Comparison of three different DNA extraction methods from a highly degraded biological material.

The identification of unknown victims is one of the most challenging tasks faced by forensic medicine. This is due to the rapid decomposition of tissues, beginning at the moment of death and caused by released enzymes and microbial activity. Decay is directly associated with the decomposition of soft tissues and also the degradation of genetic material inside cells. Decomposition rates vary depending on a number of environmental factors, including temperature, humidity, season, and soil properties. Decomposition also differs between bodies left in the open air or buried. To date, forensic medicine has identified mainly people who were the victims of various types of criminal offences. However, with advances in identification methods, increasingly frequent attempts are made to identify the victims of armed conflicts, crimes of totalitarian regimes, or genocide. The aim of the study was to compare three different methods for the extraction of nuclear DNA from material considered in forensic medicine as difficult to handle, i.e. fragments of bones and teeth, and to determine the performance of these methods and their suitability for identification procedures.

The Polish Genetic Database of Victims of Totalitarianisms.

This paper describes the creation of the Polish Genetic Database of Victims of Totalitarianism and the first research conducted under this project. On September 28th 2012, the Pomeranian Medical University in Szczecin and the Institute of National Remembrance-Commission for Prosecution of Crimes against the Polish Nation agreed to support the creation of the Polish Genetic Database of Victims of Totalitarianism (PBGOT, www.pbgot.pl). The purpose was to employ state-of-the-art methods of forensic genetics to identify the remains of unidentified victims of Communist and Nazi totalitarian regimes. The database was designed to serve as a central repository of genetic information of the victim's DNA and that of the victim's nearest living relatives, with the goal of making a positive identification of the victim. Along the way, PGBOT encountered several challenges. First, extracting useable DNA samples from the remains of individuals who had been buried for over half a century required forensic geneticists to create special procedures and protocols. Second, obtaining genetic reference material and historical information from the victim's closest relatives was both problematic and urgent. The victim's nearest living relatives were part of a dying generation, and the opportunity to obtain the best genetic and historical information about the victims would soon die with them. For this undertaking, PGBOT assembled a team of historians, archaeologists, forensic anthropologists, and forensic geneticists from several European research institutions. The field work was divided into five broad categories: (1) exhumation of victim remains and storing their biological material for later genetic testing; (2) researching archives and historical data for a more complete profile of those killed or missing and the families that lost them; (3) locating the victim's nearest relatives to obtain genetic reference samples (swabs), (4) entering the genetic data from both victims and family members into a common database; (5) making a conclusive, final identification of the victim. PGBOT's first project was to identify victims of the Communist regime buried in hidden mass graves in the Powazki Military Cemetery in Warsaw. Throughout 2012 and 2013, PGBOT carried out archaeological exhumations in the Powazki Military Cemetery that resulted in the recovery of the skeletal remains of 194 victims in several mass graves. Of the 194 sets of remains, more than 50 victims have been successfully matched and identified through genetic evidence.

[On the Differential Diagnosis of Intractable Psychogenic Chronic Cough: Neuropathic Larynx Irritable - Gabapentin's Antitussive Action]. / Zur Differenzialdiagnose des quälenden psychogenen chronischen Hustens: neuropathischer Larynx irritable - Gabapentin als Antitussivum.

We present the case of a 76 year old female inpatient who suffered from a chronic intractable cough which arose simultaneously to a severe major depression and was secondary to an exorbitant psychological distress. Chronic cough had never been experienced before and was initially considered to have a mere psychogenic origin since a comprehensive and guideline-based diagnostic screening did not reveal any underlying somatic cause. However, several factors cast doubt on the solitary psychic genesis of the chronic cough: i) occurrence immediately after a penetrant cold, ii) embedding in other complaints of laryngeal hyperreagibility (larynx irritable), such as persistent globus pharyngeus sensation, throat clearing and episodic dysphonia, iii) first occurrence on old life, iv) erupting from sleep as well, v) persistence despite remission of the major depression, and v) no sustaining benefit from specific psychotherapy and speech therapy. Therefore, diagnostics were extended to apparative tools for objective evaluation of swallowing by using fiberoptic videoendoscopic (FEES) and videofluoroscopic (VFS) techniques, which revealed signs of laryngeal neuropathy but without evidence of penetration or aspiration. A co-existing small goiter and an impaired glucose tolerance along with a putative intracellular vitamin B12 or folate deficiency (as indirectly derived from an apparent hyperhomocysteinemia) were assumed to be responsible for the neuropathy and underwent specific treatments. The impaired glucose tolerance and putative vitamin deficit were compatible with a distal symmetric sensorimotoric, even subclinical polyneuropathy of the lower extremities. The larynx irritable improved under gabapentin being confirmed by drug removals several times, and finally calmed down almost completely under gabapentin, which was in line with the scant literature of this topic. Re-examination of the larynx per FEES nine months later showed no deficits any more under the well-tolerated treatment (gabapentin, levothyroxine, vitamin B12 and folic acid substitution, weight reduction and physical training). All in all, the larynx irritable as well as the chronic cough were most probably induced by a laryngeal neuropathy and were not solely of psychic origin. Due to good treatment options a larynx irritable should be regularly taken into consideration of the investigation of intractable chronic cough. Therefore, an apparative evaluation of deglutition is recommended in the diagnostic toolbox of chronic cough - even if embedded in a psychiatric disorder or distress - before diagnosing a sole psychic origin. An hypothetical scheme of the development of a larynx irritable caused by neuropathic and non-neuropathic ("nociceptive") conditions is proposed.

Estimation of BDNF gene polymorphism and predisposition to dependence development for selected psychoactive compounds: genetic aspects of addiction with the selected drugs, amphetamine, tetrahydrocannabinol and opiates.

The etiology of drug addiction, a central nervous system (CNS) disease, is not fully known. This complex problem is believed to be connected with concurrently affecting genetic, psychological and environmental factors. The development of addiction is connected with CNS reinforcement system and dopaminergic neurotransmission. Molecular processes are postulated to be of universal character and allow to presume a similar mechanism of dependence for both ethanol and other substances. Therefore, elements of dopaminergic transmission become excellent candidates for the examination of genetic influence on the development of addiction. A relationship between alcoholic disease and the presence of TaqIA1 and DRD2 alleles permits to initiate another investigation of gene-coding DRD2 dopamine receptor. The latest results indicate the importance of brain-derived neurotrophic factor (BDNF) in the regulation of dopaminergic route. The purpose of this research was to reveal the relationship between the Val66Met BDNF gene polymorphism and dependence of psychoactive agent. The examinations were performed with the Local Research Ethics Committee approval and patient's consent. The study group consisted of 100 patients (88 men and 12 women) aged 18-52 years, qualified for research program according to the International Classification of Diseases, Tenth Revision (ICD-10) requirements, medical examination and detailed questionnaire.

Population genetics of 15 autosomal STR loci in the population of Pomorze Zachodnie (NW Poland).

Allele frequency data and forensic efficiency parameters for 15 STR loci: D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, FGA were estimated from a sample of 600 unrelated individuals from the Pomorze Zachodnie (NW Poland). The combined MP and PE for all 15 loci are 3.9x10(-18) and 0.9999988, respectively. Pairwise comparisons between Northwestern Poland and other Polish populations were performed.

Honeydew sugars in wild-caught female horse flies (Diptera: Tabanidae).

Three species of horse fly, Hybomitra illota (Osten Sacken), H. affinis (Kirby), and H. zonalis (Kirby), were collected by sweep-netting around human bait at 2 sites (abandoned airfield and Davies Bog) in Algonquin Provincial Park, Ontario. There were 3 times more horse flies collected at Davies Bog than at the abandoned airfield. The crop contents of all specimens were identified by thin-layer chromatography. Using melezitose and stachyose as honeydew indicator sugars, the relative importance of homopteran honeydew and floral nectar as carbohydrate sources for these flies was determined. Of flies testing sugar-positive, 42.0% of H. illota (n = 50), 52.8% of H. affinis (n = 36), and 100% of H. zonalis (n = 4) had recently fed on honeydew. Pooled data for the 3 species showed that 38.9% of flies at the abandoned airfield (n = 18) and 51.4% of flies at Davies Bog (n = 72) were honeydew-fed. There were no significant differences among species or between sites. These findings are compared with recent black fly and deer fly studies from the same sites.

The effect of the additional use of grommets in silicone implant arthroplasty of the metacarpophalangeal joints.

After silicone arthroplasty of the metacarpophalangeal (MP) joint there is increasing osteolysis, subsidence and fracture of the implants in the longer postoperative term. In 44 patients with rheumatoid arthritis (54 hands) 151 arthroplasties of the metacarpophalangeal joint were assessed at a mean of 3.9 years postoperatively. In 57 arthroplasties titanium protectors (grommets) were used. There were no significant differences in the clinical outcomes with respect to swelling, correction of ulnar deviation, range of active movement and grip strength. The additional use of grommets in MP joint arthroplasty slightly reduced reactive osteolysis, protected the spacers from breakage and slightly reduced the amount of pain with only a few additional complications in the midterm follow-up.

[Therapy of rheumatoid destruction of the middle finger metacarpophalangeal joint with a Swanson silastic implant stabilized resection arthroplasty: comparative study of long and intermediate term results with and without implantation of titanium grommets]. / Die Therapie des rheumatisch destruierten Langfingergrundgelenkes mit der durch das Silastik-Interponat nach Swanson stabilisierten Resektionsarthroplastik: Vergleichende Studie lang- und mittelfristiger Ergebnisse mit und ohne Einsatz von Titangrommets.

Resection arthroplasty of the metacarpophangeal joints stabilized by a Swanson-Silastic-spacer is the golden standard in MP-joint destruction in R.A. 57 (Gr. I), 91 (Gr. II), and 102 (Gr. III) arthroplasties of the metacarpophalangeal joint were assessed in three groups of patients 3.5 years (Gr. I), 4.3 years (Gr. II), and 10.1 years (Gr. III), respectively, postoperatively on the average. In patients of group I titanium protectors, called grommets, were used additionally. Nearly all patients reported a marked relief of pain. Ulnar drift was corrected from an average of 22 degrees (Gr. I), 23 degrees (Gr. II), and 34 degrees (Gr. III) preoperatively. to 8 degrees (Gr. I), 7 degrees (Gr. II), and 12 degrees (Gr. III), respectively, postoperatively. Active range of motion remains unchanged with an average of 33 degrees (Gr. I), 38 degrees (Gr. II), and 42 degrees (Gr. III) ROM preoperatively and 42 degrees (Gr. I), 37 degrees (Gr. II), and 36 degrees (Gr. III) ROM, respectively, postoperatively. The average extension deficit had improved from 45 degrees (Gr. I), 32 degrees (Gr. II), and 33 degrees (Gr. III) at surgery to 18 degrees (Gr. I), 11 degrees (Gr. II), and 11 degrees (Gr. III), respectively, at the time of follow-up. Functional improvement of the hand was found in the medium term in 82% and in the longterm in 75% of the patients. The radiographical findings showed surrounding osteolysis in 45.7% (Gr. I), 63.5% (Gr. II), and 89.4% (Gr. III) of the implants and 0% (Gr. I), 16.5% (Gr. II), and 26.9% (Gr. III) broken spacers. From the medium to the longterm run there is an increase in radiographical deterioration. The additional use of titanium grommets in MP-joint arthroplasty seems to reduce reactive osteolysis and protects Swanson Silastic-spacers from breakage without substantial influence on the clinical outcome.