Diversity, biogeography, evolutionary relationships, and conservation of Eastern Mediterranean freshwater mussels (Bivalvia: Unionidae)

Located at the junction between Europe, Africa, and Asia, with distinct evolutionary origins and varied ecological and geographical settings, together with a marked history of changes in orogeny and configuration of the main river basins, turned the Eastern Mediterranean into a region of high diversity and endemism of freshwater taxa. Freshwater mussels (Bivalvia, Unionidae) from the Western Palearctic have been widely studied in their European range, but little attention has been dedicated to these taxa in the Eastern Mediterranean region and their diversity and phylogeography are still poorly understood. The present study aims to resolve the diversity, biogeography, and evolutionary relationships of the Eastern Mediterranean freshwater mussels. To that end, we performed multiple field surveys, phylogenetic analyses, and a thorough taxonomic revaluation. We reassessed the systematics of all Unionidae species in the region, including newly collected specimens across Turkey, Israel, and Iran, combining COI+16S+28S phylogenies with molecular species delineation methods. Phylogeographical patterns were characterized based on published molecular data, newly sequenced specimens, and species distribution data, as well as ancestral range estimations. We reveal that Unionidae species richness in the Eastern Mediterranean is over 70% higher than previously assumed, counting 19 species within two subfamilies, the Unioninae (14) and Gonideinae (5). We propose two new species, Anodonta seddoni sp. nov. and Leguminaia anatolica sp. nov. Six additional taxa, Unio delicatus stat. rev., Unio eucirrus stat. rev., Unio hueti stat. rev., Unio sesirmensis stat. rev., Unio terminalis stat. rev. removed from the synonymy of Unio tigridis, as well as Unio damascensis stat. rev. removed from the synonymy of Unio crassus, are re-described. The nominal taxa Unio rothi var. komarowi O. Boettger, 1880 and Unio armeniacus Kobelt, 1911 are proposed as new synonyms of Unio bruguierianus, and Anodonta cyrea Drouët, 1881 and Anodonta cilicica Kobelt & Rolle, 1895 as new synonyms of Anodonta anatina. Also, the presence of Unio tumidus in the Maritza River is confirmed. The phylogeographic patterns described here are interpreted concerning major past geological events. Conservation needs and implications are presented, together with populations and species conservation priorities

Integrative phylogenetic, phylogeographic and morphological characterisation of the Unio crassus species complex reveals cryptic diversity with important conservation implications

The global decline of freshwater mussels and their crucial ecological services highlight the need to understand their phylogeny, phylogeography and patterns of genetic diversity to guide conservation efforts. Such knowledge is urgently needed for Unio crassus, a highly imperilled species originally widespread throughout Europe and southwest Asia. Recent studies have resurrected several species from synonymy based on mitochondrial data, revealing U. crassus to be a complex of cryptic species. To address long-standing taxonomic uncertainties hindering effective conservation, we integrate morphometric, phylogenetic, and phylogeographic analyses to examine species diversity within the U. crassus complex across its entire range. Phylogenetic analyses were performed using cytochrome c oxidase subunit I (815 specimens from 182 populations) and, for selected specimens, whole mitogenome sequences and Anchored Hybrid Enrichment (AHE) data on ∼ 600 nuclear loci. Mito-nuclear discordance was detected, consistent with mitochondrial DNA gene flow between some species during the Pliocene and Pleistocene. Fossil-calibrated phylogenies based on AHE data support a Mediterranean origin for the U. crassus complex in the Early Miocene. The results of our integrative approach support 12 species in the group: the previously recognised Unio bruguierianus, Unio carneus, Unio crassus, Unio damascensis, Unio ionicus, Unio sesirmensis, and Unio tumidiformis, and the reinstatement of five nominal taxa: Unio desectus stat. rev., Unio gontierii stat. rev., Unio mardinensis stat. rev., Unio nanus stat. rev., and Unio vicarius stat. rev. Morphometric analyses of shell contours reveal important morphospace overlaps among these species, highlighting cryptic, but geographically structured, diversity. The distribution, taxonomy, phylogeography, and conservation of each species are succinctly described.We thank Ana-Maria Benedek, Monica Sîrbu and Jouni Leinikki for their assistance with the fieldwork, and to Jeroen Goud, Sankurie Pye, Fiona Ware, Emily Mitchell, and Aleksandra Skawina for their assistance with the taxonomic investigation. We would also like to thank the editor, Dr. Guillermo Ortí, and two anonymous reviewers for their time and effort in reviewing our manuscript and for their insightful comments and valuable improvements to our work. This publication is based upon work from COST Action CA18239: CONFREMU - Conservation of freshwater mussels: a pan-European approach, supported by COST (European Cooperation in Science and Technology), including STSMs, the interaction of the authors and the writing of the paper. This work was supported by the project ConBiomics: The Missing Approach for the Conservation of Freshwater Bivalves Project No. POCI-01-0145-FEDER-030286, co-financed by FEDER through POCI and by FCT - Fundaç˜ao para a Ciˆencia e a Tecnologia, through national funds. Strategic funding UIDB/04423/2020 and UIDP/04423/2020 was provided by FCT. FCT also supported DVG (2020.03848.CEECIND), EF (CEECINST/00027/ 2021/CP2789/CT0003) and MLL (2020.03608.CEECIND). INB, AVK and IVV were supported by the Russian Science Foundation under grants (19-14-00066-P), (21-17-00126) and (21-74-10130) respectively. BVB acknowledges the bioinformatics platform of UMR 8198 for the computing resources to perform time-calibrated phylogenetic analyses; this platform is in part funded by CPER research project CLIMIBIO through the French Minist`ere de l’Enseignement Sup´erieur et de la Recherche, the Agence Nationale de la Recherche, the European Fund for Regional Development (FEDER) and the region Hauts-de-France (HdF). Support to KD came from the Czech Science Foundation (19–05510S). TT and MT were supported by the National Science Fund of Bulgaria under the project ‘Conservation of freshwater mussels on the Balkan Peninsula’ (KP-06-COST-9/20.07.2022). Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the United States Government.info:eu-repo/semantics/publishedVersio

Analysing the effect of bcl-2 expression to local control and survival in patients with early stage laryngeal cancer treated by radiotherapy

Bu çalışmanın amacı erken evre larenks kanseri tanısıyla küratif radyoterapi uygulanan olgularda Bcl-2 ekspresyonunun lokal kontrol ve genel sağkalıma etkisini değerlendirmektir.Çalışmaya Ocak 2005-Aralık 2008 tarihleri arasında Uludağ Üniversitesi Tıp Fakültesi Muammer Ağım Radyoterapi Merkezi'ne başvuran, Tis ve Evre I-II larenks kanseri tanısıyla radyoterapi uygulanan ve parafin bloklarına ulaşılabilen 53 olgu dahil edildi. Parafin bloklardan elde edilen kesitler Bcl-2 oncoprotein mause Clone 124 Scytek kiti kullanılarak immünohistokimyasal inceleme için hazırlandı. Preparatlar parlak alan ışık mikroskopu (Olympus BX50) altında, Regitnig tarafından tanımlanan prosedüre uygun olarak (+)-(+++) arasında skorlandı. İstatiksel analizlerde SPSS-18 paket program kullanıldı.Ortanca izlem süresi 61 ay olup (7-166 ay) izlem süresi içinde 9 (%17) olguda lokorejyonel yineleme saptandı. Olguların incelenmesinde 43'ünün (%81) negatif, 5'inin (%9) (+), 3'ünün (%6) (++), 2'sinin (%4) (+++) boyandığı saptandı. Tek değişkenli analizde radyoterapi sonrası yineleme saptanmasının (p=0.000), çok değişkenli analizde ise 50 yaşından genç olmanın (p=0.021) GSK'ı anlamlı olarak düşürdüğü ve radyoterapi sırasında hemoglobin değerinin 13gr/dl'den fazla olmasının hem tek değişkenli (p=0.002), hem de çok değişkenli (p=0.009) analizlerde GSK'ı anlamlı olarak artırdığı saptandı.Lokal kontrol üzerine etkili olabilecek prognostik faktörler incelendiğinde, tek değişkenli analizde, günde 20 adetten fazla sigara kullanımı (p=0.001) ile hem tek değişkenli (p=0.001) hem de çok değişkenli (p=0.018) analizlerde 50 yaşından genç olmanın lokal kontrol oranlarını anlamlı olarak düşürdüğü saptandı. Yapılan korelasyon analizinde, Bcl-2 ekspresyonu ile diğer faktörler arasında herhangi bir ilişki saptanamadı.Sonuç olarak, çalışmamızda Bcl-2 ekspresyonu ile lokal kontrol ve sağkalım arasında ilişki saptayamadık. Prognoz üzerine etkili olabileceği düşünülen etmenlerden 50 yaşından genç olmak ve günlük yirmi adetten fazla sigara kullanımı lokal kontrolü olumsuz etkilerken, hemoglobin düzeyinin 13 gr/dl'nin altında olması ve 50 yaşından genç olmanın da genel sağkalımı olumsuz etkilediği görüldü.The purpose of this study is to evaluate the effect of Bcl-2 expression to local control and overall survival in patients with early stage laryngeal cancer treated by curative radiotherapy.We studied 53 patients with Tis and stage I-II laryngeal cancer who were treated by radiotherapy between January 2005- December 2008 in Uludag University Faculty of Medicine Muammer Ağım Radiotherapy Center and whose paraffin blocks can be accessed. Sections obtained by parafin blocks were prepared for immunohistochemical analysis by using Bcl-2 oncoprotein mause Clone 124 Scytek kit. Preparates were scored between (+)-(+++) in accordance with the procedure described by Regitnig under bright field light microscope (Olympus BX50). SPSS-18 packaged software was used for statistical analysis.Average follow-up was 61 months (7-166 months) and locoregional reccurance was observed in 9 (17%) cases. In analysis, 43 cases (81%) were determined as negative, 5 cases were (9%) (+), 3 cases (6%) were (++) and 2 cases (4%) were (+++). It is observed that in univariate analysis ,determining recurrence after radiotherapy (p=0.000) and in multivariate analysis, age less than 50 (p=0.021) decrease overall survival significantly, also hemoglobin level more than 13 gr/dl during radiotherapy increases overall survival significantly both in univariate analysis (p=0.002) and multivariate analysis (p=0.009). When analysing of prognostic factors that might be effective on local control, in univariate analysis, smoking more than 20 cigarettes per day (p=0.001) and both in univariate analysis (p=0.001) and multivariate analysis (p=0.018), age less than 50 decrease local control ratio significantly. In correlation analysis, no relationship was evaluated between Bcl-2 expression and other factors.As a result, we couldn?t evaluate any relationship between Bcl-2 expression with local control and survival in our study. Age less than 50 and smoking more than 20 cigarettes per day were found as negative effect on local control; hemoglobin level less than 13 gr/dl and age less than 50 were found as negative effect on overall survival, which are the factors might be effective on prognosis

The Role of Forensic Autopsies in Diagnosis of Cancer

Background: Death from cancer is mostly considered as natural deaths. The role of cancer on the cause of death in forensic cases like sudden deaths, negligence or malpractice claims are being investigated. In a small amount of forensic cases, cancer is found incidentally at the autopsy. The aim of this study was to reveal characteristics of the cases in which cancer was diagnosed and types of malignancies in medicolegal autopsies. In addition, the role of forensic autopsies in determining oncological diseases was discussed. Methods: Forensic autopsies were performed in the Morgue Department of Forensic Medicine Council in İzmir for eight years were retrospectively reviewed. Cases that postmortem histopathological examination performed and malignant tumors diagnosed were included in the study. Data about age, sex, location of tumors, immediate causes of death and the potential relation between tumors and the primary cause of death were investigated. Results: In eight year period (between 2001 and 2009), 3722 medicolegal autopsies were done by postmortem histopathological examination. In 86 cases such kind of tumors were observed. In seven of 86 cases the tumors were determined as benign and excluded from the study. The remaining 79 cases with malignant tumors included in the study, 63 (79.7%) were male and 16 (20.3%) were female, mean age 54.96±20.35 (range: 7-88 years). The tumors were most frequently located in respiratory system (35.4%), gastrointestinal system (19%), genitourinary system (10.1%) and central nervous system (8.9%). The males most frequently had tumors in the respiratory system (42.9%) and the females in the endocrine system (25%). According to histopathological classification of the tumors, the most frequent tumors were epithelial tumors (65.8%), followed by hemopoetic (12.7%), mesenchymal (7.6%), glial (5.1%) and neuroendocrine tumors (2.5%) and timoma (1.3%). The tumors could not be histopathologically differentiated in 5.1% of the deaths. Cause of death was considered as cancer related complications in 58 cases (73.4%), other natural causes in 6 cases (7.6%) and external factors such as traumas and intoxications in 10 cases (12.7%). The cause of could not determined following complete autopsy and postmortem investigation in 5 cases (6.3%). Conclusion: Providing accurate cancer statistics by only clinical trials or medical autopsies that are diminished nowadays and clinical cancer outputs seems to be inadequate. We participated that investigating malignancies in large series of forensic autopsies would be contributed to data about public health. Keywords: cancer, medicolegal autopsy, cause of deat

Chemoradiotherapy-Associated Myelosuppression: A Retrospective Analysis of Risk Factors for Patients with Non-Small Cell Lung Cancer

Objective: Concomitant chemoradiotherapy (CRT) is the standard treatment for patients with stage III non-small cell lung cancer (NSCLC). Myelosuppression can be a significant problem in concomitant CRT. The aim of this study was to assess the parameters obtained before concomitant CRT to define the risk factors for myelosuppression in patients with locally advanced NSCLC. Methods: We retrospectively analyzed 81 patients with NSCLC who received concomitant platinum-based chemoradiotherapy between January 2008 and December 2012. Seventy-eight patients (96.2%) received etoposide (50 mg/m2, intravenously on days 1–5, 29–33) plus cisplatin (50 mg/m2, on days 1, 8, 29, and 36), and 3 patients (3.8%) received docetaxel (20 mg/m2/w, on weeks 1–8) plus cisplatin (20 mg/m2/w, on weeks 1–8) concurrently with thoracic radiotherapy to a total dose of 40–66.6 Gy. The risk factors were examined for their association with myelosupression (grade 3 or 4 leukopenia, neutropenia, thrombocytopenia, or anemia) by logistic regression analysis. Results: Grade 3 or higher neutropenia, leukopenia, thrombocytopenia, or anemia occurred in 51.8%, 53%, 8.6%, and 7.4% of the patients, respectively. Multivariate analysis revealed that the risk factors for neutropenia were performance status (p=0.032), white blood cell count (p=0.023), and pretreatment creatinine level (p=0.018). On multivariate analysis, white blood cell count (OR, 3.311; p=0.027; 95% CI, 1.148–9.545) was found as significant risk factor for CRT-induced leukopenia Conclusion: Patients with a poor pretreatment performance status, low white blood cell count, and high creatinine level are at a risk of myelosupression. These characteristics of the patients should be considered while making treatment decisions

Phylogeny, phylogeography, and evolution in the Mediterranean region: news from a freshwater mussel (Potomida, Unionida)

The Potomida genus (Bivalvia, Unionida) has a Circum-Mediterranean distribution and like other freshwater mussel species, its populations have suffered dramatic declines. Although this genus is currently considered as monotypic, it has a long history of taxonomic revisions and presently many aspects of its systematics and evolutionary history are unclear. We sampled a total of 323 individuals from 39 different sites across the Potomida genus distribution, and sequenced two mitochondrial (16S rDNA and Cytochrome c Oxidase Subunit I) and one nuclear (28S rDNA) genes to clarify its phylogeny and phylogeographic history. Our results show that the genus includes two well-supported clades, one comprising solely the western Mediterranean species Potomida littoralis, and the other including two eastern Mediterranean species, the Greek endemic P. acarnanica and the Anatolian and Middle Eastern P. semirugata. We suggest that Potomida started radiating during the upper Miocene, and that both vicariance and dispersal events shaped the diversification and distribution of the genus along the Mediterranean region. P. littoralis is further divided in two mitochondrial lineages, one restricted to Europe and the other occurring mostly in North Africa. Moreover, some European basins present both lineages in sympatry. The conservation status of the three recognized species should be reevaluated, particularly P. acarnanica, since it is restricted to two Greek river basins presenting a high risk of extinction. Overall, our results clarify some important gaps in knowledge concerning the phylogeny, phylogeography and evolution of the Potomida genus in the Mediterranean region with important taxonomical, ecological and conservational implications.Financial support was provided by Portuguese Foundation for Science and Technology (FCT) Grant to EF (SFRH/ BPD/108445/2015), to DVG (SFRH/BD/78402/2011) and Project (PTDC/AAC-AMB/117688/2010) as well as by Biotope, Conchological Society of Great Britain and Ireland, and the Mohamed bin Zayed Species Conservation Fund (Reference 15256799).info:eu-repo/semantics/publishedVersio