Identification of human remains from the Second World War mass graves uncovered in Bosnia and Herzegovina

Aim

To present the results obtained in the identification of human remains from World War II found in two mass graves in Ljubuški, Bosnia and Herzegovina.

Methods

Samples from 10 skeletal remains were collected. Teeth and femoral fragments were collected from 9 skeletons and only a femoral fragment from 1 skeleton. DNA was isolated from bone and teeth samples using an optimized phenol/chloroform DNA extraction procedure. All samples required a pre-extraction decalcification with EDTA and additional post-extraction DNA purification using filter columns. Additionally, DNA from 12 reference samples (buccal swabs from potential living relatives) was extracted using the Qiagen DNA extraction method. Quantifiler^TM Human DNA Quantification Kit was used for DNA quantification. PowerPlex ESI kit was used to simultaneously amplify 15 autosomal short tandem repeat (STR) loci, and PowerPlex Y23 was used to amplify 23 Y chromosomal STR loci. Matching probabilities were estimated using a standard statistical approach.

Results

A total of 10 samples were processed, 9 teeth and 1 femoral fragment. Nine of 10 samples were profiled using autosomal STR loci, which resulted in useful DNA profiles for 9 skeletal remains. A comparison of established victims'' profiles against a reference sample database yielded 6 positive identifications.

Conclusion

DNA analysis may efficiently contribute to the identification of remains even seven decades after the end of the World War II. The significant percentage of positively identified remains (60%), even when the number of the examined possible living relatives was relatively small (only 12), proved the importance of cooperation with the members of the local community, who helped to identify the closest missing persons’ relatives and collect referent samples from them.DNA analysis plays a key role in the identification of missing persons and victims of mass fatality incidents, and DNA profiling could be regarded as a core method for such identification (1). The primary value of this procedure significantly increased over the last twenty years due to the introduction of short tandem repeat (STR) loci in routine forensic testing, especially their optimized versions, miniSTRs or STR markers located at the Y and X chromosomes. Data obtained by DNA typing are highly reliable and can be used as a powerful tool that produces reliable results (2).Various procedures could be used to identify human remains (3) and the choice of an appropriate procedure and its usefulness primarily depend on the condition of the remains. When it comes to skeletal remains from World War II (WWII), DNA analysis has already proved to be efficient, and in most cases, the only applicable approach to human identification (4,5). Furthermore, a seven-decade long soil deposition is a significant adverse factor, which is why most of the laboratories use optimized procedures in the analysis of these samples. Over the last 20 years, DNA identification of victims of the wars in Bosnia and Herzegovina (B&H) (6) and Croatia in the 1990s (3), the analysis of WWII skeletal remains in Slovenia (4,5), and the analysis of several centuries old archeological samples (7) successfully identified a number of human skeletal remains and overcame a number of challenges, such as optimization of DNA extraction protocols, specific laboratory procedures, communication with the relatives, etc. Here we once again utilized the experience gained from these projects to identify skeletal remains from the WWII mass graves in B&H.There are no precise official data about the number of killed and missing persons from the WWII period in B&H, although this number is roughly estimated to about 180 000 (8). This estimate is even higher if we include the victims from immediate post-war incidents and mass executions by communist authorities. Most of those crimes were hidden for almost 70 years. For example, more than 2300 persons from the Ljubuški municipality alone were killed during WWII, and immediately after it, that is, in 1945. Almost 50% of the victims are still considered missing and their burial sites are not known (9).As a consequence of continuous pressure from living relatives, the regional authorities and local communities recently put in significant efforts to identify individuals discovered in several mass graves scattered throughout the region. They commissioned a DNA analysis of skeletal remains and reference samples. Here, we report the results of the identification of the WWII victims from the city of Ljubuški, south Herzegovina.     

Risk factors of post-ERCP pancreatitis in high-risk patients despite prevention with prophylactic pancreatic stents

Abstract

Objectives: To analyze risk factors of post-ERCP pancreatitis (PEP) in high risk patients in whom prophylactic pancreatic stents (PPSs) were intended to use to prevent endoscopic retrograde cholangiography (ERCP)-related complications.

Patients and methods: Three hundred and seventeen high-risk patients for developing PEP were considered for preventive pancreatic stent placement in our endoscopy unit over 5 years. 5 Fr, 3–5?cm long PPSs were used. All data were collected in a predefined database. Development of PEP despite PPS placement was analyzed.

Results: PEP occurred in 29 of 288 successfully stented patients (10.07%; 24 mild, four moderate, one severe). PPS was protective against all risks factors except for sphincter of Oddi dysfunction (SOD). PPS related complication rate was 2.78% (n?=?8). Unsuccessful stenting occurred in 29 patients (9.15%), PEP developed in 41.38% (n?=?12; seven mild, four moderate, one severe). Those patients who had more patient related risk factors were more likely to develop PEP despite preventive measures with PPS. On the contrary, PPS placement was less successful in patients who had more procedure related risk factors.

Conclusions: PPS is protective against all risks factors of PEP except for SOD in high-risk patients. More vulnerable patients who have more patient-related risk factors are more likely to develop PEP despite PPS is used, while more complex procedures predispose to unsuccessful PPS placement in patients with more procedure-related risk factors. PPS insertion in high-risk patients is effective and safe preventive method and the procedure related complication rate is reasonably low.     

PROPHYLACTIC PANCREAS STENTING FOLLOWED BY NEEDLE‐KNIFE FISTULOTOMY IN PATIENTS WITH SPHINCTER OF ODDI DYSFUNCTION AND DIFFICULT CANNULATION: NEW METHOD TO PREVENT POST‐ERCP PANCREATITIS

Introduction: The aim of the present study was to reduce post‐endoscopic retrograde cholangiopancreatography (ERCP) complications with a combination of early needle‐knife access fistulotomy and prophylactic pancreatic stenting in selected high‐risk sphincter of Oddi dysfunction (SOD) patients with difficult cannulation. Methods: Prophylactic pancreatic stent insertion was attempted in 22 consecutive patients with definite SOD and difficult cannulation. After 10 min of failed selective common bile duct cannulation, but repeated (>5×) pancreatic duct contrast filling, a prophylactic small calibre (3–5 Fr) pancreatic stent was inserted, followed by fistulotomy with a standard needle‐knife, then a standard complete biliary sphincterotomy followed. The success and complication rates were compared retrospectively with a cohort of 35 patients, in which we persisted with the application of standard methods of cannulation without pre‐cutting methods. Results: Prophylactic pancreatic stenting followed by needle‐knife fistulotomy was successfully carried out in all 22 consecutive patients, and selective biliary cannulation and complete endoscopic sphincterotomy were achieved in all but two cases. In this group, not a single case of post‐ERCP pancreatitis was observed, in contrast with a control group of three mild, 10 moderate and two severe post‐ERCP pancreatitis cases. The frequency of post‐ERCP pancreatitis was significantly different: 0% versus 43%, as were the post‐procedure (24 h mean) amylase levels: 206 U/L versus 1959 U/L, respectively. Conclusions: In selected, high‐risk, SOD patients, early, prophylactic pancreas stent insertion followed by needle‐knife fistulotomy seems a safe and effective procedure with no or only minimal risk of post‐ERCP pancreatitis. However, prospective, randomized studies are awaited to lend to support to our approach.     

Reproducibility of erythrocyte sodium-lithium countertransport activity and ambulatory blood pressure measurements in type 1 diabetes mellitus

We aimed to study the reproducibility of sodium-lithium countertransport [SLCT] activity and ambulatory blood pressure monitoring [ABPM] in type 1 diabetes. We did this by performing repeated measurements of SLCT activity and ABPM in 11 recent-onset diabetic children and in 11 patients with longer duration of diabetes. Both parameters were related to microalbuminuria. In the older group of diabetic children a significant correlation [r = 0.78; P<0.005] in SLCT activity between the first and second study was observed [514.3±186.4 vs 491.0 ± 148.0 μmol/l erythrocytes/h]. Diurnal systolic and diastolic blood pressure were comparable at both time points within the same group of diabetic children [in group 1: 102.6±6.1 vs 108.6±7.6 mmHg N.S.; in group 2: 113.4 ± 10.6 vs 114.0±7.8 mmHg N.S. Diastolic blood pressure in group 1: 57.4±4.8 vs 65.7±6.9 mmHg N.S., in group 2: 70.6±9.1 vs 68.5±5.3 mmHg N.S.]. Moreover, there was a significant correlation in both diurnal and nocturnal systolic blood pressure between the first and second study in the whole diabetic population. Both SLCT activity and blood pressure values obtained by ABPM were found to be reproducible individual characteristic markers in type 1 diabetic children. Received: 22 November 1997 / Accepted in revised form: 30 April 1998     

Effect of puberty on the development of diabetic microangiopathy

Fluorescein angiography was used to follow the development of retinal microangiopathy at the time of puberty in 34 children with type I diabetes. When the results were compared with those for the control children who had suffered from the disease for a similar length of time, it was concluded that (in contrast with certain literature data) the process of sexual maturation does not essentially influence the development of retinal microangiopathy if the diabetes is maintained under good control for a long-lasting period. Appreciable roles in the rapid progression that is frequently observed in early adulthood are attributed to the sudden changes in the previously systematic living conditions (school, family, etc.).     

Correction to: Gap Analysis Regarding Prognostication in Neurocritical Care: A Joint Statement from the German Neurocritical Care Society and the Neurocritical Care Society

Neurocritical Care - This article was updated to correct the spelling of Karl Georg Haeusler.     

Hyperglycemia in sepsis is a risk factor for development of type II diabetes

Background

Hyperglycemia is frequent in sepsis, even in patients without diabetes or impaired glucose metabolism. It is a consequence of inflammatory response and stress, so its occurrence is related to severity of illness. However, not all severely ill develop hyperglycemia and some do even in mild disease. We hypothesized the existence of latent disturbance of glucose metabolism that contributes to development of hyperglycemia and that those patients might have increased risk for diabetes.

Methods

Patients admitted with sepsis and no history of impaired glucose metabolism were included and divided in the hyperglycemia group (glucose ≥7.8 mmol/L) and normoglycemia group. Severity of sepsis was assessed. Surviving patients without diabetes at discharge were followed-up for 5 years to investigate risk for development of diabetes.

Results

Hyperglycemia was related to severity of sepsis. Follow-up was finished for 55 patients with hyperglycemia, of which 8 (15.7%) developed diabetes, and 118 patients with normoglycemia, of which 5 (4.2%) developed diabetes (P = .002). Relative risk for developing type 2 diabetes was 4.29 (95% CI, 1.35-13.64).

Conclusion

Patients with hyperglycemia in sepsis who are not diagnosed with diabetes before or during the hospitalization should be considered a population at increased risk for developing diabetes.     

Ascorbic and dehydroascorbic acids status in rats fed diets varying in vitamin E levels

Vitamin E, ascorbic and dehydroascorbic acids were determined in plasma and selected tissues of rats fed for 2-3 months different diets varying in vitamin E content. The diets contained as low as 5 IU for group LE, a normal amount of 50 IU for group NE and as high as 250 IU of vitamin E for group HE. Small increases in total vitamin C were observed only in liver, kidney, spleen and plasma with increased dietary levels of vitamin E, however, this was not followed by a substantial increment in the ascorbate/dehydroascorbate ratio. These differences were only observed between diets LE and HE; there was no correlation between vitamin E and vitamin C levels in the tissue. These data suggest that the interactions that readily take place in vitro between these two vitamins do not occur in vivo, probably due to the complexity of natural membranes.