Der Urologe

Die Arbeitszeitbelastung von Fach- und Oberärzten in der deutschen Urologie – eine Bestandsaufnahme Zusammenfassung Hintergrund Die Arbeits- und Weiterbildungsbedingungen von urologischen Assistenzärzten in Deutschland sind bereits analysiert worden. Aber wie steht es um die urologischen Oberärzte in Deutschland? Unter welchen Bedingungen müssen diese arbeiten? Bislang existieren keine uns bekannten publizierten Daten, die diese Konditionen dieser speziellen urologischen Berufsgruppe beleuchten, daher stellen wir die Ergebnisse der vorliegenden Umfrage vor. Ziel der Arbeit Die Analyse der aktuellen Arbeitsbedingungen urologischer Fach- und Oberärzte in Deutschland möglichst umfassend zu erfragen und auszuwerten. Material und Methoden Der Arbeitskreis Angestellter Ärzte des BvDU führte eine Befragung im Rahmen des Urologischen Oberarztforums 2016 sowie online über den Email-Verteiler der Deutschen Gesellschaft für Urologie (DGU) durch. Erfragt wurden die Arbeitsbelastung, Arbeitsbedingungen und -zufriedenheit, geleistete Überstunden sowie Arbeitszeit- und Opt-out-Regelungen. Ergebnisse An der Befragung nahmen 176 Oberärzte teil, davon waren 88 % männlich und 12 % weiblich. In Deutschland gibt es 1125 Oberärzte. Das Durchschnittsalter der Befragten betrug 44,9 Jahre. Bei >80 % der Befragten lag die durchschnittliche Wochenarbeitszeit bei über 50 h, 70 % unterschrieben eine Opt-out-Regelung. Es konnte dabei ein Zusammenhang zwischen einer steigenden Jobunzufriedenheit ab einer Wochenarbeitszeit über 55 h bzw. einer durchschnittlichen Anwesenheit im Rufbereitschaftsdienst über 4 h hergestellt werden. Die Anzahl der Rufbereitschaftsdienste hatte auf die Zufriedenheit keinen Einfluss. Insgesamt gaben 43 % (70 von 162) der Befragten an, dass regelmäßig Überstunden verfallen, bei 12 % (20 von 162) verfielen alle Überstunden. Rund 30 % der Oberärzte in der deutschen Urologie sind mit ihren aktuellen Arbeitsbedingungen in der aktuellen Form unzufrieden. Schlussfolgerungen Anpassungen und Verbesserungen der Arbeitsbedingungen urologischer Oberärzte sind auf Grundlage der erhobenen Daten erforderlich, um die Attraktivität des Berufsbildes und Führungspositionen in der deutschen Urologie zu erhalten.

Konzepte zur besseren Vereinbarkeit von klinischer und wissenschaftlicher Tätigkeit – ein Thesenpapier der Arbeitsgemeinschaft „Junge Urologen" der Deutschen Gesellschaft für Urologie

Qualität statt Quantität verbessert die Weiterbildung Zusammenfassung Eine Weiterbildungsevaluation findet in Deutschland zum aktuellen Zeitpunkt zwar statt – allerdings nicht flächendeckend und regelmäßig und ohne bundesweiten Standard. Vergleiche zwischen verschiedenen Kliniken und Ausbildern können daher im Moment nicht gezogen werden. Umfragemodi, wie beispielsweise in der Schweiz seit Jahrzehnten existierend, könnten eine konstruktive Grundlage in der Erhebung des aktuellen Weiterbildungsstands urologischer Assistenzärzte sein und nachfolgend die Diskussion über Verbesserungen ermöglichen. Dazu bedarf es einer berufsgruppenübergreifenden konstruktiven Zusammenarbeit, ohne die Verantwortung lediglich den Weiterbildnern zuzuschreiben. Darüber hinaus sollten Modelle der Entlohnung guter Weiterbildung gefunden werden.

Systemwandel zu erwarten. Ein Appell an eine andere Art des Wirtschaftens aus der Sicht des Chefarztes

LUTS bei BPH-Patienten

Aktuelle Forschung pädiatrischer und adulter Keimzelltumoren

Mitteilungen der DGU

Molekulares Tumorboard Prostatakarzinom Zusammenfassung In der modernen Onkologie ist das molekulare Tumorboard die Schnittstelle zwischen Klinik und Grundlagenforschung. Durch die interdisziplinäre Zusammenarbeit von Experten verschiedener Fachdisziplinen soll die Indikation zur molekularbiologischen Tumoranalysen sinnvoll gestellt, die Untersuchungsergebnisse korrekt interpretiert und hierauf basierende personalisierte Therapieempfehlungen ausgegeben werden. Bezogen auf das metastasierte Prostatakarzinom können hiervon insbesondere Patienten mit familiärem Prostatakarzinom, jungen Manifestationsalter oder nach Ausschöpfung von zugelassenen Standardtherapien profitieren. Mit der Androgenrezeptor-Splicevariante 7 (AR-V7) steht ein prädiktiver Marker für das Ansprechen auf eine Hormontherapie mit Abirateron oder Enzalutamid zur Verfügung. Testverfahren zur Bestimmung des AR-V7-Status im Blut sind bereits kommerziell erwerblich. Mutationen in den DNA-Reparaturmechanismen stellen einen weiteren Ansatzpunkt für zielgerichtete, personalisierte Therapien dar. Defekte in der homologen Rekombination erhöhen die Empfindlichkeit von Tumorzellen gegenüber Poly(ADP-Ribose)-Polymerase-(PARP-)Inhibitoren wie Olaparib. In der TOPARP-A-Studie, eine Phase-II-Studie, lag die Ansprechrate bei Patienten mit metastasiertem Prostatakarzinom und Mutationen in DNA-Reparaturgenen bei 88 %. Mikrosatelliteninstabilität ist die Folge von Defekten in der DNA-Mismatch-Reparatur. Mit dem Immun-Checkpoint-Inhibitor Pembrolizumab steht auch für diese Patientenkollektiv eine wirksame Therapie zur Verfügung. Da weder PARP-Inhibitoren noch Pembrolizumab aktuell zur Therapie des metastasierten Prostatakarzinoms in Deutschland zugelassen sind, stärkt die Empfehlung eines molekularen Tumorboards die Erfolgsaussichten für die Genehmigung eines individuellen Heilversuches durch die Krankenkassen.

Molekulares Tumorboard – Nierenzellkarzinom Zusammenfassung Die Einführung molekularer zielgerichteter Substanzen hat die Therapie des metastasierten Nierenzellkarzinoms grundlegend verändert. Eine erste Welle der Entwicklung basiert auf dem verbesserten Verständnis der Tumorbiologie seit der Entdeckung der Bedeutung des VHL-Gens für die Progression, was den Weg für Therapieoptionen geebnet hat, die v. a. die Neoangiogenese hemmen. Relevant sind hier die Überexpression proangiogener und proliferationsfördernder Faktoren (VEGF, „vascular endothelial growth factor"; PDGF; „platelet derived growth factor") sowie eine Überaktivierung des PI3K- (Phosphatidylinositol-3-Kinase‑)Akt-Signalwegs: Zielstruktur ist hierbei das „mammalian target of rapamycin" (mTOR-)Molekül, das an der Regulation zellproliferativer Prozesse beteiligt ist. Vor allem die VEGF-, PDGF- und mTOR-Signale bzw. Signalwege sind zentrale Angriffspunkte aktueller zielgerichteter Substanzen. Eine zweite Welle ist in der Entwicklung von Therapieansätzen mit der zielgerichteten Aktivierung und Modulation des Immunsystems zu sehen, die die „Immuntherapie" wieder in den Fokus des Interesses gerückt hat. Zentrale Entwicklung ist die Anwendung von Immun-Checkpoint-Inhibitoren, mit deren Hilfe eine (Re‑)Aktivierung der zellulären Abwehr und hier v. a. von T‑Zellen erfolgt, was per se das Potenzial einer zytoreduktiven Therapie durch Abtöten der Tumorzellen in sich birgt. Auch, wenn sich die Prognose durch die rasante Entwicklung der letzten Jahre signifikant verbesserte, bleibt die Behandlung anspruchsvoll, da die meisten Patienten einen Progress erleben müssen und ein Langzeitüberleben in nur etwa 20 % der Fälle erreicht werden kann, da eben auch ein Teil der Patienten primär refraktär ist oder nicht anspricht.

Ambulante Onkologie in der Urologischen Praxis – Möglich? Machbar? Perspektiven

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Drugs

Volanesorsen: First Global Approval Abstract Volanesorsen (Waylivra®), an antisense oligonucleotide inhibitor of apolipoprotein CIII (apoCIII) mRNA, is being developed by Ionis Pharmaceuticals through its subsidiary company, Akcea Therapeutics, to treat familial chylomicronemia syndrome (FCS), hypertriglyceridemia and familial partial lipodystrophy (FPL). In May 2019, volanesorsen was approved in the EU for the treatment of adult patients with FCS based on positive results from the multinational, phase III APPROACH and COMPASS studies. Other clinical trials are ongoing to assess its utility in hypertriglyceridemia, FPL and partial lipodystrophy. This article summarizes the milestones in the development of volanesorsen leading to this first approval as an adjunct to diet in adult patients with genetically confirmed FCS and at high risk for pancreatitis, in whom response to diet and triglyceride lowering therapy has been inadequate.

Correction to: Migalastat: A Review in Fabry Disease The article Migalastat: A Review in Fabry Disease, written by Emma H. McCafferty, Lesley J. Scott, was originally published Online First without open access. After publication in volume 79, issue 5, pages 543–554 [funder] requested that the article be Open Choice to make the article an open access publication. Post-publication open access was funded by [funder]. The article is forthwith distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, duplication, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

Eltrombopag in Immune Thrombocytopenia, Aplastic Anemia, and Myelodysplastic Syndrome: From Megakaryopoiesis to Immunomodulation Abstract Eltrombopag is an orally available thrombopoietin receptor agonist indicated for the treatment of immune thrombocytopenia (ITP). Beyond the effect on megakaryopoiesis, the drug also showed a stimulating effect on the hematopoietic stem cell with consistent clinical efficacy in aplastic anemia (AA) and myelodysplastic syndromes (MDS). Eltrombopag is highly effective in ITP and less so in AA and MDS. This observation underlines the importance of residual normal hematopoiesis, which is maximal in ITP, minimal/absent in AA, and dysregulated in MDS. In ITP, the drug at 50–75 mg daily induced up to 85% responses both in clinical trials and real-life studies, with the possibility of tapering and discontinuation. In AA, eltrombopag at 150 mg daily was effective in about 40% of cases relapsed/refractory to standard immunosuppression or ineligible for bone marrow transplant. In MDS, the drug seems less effective, with responses in about a quarter of patients at various schedules. The efficacy of eltrombopag in ITP, AA, and MDS suggests the existence of common immune-pathological mechanisms in these diseases, including autoimmunity against peripheral blood cells and bone marrow precursors, as well as a possible evolution of one condition into the other. Additional mechanisms of action emerging from the clinical use of eltrombopag include modulation of T-regulatory cells, restoration of Fc-γ receptor balance in phagocytes, and an iron-mobilizing effect. In this review, we analyzed the most recent literature on eltrombopag use and efficacy in patients with ITP, AA, and MDS, exploring the basis for different dosing, combined treatments, and discontinuation in each context.

Current Treatments and New Developments in the Management of Glucocorticoid-induced Osteoporosis Abstract Glucocorticoids (GCs) are often used for improvement of quality of life, particularly in the elderly, but long-term GC use may cause harm; bone loss and fractures are among the most devastating side effects. Fracture risk is particularly high in patients with a severe underlying disease with an urgent need for treatment with high-dose GCs. Moreover, it is important to realize that these patients suffer from an augmented background fracture risk as these patients have a high presence of traditional risk factors for osteoporosis, such as high age, low body mass index (BMI), smoking and relatives with osteoporosis or hip fractures. It is thus crucial for prevention of osteoporotic fractures to use the lowest dose of GC for a short period of time to prevent fractures. Another important task is optimal treatment of the underlying disease; for instance, fracture risk is higher in patients with active rheumatoid arthritis than in patients in whom rheumatoid arthritis is in remission. Thus, fracture risk is generally highest in the early phase, when GC dosage and the disease activity of the underlying disease are high. Finally, some of the traditional risk factors can be modulated, e.g., smoking and low BMI. Life-style measures, such as adequate amounts of calcium and vitamin D and exercise therapy are also crucial. In some patients, anti-osteoporotic drugs are also indicated. In general, oral bisphosphonates (BPs) are the first choice, because of their efficacy and safety combined with the low cost of the drug. However, for those patients who do not tolerate oral BPs, alternatives ("second-line therapies") are available: BP intravenously (zoledronic acid), denosumab (Dmab), and teriparatide. Both zoledronic acid and Dmab have been proven to be superior to oral bisphosphonates like risedronate in improvement of bone mineral density. For teriparatide, vertebral fracture reduction has been shown in comparison with alendronate. Thus, to reduce the global burden of GC use and fracture risk, fracture risk management in GC users should involve at least involve life-style measures and the use of the lowest possible dose of GC. In high-risk patients, anti-osteoporotic drugs should be initiated. First choice drugs are oral BPs; however, in those with contraindications and those who do not tolerate oral BPs, second-line therapies should be started. Although this is a reasonable treatment algorithm, an unmet need is that the most pivotal (second-line) drugs are not used in daily clinical practice at the initial phase, usually characterized by high-dose GC and active underlying disease, when they are most needed. In some patients second-line drugs are started later in the disease course, with lower GC dosages and higher disease activity. As this is a paradox, we think it is a challenge for physicians and expert committees to develop an algorithm with clear indications in which specific patient groups second-line anti-osteoporotic drugs should or could be initiated as first-choice treatment.

Damoctocog Alfa Pegol: A Review in Haemophilia A Abstract Damoctocog alfa pegol (Jivi®) is approved in the USA, EU, Japan and Canada for the treatment and prophylaxis of previously treated patients aged ≥ 12 years with haemophilia A. Formulated with a 60 kDa polyethylene glycol (PEG) moiety, damoctocog alfa pegol is an intravenously (IV) administered recombinant factor VIII (rFVIII) product with a longer terminal half-life than non-PEGylated FVIII and rFVIII products. In the multinational phase II/III PROTECT VIII trial, prophylaxis with damoctocog alfa pegol reduced the likelihood of bleeding in previously treated patients aged ≥ 12 years with severe haemophilia A, with dosing schedules ranging from twice weekly to once every 7 days. Interim data from the ongoing extension phase indicated that the reduced annualized bleeding rates (ABRs) were maintained for up to 5.2 years of prophylaxis with damoctocog alfa pegol. Damoctocog alfa pegol was also effective in treating bleeding episodes and in providing haemostatic control during surgery. Damoctocog alfa pegol was generally well tolerated in adult and adolescent patients with severe haemophilia A, with most adverse events considered to be unrelated to treatment. There were no new or confirmed cases of FVIII inhibitor development and anti-PEG antibodies, observed in some patients, were of low titre and transient. Damoctocog alfa pegol extends the available treatment options in previously treated adults and adolescents with haemophilia A, offering the possibility of up to once-weekly administration for suitable patients.

Dapagliflozin: A Review in Type 2 Diabetes Abstract Dapagliflozin (Forxiga®) is a highly potent, reversible and selective sodium-glucose cotransporter-2 inhibitor indicated worldwide for the treatment of type 2 diabetes (T2D). In the EU, oral dapagliflozin once daily is approved for use as monotherapy (in patients who are intolerant of metformin) and as add-on combination therapy (with other glucose-lowering agents, including insulin) for T2D when diet and exercise alone do not provide adequate glycaemic control. In numerous well-designed clinical studies and their extensions, dapagliflozin as monotherapy and combination therapy with other antihyperglycaemic agents provided effective glycaemic control and reduced bodyweight and blood pressure (BP) across a broad spectrum of patients. Dapagliflozin reduced the rate of cardiovascular (CV) death or hospitalization for heart failure (HHF), did not adversely affect major adverse CV events (MACE) and possibly reduced progression of renal disease relative to placebo in patients with established atherosclerotic CV disease (CVD) or multiple risk factors for CVD. Dapagliflozin was generally well tolerated, with a low risk of hypoglycaemia; diabetic ketoacidosis (DKA), although rare, and genital infections were more common with dapagliflozin than placebo. Given its antihyperglycaemic, cardioprotective and possibly renoprotective properties and generally favourable tolerability profile, dapagliflozin provides an important option for the management of a broad patient population, regardless of the history of CVD.

Advances in Targeted Therapies for Triple-Negative Breast Cancer Abstract While the outcomes for patients diagnosed with hormone receptor positive (HR+) and/or human epidermal growth factor receptor 2-positive (HER2+) breast cancers have continued to improve with the development of targeted therapies, the same cannot be said yet for those affected with triple-negative breast cancer (TNBC). Currently, the mainstay of treatment for the 10–15% of patients diagnosed with TNBC remains cytotoxic chemotherapy, but it is hoped that through an enhanced characterization of TNBC biology, this disease will be molecularly delineated into subgroups with targetable oncogenic drivers. This review will focus on recent therapeutic innovations for TNBC, including poly-ADP-ribosyl polymerase (PARP) inhibitors, phosphoinositide 3-kinase (PI3K) pathway inhibitors, immune checkpoint inhibitors, and cyclin-dependent kinase (CDK) inhibitors.

Onasemnogene Abeparvovec: First Global Approval Abstract Onasemnogene abeparvovec (onasemnogene abeparvovec-xioi; formerly AVXS-101; ZOLGENSMA®) is an adeno-associated viral vector-based gene therapy designed to deliver a functional copy of the human survival motor neuron(SMN) gene to the motor neuron cells of patients with spinal muscular atrophy (SMA). It has been developed by AveXis, a Novartis company, and was approved in May 2019 in the USA for the treatment of paediatric patients aged < 2 years with SMA and bi-allelic mutations in the SMN1 gene (the primary gene encoding survival motor neuron protein). Onasemnogene abeparvovec is the first gene therapy to be approved for SMA in the USA. The recommended dose is 1.1 × 1014 vector genomes per kg of bodyweight, administered as a single intravenous infusion over 60 min. Regulatory assessments for this formulation of onasemnogene abeparvovec are underway in the EU and Japan; an intrathecal formulation is currently undergoing clinical development in the USA. This article summarizes the milestones in the development of onasemnogene abeparvovec leading to this first approval for the treatment of paediatric patients aged < 2 years with SMA and bi-allelic mutations in SMN1.

Recent Developments in LRRK2-Targeted Therapy for Parkinson's Disease Abstract Kinase activating missense mutations in leucine-rich repeat kinase 2 (LRRK2) are pathogenically linked to neurodegenerative Parkinson's disease (PD). Over the past decade, substantial effort has been devoted to the development of potent and selective small molecule inhibitors of LRRK2, as well as their preclinical testing across different Parkinson's disease models. This review outlines the genetic and biochemical evidence that pathogenic missense mutations increase LRRK2 kinase activity, which in turn provides the rationale for the development of small molecule inhibitors as potential PD therapeutics. An overview of progress in the development of LRRK2 inhibitors is provided, which in particular indicates that highly selective and potent compounds capable of clinical utility have been developed. We outline evidence from rodent- and human-induced pluripotent stem cell models that support a pathogenic role for LRRK2 kinase activity, and review the substantial experiments aimed at evaluating the safety of LRRK2 inhibitors. We address challenges still to overcome in the translational therapeutic pipeline, including biomarker development and clinical trial strategies, and finally outline the potential utility of LRRK2 inhibitors for other genetic forms of PD and ultimately sporadic PD. Collective evidence supports the ongoing clinical translation of LRRK2 inhibitors as a therapeutic intervention for PD is greatly needed.

Remogliflozin Etabonate: First Global Approval Abstract Remogliflozin, a selective sodium-glucose co-transporter subtype 2 (SGLT2) inhibitor, which is to be administered as remogliflozin etabonate (Remo™, Remozen™), the prodrug for remogliflozin, recently received its first approval as a treatment for type 2 diabetes mellitus (T2DM) in India. This article summarizes the milestones in the development of remogliflozin etabonate leading to this first approval for T2DM.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Acarology

Inter- and intraspecific variability of morphological and molecular characters in Allothrombium species, with special reference to Allothrombium fuliginosum Abstract Morphology-based identification of Allothrombium spp., in view of the limited knowledge of intraspecific variation, hinders the recognition of species borders and affects the views on the actual distribution of species. Therefore, identification will benefit from reference to molecular methods. The separate species identity of specimens putatively representing Allothrombium fuliginosum and A. pulvinum, both reported as widely distributed in the Palaearctic region and considered as potential biological control agents, was checked using morphological and molecular analyses. The representatives of various Allothrombium spp. collected in the Palaearctic were included in the analysis in order to ascertain the distance between species. The results of the morphological examination, supported by statistical inference, along with the comparison of COI and/or ITS2 sequences, weaken the hypothesis of synoccurrence of both species in the Palaearctic region. Hence, we hypothesize that A. fuliginosum is widely distributed in the Palaearctic, whereas A. pulvinum should be regarded a Nearctic species.

Demodicosis in a captive African straw-coloured fruit bat ( Eidolon helvum ) Abstract Demodicosis is most frequently observed in the domestic dog (Canis familiaris), but it has rarely been reported in bats (Chiroptera). The overpopulation of Demodex spp. that causes dermatological changes is generally associated with a compromised immune system. We describe the gross and histological features of generalized demodicosis in an adult female African straw-coloured fruit bat (Eidolon helvum) drawn from a captive research colony. The histology of the lesions revealed comedones and follicular infundubular cysts harbouring numerous Demodex spp. mites, eliciting a minimal inflammatory response in the adjacent dermis. The histological examination of a full set of tissues did not reveal clear evidence of immunosuppression, although a clinical history of recent abortion and possible stressors due to captivity could be considered risk factors for the demodicosis. Attempts to determine the Demodex species using PCR on DNA extracted from the formalin fixed paraffin embedded tissue failed. This is the first clinical and histological description of demodicosis in Eidolon helvum.

Identifying the tick Amblyomma javanense (Acari: Ixodidae) from Chinese pangolin: generating species barcode, phylogenetic status and its implication in wildlife forensics Abstract Zoonotic diseases transmitted through ticks and other ectoparasites often travel across the globe with illegally traded wildlife parts and products. In this study, we analyzed a confiscated case of pangolin scales and observed a few dead ticks attached. On genetic analysis, the pangolin scales were identified to be originated from Chinese pangolin (Manis pentadactyla), an IUCN listed Critically Endangered species, and ticks were identified as Amblyomma javanense. Here, we provide the first authentic physical record of A. javanense from India as a parasite of Chinese pangolin and also generated its species DNA barcode that may be useful for biologists working on ticks in species validation and constructing phylogenies across the globe.

Integrative taxonomy of Abacarus mites (Eriophyidae) associated with hybrid sugarcane plants, including description of a new species Abstract Phytophagous mites belonging to the Eriophyoidea are extremely diverse and highly host-specific. Their accurate morphological identification is hampered by their reduced size and simplified bodies and by the existence of cryptic species complexes. Previous studies have demonstrated the urgency of applying multisource methods to accurate taxonomic identification of eriophyoid mites, especially species belonging to the genus Abacarus. This genus comprises 65 species, of which 37 are associated with grasses and four with sugarcane Saccharum (Poaceae). Recently, Abacarus specimens very similar to Abacarus sacchari were collected from the sugarcane crop in Brazil; however, their taxonomic placement was uncertain. In this study, we used an integrative approach to determine whether A. aff. sacchari specimens belong to A. sacchari or constitute a cryptic species. Morphological data were combined with molecular phylogeny based on the nucleotide sequences of three markers, one mitochondrial (COI) and two nuclear (D2 region of 28S and ITS). Morphological differences were observed between A. aff. sacchari, A. sacchari and A. doctus. The phylogenetic relationships among these three taxa and the genetic distances separating them revealed an interspecific divergence. The results of the morphological and molecular methods were congruent and supported the existence of a new species: Abacarus neosacchari n. sp. Duarte and Navia, herein described. This species belongs to the Abacarus cryptic species complex associated with sugarcane in the Americas. The results of this study, presenting the occurrence of multiple Abacarus species associated with sugarcane, contribute to the knowledge on plants and mites diversity by adding up one more clue highlighting that plant hybridization can be an important mechanism contributing to the speciation of plant-feeding arthropods.

Proteomic analysis of saliva from partially and fully engorged adult female Rhipicephalus microplus (Acari: Ixodidae) Abstract Rhipicephalus microplus salivary gland secretes a number of complex bioactive proteins during feeding. These components are important in feeding and affect anti-coagulation, anti-inflammation and also have anti-microbial effects. In this study, tick saliva was collected from partially engorged female (PEF) and fully engorged female (FEF) ticks. Liquid chromatography tandem–mass spectrometry (LC–MS/MS) and isobaric tags for relative and absolute quantification (iTRAQ) were used to identify and quantify R. microplus salivary proteins. A total of 322 unique peptides were detected and 151 proteins were characterized in both PEF and FEF. Of these, 41 proteins are considered as high-confidence proteins. Fifteen high-confidence proteins were upregulated and six high-confidence proteins were downregulated (p < 0.05; PEF:FEF ratio ≥ 1.2 or PEF:FEF ratio ≤ 0.83); 17 high-confidence proteins are slightly changed (PEF:FEF ratio > 0.83 and < 1.2). These high-confidence proteins are involved in several physiological roles, including egg development, transportation of proteins, immunity and anti-microorganism, anti-coagulant, and adhesion. In comparison with PEF, the number of upregulated proteins exceeded the number of proteins downregulated. Salivary protein may be induced by the blood-meal and these proteins contribute to successful feeding.

Coexistence of genetically different Varroa destructor in Apis mellifera colonies Abstract The aim of this study was to investigate the genetic diversity of Varroa destructor parasitizing Apis mellifera colonies and to test for possible host–parasite association at the mitochondrial DNA (mtDNA) level. Six A. melliferahaplotypes (including a novel C2aa) and five haplotypes of V. destructor were detected in 29 analyzed colonies from eight sampling sites in Serbia. We revealed the presence of the K and S1 haplotypes as well as KS1 and KP1 heteroplasmic mite individuals in all localities, while the P1 haplotype was only found in four sampling sites. Significant differences in V. destructor genetic diversity were found at both apiary and colony levels, with mite haplotypes coexisting in almost all tested colonies. In addition, a significant correlation between the number of analyzed mites per colony and the number of identified V. destructor haplotypes was observed. However, no significant host–parasite relationship was found, suggesting that mites bearing different haplotypes as well as those heteroplasmic individuals are well adapted to the host, A. mellifera, independently of the identified haplotype present in each colony. Our results will contribute to future population and biogeographic studies concerning V. destructor infesting A. mellifera, as well as to better understanding their host–parasite relationship.

Optimizing cold storage of the ectoparasitic mite Pyemotes zhonghuajia (Acari: Pyemotidae), an efficient biological control agent of stem borers Abstract Pyemotes zhonghuajia Yu, Zhang and He is a newly discovered native ectoparasitic mite that efficiently controls stem borers in China. To provide a steady and sufficient supply, extend adult lifespan and synchronize field augmentative releases of P. zhonghuajia, we determined the optimal cold storage temperature and duration by storing 1-day-old mated females at 8, 10 and 12 °C for 10–90 days with a 10-day interval in the laboratory. We then recorded mite survival during storage and monitored the post-storage reproductive performance of mites at a control temperature of 25 °C. We found that all mites survived at 10 and 12 °C for different durations, but mortality occurred when mites were stored at 8 °C for ≥ 30 days with more than 70% of mites dead when the storage duration prolonged up to 50 days. The proportion of reproductive females was higher at 10 °C but decreased with the prolonged storage duration at all test temperatures. Storage temperature had no significant effect on the pre-reproductive period and offspring sex ratio, whereas prolonged storage induced longer pre-reproductive period and lower proportion of female offspring. The reproductive period increased with increasing storage temperature and with prolonged storage up to 50 and 60 days; however, the longer reproductive period did not directly translate into greater reproductive output. We found that compared with the control, mites stored at 10 °C for up to 30 days did not significantly reduce their survival, proportion of reproductive success and number and sex ratio of offspring, suggesting that 10 °C and ≤ 30 days were the optimal cold storage temperature and duration, respectively, for post-mass production storage before the field augmentative release of P. zhonghuajia.

Resistance incidence and presence of resistance mutations in populations of Tetranychus urticae from vegetable crops in Turkey Abstract Tetranychus urticae Koch is one of the most common and harmful pests in vegetable production areas. Similar to other countries, control of T. urticae is mainly based on acaricides in Turkey. However, T. urticae rapidly develops resistance and failures in chemical control have occurred frequently. The toxicity of various acaricides was investigated in ten T. urticae populations collected from vegetable crops in Turkey. In addition, populations were screened for the presence of currently known target-site resistance mutations. It was shown that resistance to bifenthrin was the most widespread, but also half of the populations were resistant to abamectin and hexythiazox. Resistance mutations in the voltage-gated sodium channel (VGSC) and chitin synthase 1 were found in various populations. Moreover, for the first time, F1538I and L1024V VGSC mutations were reported for Turkish populations. Mutations that confer resistance to abamectin, bifenazate and METI-I acaricides such as pyridaben were not detected. These results will contribute to the design of an effective resistance management program in Turkey.

A relapsing fever Borrelia and spotted fever Rickettsia in ticks from an Andean valley, central Chile Abstract In humans, emerging infectious diseases are mostly zoonoses with ticks playing an important role as vectors. Tick-borne relapsing fever Borrelia and spotted fever Rickettsia occur in endemic foci along tropical and subtropical regions of the globe. However, both are widely neglected etiologic agents. In this study, we performed molecular analyses in order to assess the presence of Borrelia and Rickettsia DNA in ticks infesting small-mammals within a National Reserve located in the Andes Mountains, central Chile. While hard ticks were negative for the presence of both agents, sequences of four rickettsial (gltA, htrA, ompA, ompB) and two borrelial (16S rRNA and flaB) genes were obtained from larvae of an Ornithodoros sp. morphologically related with Ornithodoros atacamensis. Phylogenetic analyses indicated that the detected Borrelia and Rickettsia spp. belong to the relapsing fever and spotted fever groups, respectively. Moreover, the agents formed monophyletic clades with Rickettsia amblyommatis and "Candidatus Borrelia johnsonii." As positive ticks parasitize rodents within a highly visited National Reserve where outdoor activities are of common practice, the risk for human parasitism should not be discarded.

Expression dynamics of key ecdysteroid and juvenile hormone biosynthesis genes imply a coordinated regulation pattern in the molting process of a spider mite, Tetranychus urticae Abstract In insects, the ecdysteroid 20-hydroxyecdysone coordinates with juvenile hormone (JH) to regulate the process of molting, development and metamorphosis; however, this interaction is still unclear in the mites. In this study, we investigated the gene related to ecdysteroid and JH biosynthesis pathways, including four ecdysteroid and 11 JH biosynthesis genes. We examined their expression patterns during molting of different developmental stages of the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), an important agricultural pest that feeds on more than 1100 plant species. The expression of ecdysteroid biosynthesis Halloween genes exhibited a positive zigzag-like pattern, with a peak after 8 h of molting and a drop 8 h after entering each quiescent stage. In contrast, JH biosynthesis genes expression displayed a negative zigzag-like pattern, with a peak at 8 h after entering each quiescent stage and a drop after 8 h of each molting. These opposite patterns imply that ecdysteroid and JH expression is coordinated during the developmental transition. Our data provide an initial perspective on the co-expression of ecdysteroid and JH biosynthesis genes to regulate this important developmental process in the two-spotted spider mite.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Histochemistry and Cell Biology

Immune response profile of primary tumour, sentinel and non-sentinel axillary lymph nodes related to metastasis in breast cancer: an immunohistochemical point of view Abstract Approximately 1.67 million new cases of breast cancer (BC) are diagnosed annually, and patient survival significantly decreases when the disease metastasizes. The axillary lymph nodes (ALNs) are the main doorway for BC tumoral cell escape, through which cells can disseminate to distant organs. The immune response, which principally develops in the lymph nodes, is linked to cancer progression, and its efficacy at controlling tumoral growth is compromised during the disease. Immunohistochemistry (IHC) is one of the most widely used research techniques for studying the immune response. It allows the measurement of the expression of particular markers related to the immune populations. This review focuses on the role of the immune populations in the primary tumour in the locoregional metastasis of the ALN, and the relationship of the immune response in these regions to distant metastasis. We considered only studies of immune cells using IHC techniques. In particular, lymphocytes, macrophages and dendritic cells all play important roles in BC and have been extensively studied. Although further research is needed, there is much evidence of their role in the invasion of the ALN and distant organs. Their association with tumoral growth or protection has not yet been demonstrated decisively and is very likely to be determined by a combination of factors. Moreover, even though IHC is a widely used technique in cancer diagnosis and research, there is still room for improvement, since its quantification needs to be properly standardized.

In focus in HCB

Correction to: Similarities and differences in tissue distribution of DLK1 and DLK2 during E16.5 mouse embryogenesis In the original publication of the article, some symbols in Figure 3 were not correctly aligned with the image.

Regulatory effects of PRF and titanium surfaces on cellular adhesion, spread, and cytokine expressions of gingival keratinocytes Abstract Dental implant material has an impact on adhesion and spreading of oral mucosal cells on its surface. Platelet-rich fibrin (PRF), a second-generation platelet concentrate, can enhance cell proliferation and adhesion. The aim was to examine the regulatory effects of PRF and titanium surfaces on cellular adhesion, spread, and cytokine expressions of gingival keratinocytes. Human gingival keratinocytes were cultured on titanium grade 4, titanium grade 5 (Ti5), and HA discs at 37 °C in a CO2 incubator for 6 h and 24 h, using either elutes of titanium-PRF (T-PRF) or leukocyte and platelet-rich fibrin (L-PRF), or mammalian cell culture medium as growth media. Cell numbers were determined using a Cell Titer 96 assay. Interleukin (IL)-1β, IL-1Ra, IL-8, monocyte chemoattractant protein (MCP)-1, and vascular endothelial growth factor (VEGF) expression levels were measured using the Luminex® xMAP™ technique, and cell adhesion and spread by scanning electron microscopy. Epithelial cell adhesion and spread was most prominent to Ti5 surfaces. L-PRF stimulated cell adhesion to HA surface. Both T-PRF and L-PRF activated the expressions of IL-1 β, IL-8, IL-1Ra, MCP-1, and VEGF, T-PRF being the strongest activator. Titanium surface type has a regulatory role in epithelial cell adhesion and spread, while PRF type determines the cytokine response.

Histochemistry and Cell Biology : 61 years and not tired at all

Region-specific changes in brain kisspeptin receptor expression during estrogen depletion and the estrous cycle Abstract Kisspeptin acts as a potent neuropeptide regulator of reproduction through modulation of the hypothalamic–pituitary–gonadal axis. Previous studies revealed sex differences in brain expression patterns as well as regulation of expression by estrogen. Alternatively, sex differences and estrogen regulation of the kisspeptin receptor (encoded by Kiss1r) have not been examined at cellular resolution. In the current study, we examined whether Kiss1r mRNA expression also exhibits estrogen sensitivity and sex-dependent differences using in situ hybridization. We compared Kiss1r mRNA expression between ovariectomized (OVX) rats and estradiol (E2)-replenished OVX rats to examine estrogen sensitivity, and compared expression between gonadally intact male rats and female rats in diestrus or proestrus to examine sex differences. In OVX rats, E2 replenishment significantly reduced Kiss1r expression specifically in the hypothalamic arcuate nucleus (ARC). A difference in Kiss1r expression was also observed between diestrus and proestrus rats in the hypothalamic paraventricular nucleus (PVN), but not in the ARC. Thus, estrogen appears to have region- and context-specific effects on Kiss1r expression. However, immunostaining revealed minimal colocalization of estrogen receptor alpha (ERα) in Kiss1r-expressing neuronal populations of ARC and PVN, indicating indirect or ERα-independent regulation of Kiss1r expression. Surprisingly, unlike the kisspeptin ligand, no sexual dimorphisms were observed in either the brain distribution of Kiss1r expression or in the number of Kiss1r-expressing neurons within enriched brain nuclei. The current study reveals marked differences in regulation between kisspeptin and kisspeptin receptor, and provides an essential foundation for further study of kisspeptin signaling and function in reproduction.

A temporal and spatial map of axons in developing mouse prostate Abstract Prostate autonomic and sensory axons control glandular growth, fluid secretion, and smooth muscle contraction and are remodeled during cancer and inflammation. Morphogenetic signaling pathways reawakened during disease progression may drive this axon remodeling. These pathways are linked to proliferative activities in prostate cancer and benign prostate hyperplasia. However, little is known about which developmental signaling pathways guide axon investment into prostate. The first step in defining these pathways is pinpointing when axon subtypes first appear in prostate. We accomplished this by immunohistochemically mapping three axon subtypes (noradrenergic, cholinergic, and peptidergic) during fetal, neonatal, and adult stages of mouse prostate development. We devised a method for peri-prostatic axon density quantification and tested whether innervation is uniform across the proximo–distal axis of dorsal and ventral adult mouse prostate. Many axons directly interact with or innervate neuroendocrine cells in other organs, so we examined whether sensory or autonomic axons innervate neuroendocrine cells in prostate. We first detected noradrenergic, cholinergic, and peptidergic axons in prostate at embryonic day (E) 14.5. Noradrenergic and cholinergic axon densities are uniform across the proximal–distal axis of adult mouse prostate while peptidergic axons are denser in the periurethral and proximal regions. Peptidergic and cholinergic axons are closely associated with prostate neuroendocrine cells whereas noradrenergic axons are not. These results provide a foundation for understanding mouse prostatic axon development and organization and, provide strategies for quantifying axons during progression of prostate disease.

High-throughput quantification of the effect of DMSO on the viability of lung and breast cancer cells using an easy-to-use spectrophotometric trypan blue-based assay Abstract One of the main aspects investigated in potential therapeutic compounds is their effect on cells viability and proliferative ability. Although various methods have been developed to investigate these aspects, these methods present with shortcomings in terms of either cost, availability, accuracy, precision, or throughput. This study describes a simple, economic, reproducible, and high-throughput assay to quantify cell death and proliferation. In this assay, adherent cells are fixed, stained with trypan blue, and measured for trypan blue internalization using a spectrophotometric absorbance plate reader. Corresponding cell counts to the absorbance measurements are extrapolated from a standard curve. This assay was used to measure the effect of dimethyl sulfoxide (DMSO) on the viability of breast and lung cancer cells. Decrease in cell count associated with the increase in DMSO percentage and exposure time. The assay's results closely correlated with the conventional trypan blue exclusion assay (Pearson correlation coefficient (r) > 0.99; p < 0.0001), but with higher precision. The assay developed in this study can be used for various applications such as optimization, cell treatment investigations, proliferation, and cytotoxicity studies.

Similarities and differences in tissue distribution of DLK1 and DLK2 during E16.5 mouse embryogenesis Abstract DLK1 and DLK2 are transmembrane proteins belonging to the EGF-like repeat-containing family that function as non-canonical NOTCH inhibitory ligands. DLK1 is usually downregulated after embryo development and its distribution in some adult and embryonic tissues has been described. However, the expression and role of DLK2 in embryo and adult tissues remains unclear. To better understand the relevance of both proteins during embryo development, we analyzed the expression pattern of DLK1 and DLK2 in 16.5-day-old mouse embryos (E16.5) and evaluated the possible relationship between these two proteins in embryo tissues and cell types. We found that DLK1 and DLK2 proteins exhibited a broad distribution pattern, which was detected in developing mouse organs from each of the three germ layers: ectoderm (brain, salivary glands), mesoderm (skeletal muscle, vertebral column, kidney, cartilage), and endoderm (thymus, lung, pancreas, intestine, liver). The expression pattern of DLK1 and DLK2 indicates that both proteins could play a synergistic role during cell differentiation. This study provides additional information for understanding temporal and site-specific effects of DLK1 and DLK2 during embryo morphogenesis and cell differentiation.

Quantitative pixel intensity- and color-based image analysis on minimally compressed files: implications for whole-slide imaging Abstract Current best practice in the quantitative analysis of microscopy images dictates that image files should be saved in a lossless format such as TIFF. Use of lossy files, including those processed with the JPEG algorithm, is highly discouraged due to effects of compression on pixel characteristics. However, with the growing popularity of whole-slide imaging (WSI) and its attendant large file sizes, compressed image files are becoming more prevelent. This prompted us to perform a color-based quantitative pixel analysis of minimally compressed WSI images. Sections from three tissues stained with one of three reagents representing the colors blue (hematoxylin), red (Oil-Red-O), and brown (immunoperoxidase) were scanned with a whole slide imager in triplicate at 20x, 40x, and 63x magnifications. The resulting files were in the form of a BigTIFF with a JPEG compression automatically applied during acquisition. Images were imported into analysis software, six regions of interest were applied to various morphological locations, and the areas assessed for the color of interest. Whereas the number of designated weakly or strongly positive pixels was variable across the triplicate scans for the individual regions of interest, the total number of positive pixels was consistent. These results suggest that total positivity for a specific color representing a histochemical or immunohistochemical stain can be adequately quantitated on compressed images, but degrees of positivity (e.g., weak vs. strong) may not be as reliable. However, it is important to assess individual whole-slide imagers, file compression level and algorithm, and analysis software for reproducibility.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Toxicology and Environmental Health Sciences

Level of Petroleum Hydrocarbons in Water and Sediments of Ikoli Creek Bayelsa State Nigeria Abstract Objective This study investigated the level Petroleum hydrocarbon in Ikoli creek, Bayelsa state Nigeria with focus on water and sediments. Methods Water and sediments were assessed using standard analytical procedures. Water from surface levels and sediment samples were collected from five locations in the creek. Soxhlet extraction method and liquid-liquid techniques were used in extraction of the petroleum hydrocarbons (Total Hydrocarbon Content and Total Petroleum Hydrocarbon) from the water and sediment samples collected respectively followed by column clean up. Gas Chromatography-Flame Ionization Detector (GC-FID) was use to analyzed the Target compounds and quantified by integrating the areas of both the resolved and unresolved components. Results Results of analysis of THC and TPH in water ranges between 0.010 to 0.254 mg/L and Total Petroleum Hydrocarbon (TPH) values ranges between 0.001 to 0.437 mg/L in water respectively. While in sediments the Total Hydrocarbon Content (THC) values ranges between 0.01 to 1.43 mg/kg and Total Petroleum Hydrocarbon (TPH) ranges between 0.001 to 0.44 mg/kg. The THC and TPH value of sediments increases across each sampling station in the study area and these hydrocarbons are repository in nature. Conclusions The results showed that some sampling station in Ikoli creek were slightly polluted with petroleum hydrocarbons and does not conform to acceptable international guideline, which can cause adverse effects to humans and aquatic life over time.

Ameliorating Effects of Quercetin in a Diabetic Rat Model with Social Anxiety Disorder Abstract Objective The present study aimed to investigate the effects of quercetin on diabetes-induced behavioural, metabolic and histopathological alterations in a rat model that mimics the development of social anxiety disorder in diabetic humans. Methods Diabetes was induced by an intraperitoneal injection of alloxan (150 mg/kg bw). Three weeks later, the initial animal groups (control and diabetic) were assigned to three paralleled experiments during which quercetin was orally administered at a relatively high dose (100 mg/kg bw). The behavioural assessment included the forced swim test, the elevated plus-maze, the open field and the social interaction tests. The plasma glucose, triglycerides, cholesterol, urea and creatinine levels were evaluated, whereas the histopathology of the prefrontal cerebral cortex, liver and kidneys were checked out. Results Diabetes was associated with anxiety-like, depressive-like and defective social behaviours. The metabolic parameters were significantly elevated along with a histological damage to hepatic, renal and pre-frontal cerebral tissues. Quercetin treatment in diabetic rats alleviated the behavioural disorders while improving the social interactions. A significant decrease in all metabolic parameters was found following quercetin intake, which were accompanied by a better histo-pathological profile of the studied tissues. Conclusion Our findings suggest that the pathophysiology of diabetes is tightly linked to anxiety-like, depression-like, and deteriorated social behaviours, which may originate from a severe damage to the prefrontal cerebral cortex integrity. Interestingly, a high-dose short treatment with quercetin may mitigate the diabetes-related neurobehavioural, metabolic and histological disturbances, and prevent the defective sociability in long-term diabetic subjects.

Increased Blood Lead Level Alters the Heme Biosynthesis and Haematological Parameters of Spray Painters from Western Maharashtra, India Abstract Objective To assess the present status of blood lead levels and their effect on heme biosynthesis and hematological parameters of spray painters. Methods Forty two spray painters having the age range of 25–50 years were recruited for the study from Western Maharashtra, India and compared with age matched 50 healthy male subjects. Blood lead (PbB), erythrocytes d-ALDH, Urinary d-ALA, PBG and hematological parameters were measured. Results PbB levels (p<0.001, 458%) of spray painters were significantly increased and Non-activated δ-ALAD (p<0.001, -20.21%), activated δ-ALAD (p<0.001, -28.25%) were significantly decreased and ratio of activated to non-activated δ-ALAD (p<0.001, 16%) was significantly increased in spray painters as compared to control subjects. Urinary δ-ALA (p<0.001, 160%) and PBG (p<0.001, 62.75%) levels were significantly increased in spray painters as compared to the control group. Hematological parameters of spray painters i.e. Hb (p<0.001, -21.58%) PCV (p<0.05, -4.53%) MCV (p<0.01, -5.09%) MCH (p<0.01, -6.04), MCHC (p<0.01, - 2.31) and RBC (p<0.001, - 8.06 %) were significantly decreased while total WBC count (p<0.001, 34.63%) was significantly increased as compared to the controls. Conclusion Increased PbB level in spray painters may be due to more absorption of lead thereby inhibiting heme biosynthesis enzymes and resulting in more excretion of urinary δ-ALA, PBG and alteration of hematological parameters. Therefore it is essential to monitor these spray painters and also it is necessary to create awareness for using adequate protective measures. Also necessary training and regular screening is required to minimize adverse impact of lead exposure to them.

Effect of Food Colorants and Additives on the Hematological and Histological Characteristics of Albino Rats Abstract Objective Food colorants (synthetic, inorganic and natural) represent one of the major categories of food additives. Synthetic colorants were banned due to their adverse effects on animals and human. However, synthetic dyes are still in use because they are cheap, and stable. Methods In the present in vivo study conducted on female albino rats (Rattus norvegicus), the effect of sunset yellow (SY) and sodium benzoate (NaB) combinations on the hematological and histological profile was assessed. Different combinations of SY plus NaB were dissolved in water and administered daily to experimental rat groups for 12 weeks. Group 1 (control) received only water, group 2 received 5 mg SY plus 10 mg NaB, group 3 received 5 mg SY plus 100 mg NaB, group 4 received 50 mg SY plus 100 mg NaB, group 5 received 50 mg SY plus 10 mg NaB, group 6 received 200 mg SY plus 750 mg NaB, and group 7 received 20 mg SY plus 75 mg NaB. Histopathological examinations were performed on liver and kidney of rats at the end of the experiment. Results The results revealed a decrease in RBCs count, hematocrit, WBCs, MCV and Hb levels upon the administration of SY plus NaB. The results also showed no increase in MCH, MCHC and platelet count. Liver and kidney tissues showed some lesions due to the administration of the tested compounds in comparison to the control animals. Conclusion The chemical stress caused by the SY and NaB combinations caused some degenerative changes in the liver and kidneys of rats. It could be concluded that SY and NaB combinations causes some damage in liver and kidney tissues of experimental animals. Therefore, using SY and NaB combinations should be limited.

Effects of Repeated Pulmonary Exposure to Carbon Nanotubes on Lung function Abstract Objective The development of nanotechnology has increased the risk of exposure to types of particles other than combustion-derived ones in the environment, namely, industrial nanomaterials. However, their health impacts have not been fully elucidated yet. This study examined the effects of pulmonary exposure to two types of carbon nanotube (single-walled carbon nanotubes: SWCNT, and multi-walled carbon nanotube: MWCNT) on lung physiology in mice. Methods ICR mice were divided into three experimental groups to repeatedly administer vehicle, SWCNT (50 μg/animal), or MWCNT through intratracheal route; thereafter lung function was examined. Results The values of total respiratory system resistance (R: P<0.05), elastance (E), Newtonian resistance (Rn), tissue damping (G: P<0.05), and tissue elastance (H) were higher and the value of compliance (C) was lower in the SWCNT group than in the vehicle group. On the other hand, MWCNT tended to augment the values. Conclusions Taken together, CNT themselves inhaled, can impair airway function.

Ameliorative Effect of Zinc Oxide Nanoparticles on Nicotine Induced Testicular Dysfunction; Biochemical and Histological Study Abstract Objective The aim of this study was to evaluate potential impact of zinc oxide nanoparticle on the possible testicular toxicity induced by nicotine. Methods 24 adult male albino rats were divided into four groups; negative control group, zinc oxide nanoparticle group: rats were received 5 mg/kg/day zinc oxide nanoparticles orally, nicotine treated group: rats were administered nicotine (1 mg/kg/day) intraperitoneally and nicotine with zinc oxide nanoparticles: rats were treated with nicotine and zinc oxide nanoparticles daily for 28 days. Serum testosterone level, testicular malondialdehyde, SOD and catalase activities were measured. Testes were examined by light and electron microscopes as well. Results Significant decreases of serum testosterone level, testicular SOD and catalase activity and significant increase of testicular MDA in nicotine treated group when compared with control group were detected. Zinc oxide nanoparticles administration reversed these changes. Conclusion Zinc oxide nanoparticles ameliorated nicotine induced testicular dysfunction.

Enhancement of Alcohol Metabolism by Ginseng Berry Extract and its Mixed Herbal Beverages: In vitro and in vivo Experiments Abstract Objective To investigate the enhancement of alcohol metabolism by two ginseng berry (GB) extracts and their two types of mixed herbal beverages through in vitro and in vivo experiments. Methods Two GB extract solutions and their two herbal beverages were evaluated as enhancers of alcohol metabolism in normal human embryonic liver cells (CL-48 cell line) through assays of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity. Cytotoxicity was also assessed in the same cell line using an MTT assay. Effects on alcohol metabolism were also observed in vivo through measurement of serum alcohol, acetaldehyde, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels in alcohol treated rats. Blood samples were serially collected at 1, 2, 3, 4, 6 and 8 hrs after a single ethanol (EtOH) treatment. A single treatment with the test samples was administered orally 30 minutes after EtOH treatment. Results GB extract solutions effectively elevated the ADH and ALDH activity observed in vitro, while no treatment- related cytotoxic effects were found with test samples t concentrations up to 100 mg/mL. Significantly lower (p<0.01 or p<0.05) serum alcohol and acetaldehyde content was observed in samples from treated rats than in those from control rats (EtOH only) 1 or 2 hrs after EtOH treatment. In addition, noticeable decreases were observed in serum AST and ALT levels in treated samples 8 hrs after EtOH administration. HM40, an herbal mixture containing GB extract (40 mg/75 mL of ginsenoside Re), showed betters enhancement of alcohol metabolism through ADH/ALDH activation, as well as related hepatoprotective effects. Results GB extracts effectively enhanced alcohol metabolism without cytotoxicity while also providing possible hepatoprotective effects that could serve as a functional ingredient in anti-hangover alternative therapies. These extracts are expected to be more effective when made into herbal mixture beverages.

In vitro Models for Chemical Toxicity: Review of their Applications and Prospects Abstract Since it has been considered acceptable to expose animal to conditions and procedures that would not be considered acceptable for human, animals have long been used as surrogates for human beings for in vivo toxicology. This has focused mainly on gaining a better understanding of how the body's cells, organs, and systems function and are controlled, which failures could lead to pathological conditions. However, attitudes that rely on this approach are changing as the know ledge gained from animal studies is not directly related to humans, and there is a growing perception that it can also have dangerous consequences. In this re view, I collected the references about not only in vitro models for chemical toxicity, but also their applications and prospects focused on chemical hazard evaluations. In this review, I try to gather the issues and considerations that are considered necessary to develop a viable and reliable testing strategy to replace animals in toxicity testing. Most of the authors of the papers cited here share the opinion that animals will no longer be used for safety assessment, but that a more predictable human-derived in vitrosystem and in silico approach will be developed step-by-step. I collected information from peer-reviewed literature on the in vitro models for toxicological investigation of chemicals that may impact environmental or occupational diseases.

Chemical Composition and Biological Activity of Essential Oils of Centella asiatica (L.) Abstract Objective Centella asiatica (L.) is belonging the Apiaciaea family that is very popular for its culinary and medicinal use. In recent years, due to overexploitation, and ineffectual efforts to revitalize this valuable herbal source, the abundance of C. asiatica (L.) in its habitats has suffered significantly, cause to the risk of its extinction. Therefore, its in vitro production interested considerably. Methods In this study, mass production of C. asiatica (L.) was carried out through a cost effective micropropagation protocol and the chemical content of essential oils from micropropagated plants were analyzed by Gas chromatography (GC) and Gas chromatography-mass spectrometry (GC/MS) and biological properties compared with wild plants. Results In total, 45 compounds were identified and quantified in wild and micropropagated plants, representing 97.5% and 97.7% of the total oil, respectively. Also the oils were dominated by oxygenated sesquiterpenes, 70.8% and 42.6%, in the wild and micropropagated plants, respectively. Antibacterial assay showed the oil is effective regarding bacterial growth inhibition. The inhibition zones against some bacterial strains (Bacillus megaterium, Staphylococcus aureus, Klebsiella aerogenes) were even greater than those of chloramphenicol. The MIC of the oils, against the tested organisms, varied between 1 and 4μL mL−1. The lowest MIC (1 μL mL−1) was detected for the oil against Enterobacter aerugenes and Staphylococcous subrogation. Results were also indicated that the oil of micropropagated plants was more effective than the oils of wild ones. Cytotoxicity assay showed that the toxicity of the studied essential oils are not significant at concentrations of 5–15% (v/v). Conclusions It could concluded that the essential oil of C. asiatica, in micropropagated plants, have the good antibacterial activities without significant toxicity, at the applied concentration, and thus have great potentiality to be used as a resource for natural health products.

The Role of Aptamer Loaded Exosome Complexes in the Neurodegenerative Diseases Abstract Extracellular exosomes formed inside the cytoplasm of cells are small membranous vesicles that provide an important mode of cell-to-cell communication by delivering proteins, lipids and RNA to target cells. Exosomes are found associated with neurodegenerative diseases mainly Parkinson's disease (PD) and Alzheimer's diseases (AD), characterized by progressive degeneration of neurons and often linked with misfolded protein. Aptamers are single stranded DNA- and RNA-based oligonucleotides that bind target protein with high affinity and specificity and block or prevent the process of neurodegenerative diseases. In this review, we discuss the role of aptamer loaded exosomes in the neurode-generative diseases.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Der Nephrologe

Katheterinterventionen und interventionelle Shuntanlagen Zusammenfassung Hintergrund Das Leben der Dialysepatienten hängt unbestritten vom vaskulären Zugang ab. Standard ist die chirurgische Schaffung eines Shunts (arteriovenöse Fistel, AVF), was besondere Vorbereitung und Expertise erfordert. Eine besondere Herausforderung sind die Shuntreifung und die Versorgung der shuntassoziierten Probleme mit dem Ziel, den Patienten eine möglichst reibungslose Dialyse langfristig zu ermöglichen und zentrale Venenkatheter zu ersparen. Dies erfordert die Versorgung mittels endovaskulärer Techniken und chirurgischer Revisionen. Es existieren multiple Instrumente, die für die AVF-Versorgung zur Verfügung stehen. Des Weiteren wurden über die letzten Jahre Instrumentation und Techniken entwickelt, die eine endovaskuläre AVF-Anlage ermöglichen. Ziel der Arbeit Darstellung aktuell genutzter Katheterinterventionen und Beleuchtung verfügbarer Methoden der endovaskulären Shuntanlage. Material und Methoden In diesem Übersichtsartikel werden die Katheterinterventionen, Techniken, Komplikationen und Erfolge der jeweiligen Verfahren dargestellt. Ergebnisse Perkutane Angioplastien (PTA) mit verschiedenen Ballons, Stent- und Stentgraftanlage sowie endovaskuläre Thrombektomie mittels verschiedener Methoden haben das Armamentarium der Shuntversorgung über die letzten Jahre erweitert und verbessert und sind eine wertvolle Addition zu den bisherigen chirurgischen Verfahren zur Behandlung der Shuntdysfunktion. Die Möglichkeit der endovaskulären Shuntanlage eröffnet eine ganz neue Perspektive in der Kreierung des Dialysezugangs und weist bereits gute Früherfolge auf. Diskussion AVF-Anlage und -Versorgung sind ein zentrales und longitudinales Problem für Dialysepatienten. Es erfordert eine multidisziplinäre Zusammenarbeit. Kenntnis aller Verfahren und sorgfältige Selektion einer Methode für den individuellen Patienten bleiben von immenser Bedeutung.

Charakterisierung von Patienten mit atypischem hämolytisch-urämischen Syndrom (aHUS) in Deutschland

Physiologie und Pathophysiologie von FGF23 und Klotho

Von der chronischen Niereninsuffizienz zur Hämodialyse Zusammenfassung Jährlich beginnt in Deutschland eine hohe Anzahl von Patienten eine chronische Nierenersatztherapie, überwiegend in Form einer Hämodialyse(HD)-Therapie. Über diese Transitionsphase sind nur wenige gesicherte Daten verfügbar. Die ersten Monate einer HD-Therapie sind eine medizinische Hochrisikophase mit im Vergleich zu prävalenten HD-Patienten deutlich gesteigerter Mortalität. Risikofaktoren hierfür sind unter anderem Gefäßzugang via zentraler Venenkatheter und Malnutrition. Vorhandensein, längere Dauer und höhere Intensität einer nephrologischen Betreuung bei fortgeschrittener chronischer Nierenerkrankung (CKD) resultieren in einem besseren Outcome nach Transition zur HD. Studiendaten implizieren, dass ein späterer Beginn der Dialysetherapie (eGFR <10 ml/min/m2) im Vergleich zu einem früheren Beginn (eGFR <10 ml/min/m2) nicht mit einer schlechteren Prognose assoziiert ist; in die Entscheidung zur Einleitung einer Nierenersatztherapie müssen jedoch auch patientenindividuelle Aspekte einfließen. Welche weiteren Faktoren, Behandlungspraktiken und Patientencharakteristika bei fortgeschrittener CKD Einfluss auf die Prognose nach Dialysebeginn haben, ist nicht ausreichend bekannt; bisher liegen nur wenige Daten hierzu vor. Unser Ziel muss es sein, Therapiemodifikationen in der Prädialysephase zu identifizieren, die das Outcome nach Transition zur Dialysetherapie verbessern, um die mit dieser Phase verbundene exzeptionell hohe Mortalität zu reduzieren.

Genetische Nierenerkrankungen

„Übergänge" in der Nephrologie

29-jährige Patientin mit Syndrom der inadäquaten ADH-Sekretion unklarer Ursache

Der Sprung auf die Warteliste Zusammenfassung Der terminalen Niereninsuffizienz liegen ganz verschiedene Erkrankungen zugrunde, entsprechend unterscheiden sich die Dialysepatienten, die auf eine Transplantation hoffen, in weiten Bereichen. Per Gesetz ist vorgeschrieben, dass bei jedem Dialysepatienten die Eignung für eine Nierentransplantation geprüft werden muss. Aufgrund der großen bürokratischen Anforderungen des Gesetzgebers an diesen Prozess und der individuellen sehr hohen Krankheitsschwere von Dialysepatienten ist die Überprüfung der Eignung nicht leicht zu standardisieren, sehr zeitaufwändig und komplex. Zusätzlich hat ein Dialysepatient ein sehr hohes Risiko, Folgekrankheiten zu entwickeln, die eine Eignung zur Transplantation gefährden. Damit kommt der konstanten und konsequenten Überprüfung der Eignung der Patienten auch nach der Aufnahme auf die Warteliste (Wartelistenführung) eine große Bedeutung zu. Der Schlüssel für den Sprung auf die Warteliste und damit die erfolgreiche Transplantation ist eine enge und vertrauensvolle Kooperation zwischen dem Dialyse- und Transplantationszentrum. Leider führt die in Deutschland außerordentlich lange Wartezeit für ein Organ deswegen nicht nur zu einer Gefährdung des Transplantationserfolgs, sie erhöht gleichzeitig auch den Aufwand der Transplantationszentren durch eine verlängerte Wartelistenführung beträchtlich.

Einfluss des Spendenalters auf das Transplantatüberleben bei Kindern und Jugendlichen

Von der Kindersprechstunde in die Erwachsenensprechstunde Zusammenfassung Der Übergang von der Pädiatrie in die Erwachsenenmedizin stellt in der Nephrologie eine wichtige, aber auch schwierige Schnittstelle dar. Dies gilt insbesondere für Jugendliche mit chronischer Niereninsuffizienz und nach Nierentransplantation. Dieser Übersichtsartikel stellt Transitionsrichtlinien und -hürden dar. Psychosoziale und finanzielle Aspekte der Transition sowie Studien- und Berufswahl werden diskutiert. In den letzten Jahren der Betreuung in der Jugendmedizin müssen die Grundlagen für den Übergang, wie Schulungen und Förderung der Selbstständigkeit der Patienten, gelegt werden. In der Erwachsenennephrologie müssen zukünftig noch mehr Ressourcen für die Besonderheiten der transferierten Patienten geschaffen werden. Es ist insbesondere eine politische Aufgabe, die Finanzierung einer guten Transition zu ermöglichen.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com