Comparative proteome analysis identified CD44 as a possible serum marker for docetaxel resistance in castration-resistant prostate cancer.

Baseline or acquired resistance to docetaxel (DOC) represents a significant risk for patients with metastatic prostate cancer (PC). In the last years, novel therapy regimens have been approved providing reasonable alternatives for DOC-resistant patients making prediction of DOC resistance of great clinical importance. We aimed to identify serum biomarkers, which are able to select patients who will not benefit from DOC treatment. DOC-resistant PC3-DR and DU145-DR sublines and their sensitive parental cell lines (DU145, PC3) were comparatively analyzed using liquid chromatography-coupled tandem mass spectrometry (LC-MS/MS). Results were filtered using bioinformatics approaches to identify promising serum biomarkers. Serum levels of five proteins were determined in serum samples of 66 DOC-treated metastatic castration-resistant PC patients (mCRPC) using ELISA. Results were correlated with clinicopathological and survival data. CD44 was subjected to further functional cell culture analyses. We found at least 177 two-fold significantly overexpressed proteins in DOC-resistant cell lines. Our bioinformatics method suggested 11/177 proteins to be secreted into the serum. We determined serum levels of five (CD44, MET, GSN, IL13RA2 and LNPEP) proteins in serum samples of DOC-treated patients and found high CD44 serum levels to be independently associated with poor overall survival (p = 0.001). In accordance, silencing of CD44 in DU145-DR cells resulted in re-sensitization to DOC. In conclusion, high serum CD44 levels may help identify DOC-resistant patients and may thereby help optimize clinical decision-making regarding type and timing of therapy for mCRPC patients. In addition, our in vitro results imply the possible functional involvement of CD44 in DOC resistance.

[Association between the 5-HTTLPR polymorphism of the serotonin transporter gene and suicide: a case-control pilot study]. / A szerotonin transzporter gén 5-HTTLPR polimorfizmusa és az öngyilkosság összefüggése: eset-kontroll elrendezésu pilot vizsgálat.

OBJECTIVES: Besides psychological and social effects, biological, biochemical and genetic factors also play a role in the background of suicide. The aim is developing a complex model incorporating all the above factors so that suicide could be predicted and prevented in those at risk. Based on several studies 5-HTTLPR s allele frequency is increased in case of violent completed suicides. The aim of the present study was to validate this association in a sample of completed suicides. METHODS: During autopsy sample DNA samples were obtained for 5-HTTLPR genotyping from 200 subjects deceased due to suicide and 200 controls deceased due to other causes. Chi square tests and logistic regressions were performed according to additive, dominant and recessive models to analyse the possible association between 5-HTTLPR genotype distribution and suicide. RESULTS: Ratio of violent and non-violent suicides was 81% and 19% respectively. No significant difference was found in the distribution of 5-HTTLPR genotypes between the suicide and controls samples. No difference was found between violent and nonviolent suicides with respect to genotype distribution. A significant association was found between sl genotype and suicide at a younger age. CONCLUSIONS: Our pilot study did not support the supposed association between 5-HTTLPR and completed suicides or with violent completed suicides. However we found a significant association between sl genotype and suicide in young suicidals.

Digital photo documentation of forensically relevant injuries as part of the clinical first response protocol.

BACKGROUND: A problem encountered by medical examiners is that they have to assess injuries that have already been medically treated. Thus, they have to base their reports on clinical forensic examinations performed hours or days after an injury was sustained, or even base their assessment solely on information gleaned from medical files. In both scenarios, the forensic examiner has to rely heavily on the first responder's documentation of the original injury pattern. Medical priority will be to immediately treat a patient's injuries, and the first responder may, in addition, initially be unaware of a possibly criminal origin of an injury. As a result, the documentation of injuries is frequently of limited value for forensic purposes. This situation could be improved if photographic records were briefly made of injuries before they were treated. METHODS: German-language medicolegal, criminal, and photography journals and books were selectively searched with the help of PubMed and other databases. In addition, the authors' experiences in creating and evaluating photographic records for clinical forensic use were assessed. RESULTS: This paper is an aid to creating photographic records of sufficient quality for forensic purposes. The options provided by digital photography in particular make this endeavor feasible even in a clinical setting. In addition, our paper illuminates some technical aspects of creating and archiving photographic records for forensic use, and addresses possible error sources. CONCLUSION: With the requisite technical background knowledge, injuries can be photographically recorded to forensic standards during patient care.

Star-crossed? The association of the 5-HTTLPR s allele with season of birth in a healthy female population, and possible consequences for temperament, depression and suicide.

BACKGROUND: Birth season has well-known effects on neuropsychiatric disorders, and may also influence genotype distribution by possibly influencing chance of conception via parental idiosyncratic conception patterns or survival of foetuses or infants. The 5-HTTLPR is associated with phenomena including affective temperaments or suicide which are also associated with birth season. Our aim was to investigate the association of 5-HTTLPR genotype and birth season in a healthy female population. METHODS: Birth date and 5-HTTLPR genotype was determined for 327 psychiatrically healthy women. The association between presence of s allele and time of birth was analysed using generalized linear models. RESULTS: A significant association between s allele frequency and time of birth was detected. S allele carrier frequency was marginally significantly higher in July borns and significantly lower in autumn borns. LIMITATIONS: We investigated an adult sample so genotype frequency data do not reflect birth frequencies. Our sample consisted exclusively of females. CONCLUSIONS: There is no clear explanation for the observed association, although idiosyncratic parental conception patterns, the association of 5-HTTLPR with sudden infant/intrauterine death, or other s allele-mediated behaviours may play a role. Our results are strikingly parallel with earlier data reporting a higher risk of completed suicide in July borns, and higher scores of July borns and lower scores of autumn borns on certain affective temperament scales, both of which are also associated with the s allele of 5-HTTLPR. Thus our results may add to the growing body of evidence regarding the etiological background of affective disorders.

No mutation but high mRNA expression of Coxsackie-Adenovirus Receptor was observed in both dilated and ischemic cardiomyopathy.

The most common causes of acute myocarditis and the possible consequence of dilated cardiomyopathy are virus infections. The receptor of the two most common viruses connected to these myocardial diseases was identified as Coxsackie-Adenovirus Receptor. The purpose of this study was to assess Coxsackie-Adenovirus Receptor mRNA expression in the myocardium and search for mutations in the Coxsackie-Adenovirus Receptor gene to compare dilated, inflammatory and ischemic cardiomyopathy with control group. All the myocardial samples were obtained from 35 explanted hearts during heart transplantation, than DNA and RNA were isolated from the muscle samples. cDNA was generated from RNA using reverse transcription, and real-time PCR was performed with relative quantification by ß-actin gene as endogenous control. Using DNA extracted from the myocardial samples, we sequenced all the seven exons of the Coxsackie-Adenovirus Receptor gene. Coxsackie-Adenovirus Receptor mRNA expression was higher in both ischemic and dilated cardiomyopathy groups than in inflammatory cardiomyopathy and healthy control groups. Sequencing of CAR gene showed no sign of mutation. Therefore, the sequences result of CAR exons did not show any mutation or polymorphism, that explains a determinant role of CAR in dilated or ischemic CM. Our results suggest that high mRNA expression of Coxsackie-Adenovirus Receptor may support its role in regeneration of the damaged myocardium rather than having any role in viral mediated heart disease.

Interaction of 5-HTTLPR genotype and unipolar major depression in the emergence of aggressive/hostile traits.

OBJECTIVE: The 5-HTTLPR polymorphism has been associated both with depression and aggression/hostility. The multidirectional association between depression, aggression and the s allele may be important, since all these phenomena are related to suicidal behavior. Our aim was to investigate the association between 5-HTTLPR and aggressive/hostile traits in depressed patients and controls. METHODS: 137 depressive and 118 control women completed the Buss-Durkee Hostility Inventory and were genotyped for 5-HTTLPR. BDHI scores in the different groups were investigated by Generalized Linear Model Analysis. Association between dependent and independent variables in the model was tested by the likelihood ratio Chi-square statistic. RESULTS: Diagnosis and genotype showed a significant association with several aggressive/hostile traits. Interaction of the two main effects was also significant in case of several subscales. Post hoc analyses indicated a significant association between BDHI subscales and s allele only in the depressed group. LIMITATION: Only women were studied and since gender differences are present both in aggressive behavior and putatively in the behavioral effects of 5-HTTLPR genotype, our findings pertain only to females. CONCLUSION: Our results indicate a robust relationship between aggression/hostility and 5-HTTLPR genotype, but this association is more marked in the presence of depression. The presence of the s allele thus not only contributes to a higher risk of depression, but in depressives also leads to higher aggression/hostility. Our results have important implications for suicide research, since the s allele is associated with violent suicide, and this association may be mediated through the emergence of increased aggression/hostility in depressed patients carrying the s allele.

The role of viral infections in the development of dilated cardiomyopathy.

Enteroviruses (EVs) are the most frequent pathogens in myocarditis and in the subsequently developing dilated cardiomyopathy as well. Furthermore, persistence of other viruses might play a pathogenic role in the evolution from myocarditis to dilated cardiomyopathy. Explanted heart of 28 patients, who underwent heart transplantation were screened for EV, AdV3 and HHV6 sequences in order to assess the incidence of cardiac viral infection that may be implicated in the pathogenesis of cardiomyopathy, and estimate viral distribution in the myocardium. Viral sequences were extracted from five different regions of the hearts. Nested PCR was used to amplify conservative regions of AdV3, HHV6 and EVs. Histological examination was performed on routinely processed myocardial samples. AdV3 was verified in one fourth of the patients. ADV3 and HHV6 sequences coexisted in one case with inflammatory cardiomyopathy. Some patients had more than one positive area of their heart. AdV3 positive right ventricular samples were double in amount compared to the left ones. None of the patients had positive result for EV. This is the first occasion to identify AdV3 (a mainly respiratory infective virus) sequence in explanted hearts of cardiomyopathy patients. Though the clinical importance of our results is still unclear, AdV3 could be a new member of the viral group with possible pathogenic effect on the myocardium. Regional distribution of viral sequence location confirmed that the right ventricular wall as a biopsy sampling site might be adequate for endomyocardial biopsy pro diagnostic purposes.

How possible is the development of an operational psychometric method to assess the presence of the 5-HTTLPR s allele? Equivocal preliminary findings.

OBJECTIVE: The s allele of the 5-hydroxytryptamine transporter-linked promoter region (5-HTTLPR) polymorphism of the serotonin transporter gene has been found to be associated with neuroticism-related traits, affective temperaments and response to selective serotonin reuptake inhibitor (SSRI) treatment. The aim of the current study was to develop a psychometric tool that could at least partially substitute for laboratory testing and could predict the presence of the s allele. METHODS: The study included 138 women of Caucasian origin, mean 32.20 +/- 1.02 years old. All subjects completed the Hungarian standardised version of the Temperament Evaluation of the Memphis, Pisa, Paris, and San Diego Autoquestionnaire (TEMPS-A) instrument and were genotyped for 5-HTTLPR using PCR. The statistical analysis included the calculation of the Index of Discrimination (D), Discriminant Function Analysis, creation of scales on the basis of the above and then item analysis and calculation of sensitivity and specificity. RESULTS: Four indices were eventually developed, but their psychometric properties were relatively poor and their joint application did not improve the outcome. CONCLUSIONS: We could not create a scale that predicts the 5-HTTLPR genotype with sufficient sensitivity and specificity, therefore we could not substitute a psychometric scale for laboratory genetic testing in predicting genotype, and also possibly affective disorder characterisation and treatment.

Association of the s allele of the 5-HTTLPR with neuroticism-related traits and temperaments in a psychiatrically healthy population.

INTRODUCTION: Research concerning the genetic background of traits, temperaments and psychiatric disorders has been rapidly expanding. One of the most frequently studied genetic polymorphisms in the background of psychological and psychiatric phenomena is the 5-HTTLPR polymorphism of the serotonin transporter gene which has earlier been found to be associated with neuroticism and neuroticism-related traits and disorders. However, both the neuroticism trait and psychiatric disorders are complex and composed of several subfacets. The aim of our study was to investigate the association of the 5-HTTLPR polymorphism with several smaller, distinct and better characterisable phenomena related to the neuroticism trait. METHODS: 169 healthy females participated in the study. All participants completed the Buss-Durkee Hostility Inventory (BDHI), the State-Trait Anxiety Inventory (STAI), The Zung Self-rating Depression Scale (ZSDS), the Beck Hopelessness Scale, the SCL-51, the Temperament and Character Inventory (TCI) and the Temperament Evaluation of Memphis, Pisa, Paris and San Diego (TEMPS-A) questionnaire. All subjects were genotyped for the 5-HTTLPR using PCR. Data were analysed with ANOVA and MANCOVA with age as a covariate. RESULTS: We found that the presence of the s allele was significantly associated with anxiety, depression, hopelessness, guilt, hostility, aggression, presence of neurotic symptoms, self-directedness and affective temperaments carrying a depressive component even when controlling for age. CONCLUSIONS: Our study is the first that confirms that traits and characteristics related to neuroticism, such as increased anxiety, depression, hopelessness, somatization, feeling of guilt, hostility, aggression, lack of self-directedness and affective temperament are consistently and independently associated with the 5-HTTLPR polymorphism of the serotonin transporter gene. Our study therefore suggests that neuroticism can be considered a unified construct not only from a phenotypical but also from a genetic point of view and 5HTTLPR can be considered one component of its polygenic background. Our results thus yield further insight into the role of the 5-HTTLPR in the background of neuroticism and neuroticism-related psychiatric disorders.

[Molecular biological virus identification in dilated cardiomyopathy]. / Víruskimutatás molekuláris biológiai vizsgálattal cardiomyopathiás betegek szívizommintáiból.

UNLABELLED: Enteroviruses have been considered to be the most common cause of acute myocarditis and possible consequence of dilated cardiomyopathy. Some publications shed light to the role of other viruses in this disease as well. Our molecular investigation has demonstrated that adeno- and herpes viruses might also frequently occur in dilated cardiomyopathy. AIM: The aim of our study was to screen virus genomes in heart tissues from heart-transplanted patients to prove their possible role in the pathogenesis of dilated cardiomyopathy. METHODS: DNA and RNA were isolated from five regions of the heart muscle. Amplification for Adenovirus Type 3, Human Herpes Virus Type 6 and Enterovirus genomes were performed by nested-Polymerase Chain Reaction. Finally the virus-positive samples were direct sequenced. RESULTS: In 2 patients Adenovirus Type 3 and in 1 patient both Adenovirus Type 3 and Human Herpes Virus Type 6 were detected. No enteroviruses were found in any heart tissue. CONCLUSIONS: In our study the adenovirus genome was found to be the most frequent virus genome in explanted heart tissues. The identified viral sequences proved previous viral infection, which could have played a role in the development of dilated cardiomyopathy. Detection of different viruses in the myocardium by molecular biological examinations might contribute to adequate treatment of these patients.

[Changes of the different neuropeptide containing nerve elements in the inflamed human gall bladder]. / A küllözo neuropeptidtartalmú idegelemek számának változása humán gyulladt epehólyagban.

BACKGROUND, AIMS: The changes of different neuropeptide containing nerve elements might play a role in the pathogenesis of cholecystitis and the formation of gallstones, therefore the authors have investigated the density of the neuropeptide containing nerve fibres and immunocompetent cells in human gallbladder (control and cholecystitis). METHODS: The different neuropeptide containing nerve elements and immunocytes were detected by avidin-biotin-peroxidase (ABC) immunohistochemistry. RESULTS: In the control gallbladder the density of the different neuropeptide containing nerve fibres showed different pattern in all layers. In the inflamed gallbladder the number of the vasoactive intestinal polypeptide (VIP) positive nerve fibres increased significantly, very dense immunoreactive (IR) nerve fibres were located mainly in the tunica mucosa just below the epithelial lining. The number of the VIP IR nerve cell bodies was also increased. However, the number of the substance P (SP) IR nerve fibres was decreased significantly in the cholecystitis. The number of the neuropeptide Y (NPY) nerve fibres showed no changes, while their distribution was altered compared to the control. In the inflamed area the number of immunocompetent cells was strongly increased (being granulocytes, lymphocytes, plasma cells and mast cells) and some of them were also immunoreactive for SP, calcitonin gene-related peptide (CGRP) and VIP. Close contacts were detected between IR nerve fibres and the immunocytes in several cases. CONCLUSIONS: During inflammation the changes of the neuropeptide containing nerve fibres might alter the function (causing dilation) of the gall bladder, the activated immunocytes can also synthesize neuropeptides (SP, CGRP, VIP), so the released materials (cytokines, chemokines, histamine, as well as neuropeptides) might act in an autocrine and/or paracrine way influencing the function of the organ and of the immune system.

Subthreshold depression is linked to the functional polymorphism of the 5HT transporter gene.

BACKGROUND: The functional polymorphism of the serotonin transporter gene (5HTTLPR) has been earlier associated with affective disorders. No research has however been carried out to identify the relationship between this polymorphism and depressive traits and subclinical depressive symptoms within a psychiatrically healthy population. METHODS: One hundred and twenty-eight female subjects with no lifetime or current history of DSM-IV Axis I disorders participated in the study. All subjects completed the Zung Self-Rating Depression Scale and were genotyped for the 5HTTLPR polymorphism. RESULTS: Significant differences were found on the Zung SDS and also on the physical-vegetative subscale of the Zung SDS according to both phenotype and genotype. Subjects carrying the s allele scored significantly higher on the Zung SDS and had also significantly higher scores on the physical-vegetative symptom subscale. Furthermore, subjects carrying the ss genotype scored highest and subjects carrying the ll genotype scored the lowest on both scales. CONCLUSION: Subclinical depressive symptoms (i.e. DSM-IV subthreshold depression) are associated with the functional polymorphism of the serotonin transporter gene. The main association between this polymorphism and subclinical depression is primarily carried by the physical symptoms of depression. The s allele of the 5HTTLPR gene is associated with a "low mood endophenotype".

Sudden suffocation by surgical sponge retained after a 23-year-old thoracic surgery.

A 70-year-old male patient with a 23-year-old history of right lower lung lobectomy for primary pulmonary adenocarcinoma (T1 N0 M0) presented with recurrent bronchopneumonia and purulent sputum. Pleural callus, lung abscess, bronchopleural fistula, and stitch granulomas were confirmed by chest x-ray, computed tomography scan, and bronchoscopy in the background of his complaints. An attempt to remove the bronchial purulent discharge and tissue sampling was made by using a flexible bronchoscope. The area of the lower trachea suddenly became clogged during bronchoscopic removal of the suspected piece of tissue (which later turned out to be organizing surgical gauze). The resuscitation following ventricular fibrillation failed to save the patient's life. The forensic postmortem examination confirmed the position of the foreign body extending from the abscess cavity, crossing the midline at carina and obstructing the lower trachea. This foreign body was a remnant of the surgical gauze left behind during a thoracic surgery 23 years ago.

Arrhythmias of a sudden traumatic death.

A 73-year-old man was treated because of his paroxysmal palpitations became persistent. At the time of hospital admission atrial flutter was found. Antiarrhythmic drug treatment was unsuccessful therefore electrical cardioversion was indicated which restored his sinus rhythm. After one year of uneventful follow up Holter monitoring was indicated again. When the machine was hooked up sinus rhythm was observed. After 53 minutes a tram knocked down the patient, and he died immediately. During autopsy brain laceration, multiple cranial fractions, mediastinal hemorrhage were found. The Holter recording time was 66 minutes. Before the accident sinus rhythm was recorded. At the time of accident an electrical noise was found, followed by long sinus arrest, atrial fibrillation, nodal escape rhythm, sinus bradycardia, ventricular flutter, tachycardia, fibrillation and "dying heart" rhythm. According to our knowledge this is the first Holter monitoring observation during a sudden traumatic death.

Association analysis of 5-HTTLPR variants, 5-HT2a receptor gene 102T/C polymorphism and migraine.

It is well known that migraine has a strong genetic component, although the type and number of genes involved is not yet clear. There is evidence to suggest that serotonin-related genes participate in the pathogenesis of migraine. Previous studies have shown that gender differences influence the serotonergic neurotransmission and, in addition, the migraine prevalence is higher in females than males. Therefore, we investigated the functional polymorphism in the upstream regulatory region of the serotonin transporter gene (5-HTTLPR) and the 102T/C polymorphism of the 5-HT2A receptor gene in the Hungarian female population. These genes were analysed in 126 migraine sufferers (with or without aura)and 101 unrelated healthy controls using case control design. A borderline association (chi2 = 3.84, df = 1, p = 0.049; OR = 1.45, 95% CI = 1.00-2.12) between 5-HTTLPR short (S) allele and migraine was found. No significant difference between migraine sufferers and controls was observed for the 102T/C polymorphism of 5-HT2A receptor gene. Furthermore, there was no significant interaction between5-HTTLPR and 102T/C polymorphisms in our study population. In conclusion, our results support that the genetic susceptibility of migraine may be associated with a locus at or near the 5-HT transporter gene.

Despite the general correlation of the serotonin transporter gene regulatory region polymorphism (5-HTTLPR) and platelet serotonin concentration, lower platelet serotonin concentration in migraine patients is independent of the 5-HTTLPR variants.

Platelet serotonin (5-HT) concentrations in a headache-free period during the mid-follicular phase and the serotonin transporter gene regulatory region polymorphism (5-HTTLPR) were measured in female migraine patients without aura (n = 64) and healthy controls (n = 42). High-pressure liquid chromatography (HPLC) was used to determine the platelet 5-HT concentration and genetic polymorphism was determined by polymerase chain reaction. Significantly lower platelet 5-HT concentrations were found in migraine patients compared to controls. Concerning the 5-HTTLPR polymorphism, the S/S genotype was associated with a significantly higher platelet 5-HT concentration (P = 0.027) in the whole study population. This association between the 5-HTTLPR genotypes and platelet 5-HT concentrations was independent of the diagnosis. In addition, the platelet 5-HT concentration was lower in migraineurs in all genotypes (S/S, S/L, L/L). In conclusion, 5-HTTLPR variants may have an effect on the platelet 5-HT concentrations, but the lower 5-HT concentrations in migraine patients seem to be determined by other factors.

[Genetic investigations in forensic medicine]. / DNS vizsgálatok az igazságügyi orvostanban.

Scientific research of the last decade including the introduction of new molecular biological methods and mapping of the human genome allowed the development of a revolutionary new molecular biological approach in forensic medicine. The traditional serological methods study proteins, the new DNA analysis goes further down to study DNA structures to analyze unique individual features. The two main areas of DNA application in forensic medicine are inheritance studies and personal identification in criminal cases using biological traces. Using this new, reliable and reproducible method we can answer questions they were almost impossible in the past. This article reviews how molecular techniques used to detect genetic polymorphism in forensic medicine.