Body mass is associated with hibernation length, body temperature, and heart rate in free-ranging brown bears.

BACKGROUND: Despite centuries of research, debate remains on the scaling of metabolic rate to mass especially for intraspecific cases. The high variation of body mass within brown bears presents a unique opportunity to study the intraspecific effects of body mass on physiological variables. The amplitude of metabolic rate reduction in hibernators is dependent on body mass of the species. Small hibernators have high metabolic rates when euthermic but experience a drastic decrease in body temperature during torpor, which is necessary to reach a very low metabolic rate. Conversely, large hibernators, such as the brown bear (Ursus arctos), show a moderate decrease in temperature during hibernation, thought to be related to the bear's large size. We studied body mass, abdominal body temperature, heart rate, and accelerometer-derived activity from 63 free-ranging brown bears (1-15 years old, 15-233 kg). We tested for relationships between body mass and body temperature, heart rate, and hibernation duration. RESULTS: The smallest individuals maintained lower body temperatures during hibernation, hibernated longer, and ended hibernation later than large bears. Unlike body temperature, winter heart rates were not associated with body mass. In summer, the opposite pattern was found, with smaller individuals having higher body temperature and daytime heart rates. Body mass was associated with body temperature in the winter hypometabolic state, even in a large hibernating mammal. Smaller bears, which are known to have higher thermal conductance, reached lower body temperatures during hibernation. During summer, smaller bears had higher body temperatures and daytime heart rates, a phenomenon not previously documented within a single mammalian species. CONCLUSION: We conclude that the smallest bears hibernated more deeply and longer than large bears, likely from a combined effect of basic thermodynamics, the higher need for energy savings, and a lower cost of warming up a smaller body.

Arterial oxygenation and acid-base status before and during oxygen supplementation in captive European bison (Bison bonasus) immobilized with etorphine-acepromazine-xylazine.

Chemical immobilization of captive European bison (Bison bonasus) is often required for veterinary care, transportation, or husbandry practices playing an important role in conservation breeding and reintroduction of the species. We evaluated the efficiency and physiological effects of an etorphine-acepromazine-xylazine combination with supplemental oxygen in 39 captive European bison. Animals were darted with a combination of 1.4 mg of etorphine, 4.5 mg of acepromazine, and 20 mg of xylazine per 100 kg based on estimated body mass. Arterial blood was sampled on average 20 min after recumbency and again 19 min later and analyzed immediately with a portable i-STAT analyzer. Simultaneously, heart rate, respiratory rate, and rectal temperature were recorded. Intranasal oxygen was started after the first sampling at a flow rate of 10 mL.kg-1.min-1 of estimated body mass until the end of the procedure. The initial mean partial pressure of oxygen (PaO2) was 49.7 mmHg with 32 out of 35 sampled bison presenting with hypoxemia. We observed decreased respiratory rates and pH and mild hypercapnia consistent with a mild respiratory acidosis. After oxygen supplementation hypoxemia was resolved in 21 out of 32 bison, but respiratory acidosis was accentuated. Bison immobilized with a lower initial drug dose required supplementary injections during the procedure. We observed that lower mean rectal temperatures during the immobilization event were significantly associated with longer recovery times. For three bison, minor regurgitation was documented. No mortality or morbidity related to the immobilizations were reported for at least 2 months following the procedure. Based on our findings, we recommend a dose of 0.015 mg.kg-1 etorphine, 0.049 mg.kg-1 acepromazine, and 0.22 mg.kg-1 xylazine. This dose reduced the need for supplemental injections to obtain a sufficient level of immobilization for routine management and husbandry procedures in captive European bison. Nevertheless, this drug combination is associated with development of marked hypoxemia, mild respiratory acidosis, and a small risk of regurgitation. Oxygen supplementation is strongly recommended when using this protocol.

Examining the association of eating psychopathology with suicidality: Comparing cross-sectional and longitudinal tests of interpersonal-psychological mediators.

Eating disorders (EDs) are associated with high mortality rates from suicide. Empirical tests of the Interpersonal-Psychological Theory of Suicide (IPTS) have provided preliminary cross-sectional support for its application to individuals with EDs. Because IPTS seeks to predict development and changes in suicidal ideation (SI), longitudinal investigations are ideal. The purpose of this study was to conduct cross-sectional and longitudinal mediational tests of the effect of ED psychopathology on SI as explained by perceived burdensomeness, thwarted belongingness, and hopelessness. Participants were undergraduate students (N = 738) who completed self-report measures of ED symptoms and IPTS variables at up to three time points across 10 weeks. Multiple mediation analyses were conducted on cross-sectional and longitudinal data. Cross-sectional analyses indicate mostly consistent findings with existing literature; however, results from the longitudinal analyses failed to identify any mediational effects of ED psychopathology on SI. These differences emphasize the importance of empirical tests in both cross-sectional and longitudinal data. Given the inconsistent results, the utility of IPTS features in explaining the association between ED psychopathology and SI is unclear. Future studies should seek to replicate these findings using other methods of measurement across time (e.g., ecological momentary assessment) and within clinical ED samples.

The cost of research: Lasting effects of capture, surgery and muscle biopsy on brown bear (Ursus arctos) movement and physiology.

Animal models are a key component of translational medicine, helping transfer scientific findings into practical applications for human health. A fundamental principle of research ethics involves weighing the benefits of the research to society against the burden imposed on the animals used for scientific purposes. The utilisation of wild animals for research requires evaluation of the effects of capture and invasive sampling. Determining the severity and duration of these interventions on the animal's physiology and behaviour allows for refining study methodology and for excluding or accounting for biased data. In this study, 39 Scandinavian brown bears (Ursus arctos) captured either while hibernating in winter or via helicopter in summer and that underwent surgery as part of a human health project had their movement, body temperature and timing of onset of hibernation compared with those of 14 control bears that had not been captured during the same period. Bears captured in winter and summer showed decreased movement from den exit until late summer, compared to those in the control group. Bears captured in summer showed reduced movement and body temperature for at least, respectively, 14 and 3 days, with an 11% decrease in hourly distance, compared to pre-capture levels, but did not differ in the timing of hibernation onset. We reveal that brown bear behaviour and physiology can be altered in response to capture and surgery for days to months, post-capture. This has broad implications for the conclusions of wildlife studies that rely upon invasive sampling.

Body temperature patterns during pregnancy and parturition in moose.

Gestation and lactation have high energetic requirements. Up to three-fourths of the gestation period in moose (Alces alces) overlaps with the food-scarce period in winter. During this period, moose deal with the limited forage resources available through hypometabolism with decreased heart rate and body temperature (Tb). Body temperature is also an indicator of oestrus, pregnancy and parturition, which is well documented in several domestic species. In this study, we sought to determine if moose displayed a similar Tb pattern during pregnancy and parturition to domesticated ruminants, and if we could detect parturition by combining Tb and activity data. We studied the Tb pattern of 30 free-ranging adult female moose (≥1.5 years old), equipped with ruminal temperature loggers and GPS collars. We documented a 0.13-0.19°C higher Tb in pregnant compared to non-pregnant moose, depending on the study area with the Tb difference increasing along a south-north gradient, and a drop in Tb and in activity when parturition was imminent. Detection of parturition was highly successful when combining Tb and activity data with an accuracy of 91.5%. Our findings demonstrate that Tb responses to pregnancy and parturition in a wild capital-breeding ruminant are similar to those of domesticated ruminants.

Seasonality in Biological Rhythms in Scandinavian brown Bears.

Biological rhythms, such as rhythms in activity and body temperature, are usually highly synchronized and entrained by environmental conditions, such as photoperiod. However, how the expression of these rhythms changes during hibernation, when the perception of environmental cues is limited, has not yet been fully understood for all hibernators, especially in the wild. The brown bear (Ursus arctos) in Scandinavia lives in a highly seasonal environment and adapts to harsh winter conditions by exhibiting hibernation, characterized by reduced metabolism and activity. In this study, we aimed to explore the expression of biological rhythms in activity, body temperature and heart rate of free-ranging brown bears over the annual cycle, including active, hibernation and the transition states around den entry and exit. We found that rhythms in physiology and activity are mostly synchronized and entrained by the light-dark cycle during the bears' active state with predominantly diel and ultradian rhythms for body temperature, activity and heart rate. However, during hibernation, rhythms in body temperature and heart rate were considerably slowed down to infradian rhythms, influenced by the amount of snow in the denning area, whereas rhythms in activity remained diel. Rhythms in the transition states when bears prepared for entering or coming out of hibernation state displayed a combination of infradian and diel rhythms, indicating the preparation of the body for the change in environmental conditions. These results reveal that brown bears adjust their biological rhythms to the seasonal environment they inhabit. Rhythms in physiology and activity show simultaneity during the active state but are partly disconnected from each other during hibernation, when bears are most sheltered from the environment.

High concentrations of lead (Pb) in blood and milk of free-ranging brown bears (Ursus arctos) in Scandinavia.

Exposure to lead (Pb) is a global health problem for both humans and wildlife. Despite a dramatic decline in human Pb exposure following restrictions of leaded gasoline and industry and thereby an overall reduction of Pb entering the environment, Pb exposure continues to be a problem for wildlife species. Literature on scavenging terrestrial mammals, including interactions between Pb exposure and life history, is however limited. We quantified Pb concentration in 153 blood samples from 110 free-ranging Scandinavian brown bears (Ursus arctos), 1-25 years old, using inductively coupled plasma sector field mass spectrometry. We used generalized linear models to test effects of age, body mass, reproduction status and spatial distribution on the blood Pb concentrations of 56 female bears. We sampled 28 females together with 56 dependent cubs and paired their blood Pb concentrations. From 20 lactating females, we measured the Pb concentration in milk. The mean blood Pb concentration was 96.6 µg/L (range: 38.7-220.5 µg/L). Both the mean and range are well above established threshold concentrations for developmental neurotoxicity (12 µg/L), increased systolic blood pressure (36 µg/L) and prevalence of kidney disease in humans (15 µg/L). Lactating females had higher Pb blood concentrations compared to younger, non-lactating females. Blood Pb concentrations of dependent cubs were correlated with their mother's blood Pb concentration, which in turn was correlated with the Pb concentration in the milk. Life-long Pb exposure in Scandinavian brown bears may have adverse effects both on individual and population levels. The high blood Pb concentrations found in brown bears contrast the general reduction in environmental Pb contamination over the past decades in Scandinavia and more research is needed to identify the sources and pathways of Pb exposure in the brown bears.

Physiological and behavioural responses of moose to hunting with dogs.

Optimal management of hunted species requires an understanding of the impacts of hunting on both individual animal and population levels. Recent technological advancements in biologging enable us to obtain increasingly detailed information from free-ranging animals, covering longer periods of time, and providing the data needed to assess such impacts. In Sweden, more than 80 000 moose are harvested annually, mostly hunted with the use of baying dogs. The effects of this hunting method on animal welfare and stress are understudied. Here, we evaluated 6 real and 17 experimental hunting approaches with baying dogs [wearing global positioning system (GPS) collars] on 8 adult female moose equipped with ruminal temperature loggers, subcutaneous heart rate (HR) loggers and GPS collars with accelerometers. The obtained data were used to analyse the behavioural and physiological responses of moose to hunting with dogs. Successful experimental approaches (moose and dog were within 240 m for >10 min) resulted in higher maximum body temperature (Tb, 0.88°C higher) and a mean increase in HR of 24 bpm in moose at the day of the approach compared to the day after. The moose rested on average >90 min longer the day after the approach compared to the day of the approach. The moose travelled on average 4.2 km longer and had a 1.3 m/s higher maximum speed the day of the approach compared to the day after. Our results demonstrate that hunting with dogs increase moose energy expenditure and resting time (and consequently decrease time available for foraging) on an individual level. This could possibly affect body condition and reproduction rates if the hunting disturbances occur frequently.

Assessment of the LeadCare® Plus for Use on Scandinavian Brown Bears (Ursus arctos).

Lead (Pb) exposure is associated with adverse health effects in both humans and wildlife. Blood lead levels (BLL) of sentinel wildlife species can be used to monitor environmental lead exposure and ecosystem health. BLL analyzers, such as the LeadCare®, are validated for use in humans, assessed for use in some avian species and cattle, and are increasingly being used on wildlife to monitor lead exposure. The LeadCare® analyzers use a technique called anodic stripping voltammetry (ASV). Species-specific conversion equations have been proposed to approximate the levels found with gold standard measuring methods such as inductively coupled plasma mass spectrometry (ICP-MS) because the ASV method has been shown to underestimate BLL in some species. In this study we assessed the LeadCare® Plus (LCP) for use on Scandinavian brown bears (Ursus arctos). LCP measurements were correlated with ICP-MS with a Bland-Altman analyzed bias of 16.3-22.5%, showing a consistent overestimation of BLL analyzed with LCP. Based on this analysis we provide conversion equations for calculating ICP-MS BLL based on the LCP results in Scandinavian brown bears. Our study shows that the LeadCare® Plus can be used for monitoring of lead exposure by approximating gold standard levels using conversion equations. This enables comparison with other gold standard measured BLL within the observed range of this study (38.20-174.00 µg/L). Our study also found that Scandinavian brown bears are highly exposed to environmental lead.

Effects of reproduction and environmental factors on body temperature and activity patterns of wolverines.

BACKGROUND: Mammals in the far north are exposed to extreme seasonal changes in environmental conditions, such as temperature and photoperiod, which have notable effects on animal physiology and behaviour. The wolverine (Gulo gulo) is a carnivore with a circumpolar distribution and well-adapted to extreme environmental conditions. Still, ecophysiological studies on free-ranging wolverines are lacking. In this study, we used abdominally implanted body temperature loggers in combination with GPS collars with acceleration sensors on 14 free-ranging wolverines in northern Sweden to study daily and seasonal variation in body temperature and activity patterns. We used generalized additive mixed modelling to investigate body temperature patterns over time and Lomb-Scargle periodogram analysis to analyse circadian rhythms. RESULTS: We found that wolverines have an average core body temperature of 38.5 ± 0.2 °C with a daily variation of up to 6 °C. Body temperature patterns varied between reproductive states. Pregnant females showed a distinct decrease in body temperature during gestation. Wolverines were active both in day and night, but displayed distinct activity peaks during crepuscular hours. However, body temperature and activity patterns changed seasonally, with a gradual change from a unimodal pattern in winter with concentrated activity during the short period of day light to a bimodal pattern in autumn with activity peaks around dusk and dawn. Wolverines were less likely to display 24-h rhythms in winter, when hours of day light are limited. CONCLUSIONS: The combination of different biologging techniques gave novel insight into the ecophysiology, activity patterns and reproductive biology of free-ranging wolverines, adding important knowledge to our understanding of animals adapted to cold environments at northern latitudes.

Differentiating motives for nonmedical prescription stimulant use by personality characteristics.

Nonmedical prescription stimulant use (NPS) is increasing, particularly among college students. College students typically engage in NPS for cognitive enhancement, recreational, and appetite/weight-related purposes; however, little research has used these motives to identify specific risk for, or consequences of, NPS. Moreover, there may be unique risk factors for motive-specific NPS that have yet to be explored, such as relevant personality traits (i.e., distress tolerance, impulsivity, and perfectionism) that are associated with NPS in general. Therefore, this study aimed to examine whether NPS users and nonusers could be differentiated via facets of impulsivity, perfectionism, and distress tolerance, and whether users could be further differentiated by reported motive for use based on these traits. Midwestern university undergraduate students (N = 668) who were enrolled in a psychology research pool completed an online survey assessing demographics, NPS and motives, and measures of distress tolerance, impulsivity, and perfectionism. Participants were primarily female (78%) and aged 18-54 (M = 20.10, SD = 3.19) years. Univariate and multivariate analysis of variance tests revealed associations between lifetime NPS and higher impulsivity, higher perfectionism, and lower distress tolerance. Further tests revealed NPS for appetite/weight-related purposes was associated with lower distress tolerance, while NPS for recreational purposes was associated with higher impulsivity. These findings contribute novel information regarding NPS motives and personality constructs. This information may aid in comprehensive identification of high-risk individuals for NPS and inform the development of specialized prevention and intervention efforts.

Higher Fit-fOR-The-Aged (FORTA) Scores Comprising Medication Errors are Associated with Impaired Cognitive and Physical Function Tests in the VALFORTA Trial.

BACKGROUND: The Fit fOR The Aged (FORTA) list, a drug classification combining positive and negative labelling of drugs, has been clinically (VALFORTA-trial) validated to improve medication quality and clinical endpoints. OBJECTIVE: The objective of this study was to determine the association of medication quality with functional abilities tested in cognitive and physical function tests. PATIENTS AND METHODS: Data from the prospective, randomized controlled VALFORTA trial on 409 geriatric (mean age 81.53 years) in-hospital patients were tested for associations between the FORTA score (sum of over- and under-treatment errors) on admission and cognitive and physical function tests. Univariate and multivariate linear correlations corrected for age, sex, number of medications, number of chronic conditions, and body mass index as well as comparisons between high and low FORTA-score (cut-off 3) patients were performed. RESULTS: The FORTA score was significantly correlated with Instrumental Activities of Daily Living (p < 0.0001), the Tinetti test (p < 0.002), Essen Questionnaire on Age and Sleepiness (p < 0.0001), Mini-Mental State Examination (p < 0.0001), and handgrip strength (p < 0.04) in the univariate analysis, and with Instrumental Activities of Daily Living (p < 0.003), the Tinetti test (p < 0.003), and the Essen Questionnaire on Age and Sleepiness (p < 0.0001) in the multivariate analysis. Effect size was weak for Instrumental Activities of Daily Living (R-squared = 0.12) and the Tinetti test (R-squared = 0.03) and medium for the Essen Questionnaire on Age and Sleepiness (R-squared = 0.22). Significant differences between patients with high and low FORTA scores were found for Instrumental Activities of Daily Living, the Tinetti test, mini-nutritional assessments, Mini-Mental State Examination, Essen Questionnaire on Age and Sleepiness, and the Geriatric Depression Scale. All significant tests revealed that higher FORTA scores (lower medication quality) were associated with less favorable test outcomes. CONCLUSIONS: The FORTA score is associated with relevant aspects of comprehensive geriatric assessment, underlining the importance of medication quality for the functional and cognitive well-being of older patients. TRIAL REGISTRATION NUMBER: DRKS00000531.

Glycomic Profiling Highlights Increased Fucosylation in Pseudomyxoma Peritonei.

Pseudomyxoma peritonei (PMP) is a subtype of mucinous adenocarcinoma that most often originates from the appendix, and grows in the peritoneal cavity filling it with mucinous ascites. KRAS and GNAS mutations are frequently found in PMP, but other common driver mutations are infrequent. As altered glycosylation can promote carcinogenesis, we compared N-linked glycan profiles of PMP tissues to those of normal appendix. Glycan profiles of eight normal appendix samples and eight low-grade and eight high-grade PMP specimens were analyzed by mass spectrometry. Our results show differences in glycan profiles between PMP and the controls, especially in those of neutral glycans, and the most prominent alteration was increased fucosylation. We further demonstrate up-regulated mRNA expression of four fucosylation-related enzymes, the core fucosylation performing fucosyltransferase 8 and three GDP-fucose biosynthetic enzymes in PMP tissues when compared with the controls. Up-regulated protein expression of the latter three enzymes was further observed in PMP cells by immunohistochemistry. We also demonstrate that restoration of fucosylation either by salvage pathway or by introduction of an expression of intact GDP-mannose 4,6-dehydratase enhance expression of MUC2, which is the predominant mucin molecule secreted by the PMP cells, in an intestinal-derived adenocarcinoma cell line with defective fucosylation because of deletion in the GDP-mannose 4,6-dehydratase gene. Thus, altered glycosylation especially in the form of fucosylation is linked to the characteristic mucin production of PMP. Glycomic data are available via ProteomeXchange with identifier PXD010086.

Changes in prescription patterns in older hospitalized patients: the impact of FORTA on disease-related over- and under-treatments.

PURPOSE: Physicians often face difficulties in choosing appropriate medications for multimorbid older people. The FORTA (Fit for the Aged) classification (A: absolutely, B: beneficial, C: careful, D: don't) was proposed as a clinical tool for improving the quality of drug treatment in the aged. As an implicit tool, FORTA has been shown to aid medication optimization and improve clinical end points in the VALFORTA trial. In this prospective randomized controlled study, 207 older hospitalized patients received standard geriatric treatment and 202 patients received FORTA-guided treatment. METHODS: Here, changes of drug prescriptions at the anatomical-therapeutic-chemical system (ATC) level were evaluated separately for important diagnoses in descriptive analyses; over- and under-treatment rates were compared between groups. RESULTS: At the individual drug/drug class level related to all important diagnoses, the application of FORTA significantly improved under-treatments for 12 drugs/drug classes (e.g., ACE inhibitors to treat arterial hypertension) and over-treatments for 7 drugs/drug classes (e.g., proton pump inhibitors to treat gastroesophageal reflux disease). CONCLUSIONS: FORTA representing the first combined positive/negative labeling approach at the individual drug level aids the optimization of drug treatment in older people as detected for drugs/drug classes at the ATC level in important indications. FORTA is effective in addressing over- and under-treatments even if analyzed for smaller subgroups of VALFORTA.

Multiple components of PKA and TGF-ß pathways are mutated in pseudomyxoma peritonei.

Pseudomyxoma peritonei (PMP) is a subtype of mucinous adenocarcinoma mainly restricted to the peritoneal cavity and most commonly originating from the appendix. The genetic background of PMP is poorly understood and no targeted treatments are currently available for this fatal disease. While RAS signaling pathway is affected in most if not all PMP cases and over half of them also have a mutation in the GNAS gene, other genetic alterations and affected pathways are, to a large degree, poorly known. In this study, we sequenced whole coding genome of nine PMP tumors and paired normal tissues in order to identify additional, commonly mutated genes and signaling pathways affected in PMP. These exome sequencing results were validated with an ultra-deep amplicon sequencing method, leading to 14 validated variants. The validated results contain seven genes that contribute to the protein kinase A (PKA) pathway. PKA pathway, which also contains GNAS, is a major player of overproduction of mucin, which is the characteristic feature of PMP. In addition to PKA pathway, we identified mutations in six genes that belong to the transforming growth factor beta (TGF-ß) pathway, which is a key regulator of cell proliferation. Since either GNAS mutation or an alternative mutation in the PKA pathway was identified in 8/9 patients, inhibition of the PKA pathway might reduce mucin production in most of the PMP patients and potentially suppress disease progression.

Expression of CEA, CA19-9, CA125, and EpCAM in pseudomyxoma peritonei.

Pseudomyxoma peritonei is a fatal clinical syndrome with mucinous tumor cells disseminated into peritoneal cavity and secreting abundant mucinous ascites. The serum tumor markers CEA, CA19-9, and CA125 are used to monitor pseudomyxoma peritonei remission, but their expression at tissue level has not been well characterized. Herein, we analyzed expression of these proteins and the adenocarcinoma marker EpCAM in 92 appendix-derived pseudomyxoma peritonei tumors by immunohistochemistry. All tumors were found to ubiquitously express CEA and EpCAM. In the majority of the tumors (94.6%), CEA showed polarized immunostaining, but in 5 aggressive high-grade tumors containing numerous signet ring cells, a nonpolarized staining was detected. We found preoperative CEA serum values to correlate with peritoneal cancer index. However, the serum values of the advanced cases with nonpolarized staining pattern were normal, and the patients died within 5 years after diagnosis. Thus, serum CEA measurements did not reflect aggressiveness of these tumors. CA19-9 showed strong immunopositivity in most of the tumors (91.3%), and mutated enzyme FUT3 was demonstrated from the cases showing negative or weak staining. CA125 was infrequently expressed by tumor cells (focal staining in 6.5% of the cases), but in most of the cases (79.3%), adjacent nonneoplastic mesothelial cells showed immunopositivity. As a conclusion, CEA and EpCAM are invariably expressed by pseudomyxoma peritonei tumor cells and could be exploited to targeted therapies against this malignancy.

VALFORTA: a randomised trial to validate the FORTA (Fit fOR The Aged) classification.

TRIAL DESIGN: to further validate the FORTA (Fit fOR The Aged) concept, a bicentric randomised, controlled trial was run in two geriatric clinics. METHODS: patients (≥65 years, ≥3 drugs or ≥60 years, ≥6 drugs) with three relevant diseases and hospitalisation for ≥5 days were randomised. In the intervention, but not the control group, a FORTA team instructed ward physicians on FORTA. FORTA is the first positive/negative listing approach labelling medications used to treat chronic illnesses in older patients from A (indispensable), B (beneficial), C (questionable) to D (avoid). The primary end point was the FORTA score: sum of medication errors classified as over-, under- and mistreatment. Consecutive patients were randomised to the intervention and control ward; outcome assessment was blinded. RESULTS: four hundred and nine patients (age 81.5 years, 64% female, hospitalisation 17.4 days) were included. The primary end point was significantly (P < 0.0001) more reduced in the intervention versus control groups (2.7 ± 2.25 versus 1 ± 1.8, mean ± SD, intergroup comparison of admission/discharge differences). Over- and under-treatment scores and use of A (increase) and D (decrease) drugs were significantly improved (P < 0.01). The total number of adverse drug reactions (ADRs) was significantly reduced by FORTA (P < 0.05, number needed to treat is 5). Activities of daily living and renal failure improved significantly (P < 0.05). Blood pressure remained constant in the intervention, but decreased significantly in the control group. CONCLUSION: applying FORTA to hospitalised geriatric patients leads to improvement of medication quality and may improve secondary clinical end points (e.g. ADRs). The concept is amenable to successful communication and implementation. Registration (DRKS-ID): DRKS00000531. FUNDING: DFG-German Research Foundation (WE 1184/15-1).

Targeted therapy in gastric cancer.

Gastric cancer is often diagnosed at an advanced stage. Although chemotherapy prolongs survival and improves quality of life, the survival of gastric cancer patients with advanced disease is short. Thanks to recent insights into the molecular pathways involved in gastric carcinogenesis, new targeted treatment options have become available for gastric cancer patients. Trastuzumab, an antibody targeted to HER-2, was shown to improve survival of advanced gastric cancer patients harboring HER-2 overexpression due to gene amplification in their tumor cells, and is currently also explored in adjuvant and neoadjuvant settings. Another agent with promising results in clinical trials is ramucirumab, an antibody targeting VEGFR-2. No clear survival benefit, however, were experienced with agents targeting EGFR (cetuximab, panitumumab), VEGF-A (bevacizumab), or mTOR (everolimus). Drugs targeting c-MET/HGF are currently under investigation in biomarker-selected cohorts, with promising results in early clinical trials. This review will summarize the current status of targeted treatment options in gastric cancer.

Prospective immunohistochemical analysis of BRAF V600E mutation in melanoma.

The v-raf murine sarcoma viral oncogene homolog B1 (BRAF) V600E mutation is the most common activating genetic alteration of this oncogene and a predictive marker for the therapeutic use of BRAF inhibitors in melanoma. Our aim was to evaluate the performance of BRAF V600E mutation-specific monoclonal antibody (VE1) in a prospective diagnostic setting of melanoma patients (n = 102). All 41 cases (40.2%) that showed a V600E mutation in the cyclic minisequencing analysis of the DNA were also initially scored immunopositive. Two cases that were scored as BRAF V600E mutation positive by immunohistochemistry were negative in the DNA-based mutation analysis and determined to be immunonegative in a repeated staining with more representative specimens. Thus, BRAF V600E mutation detection using immunohistochemistry was 100% sensitive and 96.8% specific, when compared with the analysis of the DNA. None of the BRAF V600K mutations was detected by the VE1 antibody (n = 7). However, the VE1 antibody detected a rare V600E2 mutation. We also studied the role of BRAF V600E mutation in a set of melanoma patients who had been investigated for sentinel node metastasis. Melanoma lymph node metastases were diagnosed in 21.8% (12/55) of the sentinel nodes, and BRAF V600E immunopositivity was detected in 34.5% (19/55) of the cases. BRAF V600E mutation status did not correlate with any clinicopathological parameters. In conclusion, analysis of BRAF V600E mutation in melanoma by immunohistochemistry is a sensitive and specific method, which can be used to identify BRAF inhibitor-sensitive melanoma patients as a first-line method due to its rapid and affordable nature.

Genomic profile of pseudomyxoma peritonei analyzed using next-generation sequencing and immunohistochemistry.

Pseudomyxoma peritonei (PMP) is a relatively rare clinical syndrome characterized by neoplastic epithelial cells growing in the peritoneal cavity and secreting mucinous ascites. Our aim was to explore the molecular events behind this fatal but under-investigated disease. We extracted DNA from 19 appendix-derived PMP tumors and nine corresponding normal tissues, and analyzed the mutational hotspot areas of 48 cancer-related genes by amplicon-based next-generation sequencing (NGS). Further, we analyzed the protein expression of V600E mutated BRAF, MLH1, MSH2, MSH6 and p53 from a larger set of PMP tumors (n = 74) using immunohistochemistry. With NGS, we detected activating somatic KRAS mutations in all of the tumors studied. GNAS was mutated in 63% of the tumors with no marked difference between low-grade and high-grade tumors. Only one (5.3%) tumor showed oncogenic PIK3CA mutation, one showed oncogenic AKT1 mutation, three (15.8%) showed SMAD4 mutations and none showed an APC mutation. P53 protein was aberrantly expressed in higher proportion of high-grade tumors as compared with low-grade ones (31.3 vs. 7.1%, respectively; p = 0.012) and aberrant expression was an independent factor for reduced overall survival (p = 0.002). BRAF V600E mutation was only found in one (1.4%) high-grade tumor by immunohistochemistry (n = 74). All the studied tumors expressed mismatch repair proteins MLH1, MSH2 and MSH6. Our results indicate that KRAS mutations are evident in all and GNAS mutations in most of the PMPs, but BRAF V600E, PIK3CA and APC mutations are rare. Aberrantly expressed p53 is associated with high-grade histology and reduced survival.