1998 and 2007, 387 autopsies were ordered after external examination before cremation. In 55 cases (14.2%), the autopsy revealed a non-natural death, although a natural death had been attested on the death certificate. In descending order, a wrong manner of death was attested by clinicians, general practitioners and emergency physicians. With regard to the place where the first external post-mortem had been performed the lowest error rate was seen in nursing homes. Concerning the cause of death, discrepancies between the first post-mortem and autopsy were found in 59.4% of the cases. In this respect, general practitioners and clinicians were ranking first, whereas in nursing homes the cause of death was wrongly assessed in over 70% of cases. At present, the medical post-mortem does not meet the required quality standards, especially with regard to legal certainty. Determination of the cause of death on the basis of the external post-mortem examination is a challenging task even for the experienced medical examiner. As to the categorization of the manner of death it has to be stated that non-natural deaths are often not recognized or that the possibility to certify a death as unclear is not sufficiently used. As a result, it seems important to demand intensive, qualified, additional training in external post-mortem examinations for physicians.

[Aquatic fatalities--a systematic retrospective analysis]. / Ertrinkungstod--eine systematische retrospektive Untersuchung.

In a retrospective analysis of the autopsy material (n = 5,767) of the Institute of Legal Medicine of the Hanover Medical School covering the period of 1998-2007, all aquatic fatalities were evaluated, categorized and systematically compared under epidemiological and forensic criteria. The total of 156 cases of death by drowning (2.7 % of all autopsies) included 38 bathtub drownings and 28 deaths in the water for which no pathological anatomical cause of death could be reliably demonstrated. A control group (n = 221) was investigated for the presence of aqueous liquid in the sphenoid sinuses and compared with the findings of the drowning cases without signs of putrefaction. About 16 % of the control cases had fluid in the sphenoid sinuses compared with 57.6 % in the drowning group. Most of the drowning victims were men (60.9 %), whereas in the group of bathtub drownings the majority were women. More than half of the drowning cases (n = 89) could be classified as accidents. The individual groups showed a different incidence of findings associated with drowning.

Expression and distribution of cyclic AMP- and cyclic GMP-binding protein kinases in the human vagina- an immunohistochemical study.

INTRODUCTION: In contrast to research findings describing the localization of nitric oxide synthases (NOS), guanylyl cyclases, and cyclic adenosine monophosphate (cAMP)- and cyclic guanosine monophosphate (cGMP)-degrading phosphodiesterase isoenzymes in the human vagina, the distribution of proteins known as major targets for cyclic nucleotides has not yet been evaluated. cAMP- and cGMP-dependent protein kinases (cAK, cGKI) have been identified as important receptors for cyclic nucleotides downstream the signaling cascades. AIM: To investigate, by means of immunohistochemistry, the expression of cAK and cGKI in relation to endothelial NOS (eNOS), vasoactive intestinal polypeptide (VIP), and protein gene product 9.5 (PGP 9.5) in the human vagina. MAIN OUTCOME MEASURES: Expression and distribution of cAK and cGKI(alpha,beta) in relation to eNOS, VIP, and PGP 9.5 in human vaginal tissue. METHODS: Immunohistochemical techniques were applied to sections of human vaginal full wall specimens in order to evaluate the presence of cAK and cGKI(alpha,beta) in relation to VIP, PGP 9.5, and eNOS, respectively. Western blot analyses were conducted using cytosolic supernatants of homogenized specimens of the vaginal wall and epithelium. RESULTS: Immunostaining specific for cGKIbeta was observed in vascular and nonvascular smooth muscle of the vagina. In the endothelial layer, cGKIbeta was found colocalized with eNOS. In contrast, no signals indicating cGKIalpha were registered. cAK-positive subepithelial vessels were found to be innervated by a dense meshwork of PGP-containing varicose nerve fibers, some of which presented expression of VIP. The expression of cAK and cGKIbeta was confirmed by Western blotting. CONCLUSIONS: Our results demonstrate the expression of cAK and cGKIbeta in the human vagina. The colocalization with VIP and eNOS underlines the significance of both the cAMP and GMP pathway in the control of human vaginal vascular and nonvascular smooth muscle.

Urinary collecting system invasion reflects adverse long-term outcome and is associated with simultaneous metastatic spread at the time of surgery and with multilocular dissemination during postsurgical follow-up in renal cell cancer.

OBJECTIVES: To assess the prognostic implication of urinary collecting system invasion (CSI) in renal cell cancer (RCC). METHODS: Surveying a mean follow-up of 85 months, we investigated a cohort of 834 patients after radical (n = 710) or partial (n = 124) nephrectomy. At the time of surgery, 63 patients (7.6%) suffered from metastatic RCC. Various histopathologic parameters were analysed, and cancer specific survival (CSS) curves were individualized for each parameter. Furthermore, multivariate analysis was accomplished. RESULTS: Collecting system invasion was independently associated with a significant decline in CSS and was associated with simultaneous metastatic spread at the time of surgery and multilocular (involvement of at least two different organ systems) dissemination. CONCLUSIONS: The prognostic implication of CSI in RCC appears to be more complex than expected. Therefore, pathologists should report on CSI to enable selection of patients to be investigated in prospective studies which are needed to clarify the prognostic role of CSI in RCC.

[Causes of death among pilots: "acute myocardial infarction"--are the present examination methods for airworthiness sufficient?]. / Todesfälle bei Piloten: "Der akute Myokardinfarkt"--Sind die derzeitigen Flugtauglichkeitsuntersuchungen ausreichend?

The European Union plans to harmonize the aviation requirements, in particular the flight crew licensing requirements. On 23 May 2007, the German Federal Ministry of Transport, Building and Urban Affairs published the Flight Crew Licensing Requirements, which are based on the Joint Aviation Requirements, Flight Crew Licensing 3, Amendment 5. These guidelines also list the examination methods to be used for testing the medical fitness of pilots. In this Amendment some examinations which were part of the routine tests before JAR-FCL3 became effective have been deleted, e.g. the exercise ECG. This article presents two cases from the autopsy material of the Institute of Legal Medicine in Hanover and discusses the problems associated with the new examination guidelines.

[First report of lethal gamma-hydroxybutyrate (GHB) intoxication after consumption of gamma-butyrolactone (GBL) in Lower Saxony]. / Erster Todesfall durch gamma-hydroxybuttersäure (GHB) nach Aufnahme von gamma-Butyrolacton (GBL) in Niedersachsen.

A 25-year-old drug addict died from an overdose of GBL after mistaking it for water in preparing a dilution. He realized his error and wrote a call for help in the chat room of a drug forum, so that we know the time, type and approximate amount of drug consumed. The deceased had written numerous contributions in this drug forum before, which provide further information on his drug habits and the duration of the abuse of GHB, GBL and other drugs. According to the Criminal Investigation Department of Lower Saxony, this is the first death due to GHB/GBL reported in Lower Saxony.

[Practicability of the mobile one-finger scanner Cross Match MV5 in fingerprinting of corpses: are mobile fingerprinting scanners suitable for use in mass disasters?]. / Praktikabilität des mobilen Einzelfingerscanners Cross Match MV5 in der Leichendaktyloskopie: Sind die mobilen Einzelfingerscanner für den Einsatz bei Massenkatastrophen geeignet?

Dactyloscopy is a special field in the police records department and a suitable means for identifying unknown dead persons as well as solving crimes by taking fingerprints from living persons. Apart from the conventional methods of dactyloscopy, mobile and more compact instruments are to facilitate taking prints of fingertips and palms also at the scene of mass disasters. In the presented study, living persons and corpses were examined to find out the possible uses and limits of mobile one-finger scanners. The concrete issue of the investigation was whether the mobile one-finger scanner Cross Match MV5 is suitable for application in mass disasters. The device was used in 12 corpses aged 5 weeks to 76 years (mean postmortem interval 5.5 days) and in 28 living persons aged 6 weeks to 87 years. In summary, the scanner produced qualitatively good prints in all age groups of the living individuals. In the corpses, the prints were only partly evaluable. In particular, fingers and fingertips with soot blackening and livid discoloration were difficult to analyse. Postmortem rigidity also complicated the handling of the scanner. In fresh corpses, the scanner can be recommended without reservation. Even if the epidermis was detached, the scanner was able to produce evaluable prints of the dermis of the hypothenar.

Ethanol-induced malfunction of neutrophils respiratory burst on patients suffering from alcohol dependence.

BACKGROUND: Polymorphonuclear, neutrophil granulocytes (PMN) play a major role in the control of infections, and people who abuse alcohol are susceptible to infections. Resistance against infections ensues intracellularly following initial phagocytosis of microorganisms with the oxygen-dependent respiratory burst, the key enzyme of which is the respiratory burst oxidase, whereby oxygen radicals are produced for microbial destruction. To date there is insufficient information available in connection with the process of impaired defence against infection in patients suffering from alcohol dependence. Therefore, our investigation was carried out to determine the influence of alcohol exposition on the formation of oxygen radicals and the respiratory burst. METHODS: 4.5 ml of whole blood was taken from 10 healthy adults and 10 patients suffering from alcohol dependence. An additional 3.5 ml of whole blood was taken from the alcoholic patients for determination of the blood alcohol concentration. The respiratory burst of PMN was tested using the Four-Colour-Continuous Flow Cytometer. Each experimental procedure consisted of 4 test samples [negative controls, Escherichia coli, FMLP-supplement (N-formyl-l-methionyl-l-leucyl-l-phenylalanin), PMA-supplement (phorbol-12-myristate-13-acetate)]. Differing concentrations of ethanol were also introduced to each of the tests performed (0.20 to 4.00 g/l). RESULTS: Ethanol revealed a marked decrease of burst activity in those patients suffering from alcoholism with increased alcohol concentration. A dependence between the burst activity and the ethanol concentration was seen to be statistically significant. This effect was only evident after stimulation with E. coli and FMLP in those patients with alcohol dependence. CONCLUSION: The results presented in this study show an impairment in the function of PMN in those patients addicted to alcohol due to the decrease in burst activity. In view of the results of the different stimuli, the second-messenger effects were not evident. A clarification of this phenomenon could well be assumed as an allosteric receptor effect on the burst oxidase, namely, a direct effect on the phagocytosis interaction between circulating granulocytes and causative organisms.

Ki67 staining index and neuroendocrine differentiation aggravate adverse prognostic parameters in prostate cancer and are characterized by negligible inter-observer variability.

INTRODUCTION: This study aims to clarify whether neuroendocrine differentiation (NED) and/or proliferation activity assessed by means of Ki67 staining index (Ki67SI) might aggravate other established adverse prognostic parameters commonly used for predicting outcome in surgically treated prostate cancer, and to assess inter-observer variability in assigning NED and Ki67 SI. MATERIAL AND METHODS: A total of 528 patients surgically treated due to prostate cancer were evaluated in this study. Relevant data were retrospectively obtained by chart review. Immunostaining with antibodies directed against Chromogranin A and Ki67 was performed on archived surgical material, and was evaluated by two independent histopathologists blinded to the specimens. Surveying a median postsurgical follow-up of 46.4 months, postsurgical serum PSA-levels were regularly documented for identifying biochemical progress. Multivariate analysis was performed by means of the Cox regression hazards regression method to evaluate possible aggravations of established adverse prognostic parameters (nodal status, tumour stage, pretherapeutic PSA-level, and Gleason score) by NED and/or Ki67SI. Ki67 SI and NED were shown to significantly aggravate these established adverse prognostic parameters, and were found to be characterized by negligible inter-observer variability. CONCLUSION: Ki67 SI and NED should be advocated to be rendered by the histopathologist because both parameters can be immunohistochemically determined without much additional expense in time and cost involved. This concept is rewarded by an additional gain of prognostic accuracy in evaluating individual risk profile after surgery.

[Postmortem dismemberment/mutilation--medicolegal and criminalistic evaluation of the autopsies performed by the Institute of Legal Medicine at the Hanover Medical School]. / Leichenzerstückelungen/-verstümmelungen --differenzierte rechtsmedizinische und kriminalistische Aufarbeitung des Sektionsgutes des Institutes für Rechtsmedizin der Medizinischen Hochschule Hannover.

Among all the autopsies performed by the Institute of Legal Medicine at the Hanover Medical School between 1978 and 1998. 16 cases of postmortem dismemberment/mutilation were evaluable under criminalistic and medicolegal aspects. In the present study, particular attention was paid to the psychosocial circumstances of the perpetrator's and victim's lives and also to the classification of the different types of postmortem dismemberment/mutilation illustrated by case reports. Apart from an unfavourable personal life situation at the time of the offence, essential predisposing factors were: poor integration in society and family, occupational problems, drug abuse and mental diseases. As the interactions between these factors differ in every single case and may be combined with other factors, the analysis of such homicides should always be based on the assessment of the individual case.

Adiposity and age are statistically related to enhanced RASSF1A tumor suppressor gene promoter methylation in normal autopsy kidney tissue.

Age, adiposity, and smoking are risk factors for the development of renal cell carcinoma. Hypermethylation of the RAS association domain family 1A gene (RASSF1A) promoter belongs to the most frequently detected epigenetic alterations in human cancers including renal cell carcinoma. RASSF1A is functionally involved in cell cycle control in normal cells and depletion promotes a number of cellular changes increasing the risk for neoplastic growth. We investigated the hypothesis that age, modulated by the factors adiposity and anthracosis as a surrogate for smoking, is a predictor of RASSF1A promoter methylation in normal kidney tissue. Using a cross-sectional study design, we quantitatively analyzed RASSF1A methylation in 78 normal autopsy kidney tissues by quantitative combined bisulfite and restriction analysis and bisulfite sequencing, and statistically evaluated the degree of relative methylation for a relationship with the predictor age and study factors adiposity and state of anthracosis. Statistical analysis showed that age (regression analysis; P < 0.001), adiposity (univariate analysis; P = 0.016), and state of anthracosis (t test; P = 0.005) are each significantly associated with an increase of RASSF1A promoter methylation in normal kidney tissue. However, only age (P = 0.008) and adiposity (P = 0.008) were identified as independent predictors of RASSF1A promoter methylation using covariance analysis. This study provides statistical evidence that the common cancer risk factors age and adiposity enhance RASSF1A promoter methylation in nonmalignant kidney tissue.