Psychosocial and physical long-term outcome in patients with a history of takotsubo cardiomyopathy or myocardial infarction - a multi-centered case control study.

Physical long-term impacts of Takotsubo Cardiomyopathy (TTC) remain controversial and an underestimation of their severity becomes increasingly evident. Even less is known about mental long-term impacts of TTC. This study aims at a better understanding of the physical and mental long-term effects of TTC in comparison to myocardial infarctions (MI). On average 5 years after disease onset, 68 TTC patients and 68 age- and sex-matched MI patients were assessed for disease-related quality of life, depression, anxiety, chronic stress, social support, resilience, and life events prior to disease onset. Scores of TTC and MI patients were compared to each other and to normative references values. Regression analyses were used to evaluate the predictive value of the number of life events prior to disease onset for physical and mental long-term outcomes. Both groups displayed higher scores in depression and anxiety, higher levels of chronic stress, and lower scores in physical and mental quality of life in comparison to norm samples, while social support did not differ from norms. No differences between the two patient groups were observed. Within both groups, the majority of patients (TTC: 69.1%; MI: 60.3%) reported stressful life events prior to disease onset. In TTCs and MIs, the number of events had a significant impact on long-term mental health and chronic stress. Notably, both patient collectives scored higher in resilience than healthy controls. Results suggest negative long-term impacts of TTC on mental and physical wellbeing, comparable to those of MI. Besides a good somatic-medical care, psychotherapeutic support, including the development of functional coping strategies, might be warranted for TTC patients. The long-term impact of TTC should be taken as serious as that of MI.

Akkermansia muciniphila: a novel functional microbe with probiotic properties.

Akkermansia muciniphila is an intestinal anaerobe which has been proposed as a new functional microbe with probiotic properties. However, the species is not included in the European Union qualified presumption of safety (QPS) list and has not yet been assessed. Moreover, products containing A. muciniphila are not on the market and are thus controlled by the Novel Foods Regulation, which requires extensive safety assessment. This review addresses the safety aspects of the use of A. muciniphila based on published information on its functions in humans and predictions based on its activity in model animals. Further, comprehensive studies related to A. muciniphila and its safety properties have gradually appeared and are summarised here. Many of the criteria required for novel food safety assessment in Europe can thus be fulfilled. However, studies focusing on the toxicological properties of A. muciniphila, including long-term and reproduction studies, have not so far been reported and are discussed in the light of the observation that most, if not all, healthy subjects are known to carry this intestinal anaerobe. As this also applies to other beneficial bacteria found in the human intestinal tract, the A. muciniphila case can be seen as a model for the comprehensive safety evaluations required by the European authorities.

Biofouling on polymeric heat exchanger surfaces with E. coli and native biofilms.

The biofouling affinity of different polymeric surfaces (polypropylene, polysulfone, polyethylene terephthalate, and polyether ether ketone) in comparison to stainless steel (SS) was studied for the model bacterium Escherichia coli K12 DSM 498 and native biofilms originating from Rhine water. The biofilm mass deposited on the polymer surfaces was minimized by several magnitudes compared to SS. The cell count and the accumulated biomass of E. coli on the polymer surfaces showed an opposing linear trend. The promising low biofilm formation on the polymers is attributed to the combination of inherent surface properties (roughness, surface energy and hydrophobicity) when compared to SS. The fouling characteristics of E. coli biofilms show good conformity with the more complex native biofilms investigated. The results can be utilized for the development of new polymer heat exchangers when using untreated river water as coolant or for other processes needing antifouling materials.

What Makes Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells Superior Immunomodulators When Compared to Bone Marrow Derived Mesenchymal Stromal Cells?

MSCs derived from the umbilical cord tissue, termed UCX, were investigated for their immunomodulatory properties and compared to bone marrow-derived MSCs (BM-MSCs), the gold-standard in immunotherapy. Immunogenicity and immunosuppression were assessed by mixed lymphocyte reactions, suppression of lymphocyte proliferation and induction of regulatory T cells. Results showed that UCX were less immunogenic and showed higher immunosuppression activity than BM-MSCs. Further, UCX did not need prior activation or priming to exert their immunomodulatory effects. This was further corroborated in vivo in a model of acute inflammation. To elucidate the potency differences observed between UCX and BM-MSCs, gene expression related to immune modulation was analysed in both cell types. Several gene expression profile differences were found between UCX and BM-MSCs, namely decreased expression of HLA-DRA, HO-1, IGFBP1, 4 and 6, ILR1, IL6R and PTGES and increased expression of CD200, CD273, CD274, IL1B, IL-8, LIF and TGFB2. The latter were confirmed at the protein expression level. Overall, these results show that UCX seem to be naturally more potent immunosuppressors and less immunogenic than BM-MSCs. We propose that these differences may be due to increased levels of immunomodulatory surface proteins such as CD200, CD273, CD274 and cytokines such as IL1ß, IL-8, LIF and TGFß2.

In-vivo monitoring of acute DSS-Colitis using Colonoscopy, high resolution Ultrasound and bench-top Magnetic Resonance Imaging in Mice.

OBJECTIVE: The aim of this study was to establish and evaluate (colour Doppler-) high-resolution-ultrasound (hrUS) and bench-top magnetic resonance imaging (btMRI) as new methods to monitor experimental colitis. MATERIALS AND METHODS: hrUS, btMRI and endoscopy were performed in mice without colitis (n = 15), in mice with acute colitis (n = 14) and in mice with acute colitis and simultaneous treatment with infliximab (n = 19). RESULTS: Determination of colon wall thickness using hrUS (32 MHz) and measurement of the cross-sectional colonic areas by btMRI allowed discrimination between the treatment groups (mean a vs. b vs. c - btMRI: 922 vs. 2051 vs. 1472 pixel, hrUS: 0.26 vs. 0.45 vs. 0.31 mm). btMRI, endoscopy, hrUS and colour Doppler-hrUS correlated to histological scoring (p < 0.05), while endoscopy and btMRI correlated to post-mortem colon length (p < 0.05). CONCLUSIONS: The innovative in vivo techniques btMRI and hrUS are safe and technically feasible. They differentiate between distinct grades of colitis in an experimental setting, and correlate with established post-mortem parameters. In addition to endoscopic procedures, these techniques provide information regarding colon wall thickness and perfusion. Depending on the availability of these techniques, their application increases the value of in vivo monitoring in experimental acute colitis in small rodents. KEY POINTS: • Improved in vivo monitoring might balance interindividual differences in murine colitis. • In monitoring murine colitis, btMRI and hrUS are safe and technically feasible. • Very short examination times underline the usefulness especially of hrUS. • Results of btMRI and hrUS correlate with endoscopic and post-mortem findings.

Effects of reduced nocturnal temperature on pig performance and energy consumption in swine nursery rooms.

The objective of this investigation was to determine the effect of a reduced nocturnal temperature (RNT) regimen on performance of weaned pigs and energy consumption during the nursery phase of production. The age of weaned pigs assigned to experiments ranged from 16 to 22 d. In Exp. 1, 3 stations conducted 2 trials under a common protocol that provided data from 6 control rooms (CON; 820 pigs) and 6 RNT rooms (818 pigs). Two mirror-image nursery rooms were used at each station. Temperature in the CON room was set to 30°C for the first 7 d, then reduced by 2°C per week through the remainder of the experiment. Room temperature settings were held constant throughout the day and night. The temperature setting in the RNT room was the same as CON during the first 7 d, but beginning on the night of d 7, the room temperature setting was reduced 6°C from the daytime temperature from 1900 to 0700 h. The use of heating fuel and electricity were measured weekly in each room. Overall, ADG (0.43 kg), ADFI (0.62 kg), and G:F (0.69) were identical for CON and RNT rooms. Consumption of heating fuel [9,658 vs. 7,958 British thermal units (Btu)·pig(-1)·d(-1)] and electricity (0.138 vs. 0.125 kilowatt-hour (kWh)·pig(-1)·d(-1)] were not statistically different for CON and RNT rooms, respectively. In Exp. 2, 4 stations conducted at least 2 trials that provided data from 9 CON rooms (2,122 pigs) and 10 RNT rooms (2,176 pigs). Experimental treatments and protocols were the same as Exp. 1, except that the RNT regimen was imposed on the night of d 5 and the targeted nighttime temperature reduction was 8.3°C. Neither final pig BW (21.8 vs. 21.5 kg; SE = 0.64), ADG (0.45 vs. 0.44 kg; SE = 0.016), ADFI (0.61 vs. 0.60 kg; SE = 0.019), nor G:F (0.75 vs. 0.75; SE = 0.012) were different for pigs housed in CON or RNT rooms, respectively. Consumption of heating fuel and electricity was consistently reduced in RNT rooms for all 4 stations. Consumption of heating fuel (10,019 vs. 7,061 Btu·pig(-1)·d(-1); SE = 1,467) and electricity (0.026 vs. 0.021 kWh·pig-1·d-1; SE = 0.004) were lower (P < 0.05) in the RNT rooms compared with CON rooms. This represents a 30% reduction in heating fuel use and a 20% reduction in electrical use with no differences in pig growth performance or health. From these experiments, we conclude that imposing a RNT regimen from 1900 to 0700 h is effective in reducing energy costs in the nursery without compromising pig performance, which will reduce production costs and decrease emissions of greenhouse gases.

Beyond the tradition: test of an integrative conceptual model on nurse turnover.

AIM: This paper aimed to extend research on nurse turnover by developing and testing a theoretical model of turnover intention that includes two emergent key off-the-job constructs, work-family conflict (WFC) and community embeddedness (CE). BACKGROUND: Nurse turnover is considered one of the most significant issues in health care. There is a considerable body of knowledge that has focused on the study of the on-the-job factors of nurse turnover, showing the important role of job attitudes. Recently, WFC and job embeddedness (JE) have been identified as variables that could help explain levels of nurse turnover. METHODS: Using structural equation modelling from a cross-sectional survey, the relationships between the variables were explored in a sample of 440 nurses from an Italian public hospital. The questionnaire measures demographic data and psychosocial factors such as job satisfaction, organizational commitment, WFC, CE and turnover intentions. RESULTS: The findings supported the importance of non-work dimensions in turnover models. CONCLUSIONS: The results suggest that when studying turnover phenomena in health organizations, the extra-work domains (WFC and JE) can contribute to a decrease in the intention to leave, in addition to the more typically emphasized attitude dimension.

Lysosomal dysfunction causes neurodegeneration in mucolipidosis II 'knock-in' mice.

Mucolipidosis II is a neurometabolic lysosomal trafficking disorder of infancy caused by loss of mannose 6-phosphate targeting signals on lysosomal proteins, leading to lysosomal dysfunction and accumulation of non-degraded material. However, the identity of storage material and mechanisms of neurodegeneration in mucolipidosis II are unknown. We have generated 'knock-in' mice with a common mucolipidosis II patient mutation that show growth retardation, progressive brain atrophy, skeletal abnormalities, elevated lysosomal enzyme activities in serum, lysosomal storage in fibroblasts and brain and premature death, closely mimicking the mucolipidosis II disease in humans. The examination of affected mouse brains at different ages by immunohistochemistry, ultrastructural analysis, immunoblotting and mass spectrometric analyses of glycans and anionic lipids revealed that the expression and proteolytic processing of distinct lysosomal proteins such as α-l-fucosidase, ß-hexosaminidase, α-mannosidase or Niemann-Pick C2 protein are more significantly impacted by the loss of mannose 6-phosphate residues than enzymes reaching lysosomes independently of this targeting mechanism. As a consequence, fucosylated N-glycans, GM2 and GM3 gangliosides, cholesterol and bis(monoacylglycero)phosphate accumulate progressively in the brain of mucolipidosis II mice. Prominent astrogliosis and the accumulation of organelles and storage material in focally swollen axons were observed in the cerebellum and were accompanied by a loss of Purkinje cells. Moreover, an increased neuronal level of the microtubule-associated protein 1 light chain 3 and the formation of p62-positive neuronal aggregates indicate an impairment of constitutive autophagy in the mucolipidosis II brain. Our findings demonstrate the essential role of mannose 6-phosphate for selected lysosomal proteins to maintain the capability for degradation of sequestered components in lysosomes and autophagolysosomes and prevent neurodegeneration. These lysosomal proteins might be a potential target for a valid therapeutic approach for mucolipidosis II disease.

Washout of sevoflurane from the GE Avance and Amingo Carestation anesthetic machines.

BACKGROUND: Inhalational anesthetics must be removed from anesthetic machines to prevent malignant hyperthermia (MH) in susceptible patients or to treat MH occurring during inhalational general anesthesia. This study examines the sevoflurane washout from the GE Avance and Amingo Carestations™. METHODS: The care stations were contaminated with sevoflurane during general anesthesia. Then, the vaporizer was removed, the CO2 absorber was exchanged against an empty one and the breathing tubes were substituted by clean ones. In the first part, the fresh gas flow was 10 l/min. In the second part, the Advanced Breathing System™ (ABS™), the internal breathing circuit, was replaced by a laundered component. The fresh gas flow was set to 10 l/min for 10 min and to 5 l/min for the following 20 min. RESULTS: In the 25 measurements of the first part, the sevoflurane concentration decreased from a median of 31.60 ppm [interquartile range (IQR) 130.12 ppm] within 22 min in every case to values below 5 ppm and stayed there for the last 8 min of the measuring (P < 0.0001). In the 15 measurements of the second part, the sevoflurane concentration fell from the median of 8.56 ppm (IQR 8.99 ppm) within 5 min to values being significantly below 5 ppm and stayed there for the following 25 min (P < 0.0001). CONCLUSIONS: In case of sudden onset of MH, the Avance or Amingo Carestation™ can stay in place, if the fresh gas flow is increased to 10 l/min or more. To prepare these machines for MH-susceptible patients, the ABS™ should be substituted by a laundered component.

Eimeria tenella oocyst shedding and output in cecal or fecal contents following experimental challenge in broilers.

Two experiments were conducted to determine whether Eimeria tenella oocyst output in cecal and fecal contents, lesion development, and performance characteristics were affected by ad libitum versus restricted feeding and challenge level. In experiment 1, 144 Cobb 500 males were placed in battery cages with 6 chicks/pen. On d 20, half of the battery pens were placed on feed restriction and all broilers were orally challenged with Eimeria tenella oocysts at one of 3 challenge levels (0, 5,000, or 20,000 sporulated oocysts). Cecal and fecal material were collected separately from d 4 postchallenge through d 10 postchallenge for oocysts output (oocysts shed/g) determination. Six days postchallenge, 3 broilers from each pen were removed and subjected to necropsy for lesion assessment. In experiment 2, 96 Cobb 500 males were placed in identical battery pens with 8 chicks/pen. On d 14, restricted feeding was initiated and broilers were challenged with Eimeria tenella oocysts at one of 3 challenge levels (1,000, 5,000, or 20,000 oocysts). Twenty-four hour collections of cecal and fecal material were obtained separately from d 4 postchallenge through d 10 postchallenge for oocysts per gram and total output determination. Six days postchallenge, 4 broilers from each pen were removed and subjected to necropsy for lesion assessment. In both experiments, BW gain was not affected by challenge dose in either the ad libitum-fed or restrict-fed broilers. Increased lesion development was observed with increasing challenge levels, and oocyst shedding peaked between d 7 and 9 postchallenge in both experiments. Oocyst concentration was higher in cecal droppings compared with fecal material throughout peak shedding; however, total oocyst output for the challenge period was similar between fecal material and cecal droppings.

Use of Encapsulated Stem Cells to Overcome the Bottleneck of Cell Availability for Cell Therapy Approaches.

Nowadays cell-based therapy is rarely in clinical practice because of the limited availability of appropriate cells. To apply cells therapeutically, they must not cause any immune response wherefore mainly autologous cells have been used up to now. The amount of vital cells in patients is limited, and under certain circumstances in highly degenerated tissues no vital cells are left. Moreover, the extraction of these cells is connected with additional surgery; also the expansion in vitro is difficult. Other approaches avoid these problems by using allo-or even xenogenic cells. These cells are more stable concerning their therapeutic behavior and can be produced in stock. To prevent an immune response caused by these cells, cell encapsulation (e.g. with alginate) can be performed. Certain studies showed that encapsulated allo- and xenogenic cells achieve promising results in treatment of several diseases. For such cell therapy approaches, stem cells, particularly mesenchymal stem cells, are an interesting cell source. This review deals on the one hand with the use of encapsulated cells, especially stem cells, in cell therapy and on the other hand with bioreactor systems for the expansion and differentiation of mesenchymal stem cells in reproducible and sufficient amounts for potential clinical use.

Immune response of broiler chickens fed different levels of arginine and vitamin E to a coccidiosis vaccine and Eimeria challenge.

One-day-old broiler chicks (n = 300) were orally vaccinated (Coccivac-B) and divided into 6 groups to evaluate Arg at 3 levels of supplementation, 0, 0.3, or 0.6% [normal level (NARG), medium level (MARG), or high level (HARG), respectively], and 2 levels of vitamin E (VE), 40 or 80 IU/kg of feed (VE40 or VE80, respectively), in a factorial experiment. Birds were reared in floor pens with fresh pine shavings and provided a corn-soybean-based diet and water ad libitum. At d 14, all chickens were orally challenged with a mixture of Eimeria field isolates (Eimeria acervulina, Eimeria maxima, and Eimeria tenella). In vitro heterophil and monocyte oxidative burst (HOB and MOB, respectively) was measured at d 21 from cells isolated from peripheral blood. Antibody levels (IgG, IgM, and IgA isotypes, ELISA) and NO were measured at d 14 and 28. The HOB was lower in birds fed the VE40 diets but was increased with the MARG and HARG treatments, whereas birds fed the VE80 diet had a higher HOB irrespective of Arg level. Birds fed the VE80 diet had high levels of MOB, which was not further improved by Arg, whereas birds fed the VE40-MARG diet had the highest MOB response. Plasma NO was not affected by diet at d 14, but at d 28, plasma NO was higher in birds fed the VE80-MARG or the VE40-NARG diet and lower in birds fed the VE80-NARG or the VE40-MARG diet. Birds fed the VE40-HARG or VE80-MARG diet had the highest IgG levels at d 14, but at d 28, birds fed the VE80-MARG diet had the highest IgG levels. The IgM concentration was lower in birds fed NARG levels irrespective of VE levels at d 14, but at d 28, IgM levels were higher in birds fed the VE40-HARG or the VE80-MARG feed. The IgA concentration was not consistently affected at d 14 or 28. These results suggest that Arg and VE fed at levels higher than those recommended by the NRC may play complementary roles on the innate and humoral immune response against an Eimeria challenge, potentially improving vaccine efficacy and response to field infections.

Production process for stem cell based therapeutic implants: expansion of the production cell line and cultivation of encapsulated cells.

Cell based therapy promises the treatment of many diseases like diabetes mellitus, Parkinson disease or stroke. Microencapsulation of the cells protects them against host-vs-graft reactions and thus enables the usage of allogenic cell lines for the manufacturing of cell therapeutic implants. The production process of such implants consists mainly of the three steps expansion of the cells, encapsulation of the cells, and cultivation of the encapsulated cells in order to increase their vitality and thus quality. This chapter deals with the development of fixed-bed bioreactor-based cultivation procedures used in the first and third step of production. The bioreactor system for the expansion of the stem cell line (hMSC-TERT) is based on non-porous glass spheres, which support cell growth and harvesting with high yield and vitality. The cultivation process for the spherical cell based implants leads to an increase of vitality and additionally enables the application of a medium-based differentiation protocol.

Observations on unaided smoking cessation after deep brain stimulation of the nucleus accumbens.

AIMS: We explore whether clinical research on deep brain stimulation (DBS) of the nucleus accumbens (NAc) to treat addiction is justified besides theoretical speculation. METHODS: Since 2004, 10 patients who were also smokers were treated at the University of Cologne for Tourette's syndrome (TS), obsessive-compulsive disorders (OCD) or anxiety disorders (AD) by DBS of the NAc. We assessed their smoking behavior after DBS and (in retrospection) before by the Fagerstrom Test for Nicotine Dependence (FTND) and additional items. RESULTS: Three male patients were able to quit smoking after DBS. They were less dependent and higher motivated compared to the rest of the sample. They are stimulated with a higher voltage. During 1-year, 2-year, and 30-month follow-ups, we found a higher rate of successful smoking cessation (20, 30 and 30%) compared to unaided smoking cessation in the general population (13, 19 and 8.7%). CONCLUSIONS: Albeit the results of the study are severely limited by the method of retrospective self-assessment of psychiatric patients, further research of DBS of the NAc to treat addiction seems justified. In addition to biological mediators, psychosocial factors should be assessed in further prospective studies.

[Deep brain stimulation in the context of addiction--a literature-based systematic evaluation]. / Abhängigkeitserkrankungen im Kontext der Tiefen Hirnstimulation--eine literaturgestützte systematische Auswertung.

BACKGROUND: Deep Brain Stimulation (DBS) is established as option to treat Parkinson's Disease and essential tremor by now. Successful treatments of small samples suffering from dystonia, cluster headache or Tourette's Syndrome or from obessive-compulsive disorders and depression make a future extension of indication seem probable. In this context the efficiency of DBS for the treatment of addiction is being discussed. AIM AND METHODS: To assess the potential effects of DBS of different target areas on addiction a keyword-related research in Pubmed (National Library of Medicine, Washington) was undertaken and own research was integrated. FINDINGS: Only case reports and case series were being found, describing in total n = 28 patients. Dopamine replacement therapy dependence (DRTD) and pathological gambling (PG) were reported in conjunction with DBS of the nucleus subthalamicus (STN). Addiction to alcohol, nicotine and heroin were reported in conjunction with DBS of the nucleus accumbens (NAc). These findings were collected in a spreadsheet and discussed. CONCLUSIONS: For STN DBS remissions of PG and DRTD are only reported during the underlying treatment of Parkinson's disease. As method of action therefore the reduction of Parkinsonian medication seems more probable. For NAc DBS remissions of addiction to alcohol, nicotine and heroin are reported during the underlying treatment of heterogonous psychiatric disorders. In contrast to STN DBS this refers to possible, maybe supportive effects of NAc DBS. The exact methods of action are not well understood, but an high motivation of the patients to stay abstinent seems to be of relevance for the effect of NAc DBS.

Molecular analysis of the GlcNac-1-phosphotransferase.

Modification of the carbohydrate chains of soluble lysosomal enzymes with mannose 6-phosphate residues is a prerequisite for their mannose 6-phosphate receptor-dependent transport to lysosomes. GlcNac-1-phosphotransferase localized in the Golgi apparatus represents a hexameric alpha(2)beta(2)gamma(2) subunit complex and plays a key role in the formation of the mannose 6-phosphate recognition marker. Defects in the GlcNac-1-phosphotransferase complex cause two diseases, mucolipidosis type II and III, which are characterized by missorting and cellular loss of lysosomal enzymes, and lysosomal accumulation of storage material. The recent identification of two genes, GNPTAB and GNPTG, encoding the three subunits of GlcNac-1-phosphotransferase leads to an improvement of both pre- and postnatal diagnosis of affected individuals, and permits the analysis of structural requirements for efficient formation of mannose 6-phosphate residues on lysosomal enzymes. The alpha/beta subunits precursor matures by proteolytic cleavage and contains the catalytic activity as well as the capability to recognize lysosomal enzymes. The role of the gamma-subunits for activity, stability and oligomerization of the GlcNac-1-phosphotransferase subunits is still unclear.

Evaluation of a single-locus real-time polymerase chain reaction as a screening test for specific detection of methicillin-resistant Staphylococcus aureus in ICU patients.

The aim of the present study was to determine the diagnostic value of a single-locus real-time polymerase chain reaction (PCR) recently proposed for rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) from clinical samples (IDI-MRSA; Infectio Diagnostic, Sainte-Foy, Québec, Canada). This test, which was developed on the basis of studies of the sequence analysis of the mecA gene carried by staphylococcal cassette chromosome mec (SCCmec), was used to screen nasal swabs of 320 intensive care unit (ICU) patients at admission. The results were compared with those of conventional culture of swabs from several body sites. When compared with culture of swabs from the nose, throat, and wounds, the diagnostic values of the real-time PCR test from nasal swabs were as follows: 92.3% sensitivity, 98.6% specificity, 75.0% positive predictive value, and 99.6% negative predictive value. Fifteen (4.7%) samples could not be evaluated because the PCR reaction was inhibited, even after the samples were frozen and thawed for retesting. Culture of nasal swabs showed that 78 of the patients were colonized with methicillin-susceptible S. aureus. Unexpectedly, 4 (5.1%) of these samples gave false-positive results in the IDI-MRSA. These isolates were all single clones, as shown by pulsed-field gel electrophoresis and spa typing. Reliable results were obtained with the IDI-MRSA assay, even in a patient population with a low prevalence (approximately 4%) of MRSA and even when compared with swabs of different body sites. Nevertheless, further work is needed to reduce the inhibition rate of the PCR and to explain why false-positive results were obtained with methicillin-susceptible S. aureus.

Cloning and functional pharmacology of two corticotropin-releasing factor receptors from a teleost fish.

Although it is well established that fish possess corticotropin-releasing factor (CRF) and a CRF-like peptide, urotensin I, comparatively little is known about the pharmacology of their cognate receptors. Here we report the isolation and functional expression of two complementary DNAs (cDNAs), from the chum salmon Oncorhynchus keta, which encode orthologues of the mammalian and amphibian CRF type 1 (CRF(1)) and type 2 (CRF(2)) receptors. Radioligand competition binding experiments have revealed that the salmon CRF(1) and CRF(2) receptors bind urotensin I with approximately 8-fold higher affinity than rat/human CRF. These two peptides together with two related CRF-like peptides, namely, sauvagine and urocortin, were also tested in cAMP assays; for cells expressing the salmon CRF(1) receptor, EC(50) values for the stimulation of cAMP production were between 4.5+/-1.8 and 15.3+/-3.1 nM. For the salmon CRF(2) receptor, the corresponding values were: rat/human CRF, 9.4+/-0.4 nM; urotensin I, 21.2+/-2.1 nM; sauvagine, 0.7+/-0.1 nM; and urocortin, 2.2+/-0.7 nM. We have also functionally coupled the O. keta CRF(1) receptor, in Xenopus laevis oocytes, to the endogenous Ca(2+)-activated chloride conductance by co-expression with the G-protein alpha subunit, G(alpha16). The EC(50) value for channel activation by rat/human CRF (11.2+/-2.6 nM) agrees well with that obtained in cAMP assays (15.3+/-3.1 nM). We conclude that although sauvagine is 13- and 30-fold more potent than rat/human CRF and urotensin I, respectively, in activating the salmon CRF(2) receptor, neither receptor appears able to discriminate between the native ligands CRF and urotensin I.

Combined mechanical and antibiotic periodontal therapy in a case of Papillon-Lefèvre syndrome.

BACKGROUND: Papillon Lefèvre syndrome (PLS) is a rare entity and, as such, it is almost impossible to evaluate an effective therapy in a randomized controlled study. The amount of success reported after therapy for prepubertal periodontitis (PP) in PLS is highly variable from case to case. The goal of this case report is to evaluate the effects of a combined mechanical and antibiotic periodontal therapy regimen in the management of PLS. METHODS: A male patient was diagnosed as suffering from PP associated with PLS at the age of 7 years. He showed hyperkeratosis of the palms and soles, as well as advanced periodontal disease already affecting permanent teeth with maximal probing depth and vertical attachment loss of 12 mm and 11 mm, respectively. Subgingival debridement was performed with simultaneous administration of oral 250 mg amoxicillin 3 times daily and 250 mg metronidazole twice daily for one week. Clinical parameters were assessed and subgingival plaque was collected from all teeth prior to therapy and 7 and 26 months after treatment. Selective cultures for A. actinomycetemcomitans were incubated for each individual tooth and DNA probe analysis was performed for various periodontal pathogens. RESULTS: Prior to combined mechanical and antibiotic treatment, all teeth but one harbored Actinobacillus actinomycetemcomitans subgingivally. However, at 7 and 26 months after therapy A. actinomycetemcomitans could be detected neither by culture nor by DNA probes. Clinical parameters improved markedly and teeth erupting after therapy did not exhibit attachment loss of more than 1.5 mm during the observation period. CONCLUSIONS: Eradication (suppression beneath detection levels) of A. actinomycetemcomitans seems to play a significant role in the successful treatment of localized prepubertal periodontitis in PLS.

Active hair growth (anagen) is associated with angiogenesis.

After the completion of skin development, angiogenesis, i.e., the growth of new capillaries from pre-existing blood vessels, is held to occur in the skin only under pathologic conditions. It has long been noted, however, that hair follicle cycling is associated with prominent changes in skin perfusion, that the epithelial hair bulbs of anagen follicles display angiogenic properties, and that the follicular dermal papilla can produce angiogenic factors. Despite these suggestive observations, no formal proof is as yet available for the concept that angiogenesis is a physiologic event that occurs all over the mature mammalian integument whenever hair follicles switch from resting (telogen) to active growth (anagen). This study uses quantitative histomorphometry and double-immunohistologic detection techniques for the demarcation of proliferating endothelial cells, to show that synchronized hair follicle cycling in adolescent C57BL/6 mice is associated with substantial angiogenesis, and that inhibiting angiogenesis in vivo by the intraperitoneal application of a fumagillin derivative retards experimentally induced anagen development in these mice. Thus, angiogenesis is a physiologic event in normal postnatal murine skin, apparently is dictated by the hair follicle, and appears to be required for normal anagen development. Anagen-associated angiogenesis offers an attractive model for identifying the physiologic controls of cutaneous angiogenesis, and an interesting system for screening the effects of potential antiangiogenic drugs in vivo.