Translational outcomes in a full gene deletion of ubiquitin protein ligase E3A rat model of Angelman syndrome.

Angelman syndrome (AS) is a rare neurodevelopmental disorder characterized by developmental delay, impaired communication, motor deficits and ataxia, intellectual disabilities, microcephaly, and seizures. The genetic cause of AS is the loss of expression of UBE3A (ubiquitin protein ligase E6-AP) in the brain, typically due to a deletion of the maternal 15q11-q13 region. Previous studies have been performed using a mouse model with a deletion of a single exon of Ube3a. Since three splice variants of Ube3a exist, this has led to a lack of consistent reports and the theory that perhaps not all mouse studies were assessing the effects of an absence of all functional UBE3A. Herein, we report the generation and functional characterization of a novel model of Angelman syndrome by deleting the entire Ube3a gene in the rat. We validated that this resulted in the first comprehensive gene deletion rodent model. Ultrasonic vocalizations from newborn Ube3am-/p+ were reduced in the maternal inherited deletion group with no observable change in the Ube3am+/p- paternal transmission cohort. We also discovered Ube3am-/p+ exhibited delayed reflex development, motor deficits in rearing and fine motor skills, aberrant social communication, and impaired touchscreen learning and memory in young adults. These behavioral deficits were large in effect size and easily apparent in the larger rodent species. Low social communication was detected using a playback task that is unique to rats. Structural imaging illustrated decreased brain volume in Ube3am-/p+ and a variety of intriguing neuroanatomical phenotypes while Ube3am+/p- did not exhibit altered neuroanatomy. Our report identifies, for the first time, unique AS relevant functional phenotypes and anatomical markers as preclinical outcomes to test various strategies for gene and molecular therapies in AS.

[How do residents in urology evaluate their daily routine at work-a survey analysis]. / Wie wird der Berufsalltag von Weiterbildungsassistenten in der Urologie in Deutschland bewertet ­ eine Fragebogenanalyse.

BACKGROUND: The changing conditions in German hospitals is causing a shortage of young people. In order to identify starting point for improvements, the Bavarian association of urologists in collaboration with the German Society of Residents in Urology (GeSRU) conducted an online survey among residents in urology in summer of 2017. MATERIALS AND METHODS: A standardised survey composed of 38 questions was distributed to participants through a mailing list of the GeSRU. Most questions were closed-ended; however, some did allow participants to respond by means of an open-ended answer. RESULTS: A total of 218 participants provided a total of 11,764 responses: 58% were female and 42% were male. Over 70% were aged between 31 and 35 years. In all, 29% of participants responded negatively to the question asking whether they feel like their medical studies at university prepared them well for the daily routine in their workplace. Participants particularly demanded a higher degree of practical experience during their studies, as well as more teaching of soft skills. In relation to choosing their specialisation, participants considered the intern year and their clinical traineeships as crucial factors. Participants did express appreciation of their field of specialization in relation to the broad range of available treatments, the opportunity of further specialising, the clientele of patients, the opportunity of working in a small team, innovations, and the high possibilities of opening their own medical practice. On a personal level, participants specifically wished for a more structured plan relating to their further internship, involving regular meetings. They also expressed the wish for more personalised career plans, more flexible work hours, and improved advanced training, both internally and externally. CONCLUSIONS: With the aim of making urology even more attractive, participants' wishes and suggestions should be taken into consideration. These, in general, involve a more structured training plan, better working conditions with part-time programmes, and improved surrounding conditions at the workplace, in particular for families.

[What do prostate cancer patients know about smoking? : Results of a bicentric questionnaire study (KRAUT study)]. / Was wissen Patienten mit einem Prostatakarzinom über das Rauchen? : Ergebnisse einer bizentrischen Fragebogenstudie (KRAUT-Studie).

BACKGROUND: According to the results of a recent meta-analysis, cancer-specific mortality of prostate cancer (PCA) patients is enhanced by 24 % in case of a positive smoking history with a dose-dependent impact of smoking. Until now it is unknown whether this information actually reaches the patients and how extensively an informational discussion about this topic is pursued by physicians. OBJECTIVE: Three study hypotheses were defined: (1) the knowledge of PCA patients about the potential relationship between tumor progression and cigarette consumption is low, (2) only in rare cases has a clear statement been provided by treating physicians including the explicit advice to stop smoking, and (3) there was a direct association between tumor stage and the extent of cigarette consumption. MATERIALS AND METHODS: A questionnaire comprising 23 items was developed and validated with 25 uro-oncological patients prior to study start. Between September 2013 and December 2014 a total of 124 PCA patients (median age 65 years) from two urology departments were included in this questionnaire-based survey. RESULTS: The study population comprised 43 % (n = 54), 39 % (n = 48), and 18 % (n = 22) nonsmokers, former smokers and active smokers, respectively. Active and former smokers differed insignificantly in the number of pack-years only (24.8 vs. 23.7 years, p = 0.995). Of the patients, 56 % regarded an influence of cigarette consumption on the PCA-specific prognosis as possible. However, because a significant (p < 0.001) number of patients wrongly suspected smoking to be causative for PCA development, their knowledge about PCA prognosis is supposedly not based on adequate knowledge. Two of 22 active smokers (9.1 %), 5 of 48 former smokers (10.4 %), and 2 of 54 nonsmokers (3.7 %) had an informational discussion with their urologist about the association of cigarette consumption and PCA-related prognosis (a further 9.1, 4.2 and 3.7 %, respectively, received this information solely from other medical specialties). Only 1 of 22 active smokers (4.5 %) was offered medical aids for smoking cessation by the general practitioner; none of the patients received such support by an urologist. There was no association between a positive smoking history or number of pack-years and PCA tumor stage. CONCLUSIONS: Education of PCA patients about the relationship between cigarette consumption and cancer-related prognosis is currently inadequate. Following the latest findings on this topic, urologists should pursue informational discussions with their patients, thereby strengthening their position as the primary contact person for decision making in PCA management.

Rethinking psychopharmacotherapy: The role of treatment context and brain plasticity in antidepressant and antipsychotic interventions.

Emerging evidence indicates that treatment context profoundly affects psychopharmacological interventions. We review the evidence for the interaction between drug application and the context in which the drug is given both in human and animal research. We found evidence for this interaction in the placebo response in clinical trials, in our evolving knowledge of pharmacological and environmental effects on neural plasticity, and in animal studies analyzing environmental influences on psychotropic drug effects. Experimental placebo research has revealed neurobiological trajectories of mechanisms such as patients' treatment expectations and prior treatment experiences. Animal research confirmed that "enriched environments" support positive drug effects, while unfavorable environments (low sensory stimulation, low rates of social contacts) can even reverse the intended treatment outcome. Finally we provide recommendations for context conditions under which psychotropic drugs should be applied. Drug action should be steered by positive expectations, physical activity, and helpful social and physical environmental stimulation. Future drug trials should focus on fully controlling and optimizing such drug×environment interactions to improve trial sensitivity and treatment outcome.

Brain serotonin deficiency leads to social communication deficits in mice.

A deficit in brain serotonin is thought to be associated with deteriorated stress coping behaviour, affective disorders and exaggerated violence. We challenged this hypothesis in mice with a brain-specific serotonin depletion caused by a tryptophan hydroxylase 2 (TPH2) deficiency. We tested TPH2-deficient (Tph2(-/-)) animals in two social situations. As juveniles, Tph2(-/-) mice displayed reduced social contacts, whereas ultrasonic vocalizations (USVs) were unchanged within same-sex same-genotype pairings. Interestingly, juvenile females vocalized more than males across genotypes. Sexually naive adult males were exposed to fresh male or female urine, followed by an interaction with a conspecific, and re-exposed to urine. Although Tph2(-/-) mice showed normal sexual preference, they were hyper-aggressive towards their interaction partners and did not vocalize in response to sexual cues. These results highlight that central serotonin is essential for prosocial behaviour, especially USV production in adulthood, but not for sexual preference.

Lack of parvalbumin in mice leads to behavioral deficits relevant to all human autism core symptoms and related neural morphofunctional abnormalities.

Gene mutations and gene copy number variants are associated with autism spectrum disorders (ASDs). Affected gene products are often part of signaling networks implicated in synapse formation and/or function leading to alterations in the excitation/inhibition (E/I) balance. Although the network of parvalbumin (PV)-expressing interneurons has gained particular attention in ASD, little is known on PV's putative role with respect to ASD. Genetic mouse models represent powerful translational tools for studying the role of genetic and neurobiological factors underlying ASD. Here, we report that PV knockout mice (PV(-/-)) display behavioral phenotypes with relevance to all three core symptoms present in human ASD patients: abnormal reciprocal social interactions, impairments in communication and repetitive and stereotyped patterns of behavior. PV-depleted mice also showed several signs of ASD-associated comorbidities, such as reduced pain sensitivity and startle responses yet increased seizure susceptibility, whereas no evidence for behavioral phenotypes with relevance to anxiety, depression and schizophrenia was obtained. Reduced social interactions and communication were also observed in heterozygous (PV(+/-)) mice characterized by lower PV expression levels, indicating that merely a decrease in PV levels might be sufficient to elicit core ASD-like deficits. Structural magnetic resonance imaging measurements in PV(-/-) and PV(+/-) mice further revealed ASD-associated developmental neuroanatomical changes, including transient cortical hypertrophy and cerebellar hypoplasia. Electrophysiological experiments finally demonstrated that the E/I balance in these mice is altered by modification of both inhibitory and excitatory synaptic transmission. On the basis of the reported changes in PV expression patterns in several, mostly genetic rodent models of ASD, we propose that in these models downregulation of PV might represent one of the points of convergence, thus providing a common link between apparently unrelated ASD-associated synapse structure/function phenotypes.

On the relationships between ultrasonic calling and anxiety-related behavior in rats

In the present review, the phenomenon of ultrasonic vocalization in rats will be outlined, including the three classes of vocalizations, namely 40-kHz calls of pups, and 22- and 50-kHz calls of juvenile and adult rats, their general relevance to behavioral neuroscience, and their special relevance to research on anxiety, fear, and defense mechanisms. Here, the emphasis will be placed on 40- and 22-kHz calls, since they are typical for various situations with aversive properties. Among other topics, we will discuss whether such behavioral signals can index a certain affective state, and how these signals can be used in social neuroscience, especially with respect to communication. Furthermore, we will address the phenomenon of inter-individual variability in ultrasonic calling and what we currently know about the mechanisms, which may determine such variability. Finally, we will address the current knowledge on the neural and pharmacological mechanisms underlying 22-kHz ultrasonic vocalization, which show a substantial overlap with mechanisms known from other research on fear and anxiety, such as those involving the periaqueductal gray or the amygdala.

On the relationships between ultrasonic calling and anxiety-related behavior in rats.

In the present review, the phenomenon of ultrasonic vocalization in rats will be outlined, including the three classes of vocalizations, namely 40-kHz calls of pups, and 22- and 50-kHz calls of juvenile and adult rats, their general relevance to behavioral neuroscience, and their special relevance to research on anxiety, fear, and defense mechanisms. Here, the emphasis will be placed on 40- and 22-kHz calls, since they are typical for various situations with aversive properties. Among other topics, we will discuss whether such behavioral signals can index a certain affective state, and how these signals can be used in social neuroscience, especially with respect to communication. Furthermore, we will address the phenomenon of inter-individual variability in ultrasonic calling and what we currently know about the mechanisms, which may determine such variability. Finally, we will address the current knowledge on the neural and pharmacological mechanisms underlying 22-kHz ultrasonic vocalization, which show a substantial overlap with mechanisms known from other research on fear and anxiety, such as those involving the periaqueductal gray or the amygdala.

Stress revisited: a critical evaluation of the stress concept.

With the steadily increasing number of publications in the field of stress research it has become evident that the conventional usage of the stress concept bears considerable problems. The use of the term 'stress' to conditions ranging from even the mildest challenging stimulation to severely aversive conditions, is in our view inappropriate. Review of the literature reveals that the physiological 'stress' response to appetitive, rewarding stimuli that are often not considered to be stressors can be as large as the response to negative stimuli. Analysis of the physiological response during exercise supports the view that the magnitude of the neuroendocrine response reflects the metabolic and physiological demands required for behavioural activity. We propose that the term 'stress' should be restricted to conditions where an environmental demand exceeds the natural regulatory capacity of an organism, in particular situations that include unpredictability and uncontrollability. Physiologically, stress seems to be characterized by either the absence of an anticipatory response (unpredictable) or a reduced recovery (uncontrollable) of the neuroendocrine reaction. The consequences of this restricted definition for stress research and the interpretation of results in terms of the adaptive and/or maladaptive nature of the response are discussed.

Reduced scent marking and ultrasonic vocalizations in the BTBR T+tf/J mouse model of autism.

Qualitative impairments in communication, such as delayed language and poor interactive communication skills, are fundamental to the diagnosis of autism. Investigations into social communication in adult BTBR T+tf/J (BTBR) mice are needed to determine whether this inbred strain incorporates phenotypes relevant to the second diagnostic symptom of autism, communication deficits, along with its strong behavioral phenotypes relevant to the first and third diagnostic symptoms, impairments in social interactions and high levels of repetitive behavior. The aim of the present study was to simultaneously measure female urine-elicited scent marking and ultrasonic vocalizations in adult male BTBR mice, in comparison with a standard control strain with high sociability, C57BL/6J (B6), for the assessment of a potential communication deficit in BTBR. Adult male BTBR mice displayed lower scent marking and minimal ultrasonic vocalization responses to female urine obtained from both B6 and BTBR females. Lower scent marking and ultrasonic vocalizations in a social setting by BTBR, as compared with B6, are consistent with the well-replicated social deficits in this inbred mouse strain. Our findings support the interpretation that BTBR incorporate communication deficits, and suggest that scent marking and ultrasonic vocalizations offer promising measures of interest in social cues that may be widely applicable to investigations of mouse models of autism.

Alpha-synuclein deficiency affects brain Foxp1 expression and ultrasonic vocalization.

Alpha-synuclein is an abundant protein implicated in synaptic function and plasticity, but the molecular mechanism of its action is not understood. Missense mutations and gene duplication/triplication events result in Parkinson's disease, a neurodegenerative disorder of old age with impaired movement and emotion control. Here, we systematically investigated the striatal as well as the cerebellar transcriptome profile of alpha-synuclein-deficient mice via a genome-wide microarray survey in order to gain hypothesis-free molecular insights into the physiological function of alpha-synuclein. A genotype-dependent, specific and strong downregulation of forkhead box P1 (Foxp1) transcript levels was observed in all brain regions from postnatal age until old age and could be validated by qPCR. In view of the co-localization and heterodimer formation of FOXP1 with FOXP2, a transcription factor with a well established role for vocalization, and the reported regulation of both alpha-synuclein and FOXP2 expression during avian song learning, we performed a detailed assessment of mouse movements and vocalizations in the postnatal period. While there was no difference in isolation-induced behavioral activity in these animals, the alpha-synuclein-deficient mice exhibited an increased production of isolation-induced ultrasonic vocalizations (USVs). This phenotype might also reflect the reduced expression of the anxiety-related GABA-A receptor subunit gamma 2 (Gabrg2) we observed. Taken together, we identified an early behavioral consequence of alpha-synuclein deficiency and accompanying molecular changes, which supports the notion that the neural connectivity of sound or emotion control systems is affected.

New insights into the relationship of neurogenesis and affect: tickling induces hippocampal cell proliferation in rats emitting appetitive 50-kHz ultrasonic vocalizations.

Adult hippocampal cell proliferation (HCP) has been associated with psychopathology, especially depression. However, it is controversial whether a constitutively low rate of HCP is a trait predisposing an individual to psychopathology or whether HCP varies with the subject's affective state. We made use of a so-far neglected measure of affect, namely ultrasonic vocalizations, to gain new insights into the relationship of HCP and affect. Rats emit distinct types of ultrasonic vocalizations, which serve as situation-dependent affective signals. In appetitive situations, rats produce 50-kHz-calls, whereas 22-kHz-calls occur in aversive situations. We applied a standardized protocol of repeated tickling and assessed tickling-induced ultrasonic vocalizations as an index of the animals affect. Stereological quantifications of 5-bromo-2'-deoxyuridine (BrdU) and proliferating-cells-nuclear-antigen (PCNA) immunolabeled cells were used to estimate the rate of cell proliferation in the subventricular zone and the subgranular zone of the dentate gyrus in the hippocampus. The rate of cell proliferation was compared between the groups of tickled vs. non-tickled rats and between subgroups of tickled rats defined by the effect of tickling on ultrasonic vocalizations. Tickling induced ultrasonic vocalizations in a subject-dependent manner. HCP correlated positively with appetitive 50-kHz-calls, but negatively with aversive 22-kHz-calls in individual animals, while cell proliferation in the subventricular zone was not associated with the emission of ultrasonic vocalizations. Repeated tickling did not change HCP in all rats, but increased HCP in the subgroup of rats, which experienced this procedure as appetitive, i.e. in rats emitting high numbers of 50-kHz-calls or low numbers of 22-kHz-calls. Together, these data indicate that the effect of tickling on HCP depends on an interaction between a predisposing trait and stimulation-dependent variations of the subject's affective state.

Rat ultrasonic vocalization in aversively motivated situations and the role of individual differences in anxiety-related behavior.

Our previous work has shown that male Wistar rats, although identical in breeder, age and housing conditions, can differ systematically in their anxiety-related behavior as measured in the elevated plus-maze. Since such individual dispositions can affect the responsiveness in other aversively motivated situations, we asked in a 1st experiment whether such rats might also differ in a test of conditioned fear. Based on their levels of spontaneous open arm avoidance in the elevated plus-maze, 20 adult male Wistar rats were divided into those with "high open arm" (HOA) versus "low open arm" (LOA) time. These rats were then tested in a standard fear conditioning paradigm. During the conditioning procedure, they received 6 tone (3 kHz, 20 s) and shock (0.5 mA, 0.5 s) pairings, each followed by a 60 s inter-stimulus interval. Conditioned responses to the tone were tested 24 h thereafter. During both days, freezing behavior and ultrasonic vocalization were measured. Differences in ultrasonic vocalization between HOA and LOA rats were detected during the conditioning day, where vocalization was more likely in LOA rats. Furthermore, LOA rats emitted calls with higher frequency components than HOA rats. On the subsequent day of testing, the number of animals vocalizing, and the rate of vocalization was decreased, and call differences between groups were no longer detectable. In freezing, differences between HOA and LOA rats were observed on the conditioning day, where LOA rats showed more freezing behavior during the tone/shock intervals. Also, on the test day, they showed more freezing behavior during the tone intervals compared to HOA rats. These results indicate that acute and conditioned responses of rats in a conventional fear conditioning paradigm can depend on individual dispositions of anxiety-related behavior as measured with the elevated plus-maze. In a 2nd experiment, we asked whether exposure to an elevated plus-maze would lead to ultrasonic vocalization, which we tested in rats which had been handled or non-handled prior to testing. Most importantly, we found that none of the animals displayed any vocalization in the plus-maze, neither during a 1st nor a repeated test 1 day later. These data are discussed with respect to the presumed role of ultrasonic vocalization in aversively motivated situations, and the mechanisms, which may account for the behavioral differences between HOA and LOA rats in such tests.

Simplified orthotopic ileocecal pouch (Mainz pouch) for bladder substitution.

OBJECTIVE: To report operative modifications that render construction of the orthotopic Mainz pouch more simple, namely formation of the pouch using absorbable staples and utilization of the ileocecal valve as an antireflux mechanism. PATIENTS AND METHODS: Between January 2001 and May 2002 out of 92 cystectomy cases, 21 patients (20 males and 1 female) underwent a simplified orthotopic Mainz pouch. The ureters were implanted via Wallace or Nesbit technique in the supravalvular portion of the ileocecal valve. The mean (range) age was 58 (43 - 74) years and the mean follow-up is 11.6 months (range 1 - 27). RESULTS: No staple-related complications were encountered. Four renal units (8.8 %) were dilated, one secondary to an impacted ureteral stone and three secondary to ureteral stenosis requiring reimplantation. At 6 months follow-up, 5 renal units showed reflux (11.1 %) but without any clinical symptoms. 86.5 % of patients void spontaneously without significant residual urine. Continence was achieved in 74 % of cases during day and in 74 % at night. CONCLUSION: The simplified Mainz pouch is a versatile form of continent urinary diversion. The use of absorbable staples has simplified the pouch creation and yields functional results which favorably compare with other pouch forms. The ileocecal valve serves as an anti-reflux-mechanism.

[Ischemic colitis as a primary manifestation of an embolizing small-cell bronchial carcinoma infiltrating a pulmonary vein]. / Ischämische Kolitis als Primärmanifestation eines in eine Pulmonalvene eingewachsenen, embolisierenden kleinzelligen Bronchialkarzinoms.

CASE REPORT: A 59-year-old male patient was admitted to hospital for general malaise with muscle and bone pain. Some weeks ago he had been treated with roxithromycin because of pneumonia. Physical examination revealed melena. Hemoglobin was 7.6 g/l. Endoscopy of the upper gastrointestinal tract did not show a bleeding source. Colonoscopy and histologic examination revealed hemorrhage caused by ischemic colitis. To rule out cardiac embolies in the presence of paroxysmal tachyarrhythmia absoluta a transesophageal echocardiogram was performed. A mediastinal mass infiltrating the upper left pulmonary vein was detected. The mass had not been visible on routine chest X-ray. Histologic examination of a bronchoscopically taken specimen revealed an oat-cell bronchial carcinoma. CONCLUSION: In this patient invasion of a pulmonary vein by the bronchial carcinoma lead to embolies which caused mesenterial ischemia. According to our researches this is the first report of ischemic colitis as a manifestation of bronchial carcinoma.