An integrative approach highlights the discrepancy in the genetic, phenotypic, and presumptive taxonomic structure of Phoxinus (Actinopterygii, Leuciscidae, Phoxininae) in Bulgaria.

The present drainage network of Bulgaria is the result of a complex Neogene and Quaternary evolution. Karst, which has developed on 23% of the territory, further complicates the hydrological pattern. Fresh waters of Bulgaria drain into the Black Sea and the Aegean Sea basins and can be roughly divided into the Danube (Middle and Lower Danube), non-Danube Black Sea, East Aegean, and West Aegean hydrological regions. Phoxinus, a small leuciscid fish, has a mosaic distribution in all four of these regions, inhabiting small mountainous and semi-mountainous streams. Based on morphology, it was identified as three species, Phoxinus phoxinus in the Danube, Phoxinus strandjae in the non-Danube, and Phoxinus strymonicus in West Aegean region. Later, molecular data revealed Phoxinus csikii and Phoxinus lumaireul in the Middle Danube and P. csikii in the Lower Danube. Phoxinus has been the focus of many studies, showing a high molecular and morphological diversity, which is not entirely consistent with previous morphology-only-based taxonomic concepts. In this study, molecular (a mitochondrial marker and a nuclear marker) and morphological data from both historical and recently sampled collections were analysed to assess the applicability of the integrative approach in Phoxinus. The results showed a significant influence of the complex paleo- and recent hydrology on the currently observed genetic structure of the considered populations and species. Furthermore, the study also demonstrated a strong influence of phenotypic plasticity on the morphological analysis of Phoxinus and the lack of a clear differentiation between P. csikii and P. strandjae. A barcoded specimen was designated as neotype to fix the species named P. strandjae in the current taxonomic concept. Finally, a significant discordance between genetically delimited clades and phenotypic groups did not allow a proper delineation of the species distributed in Bulgaria, demonstrating that more molecular markers are needed for further taxonomic study of the Phoxinus complex.

Complex visual hallucinations associated with ranolazine treatment: a case report.

Ranolazine is a piperazine derivative and is indicated for the symptomatic treatment of patients with stable angina pectoris who are inadequately controlled or intolerant to first-line antianginal therapies. Visual hallucinations together with neurological symptoms have been described with ranolazine treatment. In this case report, we describe a case of an elderly diabetic patient with chronic angina pectoris who developed visual hallucinations during an acute worsening of kidney function after tolerating ranolazine well for six years. Regular monitoring of electrocardiograms, kidney function, and psychiatric and neurological adverse effects in elderly patients on ranolazine is advised.

Hypersexuality during treatment with cariprazine in a patient with schizophrenia? A case report.

BACKGROUND: Cariprazine is a third-generation antipsychotic with a unique mechanism of action. It functions as a partial agonist with high affinity for dopamine D2 and D3 and serotonin 5-HT1A receptors, an antagonist for 5-HT2A (moderate affinity) and 5-HT2B (high affinity) receptors. It binds to histamine H1 receptors and has a low affinity for 5-HT2C and alpha 1A-adrenergic receptors and no affinity for muscarinic (cholinergic) receptors. Among the troubling side effects, symptoms related to impulse control, such as hypersexuality, pathological gambling, compulsive shopping, compulsive eating etc., have been reported with the use of antipsychotic medications. However, no reports have been published regarding impulse control symptoms associated with cariprazine. We report a case of cariprazine-induced hypersexuality in a patient with schizophrenia, which was resolved by discontinuation of the medication. CASE PRESENTATION: A 67-year-old Caucasian woman with schizophrenia was admitted to the hospital inpatient unit after she discontinued olanzapine and psychotic symptoms reappeared. Prior to that, she was in remission, taking olanzapine for approximately one year. After discontinuation, she experienced auditory hallucinations with persecutory delusions and became anergic and withdrawn, with blunted affect. Olanzapine was reintroduced, as it was proven successful in her past treatments. However, since there were no changes, especially in negative symptoms, cariprazine was added. Seven days after the introduction of cariprazine, the patient developed compulsive sexual behaviour. Therefore, cariprazine was discontinued, and the hypersexual behaviour was resolved. CONCLUSIONS: In this case report, we describe hypersexual behaviour that could potentially be induced by cariprazine. As a single case study, conclusions cannot be drawn. Controlled studies are warranted to better determine causality and the significance of this possible side-effect of cariprazine.

Psychometric Properties of the Slovenian Version of the Cardiac Depression Scale.

Introduction: The aim of this study was to translate the Cardiac Depression Scale into the Slovenian language and test its validity and reliability on Slovenian patients with heart disease. Methods: A total of 272 patients with heart disease who underwent elective coronary angiography at Celje General Hospital participated in this study. We used the Slovenian Cardiac Depression Scale (S-CDS), the Spielberger State Anxiety Inventory (STAI-S), and the Center for Epidemiologic Studies Depression Scale-20 (CES-D) to collect data. An exploratory and confirmatory factor analysis, internal consistency, test-retest reliability, and concurrent validity were performed. Results: Cronbach's alpha for the total scale was 0.92 and the test-retest reliability was 0.71. Exploratory factor analysis confirmed six factors, accounting for 61% of the total variance. The confirmatory factor analysis indicated that a two- and one-factor solution had acceptable goodness-of-fit measures. However, we kept a more parsimonious one-factor method, given a high correlation between the two factors and the theoretical background in previous studies. Concurrent validation against the CES-D and the STAI-S showed moderate to strong correlations. Conclusions: The S-CDS is a reliable and valid instrument for screening for depression in Slovenian patients with heart disease.

The minnow Phoxinus lumaireul (Leuciscidae) shifts the Adriatic-Black Sea basin divide in the north-western Dinaric Karst region.

Karst landscapes are characterized by intermittent and sinking streams. The most common method used to study underground hydrological connections in karst is tracing tests. However, a more biologically oriented approach has been suggested: analysis of the genetic structure of aquatic organisms. Biological tracers can be sought among trogloxenes, that is, surface species that occasionally enter caves and groundwater. One such example is the fish genus Phoxinus, which exhibits high genetic diversity and complex phylogeography in the Balkan Peninsula. In the north-western Dinaric Karst, the complex hydrological network was digitalized in 2020. Contemporaneously, Phoxinus lumaireul populations in the Slovenian Dinaric Karst were intensively sampled and analysed for fragments of two mitochondrial genes and one nuclear gene. The derived phylogeographic structure and data on hydrological connections were compared to evaluate support for three alternative scenarios: The genetic structure (1) is a consequence of the ongoing geneflow through underground connections, (2) reflects a previous hydrological network or (3) is an outcome of anthropogenic translocations. The results suggest that the first two scenarios seem to have played a major role, while the third has not had profound effects on the genetic composition. Comparison between the genetic structure of Slovenian Dinaric Karst sampling sites and that of hydrologically isolated reference sampling sites indicated a greater genetic connectivity in the former. Moreover, the range of Adriatic (1a) and Black Sea (1c) haplotypes does not correspond to the Adriatic-Black Sea basin divide but is shifted northwards.

Evaluating anxiety in elective coronary angiography study: rationale, design, and study methodology.

AIMS: Prevalence of anxiety disorder in coronary artery disease reaches up to 15% and about half of patients with coronary artery disease have anxiety or depression comorbidity. Prevalence of anxiety in patients undergoing percutaneous coronary intervention ranges 24-72%. Depression can often overlap with anxiety symptoms and the evaluation of anxiety in elective coronary angiography study (ANGST) aims to determine the prevalence of anxiety by excluding patients with comorbid depressive symptoms. ANGST also aims to determine how anxiety correlates with psychological parameters (personality traits, coping strategies) and with outcome of elective coronary angiography (ECA). METHODS: We will conduct a prospective single-center cross-section study in patients undergoing ECA. Anxiety will be evaluated at four time points using self-rating questionnaires: 14 days prior to ECA; 2-4 h before ECA; 24 h after ECA, but prior to discharge; and 4-6 weeks after discharge. The primary outcome of ANGST is the burden of anxiety experienced by patients without depressive symptoms and a correlation of anxiety with ECA outcome. CONCLUSION: Our study aims to provide evidence on which personality traits and coping strategies affect the levels of anxiety. We will also determine psychometric properties of the two questionnaires used in our study. The results will have implications for improvement of interventions designed to recognize anxiety and will offer future research of psychological and/or pharmacological interventions to reduce the burden of anxiety.

Neotype designation for Thymallus aeliani Valenciennes, 1848 from a museum topotype specimen and its affiliation with Adriatic grayling on the basis of mitochondrial DNA.

In 1848, the grayling Thymallus aeliani (Valenciennes) was described from Lake Maggiore, Italy, in the north Adriatic basin. Genetic analyses of the mitochondrial control region showed a unique evolutionary history of grayling inhabiting the rivers of northern Adriatic basin, from the upper reaches of the Po River and its left tributaries in the west to the Soca River in the east, which resulted in the designation of this phylogenetic lineage as Adriatic grayling. Consequently, the name T. aeliani was connected to the Adriatic lineage, re-establishing the validity of this taxon. However, the mitochondrial haplotypes belonging to Adriatic grayling were never compared with the type specimens of T. aeliani, as their whereabouts were unknown. In this study, a neotype for T. aeliani was designated using topotypical specimens stored at the Natural History Museum in Vienna. The neotype (NMW 68027:2 labelled as "Lago Maggiore, Bellotti, 1880") was designated pursuant to the conditions stipulated in Article 75.3 of the International Code of Zoological Nomenclature. Furthermore, the mitochondrial control region of the neotype was compared to haplotypes of the Adriatic lineage and showed high genetic similarity, which therefore connects the species name T. aeliani to the Adriatic grayling. This crucial step in fixing nomenclatural status of this species is very important for its protection and management.

New data on Garra makiensis (Cyprinidae, Labeoinae) from the Awash River (Ethiopia) with remarks on its relationships to congeners on the Arabian Peninsula.

On the African continent, the genus Garra consists of several species often insufficiently separated from each other by diagnostic characters. Herein, a detailed morphological redescription of Garra makiensis from the Awash River drainage is presented, together with additional data on the type specimens of G. makiensis and G. rothschildi. Mitochondrial CO1 sequence data are also provided, including the historic paralectotype of G. makiensis, with a comparison to Garra species from Africa and the Middle East. Based on these sequences, G. makiensis clusters outside the group of African congeners and is a sister lineage to species from the south-east of the Arabian Peninsula. Although morphologically variable, G. makiensis is characterised by having a single unbranched pectoral-fin ray, a short distance between vent and anal-fin origin (7.3-19.7 % of pelvic - anal distance), chest and belly covered with scales, and a prominent axillary scale at base of pelvic fin (18.8-35.5 % of pelvic-fin length).

European minnows through time: museum collections aid genetic assessment of species introductions in freshwater fishes (Cyprinidae: Phoxinus species complex).

Massive fish introductions have taken place throughout much of the world, mostly over the last 70 years, and present a major threat to the genetic diversity of native fishes. Introductions have been reported for European Phoxinus, a ubiquitous small cyprinid that populates a wide variety of habitats. Species delineation in European Phoxinus has proven difficult with one reason being ranges of distribution that often traverse drainage boundaries. The present study combines recent samples with museum samples to better understand the current distribution of Phoxinus species and their distributions prior to the massive introductions of fishes in Europe, and to evaluate the use of museum specimens for species distribution studies. For these purposes, genetic lineages from sites collected prior to 1900 (n = 14), and between 1900 and 1950 (n = 8), were analysed using two mitochondrial and nuclear markers. Although possible fish introductions were detected, our results show that the distribution of genetic lineages of museum samples is comparable to that of the extant lineages of European Phoxinus present in those areas. These observations suggest that in the studied ranges the distribution of Phoxinus lineages has been driven by natural processes.

Corrigendum to «Contrasting morphology with molecular data: an approach to revision of species complexes based on the example of European (Cyprinidae)¼ by Palandacic et al. 2017.

[This corrects the article DOI: 10.1186/s12862-017-1032-x.].

Contrasting morphology with molecular data: an approach to revision of species complexes based on the example of European Phoxinus (Cyprinidae).

BACKGROUND: Molecular taxonomy studies and barcoding projects can provide rapid means of detecting cryptic diversity. Nevertheless, the use of molecular data for species delimitation should be undertaken with caution. Especially the single-gene approaches are linked with certain pitfalls for taxonomical inference. In the present study, recent and historical species descriptions based upon morphology were used as primary species hypotheses, which were then evaluated with molecular data (including in type and historical museum material) to form secondary species hypotheses. As an example of cryptic diversity and taxonomic controversy, the European Phoxinus phoxinus species complex was used. RESULTS: The results of the revision showed that of the fourteen primary species hypotheses, three were rejected, namely P. ketmaieri, P. likai, and P. apollonicus. For three species (P. strandjae, P. strymonicus, P. morella), further investigation with increased data sampling was suggested, while two primary hypotheses, P. bigerri and P. colchicus, were supported as secondary species hypotheses. Finally, six of the primary species hypotheses (P. phoxinus, P. lumaireul, P. karsticus, P. septimanae, P. marsilii and P. csikii) were well supported by mitochondrial but only limitedly corroborated by nuclear data analysis. CONCLUSION: The approach has proven useful for revision of species complexes, and the study can serve as an overview of the Phoxinus genus in Europe, as well as a solid basis for further work.

Identification of rare genetic variation of NLRP1 gene in familial multiple sclerosis.

The genetic etiology and the contribution of rare genetic variation in multiple sclerosis (MS) has not yet been elucidated. Although familial forms of MS have been described, no convincing rare and penetrant variants have been reported to date. We aimed to characterize the contribution of rare genetic variation in familial and sporadic MS and have identified a family with two sibs affected by concomitant MS and malignant melanoma (MM). We performed whole exome sequencing in this primary family and 38 multiplex MS families and 44 sporadic MS cases and performed transcriptional and immunologic assessment of the identified variants. We identified a potentially causative homozygous missense variant in NLRP1 gene (Gly587Ser) in the primary family. Further possibly pathogenic NLRP1 variants were identified in the expanded cohort of patients. Stimulation of peripheral blood mononuclear cells from MS patients with putatively pathogenic NLRP1 variants showed an increase in IL-1B gene expression and active cytokine IL-1ß production, as well as global activation of NLRP1-driven immunologic pathways. We report a novel familial association of MS and MM, and propose a possible underlying genetic basis in NLRP1 gene. Furthermore, we provide initial evidence of the broader implications of NLRP1-related pathway dysfunction in MS.

Morphological divergence of lake and stream Phoxinus of Northern Italy and the Danube basin based on geometric morphometric analysis.

Minnows of the genus Phoxinus are promising candidates to investigate adaptive divergence, as they inhabit both still and running waters of a variety of altitudes and climatic zones in Europe. We used landmark-based geometric morphometric methods to quantify the level of morphological variability in Phoxinus populations from streams and lakes of Northern Italy and the Danube basin. We analyzed body shape differences of populations in the dorsal, lateral, and ventral planes, using a large array of landmarks and semilandmarks. As the species identification of Phoxinus on morphological characters is ambiguous, we used two mitochondrial genes to determine the genetic background of the samples and to ensure we are comparing homogenous groups. We have found significant body shape differences between habitats: Minnow populations inhabiting streams had a deeper body and caudal peduncle and more laterally inserted pectoral fins than minnows inhabiting lakes. We have also found significant body shape differences between genetic groups: Italian minnows had deeper bodies, deeper and shorter caudal peduncles, and a shorter and wider gape than both groups from the Danube. Our results show that the morphology of Phoxinus is highly influenced by habitat and that body shape variation between habitats was within the same range as between genetic groups. These morphological differences are possibly linked to different modes of swimming and foraging in the respective habitats and are likely results of phenotypic plasticity. However, differences in shape and interlandmark distances between the groups suggest that some (though few) morphometric characters might be useful for separating Phoxinus species.

Molecular data suggest a multispecies complex of Phoxinus (Cyprinidae) in the Western Balkan Peninsula.

This study aimed to evaluate the phylogeographic distribution of Phoxinus phoxinus sensu lato populations in the Balkan Peninsula using molecular methods. For the western Balkan Peninsula, two species (Danubian P. phoxinus and Adriatic P. lumaireul) have been suggested, but not confirmed by subsequent morphological studies or by genetic data. For the present study, more than 300 specimens were collected from the western Balkans. A partial sequence of the mt cyt b gene analysed using Bayesian and ML methods revealed several well-supported clades, with distances ranging from 4% to 11%. The clades were corroborated by RAG1 sequence analysis. Thus, the molecular analysis points to a multispecies complex of Phoxinus sp. with ranges of the clades extending north-west to south-east along the western Balkan Peninsula. In addition, the dispersion of fish through subterranean water connections in karst is indicated.

Fish migrate underground: the example of Delminichthys adspersus (Cyprinidae).

Complex aquatic systems of karst harbour a rich but little-investigated biodiversity. In Croatia and Bosnia-Herzegovina karst, temporal springs are inhabited by a group of minnow-like fishes that retreat to the associated ground water during dry seasons and spend several months underground. The most abundant species in this group is Delminichthys adspersus (Heckel 1843), which also has the most fragmented distribution range. To determine the population composition and dispersal patterns, and to detect potential underground migration, a large genetic data set comprising 544 specimens of D. adspersus covering most of its distribution area was analysed. Analysis of mitochondrial cytochrome b sequences (∼1000 bp) and eight microsatellite loci showed that D. adspersus comprises at least three subpopulations with gene flow occurring among them. Coalescent-based analysis revealed a complex migration pattern, with several unidirectional dispersal paths, including between temporal springs that share no surface connection. The results of this study suggest the existence of recurrent underground migration of fish in a karst environment and demonstrate the complexity of its hydrological network. The findings are relevant to conservation strategies for endemic karst organisms and karst ecosystems as a whole.