The COVID-19 pandemic has demonstrated the need for massively-parallel, cost-effective tests monitoring viral spread. Here we present SARSeq, saliva analysis by RNA sequencing, a method to detect SARS-CoV-2 and other respiratory viruses on tens of thousands of samples in parallel. SARSeq relies on next generation sequencing of multiple amplicons generated in a multiplexed RT-PCR reaction. Two-dimensional, unique dual indexing, using four indices per sample, enables unambiguous and scalable assignment of reads to individual samples. We calibrate SARSeq on SARS-CoV-2 synthetic RNA, virions, and hundreds of human samples of various types. Robustness and sensitivity were virtually identical to quantitative RT-PCR. Double-blinded benchmarking to gold standard quantitative-RT-PCR performed by human diagnostics laboratories confirms this high sensitivity. SARSeq can be used to detect Influenza A and B viruses and human rhinovirus in parallel, and can be expanded for detection of other pathogens. Thus, SARSeq is ideally suited for differential diagnostic of infections during a pandemic.

Aim:The aim of this study was to evaluate biochemical parameters in serum of women with preeclampsia and IUGR.Material and methods:A clinical prospective study was conducted and included 120 pregnant women divided in two groups: non IUGR group included healthy pregnant women (n=60) and IUGR group included pregnant women with preeclampsia and IUGR (n=60). Outcome measures were following values of biochemical parameters in serum of mother and fetuses: aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), bilirubin (indirect and direct) and cholesterol. A blood for analysis was drawn from the cubital vein of mothers and the umbilical vein of the fetuses during delivery period.Results:The mean of maternal age was 30.0±6.1 years in women with preeclampsia and IUGR and 28.1±5.1 years in healthy pregnant women, p > 0.05. The most of women with preeclampsia and IUGR had grade III of placental maturation (48.3%). There is a significant association between the placental maturation and the diagnosis, p < 0.001. There was a statistically significant difference in body mass of newborns between IUGR and non IUGR groups, p < 0.001. There was a significant statistically difference in serum value of AST, ALT, LDH and total cholesterol between women with preeclampsia and IUGR and healthy pregnant women (all p < 0.001).Conclusion:Measurement of AST, ALT, LDH, and total cholesterol in serum of pregnant women and newborns with IUGR allows the differentiation and threatening risk of perinatal complications due to hypoxia.

During a pandemic, mitigation as well as protection of system-critical or vulnerable institutions requires massive parallel, yet cost effective testing to monitor the spread of agents such as the current SARS-CoV2 virus. Here we present SARSeq, saliva analysis by RNA sequencing, as an approach to monitor presence of SARS-CoV2 and other respiratory viruses performed on tens of thousands of samples in parallel. SARSeq is based on next generation sequencing of multiple amplicons generated in parallel in a multiplexed RT-PCR reaction. It relies on a two-dimensional unique dual indexing strategy using four indices in total for unambiguous and scalable assignment of reads to individual samples. We calibrated this method using dilutions of synthetic RNA and virions to show sensitivity down to few molecules, and applied it to hundreds of patient samples validating robust performance across various sample types. Double blinded benchmarking to gold-standard quantitative RT-PCR performed in a clinical setting and a human diagnostics laboratory showed robust performance up to a Ct of 36. The false positive rate, likely due to cross contamination during sample pipetting, was estimated at 0.04-0.1%. In addition to SARS-CoV2, SARSeq detects Influenza A and B viruses as well as human rhinovirus and can be easily expanded to include detection of other pathogens. In sum, SARSeq is an ideal platform for differential diagnostic of respiratory diseases at a scale, as is required during a pandemic.

Introduction:Postmenopausal uterine bleeding is a „cancer until proven otherwise”. Endometrial cancer is a typical disease among postmenopause woman, because every bleeding in this age etiology associated with endometrial cancer (10-30%). The lifespan of women today has been extended and post menopause today last one third of a woman’s life. Early diagnosis of endometrial cancer has a very high cure rate. Screening for this cancer has limits in practice and is necessary given the definition of high-risk groups would be subject to primary and secondary prevention.Goal:Primary to evaluate the leading causes of postmenopausal uterine bleeding among patients at risk for endometrial cancer (diabetes, obesity, nulliparity, late menopause (after 55 years) and compared them with the causes of postmenopausal uterine bleeding patients without this risk.Material and methods:A retrospective, descriptive study with a targeted sample of 50 consecutive patients who had registered postmenopausal uterine bleeding in high-risk groups (cohorts) and the same number of patients with postmenopausal uterine bleeding that does not belong to the risk group (control group). Each patient underwent clinical examination, then fractionated curettements and its histopathological verification and assessment of treated clinical stage of disease with PH analysis of the resected specimens.Results:The patients of the studied risk group were significantly affected by endometrial cancer compared with the control group (RR=2.45, 95% CI 1.2 4.6, p=0.005). Endocervical pathology did not differ between groups. Clinical forms of bleeding: for those that are profuse bleeding cancer was present in 54.6% of cases. With intermittent bleeding cancer is verified in the 33.3% of patients. Risk patient groups with cancer frequently suffer from clinically more advanced stages of histologically aggressive endometrial cancer (serous adenocarcinoma–type II, low differentiated cancer).

Introduction:Hemolytic disease of the newborn was first described in the medical literature 1609, when it was diagnosed in one French housewife. In 1932 Diamond and colleagues described the mutual relationship of fetal hydrops, jaundice, anemia and erythoblastosis, which was later called fetal erytroblastosis. Hemolytic disease of the newborn (HDN) in the strict sense is considered disease whose basis is accelerated immune destruction of fetal/child erythrocytes that are bound to IgG antibodies of maternal origin. These antibodies are directed against antigens of father’s origin, which are present in the fetal/children’s erythrocytes and that the mother’s immune system recognizes them as foreign antigens.Goal:The goal is that in the period from January 1st 2011 to October 23st 2013 determine the frequency of ABO and Rh D incompatibilities in our sample of pregnant women/mothers, and to underscore the importance of regular check of ABO Rh D negative pregnant women and application specific Rh D protection.Material and methods:In the General Hospital “Prim. Dr. Abdulah Nakas” in Sarajevo by retrospective study are followed several relevant variables. Immune alloantibodies were detected in vivo by indirect Coombs test (ICT) with serum mother and O test erythrocytes, by direct Coombs test (DCT) with erythrocytes of a newborn.Results:The total number of births ABO Rh D negative was 596 (14%) and ABO Rh D positive mothers 4261 (86%). Of the total number of Rh D negative mothers there was A Rh D: negative mothers 42%; O Rh D negative 33%; B Rh D: negative 17% and AB Rh D: negative 8%. Most of immune antibodies appear in mothers with O Rh D: negative blood type. The emergence of immune antibodies in the Rh D negative mothers was 1%, the appearance of ABO incompatibilities amounted to 2.3% of our sample.Conclusion:In order to reduce the occurrence of alloimmunization of the mother to erythrocyte antigens of the newborn that can lead to major complications in subsequent pregnancies of Rh D: negative mothers and HDN constant monitoring in order to prevent them is necessary. Prevention is essential because once immunized mother will remain immunized for life.