Co-Occurrence of Sarcopenia and Frailty in Acutely Admitted Older Medical Patients: Results from the Copenhagen PROTECT Study.

BACKGROUND: Sarcopenia and frailty are often used interchangeably in clinical practice yet represent two distinct conditions and require different therapeutic approaches. The literature regarding the co-occurrence of both conditions in older patients is scarce as most studies have investigated the prevalence of sarcopenia and frailty separately. OBJECTIVES: We aim to evaluate the prevalence and co-occurrence of sarcopenia and frailty in a large sample of acutely admitted older medical patients. DESIGN: Secondary analyses using cross-sectional data from the Copenhagen PROTECT study. SETTING: Patients were included from the acute medical ward at Copenhagen University Hospital, Bispebjerg and Frederiksberg, Copenhagen, Denmark, between November 2019 and November 2021. PARTICIPANTS: Acutely admitted older medical patients (≥65 years). MEASUREMENTS: Handgrip strength (HGS) was investigated using a handheld dynamometer. Lean mass (SMI) was investigated using direct-segmental multifrequency bioelectrical impedance analyses (DSM-BIA). Low HGS, low SMI, and sarcopenia were defined according to the recent definitions from the European Working Group on Sarcopenia in Older People (EWGSOP2). The Clinical Frailty Scale (CFS) was used to evaluate frailty, with a value > 5 indicating the presence of frailty. Patients were enrolled and tested within 24 hours of admission. RESULTS: This study included 638 patients (mean age: 78.2±7.6, 55% female) with complete records of SMI, HGS, and the CFS. The prevalence of low HGS, low SMI, sarcopenia, and frailty were 39.0%, 33.1%, 19.7%, and 39.0%, respectively. Sarcopenia and frailty co-occurred in 12.1% of the patients. CONCLUSIONS: It is well-known that sarcopenia and frailty represent clinical manifestations of ageing and overlap in terms of the impairment in physical function observed in both conditions. Our results demonstrate that sarcopenia and frailty do not necessarily co-occur within the older acutely admitted patient, highlighting the need for separate assessments of frailty and sarcopenia to ensure the accurate characterization of the health status of older patients.

Effects of alternating blood flow restricted training and heavy-load resistance training on myofiber morphology and mechanical muscle function.

To investigate if short-term block-structured training consisting of alternating weeks of blood flow restricted low-load resistance training (BFR-RT) and conventional free-flow heavy-load resistance training (HL-RT) leads to superior gains in mechanical muscle function, myofiber size, and satellite cell (SC) content and myonuclear number compared with HL-RT alone. Eighteen active young participants (women/men: 5/13, 23 ± 1.2 yr) were randomized to 6 wk (22 sessions) of lower limb HL-RT [70-90% one repetition maximum (1-RM)] (HRT, n = 9) or block-structured training alternating weekly between BFR-RT (20% 1-RM) and HL-RT (BFR-HRT, n = 9). Maximal isometric knee extensor strength (MVC) and muscle biopsies (VL) were obtained pre- and posttraining to examine changes in muscle strength, myofiber cross-sectional area (CSA), myonuclear (MN) number, and SC content. MVC increased in both training groups (BFR-HRT: +12%, HRT: +7%; P < 0.05). Type II myofiber CSA increased similarly (+16%) in BFR-HRT and HRT (P < 0.05), while gains in type I CSA were observed following HRT only (+12%, P < 0.05). In addition, myonuclear number remained unchanged, whereas SC content increased in type II myofibers following HRT (+59%, P < 0.05). Short-term alternating BFR-RT and HL-RT did not produce superior gains in muscle strength or myofiber size compared with HL-RT alone. Noticeably, however, conventional HL-RT could be periodically replaced by low-load BFR-RT without compromising training-induced gains in maximal muscle strength and type II myofiber size, respectively.NEW & NOTEWORTHY The present data demonstrate that periodically substituting heavy-load resistance training (HL-RT) with low-load blood flow restricted resistance training (BFR-RT) leads to similar gains in type II myofiber CSA and muscle strength as achieved by HL-RT alone. Furthermore, we have for the first time evaluated myonuclear content and myonuclear domain size before and after training intervention across separate fiber size clusters and found no within-cluster changes for these parameters with training.

Live-cell imaging of new polyene sterols for improved analysis of intracellular cholesterol transport.

Analysis of intracellular cholesterol transport by fluorescence microscopy requires suitable fluorescent analogues of cholesterol. Most existing cholesterol analogues contain lipophilic dyes which can compromise the sterol properties in membranes. An alternative strategy is to introduce additional double bonds into the sterol ring system resulting in intrinsic fluorescence, while at the same time keeping the cholesterol-like properties of the analogues. Existing polyene sterols, such as dehydroergosterol (DHE) or cholestatrienol (CTL), however, contain only three double bonds and suffer from low brightness, significant photobleaching and excitation/emission in the ultraviolet region. Thus, special equipment is required to image such sterols. Here, we describe synthesis, characterization and intracellular imaging of new polyene sterols containing four conjugated double bonds in the sterol ring system. We show that such analogues have red-shifted excitation and emission by ∼20 nm compared to DHE or CTL. The red shift was even more pronounced when preventing keto-enol tautomer equilibration by protecting the 3'-hydroxy group with acetate. We show that the latter analogue can be imaged on a conventional wide field microscope with a DAPI/filipin filter cube. The new polyene sterols show reduced photobleaching compared to DHE or CTL allowing for improved deconvolution microscopy of sterol containing cellular membranes.

Is it necessary to perform full pathologic review of all gastric remnants following sleeve gastrectomy?

BACKGROUND: This study attempts to determine if enough pathological abnormalities in gastric remnants from sleeve gastrectomy exist to warrant full pathologic evaluation in all remnants. METHODS: Data was collected on patients undergoing sleeve gastrectomy between 08/01/2011 and 06/30/2014. Significant abnormalities were classified as any pathology that might require follow-up or treatment beyond standard follow-up. Age, comorbidities, gender, and Helicobacter pylori titers were analyzed and compared with pathology specimens using 95% confidence intervals and Phi contingency coefficients. RESULTS: Full pathologic evaluation was available for 351/387 patients (91.2%). No examples of malignancy or dysplasia were identified. Gastritis was the most common abnormality. There was a statistically significant association between preoperative H. pylori and significantly abnormal pathology (p = 0.003). Other comorbidities had no association. CONCLUSIONS: These results suggest that full pathologic evaluation of the gastric remnant following sleeve gastrectomy is unnecessary, particularly when gross pathology is not noted at initial operation.

Evaluation of a cross contamination model describing transfer of Salmonella spp. and Listeria monocytogenes during grinding of pork and beef.

In a previous study, a model was developed to describe the transfer and survival of Salmonella during grinding of pork (Møller, C.O.A., Nauta, M.J., Christensen, B.B., Dalgaard, P., Hansen, T.B., 2012. Modelling transfer of Salmonella typhimurium DT104 during simulation of grinding of pork. Journal of Applied Microbiology 112 (1), 90-98). The robustness of this model is now evaluated by studying its performance for predicting the transfer and survival of Salmonella spp. and Listeria monocytogenes during grinding of different types of meat (pork and beef), using two different grinders, different sizes and different numbers of pieces of meats to be ground. A total of 19 grinding trials were collected. Acceptable Simulation Zone (ASZ), visual inspection of the data, Quantitative Microbiological Risk Assessment (QMRA), as well as the Total Transfer Potential (TTP) were used as approaches to evaluate model performance and to access the quality of the cross contamination model predictions. Using the ASZ approach and considering that 70% of the observed counts have to be inside a defined acceptable zone of ±0.5 log10CFU per portion, it was found that the cross contamination parameters suggested by Møller et al. (2012) were not able to describe all 19 trials. However, for each of the collected grinding trials, the transfer event was well described when fitted to the model structure proposed by Møller et al. (2012). Parameter estimates obtained by fitting observed trials performed at different conditions, such as size and number of pieces of meat to be ground, may not be applied to describe cross contamination of unlike processing. Nevertheless, the risk estimates, as well as the TTP, revealed that the risk of disease may be reduced when the grinding of meat is performed in a grinder made of stainless steel (for all surfaces in contact with the meat), using a well-sharpened knife and holding at room temperatures lower than 4°C.

Initial Pseudomonas aeruginosa infection in patients with cystic fibrosis: characteristics of eradicated and persistent isolates.

Despite intensive eradication therapy, some CF patients with early Pseudomonas aeruginosa infection rapidly develop a chronic infection. To elucidate factors associated with this persistence, bacterial characteristics of early P. aeruginosa isolates were analysed that were either eradicated rapidly or persisted despite multiple antimicrobial treatments. Eighty-six early infection episodes were studied. First P. aeruginosa isolates from patients with eradication (36) or persistent infection (16) were included; isolates from patients with intermittent infection (34) were omitted from the study. Virulence assays, antimicrobial resistance, cytotoxicity and mutation frequencies were analysed in vitro. P. aeruginosa was genotyped by SNP-array. Transcriptomic profiles of two eradicated and two persistent strains were compared. Nineteen per cent of patients developed persistent infection; 42% achieved eradication. Secretion of virulence factors and mutation frequencies were highly variable among both eradicated and persistent isolates and were not different between the groups. Cytotoxicity was present in 57% of eradicated vs. 100% of persistent isolates (p <0.01). None of the isolates were resistant to antibiotics. The isolates were genotypically highly diverse. Multivariate analysis showed that in vitro determined bacterial characteristics could not predict persistence after first P. aeruginosa infection. Preliminary transcriptomic data showed increased expression of some genes related to a metabolic pathway. The early onset of chronic infection was not associated with (in vitro determined) bacterial characteristics only. Although the persistent isolates were more often cytotoxic, for the individual patient it was not possible to predict the risk of persistence based on bacterial characteristics. Unknown factors such as host-pathogen and pathogen-pathogen interactions should be further explored.

Variation near the hepatocyte nuclear factor (HNF)-4alpha gene associates with type 2 diabetes in the Danish population.

AIMS/HYPOTHESIS: The hepatocyte nuclear factor (HNF)-4alpha is an orphan nuclear receptor, which plays crucial roles in regulating hepatic gluconeogenesis and insulin secretion. The gene encoding HNF-4alpha (HNF4A) is located on chromosome 20q12-q13 in a region that in several studies has shown linkage with type 2 diabetes. Recently, two independent studies identified single nucleotide polymorphisms (SNPs) in a 90-kb region spanning HNF4A, which showed strong association with type 2 diabetes in the Finnish and Ashkenazi Jewish populations. In an attempt to replicate and extend these findings, we selected four SNPs in the same HNF4A region, which in the Finnish and Ashkenazi Jewish populations were associated with type 2 diabetes, and examined their relationships with type 2 diabetes and prediabetic phenotypes in the Danish Caucasian population. METHODS: The rs1884614, rs2425637, rs1885088 and rs3818247 were analysed in case-control studies of 1387, 1429, 1417 and 1371 type 2 diabetic patients and 4766, 4727, 4665 and 4748 glucose-tolerant subjects respectively. Genotype-quantitative trait analyses comprised 4430, 4394, 4336 and 4413 middle-aged glucose-tolerant subjects from the population-based Inter99 cohort for the rs1884614, rs2425637, rs1885088 and rs3818247 respectively. RESULTS: The risk allele of the rs1884614, which is located 4 kb upstream of the HNF4A P2 promoter, was associated with type 2 diabetes (odds ratio [OR]=1.14, p=0.02) and with a subtle increase in post-OGTT plasma glucose levels in glucose-tolerant subjects (additive model, p=0.05). CONCLUSIONS/INTERPRETATION: Consistent with results from studies of Finnish and Ashkenazi Jewish subjects, variation near the P2 region of HNF4A is associated with type 2 diabetes in the Danish population.

Studies of relationships between variation of the human G protein-coupled receptor 40 Gene and Type 2 diabetes and insulin release.

AIMS: Recently, a novel human G protein-coupled receptor 40 (GPR40), which is predominantly expressed in pancreatic islets, was shown to mediate an amplifying effect of long-chain fatty acids on glucose-induced insulin secretion. The present aim was to examine the coding region of GPR40 for variation and to assess whether identified variants confer an increased risk of Type 2 diabetes or altered insulin release. METHODS: Mutation analysis was performed in 43 patients with Type 2 diabetes, 18 normal glucose-tolerant subjects, and 3 maturity-onset of diabetes in the young (MODY) X patients using direct sequencing. Genotyping was performed using polymerase chain reaction (PCR)-generated primer extension products analysis by high throughput chip-based mass spectrometry (MALDI-TOF). The potential impact of GPR40 mutations on [(3)H]-myo-inositol turnover was estimated in COS-7 cells after stimulation with various concentrations of 5,8,11-eicosatriynoic acid. RESULTS: Two nucleotide substitutions, an Arg211His polymorphism and a rare Asp175Asn mutation, were identified. Both variants showed EC(50) values similar to the wild type. However, the maximal efficacy of the rare Asp175Asn was 39% lower compared with the wild type (P = 0.01). The Arg211His polymorphism had a similar allele frequency among 1384 Type 2 diabetic patients [MAF%; 23.4 (95% CI: 21.8-25.0)] and 4424 middle-aged glucose-tolerant subjects [24.1% (23.2-25.0)]. A genotype-quantitative trait study of 5597 non-diabetic, middle-aged subjects from the Inter99 cohort showed no significant differences in oral glucose tolerance test (OGTT)-derived estimates of insulin release between carriers of various GPR40 genotypes. CONCLUSIONS: Variations in the coding region of GPR40 do not appear to be associated with Type 2 diabetes or insulin release alterations.

Large-scale studies of the HphI insulin gene variable-number-of-tandem-repeats polymorphism in relation to Type 2 diabetes mellitus and insulin release.

AIMS/HYPOTHESIS: The class III allele of the variable-number-of-tandem-repeats polymorphism located 5' of the insulin gene (INS-VNTR) has been associated with Type 2 diabetes and altered birthweight. It has also been suggested, although inconsistently, that the class III allele plays a role in glucose-induced insulin response among NGT individuals. METHODS: We investigated the impact of the class III allele on Type 2 diabetes susceptibility in a case-control study involving 1462 Type 2 diabetic patients and 4931 NGT subjects. We also examined the potential impact of the class III allele in genotype-quantitative trait studies in three Danish study populations containing (i) 358 young healthy subjects; (ii) 4444 middle-aged NGT subjects, 490 subjects with IFG and 678 subjects with IGT; and (iii) 221 NGT subjects, of whom one parent had Type 2 diabetes. RESULTS: There was no difference in frequency of the class III allele or in genotype distribution between the 1462 Type 2 diabetic patients and the 4931 NGT subjects. Among the 358 young subjects the class III/III carriers had significantly reduced post-IVGTT acute serum insulin and C-peptide responses (p=0.04 and 0.03 respectively). However, among the 4444 middle-aged subjects we failed to demonstrate any association between the class III allele and post-OGTT serum insulin and C-peptide levels. CONCLUSIONS/INTERPRETATION: The class III allele of the INS-VNTR does not confer susceptibility to Type 2 diabetes or consistent alterations in glucose-induced insulin release in the examined populations, which consisted of Danish Caucasians.

Studies of variability in the islet amyloid polypeptide gene in relation to Type 2 diabetes.

AIMS: To explore whether the coding region of the islet amyloid polypeptide (IAPP) gene contains genetic variants associated with Type 2 diabetes and whether a previously reported association of the promoter variant -132g-->a with Type 2 diabetes could be reproduced in Danish Caucasians. METHODS: The coding region was analyzed using single strand conformation polymorphism (SSCP) and heteroduplex analysis in 192 Type 2 diabetic patients. Restriction fragment length polymorphism (RFLP) was employed to screen for the promoter variant in 414 Type 2 diabetic patients and 182 glucose-tolerant control subjects. RESULTS: The SSCP analysis identified an IVS+75a-->g variant in two patients. The frequency of heterozygous carriers of the promoter variant in the case-control study was 4.1% (17/414) and 7.1% (13/182), respectively. Odds ratio of the prevalence of Type 2 diabetes in carriers compared with non-carriers was estimated to be 0.47 (95% confidence interval 0.19, 1.15). CONCLUSION: Neither variability in the coding region of the IAPP gene nor the -132g-->a promoter variant was associated with Type 2 diabetes among the studied Danish Caucasians.

In situ detection of horizontal transfer of mobile genetic elements.

Plasmid transfer was investigated in microbial populations associated with different types of surfaces. The general strategy behind these investigations was to label the transferable plasmid with a gene encoding a fluorescent protein in order to make it a transfer reporter. This was achieved by fusing the reporter gene with a lac promoter expression cassette and combining this with a donor cell-associated lacI repressor cassette. After construction of a range of strains and plasmids with combinations of genes expressing fluorescent proteins from constitutive (cell tagging) or regulated promoters (transfer reporters) it was thus possible to detect transfer events in situ and correlate these with either the location of donor and recipient cells or with the growth activity of the cells. In some cases, expression of unstable Gfp from a growth-controlled promoter, rrnB from Escherichia coli, was used to monitor bacterial growth activity in situ. Differential tagging of mobilizing and mobilizable plasmids with different genes encoding fluorescent proteins with varying emission wavelengths allowed in situ detection of plasmid mobilization and detection of retro-transfer on agar surfaces. The obtained data show that the several different types of fluorescent reporters, which are now available, allow more informative in situ investigations of horizontal gene transfer to be carried out, and by combining these genes with various expression systems it is possible to simultaneously monitor donor/recipient positioning, cellular activity and appearance of transconjugants.

[Kawasaki syndrome in an adult]. / Kawasaki-syndrom hos en voksen.

Kawasaki syndrome (KS) is an acute, febrile, exanthemous illness characterized by vasculitis of unknown origin mostly seen in children younger than four years of age. We describe a 24-year-old white male who fulfilled the diagnostic criteria for KS. In addition to the diagnostic symptoms of KS this patient had symptoms from the liver and lungs.

House dust mite antigen exposure of patients with atopic dermatitis of psoriasis.

We studied the amount of house dust antigen in the beds of 55 patients with atopic dermatitis, eleven patients with psoriasis and ten healthy volunteers using a commercial ELISA which can determine the amount of antigen from Dermatophagoides pteronyssinus, D. farinae and D. microseras expressed as nanogram (ng) antigen per gram of house dust. The World Health Organization has indicated that 10,000 ng house dust mite antigen per gram house dust can elicit an asthma attack in IgE-sensitized patients with asthma bronchiale. There are no recommendations for patients with atopic eczema. We observed no statistical significant differences between each group concerning the amount of house dust found in the beds or the amount of house dust mite antigen. However, there were very wide variations. Twenty-seven percent (15/55) of patients with atopic dermatitis and 27% (3/11) of psoriasis patients had levels of house dust mite antigen above 10,000 ng per gram of house dust compared with healthy volunteers (1/10). Half of the patients had a type I allergy to house dust mite antigen using prick tests. This group had a total serum IgE of 2,034 kU/I (median value) compared to 301 kU/I in the group without type I allergy to house dust mite antigen (p < 0.01). The exposure to house dust mite antigen was similar in the two groups. We conclude that only 1/4 of patients with atopic dermatitis are exposed to high levels of house dust mite antigen in their bed environment equal to what is found for patients with another scaling disorder (psoriasis). Patients who have an increased serum IgE have significantly increased type I allergy to house dust mite antigen even though their exposure is not different from patients with low IgE.

Transcription properties of a cell type-specific TATA-binding protein, TRF.

Eukaryotic cells are thought to contain a single TATA-binding protein (TBP) that directs transcription by cellular RNA polymerases. Here we report a cell type-specific TBP-related factor (TRF) that can form a stable TRF/IIA/IIB TATA DNA complex and substitute for TBP in directing RNA polymerase II transcription in vitro. Transfection studies reveal that TRF can differentially mediate activation by some enhancer proteins but not others. Like TBP, TRF forms a stable complex containing multiple novel subunits, nTAFs. Antibody staining of embryos and polytene chromosomes reveals cell type-specific expression and gene-selective properties consistent with the shaker/male sterile phenotype of trf mutants. These findings suggest TRF is a homolog of TBP that functions to direct tissue- and gene-specific transcription.

Drosophila TFIIE: purification, cloning, and functional reconstitution.

We present a physical and molecular genetic characterization of Drosophila melanogaster TFIIE (dTFIIE), a component of the basal RNA polymerase II transcription apparatus. We have purified dTFIIE to near homogeneity from nuclear extracts of Drosophila embryos and found that it is composed of two subunits with apparent molecular weights of 55 and 38 kDa. Peptide sequence information derived from the two subunits was used to isolate the corresponding cDNA clones, revealing that dTFIIE and human TFIIE share extensive amino acid similarity. Functional conservation was demonstrated by the ability of bacterially expressed dTFIIE to substitute for human TFIIE in an in vitro transcription assay reconstituted from purified components. Cytological mapping analysis shows that both subunits are encoded by single copy genes located on chromosome III.

[Milker's nodules]. / Malkerknuder.

Milker's nodule is a parapox virus infection seen mostly on the hands of dairy farmers. We saw 15 cases over a period of two years in the County of North Jutland. Clinically, milker's nodule goes through a papular, a nodular and a crusted stage. Most patients were seen when the infection was in the nodular stage, an often painful condition requiring treatment. Three patients developed an erythema multiformelike secondary eruption. Lesions from nine patients were removed for histological examination. The histology of all lesions was consistent with milker's nodule. In three of seven patients parapox virus was demonstrated by electron microscopy. Treatment was commonly curettage followed by cauterization.

SREBP transcriptional activity is mediated through an interaction with the CREB-binding protein.

The sterol regulatory element binding proteins (SREBP-1 and -2) activate transcription of genes whose products are involved in the cellular uptake and synthesis of cholesterol. Although considerable effort has been exerted to define the events regulating the levels of active SREBP, little is known about the transcriptional cofactors mediating SREBP function. In an unbiased search for potential coactivators of SREBP, we isolated a protein of 265 kD from HeLa cells that directly bound SREBP-1 and SREBP-2. Peptide sequencing and Western blot analysis established that the 265-kD protein was CBP (CREB-binding protein), a recently identified transcriptional coactivator. The putative activation domain of SREBP was shown to bind specifically to amino-terminal domains of recombinant CBP and p300 (a CBP-related protein). Moreover, transfection studies demonstrated that CBP enhances the ability of SREBP to activate transcription of reporter genes in HeLa cells. Together, these data suggest that CBP mediates SREBP transcriptional activity, thus revealing a new step in the biochemical pathway regulating cholesterol metabolism.

[Use of contraception and reasons for choosing abortion among abortion applicants]. / Abortsøgendes praeventionsbrug og årsager til valg af abort.

The object of this study was to describe a group of women applying for legal abortion in relation to their use of contraception and reasons for choosing an abortion. During a period of 13 months (1991-92) a questionnaire was distributed to women applying for legal abortion at Hillerød Hospital in Denmark. Three hundred and thirty-nine women participated. Fifty-nine percent of the women had become pregnant although they had used contraception. As seen in other studies, women still state demographic factors as their most important reasons for choosing an abortion. Women with two or more children do not want to have more children. Single women do not want children without being in a stable relationship. Furthermore occupation and education were frequently stated as important reasons for having an abortion. Economy and housing were not main reasons but contributory factors. Thirty percent of the women expressed ambivalence about the choice of abortion at the time when the abort was due.