Third-generation tyrosine kinase inhibitors (TKIs) were developed to overcome T790M-mediated resistance to earlier generations of epidermal growth factor receptor (EGFR)-targeted TKIs. We compared four well-established and one in-house method for the analysis of the EGFR T790M mutation in plasma cell-free DNA (cfDNA), in hope to find a better way to select non-small cell lung cancer (NSCLC) patients appropriate for 3rd-generation TKI therapy. For sensitivity levels of each method, plasmid DNA with EGFR T790M mutations was serially diluted with cfDNA from healthy controls with wild type EGFR. The clinical performance was analyzed in a clinical cohort of EGFR mutation-positive NSCLC patients with acquired EGFR TKI resistance (n = 40). All methods except the therascreen kit (Qiagen) had a sensitivity level of 10 copies of T790M plasmid DNA in the spiked specimen. The detection rates of the EGFR T790M mutation in plasma cfDNA from the clinical cohort were 42.5, 35, 32.5, 22.5, and 17.5% for the in-house ARMS method, Bio-Rad droplet digital PCR, PANAMutyper, Therascreen EGFR Plasma RGQ PCR Kit and Cobas EGFR Mutation kit (with suboptimal template amounts), respectively. Osimertinib was given to 17 of 20 patients with EGFR T790M mutations. The best treatment responses, based on the RECIST criteria, included 6 partial responses (PR) and 7 stable diseases (SD). The PANAMutyper and the Bio-Rad droplet digital PCR were comparable, the Cobas EGFR Mutation kit required significantly more template for testing. The best combination would be the in-house ARMS method plus the PANAMutyper or Bio-Rad droplet digital PCR, which would have a detection rate of 50% (20/40) and a disease control rate of 76% (13/17).

Background: Surgical site infection (SSI) hampers the advantages of surgical management, which requires early forecast particularly in patients receiving colorectal surgery. This study is to explore potential relationship between individual abdominal anatomic characteristics including subcutaneous fat thickness (SFT), rectus abdominis thickness (RAT), and abdomen depth (AD), with the incidence of SSI in elective radical resection of colon malignancy. Materials and Methods: This retrospective case-control study has recruited 55 patients in each SSI and non-SSI group with propensity score match method. Demographics, clinical attributes, and pre- and intra-operative information were compared between groups with univariate analysis to elicit significant parameters, which were subsequently brought into logistic regression and receiver-operating characteristic (ROC) analysis. Results: Patients with SSI showed lower preoperative albumin (p = 0.0022), higher RAT (p = 0.014), AD (p = 0.029), and the multiplied value (RAT × AD) (p = 0.0026) contrasted with patients without SSI. Logistic regression demonstrated RAT × AD as an independent risk factor for SSI (OR = 1.007, p < 0.001) and a biomarker for SSI prediction (AUC = 0.83, 95% CI: 0.74 ~ 0.91). Conclusions: Preoperative RAT and abdomen depth are associated with the risk of postoperative SSI in patients receiving elective radical resection of colon cancer. Trial Registration: www.researchregistry.com, identifier researchregistry3669.

In brain/head-and-neck radiotherapy (RT), thermoplastic immobilization masks guarantee reproducible patient positioning in treatment position between MRI, CT, and irradiation. Since immobilization masks do not fit in the diagnostic MR head/head-and-neck coils, flexible surface coils are used for MRI imaging in clinical practice. These coils are placed around the head/neck, in contact with the immobilization masks. However, the positioning of these flexible coils is technician dependent, thus leading to poor image reproducibility. Additionally, flexible surface coils have an inferior signal-to-noise-ratio (SNR) compared to diagnostic coils. The aim of this work was to create a new immobilization setup which fits into the diagnostic MR coils in order to enhance MR image quality and reproducibility. For this purpose, a practical immobilization setup was constructed. The performances of the standard clinical and the proposed setups were compared with four tests: SNR, image quality, motion restriction, and reproducibility of inter-fraction subject positioning. The new immobilization setup resulted in 3.4 times higher SNR values on average than the standard setup, except directly below the flexible surface coils where similar SNR was observed. Overall, the image quality was superior for brain/head-and-neck images acquired with the proposed RT setup. Comparable motion restriction in feet-head/left-right directions (maximum motion ≈1 mm) and comparable inter-fraction repositioning accuracy (mean inter-fraction movement 1 ± 0.5 mm) were observed for the standard and the new setup.

Background:Streptococcus pneumoniae is the most common pathogen causing death in children under 5 years old. This retrospective surveillance aimed to analyze serotype distribution, drug resistance, virulence factors, and molecular characteristics of pneumonia isolates from children in Shanghai, China. Methods: A total of 287 clinical pneumococcal isolates were collected from January to December in 2018 and were divided into community-acquired pneumonia (CAP) and healthcare-associated pneumonia (HAP) two groups according to where someone contracts the infection. All isolates were serotyped by multiplex sequential PCR and antimicrobial susceptibility testing was performed using E-test or disk diffusion method. The molecular epidemiology was analyzed using multilocus sequence typing and seven housekeeping genes were sequenced to identified the sequence types (STs). In addition, we investigated the presence of virulence genes via PCR. Results: The most common serotypes were 19F, 6A, 19A, 23F, 14, and 6B, and the coverage rates of the 7-, 10- and 13-valent pneumococcal conjugate vaccines were 58.9, 58.9, and 80.5%, respectively. More PCV13/non-PCV7 serotypes and higher rate of penicillin non-susceptible S. pneumoniae were seen in HAP. Molecular epidemiological typing showed a high level of diversity and five international antibiotic-resistant clones were found, including Taiwan19F-14, Spain23F-1, Spain6B-2, Taiwan23F-15 and Sweden15A-25. No significant difference was observed in the presence of virulence genes among the isolates obtained from CAP and HAP. All of the S. pneumoniae isolates carried lytA, ply, psaA, pavA, spxB, htrA, and clpP, and the carriage rate of nanA and piaA were 96.2 and 99.0%. Conversely, cps2A, cbpA, and pspA were present in 33.8-44.3% of the isolates. Conclusions: Serotype changes and emerging multidrug-resistant international clones were found in current study. lytA, ply, psaA, pavA, spxB, htrA, and clpP may be good protein vaccine candidates. Long-term high-quality surveillance should be conducted to assess impact and effectiveness brought by vaccines, and provide a foundation for prevention strategies and vaccine policies.

Background: The main property of a successful conjugation of antibodies to nanoparticles is keeping the potency of antibody for binding the antigen, and an oriented conjugation can do that. Under such ground, this study was carried out to explore the efficiency of two conjugation methods in binding iron nanoparticles to an antibody produced against PSCA (prostate stem cell antigen) using in vitro labeling of PC3 cells. Methods: In this experimental study, we conjugated dextran-superparamagnetic iron oxide nanoparticles (dexSPIONs) to anti-PSCA antibody by two different methods, including targeting carbohydrate moieties in FC domain and the free amine group of amino acid side chains. Ultimately, Iron staining was done by anti-PSCA antibody-dexSPIONs in PC3 cells to detect antibody binding to the cells. Results: A strong blue dye was induced by iron staining in conjugated dexSPIONs on the membrane of PC3 cells by the former method than the second one. Moreover, cells treated with 20 nm diameters of dexSPIONs showed higher resolution of blue color than those treated with 100 nm nanoparticles. Conclusion: This oriented conjugation method promoted the efficiency of targeting tumor antigens, and the presence of iron particles might enhance MRI image intensity in vivo by targeting PSCA-overexpressing cells in future studies.

Background: Male and female newborns have differences in their fetal development, fetal-to-neonatal transition, and postnatal morbidity. However, the cardiovascular fetal-to-neonatal adaption is similar between sexes. No study has examined sex differences in newborns during hypoxia, asphyxia, cardio-pulmonary resuscitation, or post-resuscitation recovery. Methods: Secondary analysis (two previous publications and two studies currently under peer-review) of 110 term newborn mixed breed piglets (1-3 days of age, weighing 2.0 ± 0.2 kg), which were exposed to 30 min normocapnic hypoxia followed by asphyxia until asystole, which was achieved by disconnecting the ventilator and clamping the endotracheal tube. This was followed by cardio-pulmonary resuscitation. For the analysis piglets were divided into female and male groups. Cardiac function, carotid blood flow, and cerebral and renal oxygenation were continuously recorded throughout the experiment. Results: A total of 35/41 (85%) female and 54/69 (78%) male piglets resuscitated achieved ROSC (p = 0.881). The median (IQR) time to achieve return of spontaneous circulation in females and males was 111 (80-228) s and 106 (80-206) s (p = 0.875), respectively. The 4-h survival rate was similar between females and males with 28/35 (80%) and 49/54 (91%) piglets surviving (p = 0.241), respectively. Conclusions: No difference between female and male newborn piglets was observed during hypoxia, asphyxia, resuscitation, and post-resuscitation recovery.

Herein we introduce the constellation plots as a general approach that merges different and complementary molecular representations to enhance the information contained in a visual representation and analysis of chemical space. The method is based on a combination of a sub-structure based representation and classification of compounds with a "classical" coordinate-based representation of chemical space. A distinctive outcome of the method is that organizing the compounds in analog series leads to the formation of groups of molecules, aka "constellations" in chemical space. The novel approach is general and can be used to rapidly identify, for instance, insightful and "bright" Structure-Activity Relationships (StARs) in chemical space that are easy to interpret. This kind of analysis is expected to be especially useful for lead identification in large datasets of unannotated molecules, such as those obtained through high-throughput screening. We demonstrate the application of the method using two datasets of focused inhibitors designed against DNMTs and AKT1.

Lower-limb intersegmental coordination is a complex component of human walking. Aging may result in impairments of motor control and coordination contributing to the decline in mobility inducing loss of autonomy. Investigating intersegmental coordination could therefore provide insights into age-related changes in neuromuscular control of gait. However, it is unknown whether the age-related declines in gait performance relates to intersegmental coordination. The aim of this study was to evaluate the impact of aging on the coordination of lower limb kinematics and kinetics during walking at a conformable speed. We then assessed the body kinematics and kinetics from gait analyses of 84 volunteers from 25 to 85 years old when walking was performed at their self-selected speeds. Principal Component Analysis (PCA) was used to assess lower-limb intersegmental coordination and to evaluate the planar covariation of the Shank-Thigh and Foot-Shank segments. Ankle and knee stiffness were also estimated. Age-related effects on planar covariation parameters was evaluated using multiple linear regressions (i.e., without a priori age group determination) adjusted to normalized self-selected gait velocity. Colinearity between parameters was assessed using a variation inflation factor (VIF) and those with a VIF < 5 were entered in the analysis. Normalized gait velocity significantly decreased with aging (r = -0.24; P = 0.028). Planar covariation of inter-segmental coordination was consistent across age (99.3 ± 0.24% of explained variance of PCA). Significant relationships were found between age and intersegmental foot-shank coordination, range of motion of the ankle, maximal power of the knee, and the ankle. Lower-limb coordination was modified with age, particularly the coordination between foot, and shank. Such modifications may influence the ankle motion and thus, ankle power. This observation may explain the decrease in the ankle plantar flexor strength mainly reported in the literature. We therefore hypothesize that this modification of coordination constitutes a neuromuscular adaptation of gait control accompanying a loss of ankle strength and amplitude by increasing the knee power in order to maintain gait efficiency. We propose that foot-shank coordination might represent a valid outcome measure to estimate the efficacy of rehabilitative strategies and to evaluate their efficiency in restoring lower-limb synergies during walking.

No abstract available

Defined Daily Doses (DDD) and Defined Course Doses (DCD) have been established in both human and veterinary medicine in order to standardize the measurement of treatments in a population. In 2016 the European Medicines Agency published average defined daily dose (DDDvet) and defined course dose (DCDvet) values for antimicrobial agents used in livestock production. Similarly, national defined doses (DDDch and DCDch) for the pig sector in Switzerland have recently been determined. The aim of this study was to compare the outcome of calculating antimicrobial consumption based on either DDDvet/DCDvet or DDDch/DCDch. Data from 227 Swiss pig farms describing antimicrobial use in 2015 was collected. The numbers of treatment days and treatments were calculated using DDDvet/DCDvet and DDDch/DCDch respectively, for each farm in total and for different antimicrobial classes. Associations between calculated numbers of DDDvet/DCDvet and DDDch/DCDch on farm level were investigated. In addition, differences concerning antimicrobial use were investigated between different production types of farms (piglet-producer, finishing farm or farrow-to-finishing farm). Using DDDch/DCDch values we calculated 1,805,494 treatment days and 433,678 treatments compared to 1,456,771 treatment days (19% ratio) and 303,913 treatments (30% ratio) based on DDDvet/DCDvet. Penicillins (21.4/26.6%), polypeptides (18.6/27.6%) and fluoroquinolones (9.5/8.8%) were the most frequently used classes of antimicrobials based on calculation using both DDDch and DDDvet. Similar findings were observed for complete treatments (DCDch/vet) (penicillins: 52.8/39.6%; polypeptides: 7.8/14.2%; fluoroquinolones: 13.2/12.9%). The number of treatment days or treatments per farm was higher for piglet-producers and farrow-to-finishing farms compared to finisher farms regardless of whether Swiss or European DDD or DCD values were used for the calculation (each P < 0.001). Similar results for antimicrobial use (AMU) obtained at farm level were observed when calculated either by Swiss or European definitions. Nevertheless, marked differences could be observed in the assessment of the use of specific antimicrobial classes in the field based on DDDvet/DCDvet compared to DDDch/DCDch.

Th17/Treg balance skews towards Th17 in ITP patient. IRF4 has been highlighted for its close relationship to the immunosuppressive function of Treg cells and the IL-17 synthesis in CD4+ T cells. This study was aimed at examining the effects of IRF4 to the Th17/Treg cells in patients with ITP.

A year-long prospective study characterized the seasonality of oestrus cycles in primiparous, nonpregnant Swakara (n=8) and Damara (n=5) ewes through surveillance of plasma progesterone (P4) levels. During this period, Swakara and Damara groups evidently averaged 23 oestrus cycles with an average length of 17 days. Damara ewes showed greater mean peak plasma P4 levels (11.4±0.16ng/ml) than Swakara ewes (5.4±0.11ng/ml) (P<0.05). Oestrus cycles in Damara ewes showed relatively uniform plasma P4 peaks throughout the year ranging from 10.6±0.16 to 12.6±0.24ng/ml. In Swakara ewes, P4 peaks were highest in the autumn oestrus cycles (from 7.1±0.16 to 7.5±0.11ng/ml), rapidly declining through winter to 2.2±0.08ng/ml by midspring and then rapidly increasing to 4.9±0.37ng/ml at the commencement of summer, followed by a gradual increase from 5.7± to 7.1±ng/ml by the start of autumn. The annual mean area under the curve temporal progesterone measurements (AUCPM) in Damara ewes (115.9±18.6ng⁎day/ml) was greater than that in Swakara ewes (58.6±25.3ng⁎day/ml) (p<0.05). For Swakara ewes, the mean AUCPM in summer and autumn cycles (68.2±14.7 and 79.5±10.0ng⁎day/ml, respectively) were greater than those in spring and winter cycles (28.7±12.3 and 55.0±27.3ng⁎day/ml), respectively (P<0.05). There was no seasonal variation in the exposure of the Damara ewes to P4 in between seasons (P>0.05), though, however, the Damara ewes had greater P4 levels than the Swakara ewes (P<0.05). Progesterone profiles showed that Swakara ewes possessed 'residual' seasonality, whereas the Damara ewes were no longer seasonal. The implications of this disparity in the seasonal exposure of Swakara and Damara ewes to luteal P4 on fertility warrant further investigation.

Sri Lanka is progressing towards its goal of eliminating human rabies. This goal rests on programs designed to limit canine rabies, which in turn requires a combination of targeted dog rabies control and a better understanding of the movement of the virus between domestic animals, people, and wildlife. Coordinated and integrated surveillance of the disease between human and animal health sectors underpins successful rabies elimination. Our objective was to review surveillance data from 2005 to 2014 to assemble the first multispecies synthesis of rabies information in Sri Lanka and, in doing so, assess needs and opportunities for a One Health approach to rabies surveillance in the country. Our descriptive epidemiological findings were consistent with other studies showing a decline in human cases, endemic and unchanging numbers of dog cases, a relationship between human density and the occurrence of human and animal cases, and significant gaps in understanding trends in rabies incidences in livestock and wildlife. Assessing the trends in the data from the three government organizations responsible for rabies surveillance was difficult due to lack of information on animal population sizes, unquantified sampling biases due to inequities in access to diagnostic capacities, regulatory and administrative barriers, and a continued reliance on clinical means to establish a diagnosis. The information required for a comprehensive rabies control programme was not standardized or consistent, was not in one place, showed significant gaps in completeness, and was not amenable to routine and rapid analysis. Achieving rabies elimination in Sri Lanka would benefit from harmonization of diagnostic and information management standards across animal and human health sectors as well as equitable access to diagnostic capacity for all regions and species.

Chronic exposure to ultraviolet (UV) radiation (mainly UVA) induces a sustained increase of matrix metalloproteinases (MMPs), especially MMP1, MMP2, MMP3 and MMP9 in human skin fibroblasts. MMPs can lead to the degradation of fibrous connective tissue, which is the main cause of skin photoageing. The molecular mechanisms through which fibroblasts sense UVA and trigger the cell signalling pathways involved in the upregulation of MMPs have not been well elucidated.

Tamoxifen is very successfully used for the induction of CreERT -mediated genomic recombination in conditional mouse models. Recent studies, however, indicated that tamoxifen might also affect the fibrotic response in several disease models following administration, both in vitro and in vivo. In order to investigate a possible effect of tamoxifen on pancreatic fibrogenesis and to evaluate an optimal treatment scheme in an experimental pancreatitis mouse model, we administered tamoxifen by oral gavage to both male and female C57BL/6J mice and then waited for different periods of time before inducing chronic pancreatitis by cerulein. We observed a sex-specific and time-dependent effect of tamoxifen on the fibrotic response as measured by collagen deposition and the number of myofibroblasts and macrophages. The findings of in vitro studies, in which cerulein was administrated with or without 4-hydroxytamoxifen to stimulate primary murine female and male pancreatic stellate cells, supported our in vivo observations. Real-time PCR also indicated that this effect may be related to differences in ERα expression between female and male stellate cells. Our data demonstrate that tamoxifen administration has unignorable side effects, which affect the experimental outcome in a cerulein-based model of chronic pancreatitis in mice. We suggest a 2-week waiting period before cerulein administration to reduce side effects to a minimum for the described fibrosis model in female mice.

Bacteria exist in polymicrobial environments and compete to prevail in a niche. The type VI secretion system (T6SS) is a nanomachine employed by Gram-negative bacteria to deliver effector proteins into target cells. Consequently, T6SS-positive bacteria produce a wealth of antibacterial effector proteins to promote their survival among a prokaryotic community. These toxins are loaded onto the VgrG-PAAR spike and Hcp tube of the T6SS apparatus and recent work has started to document the specificity of effectors for certain spike components. Pseudomonas aeruginosa encodes several PAAR proteins, whose roles have been poorly investigated. Here we describe a phospholipase family antibacterial effector immunity pair from Pseudomonas aeruginosa and demonstrate that a specific PAAR protein is necessary for the delivery of the effector and its cognate VgrG. Furthermore, the PAAR protein appears to restrict the delivery of other phospholipase effectors that utilise distinct VgrG proteins. We provide further evidence for competition for PAAR protein recruitment to the T6SS apparatus, which determines the identities of the delivered effectors.

To examine the knowledge and awareness of brain death among the Saudi population and to identify what is needed to improve awareness in this community.

Long noncoding RNAs (lncRNAs) are involved in the pathology of colorectal cancer (CRC). Current efforts to eradicate CRC predominantly focused on targeting the proliferation of rapidly growing cancer epithelial cells. This is largely ineffective with resistance arising in most tumors after exposure to chemotherapy. Despite the long-standing recognition of the crosstalk between carcinoma-associated fibroblasts (CAFs) and cancer cells in the tumor microenvironment, how CAFs may contribute to drug resistance in neighboring cancer cells is not well characterized. Here, we show that lncRNA CCAL (colorectal cancer-associated lncRNA) promotes oxaliplatin (Oxa) resistance of CRC cells. RNA-ISH shows higher CCAL expressed in the tumor stroma compared to cancer nests of CRC tissues. Functional studies reveal that CCAL is transferred from CAFs to the cancer cells via exosomes, where it suppresses CRC cell apoptosis, confers chemoresistance and activates β-catenin pathway in vitro and in vivo. Mechanistically, CCAL interacts directly with mRNA stabilizing protein HuR (human antigen R) to increase β-catenin mRNA and protein levels. Our findings indicate that CCAL expressed by CAFs of the colorectal tumor stroma contributes to tumor chemoresistance and CCAL may serve as a potential therapeutic target for Oxa resistance.

Platelet-rich fibrin (PRF) membranes can preserve alveolar ridge dimension after tooth extraction. Thus, it can be presumed that PRF suppresses the catabolic events that are caused by osteoclastic bone resorption.

Bone containing tissues such as osteochondral joint are resistant to routine tissue processing, therefore require decalcification. This technique causes removal of mineral salts, but in the process may macerate the organic tissue, hence the need for tissue fixation. Such severe processing demands careful antigen retrieval to necessitate optimal staining. The aim of our study was to compare five different antigen retrieval protocols (heat retrieval and protein digestion) following decalcification of rabbit knee joints using two different techniques (20% formic acid and 10% ethylenediamine-tetra acetic acid: EDTA). Osteochondral sections were compared based on time required for decalcification, ease of sectioning, morphological integrity using HE staining and antigen preservation (Collagen type II) using immunohistochemistry. The two decalcification solutions did not impair the tissue morphology and ease of sectioning. Joints processed with formic acid decalcified four times faster than EDTA. Among the five antigen retrieval approaches, maximal collagen II uptake with minimal nonspecific staining was found with protein digestion (pronase and hyaluronidase) in both formic acid and EDTA sections. For osteo-chondral sections, we recommend using 10% EDTA for decalcification and pronase plus hyaluronidase for antigen retrieval if maintaining tissue morphology is crucial, whereas if time is of the essence, 20% FA with pronase plus hyaluronidase is the faster option while still preserving structural integrity. Clin. Anat. 33:343-349, 2020. © 2019 Wiley Periodicals, Inc.