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Abstract Background Geographical access to health care facilities is known to influence health services usage. As societies age, accessibility to health care becomes an increasingly acute public health concern. It is known that seniors tend to have lower mobility levels, and it is possible that this may negatively affect their ability to reach facilities and services. Therefore, it becomes important to examine the mobility situation of seniors vis-a-vis the spatial distribution of health care facilities, to identify areas where accessibility is low and interventions may be required. Methods Accessibility is implemented using a cumulative opportunities measure. Instead of assuming a fixed bandwidth (i.e. a distance threshold) for measuring accessibility, in this paper the bandwidth is defined using model-based estimates of average trip length. Average trip length is an all-purpose indicator of individual mobility and geographical reach. Adoption of a spatial modelling approach allows us to tailor these estimates of travel behaviour to specific locations and person profiles. Replacing a fixed bandwidth with these estimates permits us to calculate customized location- and person-based accessibility measures that allow inter-personal as well as geographical comparisons. Data The case study is Montreal Island. Geo-coded travel behaviour data, specifically average trip length, and relevant traveller's attributes are obtained from the Montreal Household Travel Survey. These data are complemented with information from the Census. Health care facilities, also geo-coded, are extracted from a comprehensive business point database. Health care facilities are selected based on Standard Industrial Classification codes 8011-21 (Medical Doctors and Dentists). Results Model-based estimates of average trip length show that travel behaviour varies widely across space. With the exception of seniors in the downtown area, older residents of Montreal Island tend to be significantly less mobile than people of other age cohorts. The combination of average trip length estimates with the spatial distribution of health care facilities indicates that despite being more mobile, suburban residents tend to have lower levels of accessibility compared to central city residents. The effect is more marked for seniors. Furthermore, the results indicate that accessibility calculated using a fixed bandwidth would produce patterns of exposure to health care facilities that would be difficult to achieve for suburban seniors given actual mobility patterns. Conclusions The analysis shows large disparities in accessibility between seniors and non-seniors, between urban and suburban seniors, and between vehicle owning and non-owning seniors. This research was concerned with potential accessibility levels. Follow up research could consider the results reported here to select case studies of actual access and usage of health care facilities, and related health outcomes.

School Absence as Functional Impairment of Juveniles with Personality Disorders Based on ICD-11 This longitudinal study focused on the question whether specialized treatment with applied transference focused psychotherapy in a day hospital program can improve school dysfunction in adolescents with borderline and other personality disorders (PD). Moreover, we investigated influential factors for the improvement of school attendance. Among the 175 juvenile patients, before treatment 60 % showed school absence. Our results indicate that the treatment could improve school attendance significantly. At the same time, we could show that some psychological variables can be considered essential predictors for the development of attending school regularly. More conduct problems and dissocial behavior diminished the likelihood to attend school increasingly regular; the higher borderline and avoidance PD criteria were fulfilled, the higher were the odds to improve school attendance. With respect to the changes regarding functional impairment of PD in the upcoming ICD-11 our results could generate knowledge that school dysfunction in adolescents with PD can be improved significantly with applied transference focused psychotherapy in a day hospital program, especially in juveniles with borderline and avoidant PD. The alteration in ICD-11 with its focus on functional impairment enables to assess and treat this problem earlier in adolescence. A specific focus for future research based on ICD-11 should address the issue how juveniles with pd, school dysfunction and conduct problems can benefit more from treatment.

Background: Metabolic syndrome (MetS) is a collection of risk factors, including hypertension, high fasting blood glucose, high fasting triglyceride and low high-density lipoprotein (HDL) cholesterol levels that may increase the risk for cardiovascular disease and type 2 diabetes. The study aimed to determine the prevalence of MetS among adults attending a Free State district hospital's outpatient department., Methods: A cross-sectional study included a consecutive sample of consenting patients 18 years and older from 18 October 2021 to 19 November 2021. Patients' waist circumference was measured, and data were extracted from patients' files., Results: The 409 participants were predominantly females (64.2%). The median age was 60 years. Triglyceride and HDL cholesterol levels were available for 27.4% and 26.9% of patients, respectively. Of the 278 (68.0%) patients with sufficient information to determine their MetS status, 187 (67.3%) had MetS. Of the males with sufficient information, 49.1% (n = 56/114) had MetS compared to 79.9% (n = 131/164) of the females with sufficient information (p  0.001). The age group 60-79 years had the highest prevalence (76.7%, p  0.001). In all race groups, at least two-thirds of patients had MetS (p = 0.831)., Conclusion: Incomplete patient notes and failure to do investigations led to a third of patients not having sufficient information to determine their MetS status. In patients with sufficient information, a high prevalence of MetS was found.Contribution: This study highlights the challenges of determining MetS retrospectively in an outpatient population and the need for completeness of medical note keeping and routine investigations in high-risk patients. It also notes the high prevalence of MetS.

To design new CARs targeting hepatitis B virus (HBV), we isolated human monoclonal antibodies recognizing the HBV envelope proteins from single B cells of a patient with a resolved infection. HBV-specific memory B cells were isolated by incubating peripheral blood mononuclear cells with biotinylated hepatitis B surface antigen (HBsAg), followed by single-cell flow cytometry-based sorting of live, CD19 + IgG + HBsAg + cells. Amplification and sequencing of immunoglobulin genes from single memory B cells identified variable heavy and light chain sequences. Corresponding immunoglobulin chains were cloned into IgG1 expression vectors and expressed in mammalian cells. Two antibodies named 4D06 and 4D08 were found to be highly specific for HBsAg, recognized a conformational and a linear epitope, respectively, and showed broad reactivity and neutralization capacity against all major HBV genotypes. 4D06 and 4D08 variable chain fragments were cloned into a 2 nd generation CAR format with CD28 and CD3zeta intracellular signaling domains. The new CAR constructs displayed a high functional avidity when expressed on primary human T cells. CAR-grafted T cells proved to be polyfunctional regarding cytokine secretion and killed HBV-positive target cells. Interestingly, background activation of the 4D08-CAR recognizing a linear instead of a conformational epitope was consistently low. In a preclinical model of chronic HBV infection, murine T cells grafted with the 4D06 and the 4D08 CAR showed on target activity indicated by a transient increase in serum transaminases, and a lower number of HBV-positive hepatocytes in the mice treated. This study demonstrates an efficient and fast approach to identifying pathogen-specific monoclonal human antibodies from small donor cell numbers for the subsequent generation of new CARs., Competing Interests: UP is a co-founder, shareholder, and SCG Cell Therapy Pte Ltd board member. UP received personal fees from Abbott, Abbvie, Arbutus, Gilead, GSK, J&J, MSD, Roche, Sanofi, Sobi, and Vaccitech. SR was a founder, has served as an advisor, and has patents licensed to Juno Therapeutics; is a founder of and holds equity in Lyell Immunopharma; and has served on the advisory boards for Adaptive Biotechnologies and Nohla. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Schreiber, Dressler, Loffredo-Verde, Asen, Färber, Wang, Groll, Chakraborty, Kolbe, Kreer, Kosinska, Simon, Urban, Klein, Riddell and Protzer.)

Background: The unknown tissue of origin in head and neck cancer of unknown primary (hnCUP) leads to invasive diagnostic procedures and unspecific and potentially inefficient treatment options for patients. The most common histologic subtype, squamous cell carcinoma, can stem from various tumor primary sites, including the oral cavity, oropharynx, larynx, head and neck skin, lungs, and esophagus. DNA methylation profiles are highly tissue-specific and have been successfully used to classify tissue origin. We therefore developed a support vector machine (SVM) classifier trained with publicly available DNA methylation profiles of commonly cervically metastasizing squamous cell carcinomas (n = 1103) in order to identify the primary tissue of origin of our own cohort of squamous cell hnCUP patient's samples (n = 28). Methylation analysis was performed with Infinium MethylationEPIC v1.0 BeadChip by Illumina., Results: The SVM algorithm achieved the highest overall accuracy of tested classifiers, with 87%. Squamous cell hnCUP samples on DNA methylation level resembled squamous cell carcinomas commonly metastasizing into cervical lymph nodes. The most frequently predicted cancer localization was the oral cavity in 11 cases (39%), followed by the oropharynx and larynx (both 7, 25%), skin (2, 7%), and esophagus (1, 4%). These frequencies concord with the expected distribution of lymph node metastases in epidemiological studies., Conclusions: On DNA methylation level, hnCUP is comparable to primary tumor tissue cancer types that commonly metastasize to cervical lymph nodes. Our SVM-based classifier can accurately predict these cancers' tissues of origin and could significantly reduce the invasiveness of hnCUP diagnostics and enable a more precise therapy after clinical validation., (© 2024. The Author(s).)

Introduction: Radiohybrid (rh) ligands, a novel class of prostate-specific membrane antigen (PSMA)-targeted radiopharmaceuticals, can be labeled either with [ 18 F]fluorine via isotopic exchange or with radiometals (such as [ 68 Ga]Gallium, [ 177 Lu]Lutetium, [ 225 Ac]Actinium). Among these, [ 18 F, nat Ga]rhPSMA-7 has recently entered clinical assessment., Aim: Since [ 18 F, nat Ga]rhPSMA-7 is composed of four stereoisomers ([ 18 F, nat Ga]rhPSMA-7.1, -7.2, -7.3 and -7.4), we initiated a preclinical selection process to identify the isomer with the most favorable pharmacokinetics for further clinical investigation., Methods: A synthetic protocol for enantiopure [ 19 F, nat Ga]rhPSMA-7 isomers has been developed. The comparative evaluation of the four isomers comprised human serum albumin binding, lipophilicity, IC 50 , internalization and classical biodistribution studies and competition experiments in LNCaP tumor-bearing CB-17 SCID mice. In addition, a radio high-performance liquid chromatography-based method was developed allowing quantitative, intraindividual comparison of [ 18 F, nat Ga]rhPSMA-7.1 to -7.4 in LNCaP tumor-bearing mice., Results: Cell studies revealed high PSMA affinity and internalization for [ 18/19 F, nat Ga]rhPSMA-7.2, -7.3 and -7.4, whereas [ 18/19 F, nat Ga]rhPSMA-7.1 showed approximately twofold lower values. Although the biodistribution profile obtained was typical of PSMA inhibitors, it did not allow for selection of a lead candidate for clinical studies. Thus, an intraindividual comparison of all four isomers in LNCaP tumor-bearing mice was carried out by injection of a diastereomeric mixture, followed by analysis of the differential uptake and excretion pattern of each isomer. Based on its high tumor accumulation and low uptake in blood, liver and kidneys, [ 18 F, nat Ga]rhPSMA-7.3 was identified as the preferred isomer and transferred into clinical studies., Conclusion: [ 18 F, nat Ga]rhPSMA-7.3 has been selected as a lead compound for clinical development of a [ 18 F]rhPSMA-based candidate. The intraindividual differential uptake and excretion analysis in vivo allowed for an accurate comparison and assessment of radiopharmaceuticals.

Genebanks hold comprehensive collections of cultivars, landraces and crop wild relatives of all major food crops, but their detailed characterization has so far been limited to sparse core sets. The analysis of genome-wide genotyping-by-sequencing data for almost all barley accessions of the German ex situ genebank provides insights into the global population structure of domesticated barley and points out redundancies and coverage gaps in one of the world's major genebanks. Our large sample size and dense marker data afford great power for genome-wide association scans. We detect known and novel loci underlying morphological traits differentiating barley genepools, find evidence for convergent selection for barbless awns in barley and rice and show that a major-effect resistance locus conferring resistance to bymovirus infection has been favored by traditional farmers. This study outlines future directions for genomics-assisted genebank management and the utilization of germplasm collections for linking natural variation to human selection during crop evolution.

Activation of the hamstrings has been discussed as a measure for reducing strain on the ACL during jump landings in alpine skiing. The current study tested the hypothesis that hamstring and quadriceps activation can be voluntarily increased by the athlete. Specifically, two different instructions - to increase hamstring activation or to increase upper-leg co-contraction - were compared to normal landings. Eight members of the German national and junior national squad in freestyle skiing (age 19.6 ± 3.8 years; weight 66.1 ± 13.2 kg; height 172.2 ± 7.7 cm) performed 12 jump landings on a prepared run, 4 with no specific instruction, 4 with the instruction to generally activate the thigh muscles, and 4 with the instruction to specifically activate the hamstrings. Electromyographic (EMG) signals were recorded on the biceps femoris (BF), semitendinosus (ST), vastus lateralis (VL), rectus femoris (RF) and vastus medialis (VM). EMG activation levels were integrated over three landing phases and analysed with a repeated measures ANOVA. The instruction produced a significant main effect in ST (p = .026), VM (p = .032) and RF (p = .001). Contrary to previous research, the current study suggests that hamstring muscle activation levels can be voluntarily increased during jump landing, particularly in co-activation with its antagonists.

The strong link between T-cell metabolism and effector functions is well characterized in the murine system but hardly investigated in human T cells. Therefore, we analyzed glycolytic and mitochondrial activity in correlation to function in activated human CD4 and CD8 T cells. Glycolysis was barely detectable upon stimulation but accelerated beyond 24 h, whereas mitochondrial activity was elevated immediately in both T-cell populations. Glucose deprivation or mitochondrial restriction reduced proliferation, had only a transient impact on "on-blast formation" and no impact on viability, IFN-γ, IL-2, IL-4, and IL-10 production, whereas TNF was reduced. Similar results were obtained in bulk T cells and T-cell subsets. Elevated respiration under glucose restriction demonstrated metabolic flexibility. Administration of the glycolytic inhibitor 2-deoxy-glucose suppressed both glycolysis and respiration and exerted a strong impact on cytokine production that persisted for IFN-γ after removal of 2-deoxy-glucose. Taken together, glycolytic or mitochondrial restriction alone compromised proliferation of human T cells, but barely affected their effector functions. In contrast, effector functions were severely affected by 2-deoxy-glucose treatment., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: One of the most insidious characteristics of cancer is its spread to and ability to compromise distant organs via the complex process of metastasis. Communication between cancer cells and organ-resident cells via cytokines/chemokines and direct cell-cell contacts are key steps for survival, proliferation and invasion of metastasized cancer cells in organs. Precision-cut liver slices (PCLS) are considered to closely reflect the in vivo situation and are potentially useful for studying the interaction of cancer cells with liver-resident cells as well as being a potentially useful tool for screening anti-cancer reagents. Application of the PCLS technique in the field of cancer research however, has not yet been well developed., Results: We established the mouse PCLS system using perfluorodecalin (PFD) as an artificial oxygen carrier. Using this system we show that the adherence of green fluorescent protein (GFP) labeled MDA-MB-231 (highly invasive) cells to liver tissue in the PCLS was 5-fold greater than that of SK-BR-3 (less invasive) cells. In addition, we generated PCLS from THOC5, a member of transcription/export complex (TREX), knockout (KO) mice. The PCLS still expressed Gapdh or Albumin mRNAs at normal levels, while several chemokine/growth factor or metalloprotease genes, such as Cxcl12, Pdgfa, Tgfb, Wnt11, and Mmp1a genes were downregulated more than 2-fold. Interestingly, adhesion of cancer cells to THOC5 KO liver slices was far less (greater than 80% reduction) than to wild-type liver slices., Conclusion: Mouse PCLS cultures in the presence of PFD may serve as a useful tool for screening local adherence and invasiveness of individual cancer cells, since single cells can be observed. This method may also prove useful for identification of genes in liver-resident cells that support cancer invasion by using PCLS from transgenic liver.

Background: THO (Suppressors of the transcriptional defects of hpr1 delta by overexpression) complex 5 (THOC5), an mRNA export protein, is involved in the expression of only 1% of all genes. Using an interferon inducible knockout mouse system, we have previously shown that THOC5 is an essential element in the maintenance of hematopoietic stem cells and cytokine-mediated hematopoiesis in adult mice. Here we interrogate THOC5 function in cell differentiation beyond the hematopoietic system and study pathological changes caused by THOC5 deficiency., Results: To examine whether THOC5 plays a role in general differentiation processes, we generated tamoxifen inducible THOC5 knockout mice. We show here that the depletion of THOC5 impaired not only hematopoietic differentiation, but also differentiation and self renewal of the gut epithelium. Depletion of the THOC5 gene did not cause pathological alterations in liver or kidney. We further show that THOC5 is indispensable for processing of mRNAs induced by Wnt (wingless/integrated) signaling which play key roles in epithelial cell differentiation/proliferation. A subset of Wnt target mRNAs, SRY-box containing gene 9 (Sox9), and achaete-scute complex homolog 2 (Ascl2), but not Fibronectin 1 (Fn1), were down-regulated in THOC5 knockout intestinal cells. The down-regulated Wnt target mRNAs were able to bind to THOC5. Furthermore, pathological alterations in the gastrointestinal tract induced translocation of intestinal bacteria and caused sepsis in mice. The bacteria translocation may cause Toll-like receptor activation. We identified one of the Toll-like receptor inducible genes, prostaglandin-endoperoxidase synthase 2 (Ptgs2 or COX2) transcript as THOC5 target mRNA., Conclusion: THOC5 is indispensable for processing of only a subset of mRNAs, but plays a key role in processing of mRNAs inducible by Wnt signals. Furthermore, THOC5 is dispensable for general mRNA export in terminally differentiated organs, indicating that multiple mRNA export pathways exist. These data imply that THOC5 may be a useful tool for studying intestinal stem cells, for modifying the differentiation processes and for cancer therapy.

Background: The transcription/export complex is evolutionarily conserved from yeast to man and is required for coupled transcription elongation and nuclear export of mRNAs. FMIP(Fms interacting protein) is a member of the THO (suppressors of the transcriptional defects of hpr1delta by overexpression) complex which is a subcomplex of the transcription/export complex. THO complex (THOC) components are not essential for bulk poly (A)+ RNA export in higher eukaryotes, but for the nuclear export of subset of mRNAs, however, their exact role is still unclear., Results: To study the role of THOC5/Fms interacting protein in vivo, we generated THOC5/Fms interacting protein knockout mice. Since these mice are embryonic lethal, we then generated interferon inducible conditional THOC5/Fms interacting protein knockout mice. After three poly injections all of the mice died within 14 days. No pathological alterations, however, were observed in liver, kidney or heart. Thus we considered the hematopoietic system and found that seven days after poly injection, the number of blood cells in peripheral blood decreased drastically. Investigation of bone marrow cells showed that these became apoptotic within seven days after poly injection. Committed myeloid progenitor cells and cells with long term reconstituting potential were lost from bone marrow within four days after poly injection. Furthermore, infusion of normal bone marrow cells rescued mice from death induced by loss of THOC5/Fms interacting protein., Conclusion: THOC5/Fms interacting protein is an essential element in the maintenance of hematopoiesis. Furthermore, mechanistically depletion of THOC5/Fms interacting protein causes the down-regulation of its direct interacting partner, THOC1 which may contribute to altered THO complex function and cell death.

Doppler vibrometers are used by many research groups to monitor the motion of the tympanic membrane (TM) and of middle ear ossicles for in vivo and in vitro studies. Power densities in these applications reach 80 W/cm(2). To determine the safe limit of exposure, a cw dye laser at a wavelength of 633 nm was used to investigate the threshold of thermal damage of TM of pigs under exposure times of 60 s. To determine the applied power density accurately, the spot size of the laser beam was monitored by an objective lens and a CCD camera. Twenty-six laser exposed samples of TM were stained by haematoxylin and eosin stain and the semi-thin sections were examined microscopically. In none of the sections was any laser induced damage observed with power densities below 7.1 kW/cm(2), whereas serious damage occurred showing coagulation, carbonisation and perforation in all cases with laser powers above 8.2 kW/cm(2). The threshold for damage and the conical shape of the damage zone is explained by photon propagation and absorption in the tissue especially by the increase of the scattering factor at higher tissue temperature. The thermal damage threshold of 8 kW/cm(2) is compared to the maximum permissible exposure given in laser safety standards for skin.

By means of concrements, which were obtained endoscopically by extraction from the bile duct, and of gallstones, which were given the patient by the surgeon on the occasion of the gallbladder operation, we performed stone analyses with the aid of infrared spectroscopy and compared them correlatively with clinical and case history data. The paper focussed on the detection of the stony structure of choledochal concrements, separated into stone core and stone mantle. A method for the reliable differentiation of stones left (residual stones) from new formations (recurrent stones) in the bile ducts has not been possible to date. Thus, the burning question of the surgeon cannot be answered with certainty. However, there is no doubt about surgical suture material as incrustation core for a pigment stone in terms of a true recurrent stone, which poses a real challenge to biliary tract surgery.