Optimal Timing for Venous Systemic Oxygen Persufflation Supplemented with Nitric Oxide Gas in Cold-Stored, Warm Ischemia-Damaged Experimental Liver Grafts.
Worldwide shortage of donor organs has increased the use of donation after cardiac death (DCD). The aim of this study was to analyze the best time point for venous systemic oxygen persufflation (VSOP) supplemented with nitric oxide (NO) gas during the 1st and 24th hour of cold storage (CS) in warm ischemia (WI)-damaged experimental liver grafts. Liver grafts (n = 5) were retrieved after 30 min of WI induced by cardiac arrest and CS in histidine-tryptophan-ketoglutarate solution at 4°C. The 1st hour group was immediately persufflated with a VSOP plus NO (VSOP+NO) mixture for 1 h followed by 23 h of static CS (DCD+NO 1st hour). The 24th hour group entailed CS for 23 h followed by 1 h of VSOP+NO persufflation (DCD+NO 24th hour). CS livers without WI but with VSOP served as controls. CS livers with WI represented the fourth group (DCD). Viability of the liver grafts was assessed by normothermic isolated reperfusion for 45 min with oxygenated Krebs-Henseleit buffer. Data are presented as mean ± SEM (control vs. DCD vs. DCD+NO 1st hour vs. DCD+NO 24th hour). After 45 min of reperfusion, the DCD+NO 1st hour group showed significantly lower aspartate aminotransferase (13.4 ± 5.3, 63.2 ± 17.3, 25.6 ± 3.9, and 82.8 ± 27.3 U/l) and lactate dehydrogenase levels (289.4 ± 41.2, 2,139.4 ± 542.7, 577.2 ± 117.2, and 2,429 ± 221.6 U/l). Malondialdehyde levels were significantly abrogated (1.0 ± 0.3, 2.7 ± 1, 1.0 ± 0, and 3.9 ± 1.2 nmol/ml). Significantly higher levels of portal venous pressure were recorded in the DCD+NO 24th hour group (12.0 ± 1, 21.2 ± 3.1, 16.1 ± 1, and 23.2 ± 3.5 mm Hg). NO levels were recorded after 5 min of reperfusion (1.42 ± 0.17, 1.8 ± 0.2, 2.7 ± 0.2, and 2.6 ± 0.1 μmol/l). Bile production levels showed no statistical significance (23.2 ± 3.8, 27.3 ± 1.8, 43.5 ± 18, and 31 ± 2.5 μl/45 min). Our results present the beneficial effects of NO combined with VSOP during the 1st hour of CS of WI-damaged experimental liver grafts.


Role of the primate flocculus in adaptation of the vestibulo-ocular reflex.
Neuronal events associated with adaptation of the horizontal vestibulo-ocular reflex (HVOR) induced by sustained vestibular-visual mismatching were investigated in the primate flocculus. The floccular area related to the HVOR (H-zone) was identified by electrical micro-stimulation which induced ipsilaterally directed horizontal eye movement. It was thus found that Purkinje cells in the H-zone consistently changed their simple spike responses to head rotation in parallel with the adaptive HVOR gain change. This was demonstrated by observing the change of simple spike firing of Purkinje cells during adaptation of HVOR either in a population study or an individual study. Since similar changes occurred even after bilateral lesioning of vestibular nuclei had extinguished the HVOR, these changes appear to represent vestibular, but not eye velocity, mossy fiber responsiveness. The complex spike discharge, on the other hand, modulated during vestibular-visual stimulation with a reciprocal pattern to the adaptive changes in the simple spike discharge. These results are consistent with the hypothesis that the flocculus Purkinje cells adaptively control the HVOR through their simple spike activity under influences of retinal error signals conveyed by visual climbing fiber pathways.


Intracellular hyperthermia. A biophysical approach to cancer treatment via intracellular temperature and biophysical alterations.
This paper introduces a new multi-disciplinary "intracellular" biophysical treatment of cancer. The basic concept uses locally induced heat energy after tumor phagocytosis of submicron particles whose composition permits magnetic excitation. The key to this process is the utilization of the cancer cell membrane to contain the energy within the cancer cell. Any magnetic or electric dipole contained within or introduced into the cell, or that is capable of being produced by an external field, can be used. Submicron particles are colloidally suspended, injected intravenously and are phagocytized by cancer cells. Application of an external high frequency or pulsed electromagnetic field then raises the particles' temperature thus generating intracellular heat in precise increments. This results in selective thermal destruction of cancer cells with little effect on normal cells. Experimental evidence is presented showing tumor cell destruction in spontaneous mammary tumors in Sprague Dawley rats. In addition, we suggest that certain biophysical properties are altered within the cancer cells and could be used to enhance this effect. Specific radioisotopes or tumor specific antibodies bound to particles or chemotherapeutic microspheres increase cancer cell sensitivity and affinity for these particles. This "intracellular" treatment of cancer has a wide potential range of applications.


Microsatellite instability in patients with gastric remnant cancer.
BACKGROUND: About 2% of patients who undergo partial distal gastrectomy for gastroduodenal diseases develop gastric remnant cancer 10 to 30 years after the gastrectomy. It is important in clinical practice to determine a molecular marker to identify patients susceptible to gastric remnant cancer.METHODS: We investigated nine gastric remnant cancers (from nine individuals who had gastrectomies for primary gastric cancer or gastroduodenal ulcer) for microsatellite instability (MSI) at six loci, using the polymerase chain reaction (PCR). A control group of ten patients with sporadic gastric cancers in the upper third of the stomach was also similarly analyzed.RESULTS: MSI was demonstrated in eight of nine cancers from the individuals who had had primary gastric cancer or gastroduodenal ulcer (88.9%) compared with two of ten cancers from the individuals with sporadic gastric cancer in the upper third of the stomach (20%).CONCLUSION: These results suggest that one or more MSI is associated with remnant gastric cancer after gastrectomy.


Biological availability of mutagenic compounds adsorbed onto diesel exhaust particulate.
Diesel particles were collected from the exhaust of a VW Golf diesel car by electrostatic precipitation. The particulate and its DCM extract were highly mutagenic in the Salmonella/microsome test in the presence and absence of metabolic activation; the highest response was observed with TA98 and TA1538 tester strains. The biological availability of particulate-associated mutagenic compounds was demonstrated by administering powder to rats and assaying, in vitro, the urine excreted within 24 h after treatment. The highest activity was obtained with TA98 in the presence of metabolic activation. Typical dose-effect responses were evident in urine of animals treated by all the administration routes tested (i.p., s.c. and per os), both in the presence and absence of a suspending vehicle. Concentration of mutagenic compounds present in urine of treated animals could be achieved by chromatography on Amberlite XAD-2 and XAD-7 resins. This study provides direct evidence for bioavailability to animal tissues of mutagens adsorbed onto diesel particulate, although part of the activity might be ascribed to nitroaromatic compounds formed during the collection of the powder. The present study is part of a more comprehensive work on diesel exhaust particulate, and results have to be considered in this light before any final conclusion can be drawn.


What is the "modified" CTAB protocol? Characterizing modifications to the CTAB DNA extraction protocol.
Cetyltrimethylammonium bromide (CTAB)-based methods are widely used to isolate DNA from plant tissues, but the unique chemical composition of secondary metabolites among plant species has necessitated optimization. Research articles often cite a "modified" CTAB protocol without explicitly stating how the protocol had been altered, creating non-reproducible studies. Furthermore, the various modifications that have been applied to the CTAB protocol have not been rigorously reviewed and doing so could reveal optimization strategies across study systems. We surveyed the literature for modified CTAB protocols used for the isolation of plant DNA. We found that every stage of the CTAB protocol has been modified, and we summarized those modifications to provide recommendations for extraction optimization. Future genomic studies will rely on optimized CTAB protocols. Our review of the modifications that have been used, as well as the protocols we provide here, could better standardize DNA extractions, allowing for repeatable and transparent studies.


Two-stage malignant transformation in hamster embryo cells.
Transformation of primary hamster embryo cells was investigated using 3-methylcholanthrene (MCA), a combination of MCA and 12-O-tetradecanoylphorbol-13-acetate (TPA), and initiation with MCA or dibenz(a,h)anthracene (DBA) followed by promotion with TPA. Evidence for transformation was (a) abnormal cellular morphology, (b) increased lifespan, (c) growth in soft agar, and (d) tumour induction by s.c. inoculation into suckling hamsters.Cells treated with either MCA or MCA+TPA showed the same latent period to morphological transformation, although their tumorigenic potential varied. Cells did not form tumours when TPA was administered 7 days after treatment with either MCA or DBA. However, when administration of TPA was delayed to 27 days after treatment with a transforming dose of MCA or a subthreshold dose of DBA, the cells transformed and produced tumours in hamsters.Our results show that TPA may act as an inhibitor or promoter, depending on the length of time between treatment of the hamster embryo cells with the carcinogen and administration of the TPA. It appears that treatment of cells with TPA before the initiating event is complete inhibits or delays the development of their ability to induce tumours in animals or grow in soft agar. However, with a sufficient interval between the application of the initiating carcinogen and the promoter, transformation occurs, and the ability of cells treated with subthreshold doses of DBA to form tumours is enhanced.


Nitrogen fixation by Myrica gale root nodules Massachusetts wetland.
Nitrogenase activity was measured by acetylene reduction in excised Myrica gale nodules collected throughout the growing season at two sites associated with a small lake in central Massachusetts. One site was in an open, weakly minerotrophic peatland dominated by M. gale and the other was on the lakeshore. Nitrogenase activity appeared in late May when the leaves unfolded, reached a peak in July when the maximum number of leaves was present, and declined until it was no longer measurable in late October several days after all leaves had fallen. Summer activities were substantially higher at the peatland site than the lakeshore. Maximum activities were 19.8±1.9 and 8.1±0.7 μmol/h x g dry weight (x±SE; N=20) at the peatland and lakeshore sites respectively.Nitrogenase activities were very low at 5° C and increased linearly from 10 to 30° C, the highest temperature examined. The maximum soil temperature measured was 20° C, and no significant diurnal fluctuation in activity was detected.Annual nitrogen fixation calculated from the seasonal nitrogenase activity curve was 34 kg N/ha x yr at the peatland site with mean dry weight nodule biomass of 104 kg/ha, and 24 kg N/ha x yr at the lakeshore with 111 kg/ha nodule biomass. These rates of nitrogen fixation are equivalent to 4-5x the amount of nitrogen contained in bulk precipitation and are major components in the nitrogen budgets of the M. gale plants and wetlands in which they grow.


In vitro activity of telavancin against Gram-positive isolates from complicated skin and skin structure infections: results from 2 phase 3 (ATLAS) clinical studies.
During phase 3 clinical studies of telavancin for treatment of complicated skin and skin structure infections, a total of 1530 aerobic Gram-positive isolates were identified at baseline. The majority of these strains were Staphylococcus aureus (n = 1214; 62% methicillin-resistant). All isolates were inhibited by ≤ 1 μg/mL of telavancin.


Comments on "Determination of mercury, cadmium, lead, zinc, selenium and iron by ICP-OES in mushroom samples from around thermal power plant in Muğla, Turkey". doi:10.1007/s00128-011-0357-1.
In several articles on trace elements in mushrooms erroneous data were published on minerals sequestered in fruiting bodies. The biased analytical data published gave a false picture on the composition and nutritional value of mushrooms with respect to minerals. Wild mushrooms are relatively rich in trace elements and some species can hyperaccumulate certain metals. Selenium as reported in the referenced article is discussed in light of typical Se concentrations determined using validated methods as reported by other authors.


Giant cell tumor of bone. Variations in patterns of appearance of different cell types.
Eleven benign giant cell tumors of bone were studied in the electron microscope, and the fine structural localization of acid phosphatase was elucidated. Three distinct cell types are always present in these tumors: stromal cells type 1; stromal cells type 2; and multinucleated giant cells. Small mononuclear cells may also occur, but are not likely to be actively participating in the neoplastic process. The range of variability in the fine structure of the different cell types constituting this tumor has been established. Variations in appearances include: a) presence of nuclear pseudoinclusions in stromal cells type 1 and multinucleated giant cells; b) aberrations in the structure of the rough surfaced endoplasmic reticulum in the same cell types; c) occurrence of ruffled borders, ectoplasmic layers and cytoplasmic labyrinths containing acid phosphatase in the giant cells. Some giant cells show evidence of marked phagocytic activity and contain large and numerous residual bodies carrying acid phosphatase. The significance of the interrelations between the different cell types are discussed and the possible role of stromal cells type 2 in immunological mechanisms directed against the tumor cells are mentioned.


Influence of ethanolic extract of avocado (Persea americana Mill.) seed flour on the estrous cycle, the serum concentrations of reproductive hormones, and the activities of oxidative stress markers in female cavies (Cavia porcellus L.).
The purpose of this study was to determine the effects of the ethanolic extract of avocado seed flour on the estrous cycle characteristics, the concentrations of reproductive hormones [luteinizing hormone (LH) and estradiol], and the activities of some tissues (ovarian and uterine) that are markers of oxidative stress in female guinea pigs (Cavia porcellus). Twenty-four female cavies with normal estrous cycles and equivalent body weights (464.25 and 71.88 gm) were randomly assigned to four groups, each with six females. The control group received 1 ml of distilled water orally, whereas the EE100, EE200, and EE400 groups received 100, 200, and 400 mg/kg bw of ethanolic extract of Persea americana seed, respectively. Following that, three consecutive estrous cycles were observed using vaginal smears. After the trial, the females were slaughtered, and blood and organs were gathered for study. The mean duration of the estrus phase is considerably (p < 0.05) longer in group EE100 animals than in control animals. LH concentrations were substantially (p < 0.05) higher in females in group EE200 than in controls. Total cholesterol levels typically dropped in females in the extract groups, but only significantly (p < 0.05) in those in group EE100 compared to the control group. Serum estradiol levels increased considerably (p < 0.05) in treated females compared to controls. Catalase activity rose considerably (p < 0.05) in the ovaries of group EE400 when compared to the control. Superoxide dismutase activity rose considerably (p < 0.05) in the uterus of female cavies given the extract compared to the control. Avocado seed ethanolic extract prolongs the estrus phase, increases estradiol and LH levels, and protects the uterus from oxidative stress in female cavies.


Contribution of the myosin binding protein C motif to functional effects in permeabilized rat trabeculae.
Myosin binding protein C (MyBP-C) is a thick-filament protein that limits cross-bridge cycling rates and reduces myocyte power output. To investigate mechanisms by which MyBP-C affects contraction, we assessed effects of recombinant N-terminal domains of cardiac MyBP-C (cMyBP-C) on contractile properties of permeabilized rat cardiac trabeculae. Here, we show that N-terminal fragments of cMyBP-C that contained the first three immunoglobulin domains of cMyBP-C (i.e., C0, C1, and C2) plus the unique linker sequence termed the MyBP-C "motif" or "m-domain" increased Ca(2+) sensitivity of tension and increased rates of tension redevelopment (i.e., k(tr)) at submaximal levels of Ca(2+). At concentrations > or =20 microM, recombinant proteins also activated force in the absence of Ca(2+) and inhibited maximum Ca(2+)-activated force. Recombinant proteins that lacked the combination of C1 and the motif did not affect contractile properties. These results suggest that the C1 domain plus the motif constitute a functional unit of MyBP-C that can activate the thin filament.


Charge neutral halogen bonding tetradentate-iodotriazole macrocycles capable of anion recognition and sensing in highly competitive aqueous media.
A series of neutral tetradentate halogen bonding (XB) macrocycles, comprising of two bis-iodotriazole XB donors were synthesised in 60-70% yields via a stepwise CuAAC-mediated cyclisation strategy. Extensive 1H NMR anion titration experiments reveal halide binding affinities are critically dependent on the substitution pattern of the xylyl spacer unit. The meta-substituted macrocycle remarkably displays cooperative tetradentate XB-halide anion recognition in highly competitive 40% aqueous-organic D2O/acetone-d6 (40 : 60, v/v) solvent mixtures. Integration of para-xylyl and naphthyl spacer units generates extended macrocyclic cavities, capable of selective oxalate recognition. Furthermore, preliminary fluorescence exeperiments reveal dicarboxylate specific sensing can be achieved through monitoring of the naphthylene centred emission.


Colovesical Fistula After Renal Transplantation: Case Report.
Colovesical fistula is a relatively rare condition that is primarily related to diverticular disease. There are few reports of colovesical fistula after renal transplantation. We report of a 53-year-old man who was diagnosed with colovesical fistula after recurrent urinary tract infection, 5 months after undergoing cadaveric renal transplantation. Laparoscopic partial resection of the sigmoid colon with the use of the Hartmann procedure was performed. Six months after that surgery, there was no evidence of recurrent urinary tract infection and the patient's renal graft function was preserved. Physicians should keep colovesical fistula in mind as a cause of recurrent urinary tract infection in renal transplant recipients, especially in those with a history of diverticular disease.


Changes in corneal autofluorescence and corneal epithelial barrier function with aging.
Corneal epithelial permeability studies using fluorophotometers were performed on 90 eyes of 51 normal subjects. After recording the autofluorescence of the cornea (AFC) and of the lens (AFL), we applied 20 microliters of 2% sodium fluorescein to the conjunctival sac. The corneal fluorescence 45 min later (F45), which paralleled the corneal epithelial permeability to fluorescein, was measured by fluorophotometer and analyzed. All parameters of the two eyes of each subject correlated well with each other. There was positive correlation between AFL and patient age and between AFC and patient age (r = 0.78, p < 0.001, and r = 0.74, p < 0.001, respectively). The F45 increased exponentially with advancing age (r = 0.67, p < 0.001). The strong correlation between AFC and AFL in each eye (r = 0.79, p < 0.001) indicated corresponding aging processes in both the cornea and the lens. The increase in epithelial permeability with age possibly represents a subclinical breakdown of barrier function, rendering the corneas more vulnerable to insults.


A widespread group of large plasmids in methanotrophic Methanoperedens archaea.
Anaerobic methanotrophic (ANME) archaea obtain energy from the breakdown of methane, yet their extrachromosomal genetic elements are little understood. Here we describe large plasmids associated with ANME archaea of the Methanoperedens genus in enrichment cultures and other natural anoxic environments. By manual curation we show that two of the plasmids are large (155,605 bp and 191,912 bp), circular, and may replicate bidirectionally. The plasmids occur in the same copy number as the main chromosome, and plasmid genes are actively transcribed. One of the plasmids encodes three tRNAs, ribosomal protein uL16 and elongation factor eEF2; these genes appear to be missing in the host Methanoperedens genome, suggesting an obligate interdependence between plasmid and host. Our work opens the way for the development of genetic vectors to shed light on the physiology and biochemistry of Methanoperedens, and potentially genetically edit them to enhance growth and accelerate methane oxidation rates.


Major Bleeding During Thrombolytic Therapy for Acute Lower Limb Ischaemia: Value of Laboratory Tests for Clinical Decision Making, 17 Years of Experience.
Regular measurement of fibrinogen as dose guidance in catheter directed thrombolysis (CDT) for acute limb ischaemia (ALI) has recently been dropped from European guidelines based on inconsistent literature. This study aimed to determine whether low fibrinogen levels and high activated partial thromboplastin time (APTT) are associated with an increased major bleeding risk during CDT. All consecutive patients treated with CDT for ALI in two Dutch hospitals between January 2004 and April 2021 were analysed retrospectively. Patients were treated with two dosing regimens (low dose: 50 000 IU/hour; high dose: 100 000 IU/hour) of urokinase and, after 2018, with a single low dose regimen of alteplase (rtPA) due to urokinase manufacturing problems. The incidence of major bleeding and associated APTT and fibrinogen levels were reviewed from patient charts. Of the 443 included cases, 277 underwent CDT with urokinase and 166 with rtPA. The incidence of major bleeding in the whole cohort was 7%. Patients with a fibrinogen levels < 1.0 g/L developed more major bleeding than those in whom the fibrinogen level did not drop below 1.0 g/L (15% vs. 6%; p = .041). Systemic heparinisation during CDT or high (> 80 seconds) APTT were not significantly associated with major bleeding. Angiographic success (47% vs. 72%; p = .003) and 30 day amputation free survival (53% vs. 82%; p < .001) were lower for cases with major bleeding. Older age (odds ratio [OR] 1.06, 95% confidence interval [CI] 1.02 - 1.11), cardiac history (OR 3.35, 95% CI 1.39 - 8.06), high dose regimens (≥ 75 000 IU/hour urokinase; OR 2.67, 95% CI 1.18 - 6.04), and fibrinogen values < 1.0 g/L (OR 5.59, 95% CI 1.98 - 15.77) were independent predictors for major bleeding during CDT. High dose thrombolytic regimens and fibrinogen levels of ≤ 1.0 g/L are associated with more major bleeding during thrombolytic therapy. Major bleeding significantly worsened the clinical outcome. A prospective comparative study is needed to assess the benefit of monitoring fibrinogen levels.


Kikuchi-Fujimoto Disease in the Axilla after COVID-19 Vaccination: A Case Report.
Kikuchi-Fujimoto disease (KFD) is a rare, self-limiting inflammatory condition of unknown etiology that is characterized by fever and painful lymphadenopathy. KFD commonly involves the posterior cervical region and very rarely occurs in the axilla. We report on a case of KFD that presented 3 weeks after receiving the messenger ribonucleic acid-based coronavirus disease 2019 (COVID-19) vaccine. In this case, we suspected the lesions as COVID-19 vaccination-related lymphadenopathy on the initial ultrasonographic examination. Through this case report, we highlight that KFD should be considered in the differential diagnosis of patients with axillary lymphadenopathy who have undergone COVID-19 vaccination, as unusual side effects of COVID-19 vaccination have been increasingly reported in the literature owing to the rapid development of various COVID-19 vaccines during the pandemic period. In addition, we emphasize the importance of clinical suspicion in diagnosing KFD due to the fact that axillary involvement of KFD is extremely rare.


Central regulation of stress-evoked peripheral immune responses.
Stress-linked psychiatric disorders, including anxiety and major depressive disorder, are associated with systemic inflammation. Recent studies have reported stress-induced alterations in haematopoiesis that result in monocytosis, neutrophilia, lymphocytopenia and, consequently, in the upregulation of pro-inflammatory processes in immunologically relevant peripheral tissues. There is now evidence that this peripheral inflammation contributes to the development of psychiatric symptoms as well as to common co-morbidities of psychiatric disorders such as metabolic syndrome and immunosuppression. Here, we review the specific brain and spinal regions, and the neuronal populations within them, that respond to stress and transmit signals to peripheral tissues via the autonomic nervous system or neuroendocrine pathways to influence immunological function. We comprehensively summarize studies that have employed retrograde tracing to define neurocircuits linking the brain to the bone marrow, spleen, gut, adipose tissue and liver. Moreover, we highlight studies that have used chemogenetic or optogenetic manipulation or intracerebroventricular administration of peptide hormones to control somatic immune responses. Collectively, this growing body of literature illustrates potential mechanisms through which stress signals are conveyed from the CNS to immune cells to regulate stress-relevant behaviours and comorbid pathophysiology.


Arterial thermodilution in burn patients suggests a more rapid fluid administration during early resuscitation.
A number of target points have been used for fluid replacement in severely burned patients. The aim of our prospective randomized study was to compare the effect of two different types of fluid resuscitation regimes on the multiple organ dysfunction score (MODS) and central venous oxygen saturation (ScvO(2)) in the first 3 days after injury. Twenty-four patients admitted to the critical care unit of a university hospital with the presence of burn injury affecting more than 15% of the body surface area and in-hospital fluid resuscitation started within 3 h after burn injury were included. Patients were randomized into two groups. Fluid resuscitation was guided by the hourly urine output (HUO Group, n=12) or by the intrathoracic blood volume index (ITBVI Group, n=12). Invasive transpulmonary hemodynamic measurements utilizing pulse contour analysis with a single dilution technique and continuous ScvO(2) measurements were performed in both groups. The mean ScvO(2) was significantly lower in the HUO Group than in the ITBVI Group (P=0.024) for the first 24 h. MODS was significantly higher in the HUO Group than in the ITBVI group 48 h (P=0.024) and 72 h after injury (P=0.014). The two main outcome parameters, i.e., MODS calculated at 48 and 72 h after injury and ScvO(2) were negatively correlated on day 1 (r=-0.684, P=0.004, r=-0.677, P=0.003). There were no statistical differences in clinical outcome parameters. Our data suggest that ITBVI may be a better target parameter than HOU in the fluid resuscitation of severely burned patients after injury.


The application of population pharmacokinetic modeling to individualized antibiotic therapy.
This paper describes applications of population pharmacokinetic modeling to the optimization of antibiotic dosing. Parametric and nonparametric pharmacokinetic modeling approaches are discussed. Population models can be important extensions of therapeutic drug monitoring (TDM) in infectious disease. The concept of population model-based individualized antimicrobial therapy is described. With the availability of population modeling for obtaining PK parameter estimates, the focus has shifted to quantifying the antimicrobial effect and linking kinetics to drug effects. Examples of integrated pharmacokinetic-pharmacodynamic (PK-PD) models to describe bacterial killing as a function of drug concentration are discussed. Application of PK-PD mathematical models that correlate with microbiological and clinical outcomes will provide us with a better rationale for the proper dose selection of anti-infective therapy in different patient populations.


Prevalence and location of nodal metastases in distal esophageal adenocarcinoma confined to the wall: implications for therapy.
The purpose of this study was to characterize the prevalence and location of regional lymph node metastases in adenocarcinoma confined to the esophagal wall, to determine the extent of dissection required, and to investigate the applicability of nonoperative therapy. Histologic evaluation of the resected specimens after en bloc esophagogastrectomy with mediastinal and abdominal lymphadenectomy was performed on 37 patients with adenocarcinoma confined to the esophageal wall. Follow-up was complete in all patients (median 24 months). Fifteen patients (41%) had intramucosal tumors. Twelve (32%) had submucosal tumors and 10 (27%) had muscular invasion. The prevalence of regional lymph node metastases (15/37 patients, 41%) increased progressively with depth of tumor invasion, with involved nodes identified in 80% of patients with muscular invasion. Lymph node metastases were also more common at distant node stations in intramuscular tumors (5/10, 50%). Actuarial survival for the entire group was 63% at 5 years. Recurrence was identified in 6 of the 37 patients (16%), with the risk of recurrence correlating with tumor depth. Tumor depth is a strong predictor of the probabilities of regional lymph node metastases, the likelihood of involvement of distant node groups, and the risk of recurrence. Patients with invasion of the muscular wall are at particularly high risk. En bloc esophagectomy with mediastinal and abdominal lymphadenectomy has the highest likelihood of achieving an R0 resection. The long-term survival and low recurrence rate achieved with an en bloc esophagectomy emphasizes the importance of an aggressive lymph node dissection to remove all potentially involved nodes.


The preventability of cancer in Europe: A quantitative assessment of avoidable cancer cases across 17 cancer sites and 38 countries in 2020.
The aim of this study is to provide an update of the estimated proportion of avoidable cases across European region for cancers associated with modifiable risk factors. Age-standardised incidence rates for 38 European countries in 2020 were derived from GLOBOCAN database. For 17 cancer sites, we estimated the number and proportion of avoidable cancer cases. The mean of the three lowest country-specific incidence (excluding rates of zero) was defined as the attainable rate for each combination of cancer site, sex, and age group. The number and proportion of avoidable cancer cases was calculated as the difference between the country-specific and attainable incidence rates. Approximately, 697,000 cancer cases (33%) of all cases in men, and 837,000 (44%) in women were potentially avoidable. Lung, colorectal, and female breast cancer contributed the largest avoidable burden from cancer, with a combined 790,000 cases, followed by mesothelioma, melanoma of the skin, laryngeal, and oesophageal cancer as major contributors. Large geographical variations were found in the estimated proportion of avoidable cancer cases by sex. A cohesive population-level preventative effort to reduce modifiable cancer risk factors could potentially have considerable impact in reducing the future burden of many cancers in Europe.


The carboxy-terminal half of nonstructural protein 3A is not essential for foot-and-mouth disease virus replication in cultured cell lines.
In foot-and-mouth disease (FMD)-endemic parts of the globe, control is mainly implemented by preventive vaccination with an inactivated purified vaccine. ELISAs detecting antibodies to the viral nonstructural proteins (NSP) distinguish FMD virus (FMDV)-infected animals in the vaccinated population (DIVA). However, residual NSPs present in the vaccines are suspected to be a cause of occasional false positive results, and therefore, an epitope-deleted negative marker vaccine strategy is considered a more logical option. In this study, employing a serotype Asia 1 FMDV infectious cDNA clone, it is demonstrated that while large deletions differing in size and location in the carboxy-terminal half of 3A downstream of the putative hydrophobic membrane-binding domain (deletion of residues 86-110, 101-149, 81-149 and 81-153) are tolerated by the virus without affecting its infectivity in cultured cell lines, deletions in the amino-terminal half (residues 5-54, 21-50, 21-80, 55-80 and 5-149) containing the dimerization and the transmembrane domains are deleterious to its multiplication. Most importantly, the virus could dispense with the entire carboxy-terminal half of 3A (residues 81-153) including the residues involved in the formation of the 3A-3B1 cleavage junction. The rescue of a replication-competent FMDV variant carrying the largest deletion ever in 3A (residues 81-153) and the fact that the deleted region contains a series of linear B-cell epitopes inspired us to devise an indirect ELISA based on a recombinant 3A carboxy-terminal fragment and to evaluate its potential to serve as a companion diagnostic assay for differential serosurveillance if the 3A-truncated virus is used as a marker vaccine.


Impact of dietary counselling on the nutritional status and quality of life among pulmonary tuberculosis patients - A randomized control trial.
Undernutrition and TB have a bidirectional relationship, which is especially relevant in the Indian context. Undernutrition is an established risk factor for the progression of latent TB infection to active TB. Undernutrition at the population level contributes to an estimated 55% of annual TB incidence in India. TB leads to weight loss, wasting, and worsening of nutritional status. Hence, the present study aimed to determine the impact of dietary counselling on the nutritional status and the health-related quality of life of PTB patients. This was an interventional study (randomized controlled trial) that involved patients with PTB. 46 patients (23 patients as experimental and 23 patients as a control group) were enrolled in the study from June 2019 to February 2020 and they were divided into 2 categories based on BMI (underweight and normal weight). The special dietary counselling was given to the experimental group patients and the normal protocol was followed with the control group. The patients were followed up, till completion of treatment i.e., 6 months from enrolment. In our study, nearly half of the patients were underweight and DM was the predominant comorbidity. The BMI increased after dietary counselling in the experimental group than the control group (P = 0.0053) in underweight individuals. Total protein (P = 0.0025), and serum albumin (P = 0.0048) levels were found to be significantly improved in the experimental group. SGRQ symptom score (P = 0.0036) has significantly reduced in the experimental group in underweight individuals than the control group. Personalized dietary counselling was found to have a positive impact on BMI, total protein, and albumin levels in the experimental group, especially in underweight individuals. Besides, the quality of life measured using SGRQ showed that symptom score were also significantly reduced in the experimental group than the control group.


The Use of Halo Gravity Traction in the Treatment of Severe Early Onset Spinal Deformity.
Retrospective Review of Prospective cohort. To describe the feasibility of preoperative halo gravity traction (HGT) with subsequent growing rod/guided growth (GR/GG) placement in early onset spinal deformity (EOSD). In children with severe EOSD, primary implantation of GR/GG constructs is not always possible. We describe a staged protocol with preoperative HGT followed by GR/GG implantation. EOSD patients treated with HGT prior to GR/GG implantation were included. HGT used traction up to 50% body weight for 4 to 29 weeks. Pulmonary function tests (PFTs) were performed before and after HGT. Coronal Cobb (CC) and Sagittal Cobb (SC) angles were measured on the Pre-HGT, Post-HGT and 6 week postop x-rays. Thirty patients were included. Average age at GR/GG implantation was 9 years. Most cases (n = 24, 80%) were idiopathic. Most pts had kyphoscoliosis (n = 16, 53.3%). Pre-HGT CC averaged 112 ± 22° and SC averaged 106 ± 26°. CC and SC improved 29% after HGT. There was a significant improvement in body mass index following HGT. CC improved further to 70 ± 14° (36% vs. pre-HGT) and SC to 63 ± 21° (41%) with GR/GG placement. HGT-related complications occurred in nine patients (30%); eight pin site infections, one cranial abscess. Most HGT complications were managed with local pin care and oral antibiotics. Halo revision was required in two pts (6.7%). There was no change in PFTs with HGT (P > 0.05). Averagely, 14 levels were spanned during GR/GG implantation; two patients required vertebral column resection. Surgical complications occurred in nine (30%) patients. At average 16 month follow-up, seven patients (23.3%) required reoperation. Preoperative HGT can make severe EOSD curves amenable to GR/GG implantation. HGT results in ∼30% correction with improvement to ∼35-40% following GR/GG. HGT has a 30% complication rate but most are pin-site infections managed with pin-site care and oral antibiotics; 6.7% of patients require revision. 4.


Comparison of rRNA depletion methods for efficient bacterial mRNA sequencing.
Current methods of high-throughput RNA sequencing of prokaryotes, including transcriptome analysis or ribosomal profiling, need deep sequencing to achieve sufficient numbers of effective reads (e.g., mapping to mRNA) in order to also find weakly expressed genetic elements. The fraction of high-quality reads mapping to coding RNAs (i.e., mRNA) is mainly influenced by the large content of rRNA and, to a lesser extent, tRNA in total RNA. Thus, depletion of rRNA increases coverage and thus sequencing costs. RiboZero, a depletion kit based on probe hybridisation and rRNA-removal was found to be most efficient in the past, but it was discontinued in 2018. To facilitate comparability with previous experiments and to help choose adequate replacements, we compare three commercially available rRNA depletion kits also based on hybridization and magnetic beads, i.e., riboPOOLs, RiboMinus and MICROBExpress, with the former RiboZero. Additionally, we constructed biotinylated probes for magnetic bead capture and rRNA depletion in this study. Based on E. coli, we found similar efficiencies in rRNA depletion for riboPOOLs and the self-made depletion method; both comparable to the former RiboZero, followed by RiboMinus, succeeded by MICROBExpress. Further, our in-house protocol allows customized species-specific rRNA or even tRNA depletion or depletion of other RNA targets. Both, the self-made biotinylated probes and riboPOOLs, were most successful in reducing the rRNA content and thereby increasing sequencing depth concerning mRNA reads. Additionally, the number of reads matching to weakly expressed genes are increased. In conclusion, the self-made specific biotinylated probes and riboPOOLs are an adequate replacement for the former RiboZero. Both are very efficient in depleting rRNAs, increasing mRNA reads and thus sequencing efficiency.


Methionine consumption by cancer cells drives a progressive upregulation of PD-1 expression in CD4 T cells.
Programmed cell death protein 1 (PD-1), expressed on tumor-infiltrating T cells, is a T cell exhaustion marker. The mechanisms underlying PD-1 upregulation in CD4 T cells remain unknown. Here we develop nutrient-deprived media and a conditional knockout female mouse model to study the mechanism underlying PD-1 upregulation. Reduced methionine increases PD-1 expression on CD4 T cells. The genetic ablation of SLC43A2 in cancer cells restores methionine metabolism in CD4 T cells, increasing the intracellular levels of S-adenosylmethionine and yielding H3K79me2. Reduced H3K79me2 due to methionine deprivation downregulates AMPK, upregulates PD-1 expression and impairs antitumor immunity in CD4 T cells. Methionine supplementation restores H3K79 methylation and AMPK expression, lowering PD-1 levels. AMPK-deficient CD4 T cells exhibit increased endoplasmic reticulum stress and Xbp1s transcript levels. Our results demonstrate that AMPK is a methionine-dependent regulator of the epigenetic control of PD-1 expression in CD4 T cells, a metabolic checkpoint for CD4 T cell exhaustion.


Outcomes and Prognostic Factors of Cataract Surgery in Adult Extreme Microphthalmos With Axial Length <18 mm or Corneal Diameter <8 mm.
To study the long-term visual outcomes, complications, and prognostic factors for cataract surgery in extreme microphthalmos. Prospective cohort study. Thirty eyes with simple microphthalmos (11 eyes, axial length [AL] <18 mm), complex microphthalmos (8 eyes, AL <18 mm), and relative anterior microphthalmos (RAM; 11 eyes, corneal diameter [CD] <8 mm) who underwent cataract surgery (phacoemulsification) in our hospital were followed for a mean of 25.3 months with at least 3 visits at early (1 day to 1 week), mid (1-3 months), and late (>6 months) stages after surgery. The main outcome measures included the best-corrected visual acuity (BCVA), intraocular pressure (IOP), and complications. Significant improvements of BCVA were observed at the mid and late postoperative visits in the entire cohort when compared with preoperative value (P < .05). When eyes were divided into 3 groups, a statistically significant improvement in late-stage BCVA was observed in the simple microphthalmos and RAM groups. The AL, preoperative anterior chamber depth (ACD), and preoperative BCVA were significant prognostic factors for late-stage BCVA after surgery. The most common complications were early corneal edema (73%), glaucoma (33%), and posterior capsular opacification (23%). Preoperative ACD was significantly associated with the incidence of glaucoma after surgery (P < .05). Severe complications included suprachoroidal hemorrhage (3%), endothelial dysfunction (7%), and retinal detachment (7%). Cataract surgery provided visual improvement in extreme microphthalmos (simple microphthalmos and RAM), with higher risks of complications than with routine cataract surgeries. Extreme microphthalmos with preoperative characteristics of relatively longer AL, deeper ACD, and better BCVA may benefit more from cataract surgery.


Perception of the Professional Knowledge of and Education on the Medical Technology Products among the Pharmacists in the Baltic and Nordic Countries-A Cross-Sectional Exploratory Study.
With increased development of medical technology (MT), new challenges emerge related to education and training of pharmacists and other healthcare specialists. Currently, only a few universities in the EU promote MT education and research. The aim of this study was to evaluate the current status, views on, and need for the education on MT for the pharmacy students and practicing pharmacists in the Baltic and Nordic countries. The representatives of higher education institutions and community/hospital pharmacists from six Baltic and Nordic countries participated in a qualitative cross-sectional exploratory internet-based study from May to October 2014. Approximately two-third of the respondents considered professional knowledge about MT products important for pharmacists, but half of them had never participated in any MT courses. More practicing pharmacists than representatives of academia underlined the need for increased MT education for pharmacy students in the future. The pharmacists in the Baltic and Nordic countries consider the professional knowledge about MT as pertinent in their education and work. The limited number and status of MT courses available today, however, is a major concern among both pharmacy students and practicing pharmacists in these countries. In the future, increasing education combining theory and practice about MT products would be one possible solution to overcome this challenge.


A case of echocardiographic evaluation of ruptured pseudoaneurysm of the mitral-aortic intervalvular fibrosa by infective endocarditis.
This report describes an unusual case of ruptured pseudoaneurysm (PSA) of mitral-aortic intervalvular fibrosa (MAIVF) caused by infective endocarditis. The PSA ruptured into the left sinus of Valsalva in addition to the left atrium, resulting in complicated shunting among the aorta, left ventricle and left atrium, leading to refractory heart failure. The transesophageal echocardiography provided the precise information concerning the anatomical detail of the PSA, which is crucial for the surgical repair. This is the first report describing a patient with PSA of MAIVF with two rupture sites.


Reaction time and rhythm of movement in Huntington's disease.
Huntington disease (HD) is characterized by several hyperkinesias though motor slowness is also another cardinal in this disease. In addition, self-paced timing movements are also disturbed in HD, which may also affect several rhythmic voluntary movements such as gait. Motor slowness can be measured with clinical scales such as the Unified Huntington's Disease Rating Scale (UHDRS) and timed tests, but also with the reaction time (RT) paradigm. We evaluated RT as a measure of motor slowness in 30 patients with genetically confirmed Huntington's disease and 24 control subjects. We also evaluated self-paced timing precision (SPTP) by applying a simple software program devised by our group. Clinical assessment was performed according to the UHDRS, including motor section, total functional capacity (TFC) and cognitive section (verbal fluency test, symbol digit, and Stroop test) The mean values obtained for RT and SPTP were statistically different in HD as compared with those from controls (p<0.0005). We observed a statistically significant correlation between RT and TFC scores (rs=-0.57, p<0.005 Spearman's correlation) and also between SPTP values and TFC scores (rs=-0.40, p<0.05 Spearman's correlation). In addition, RT and SPTP significantly correlated with cognitive scores (including digit symbol, verbal fluency and Stroop tests). Simple tests such as RT and SPTP provide an objective evaluation of motor impairment in HD yielding measures that correlate with clinical assessment and functional disability.


Putting molecules in their place.
Each class of microscope is limited to imaging specific aspects of cell structure and/or molecular organization. However, imaging the specimen by complementary microscopes and correlating the data can overcome this limitation. Whilst not a new approach, the field of correlative imaging is currently benefitting from the emergence of new microscope techniques. Here we describe the correlation of cryogenic fluorescence tomography (CFT) with soft X-ray tomography (SXT). This amalgamation of techniques integrates 3D molecular localization data (CFT) with a high-resolution, 3D cell reconstruction of the cell (SXT). Cells are imaged in both modalities in a near-native, cryopreserved state. Here we describe the current state of the art in correlative CFT-SXT, and discuss the future outlook for this method.


Linking the Estimation of Threat and COVID-19 Fear and Safety Behavior Use: Does Intolerance of Uncertainty Matter?
Research has shown threat overestimation is significantly associated with intolerance of uncertainty (IU), and both processes predict higher anxiety and safety behavior usage. However, the extent to which threat overestimation predicts subsequent COVID-19-related distress may vary as a function of IU. The present study examined IU as a moderator of the relationship between COVID-19 threat estimation and subsequent COVID-19 fear and safety behavior use. Between February 27 and March 26, 2020, participants (N = 57) completed a self-report measure of IU and estimated the number of people they believed had died from COVID-19. Four weeks later, participants completed measures of COVID-19 fear and safety behavior use. Results revealed IU significantly predicted subsequent COVID-19 fear and safety behavior use. IU also moderated the effect of threat estimation on COVID-19 fear such that those who underestimated threat and experienced low to moderate levels of IU reported experiencing lower levels of COVID-19 fear 1 month later.


Exploration into Plasma Hsa_circ_0052184 as a New Biomarker of Colorectal Cancer Prognosis.
Circular RNAs (circRNAs) are strong modulators of tumor pathology. Herein, our goal was to examine the plasma hsa_circ_0052184 content among colorectal cancer (CRC) patients, and assess its association with patient clinicopathological profile and diagnostic values. Overall, we collected 228 presurgical CRC and 146 normal plasma samples from The First People's Hospital of Wenling. Circulating hsa_circ_0052184 levels were assessed via qRT-PCR, and the diagnostic prediction was conducted with the receiver operating characteristic (ROC) curve. Relative to healthy controls, CRC patients exhibited markedly enhanced circulating hsa_circ_0052184 levels, which were closely correlated with advanced stage of disease and worse outcome. Based on our uni- (UA) and multivariate assessments (MA), elevated hsa_circ_0052184 levels were a stand-alone predictor of poor prognosis. The ROC curve depicted an area under the curve (AUC) for CRC diagnosis to be 0.9072. Circulating hsa_circ_0052184 is a potential bioindicator of CRC outcome.


Impact of therapeutic drug level monitoring on outcomes of patients with Crohn's disease treated with Infliximab: real world data from a retrospective single centre cohort study.
Therapeutic drug monitoring (TDM) by measuring infliximab (IFX) trough levels and antibodies to infliximab (ATI) is used to optimise treatment in inflammatory bowel disease. We aimed to explore the clinical outcomes of TDM for patients with Crohn's disease on IFX in real life setting. This is a retrospective observational study. Primary outcomes were the clinicians' response to each TDM result and the rate of IFX discontinuation due to secondary loss of response or serious adverse event. Secondary outcomes included the intestinal surgery rate after IFX initiation and remission 6 months after TDM. Multivariate logistic regression was performed to identify factors associated with IFX discontinuation and abdominal surgery. 291 patients were included. 238 (81.8%) patients were tested for TDM at least once during their follow-up with 672 TDM results. 95/238 patients (39.9%) had undetectable levels and 76 (31.9%) had positive ATI at least once. The median infliximab trough level was 3.4 µg/mL. IFX was discontinued in 109 patients (37.5%). 526/672 (78.3%) TDMs results were not followed by altered patient management. Treatment was discontinued in 40 (75.5%) patients never tested for TDM compared with 69 (29.0%) of those tested (p<0.01). Fewer TDM tested patients (29; 12.2%) required intestinal surgery post IFX initiation compared with TDM not-tested (15; 28.3%). Not being TDM tested was independently associated with IFX discontinuation and abdominal surgery. IFX discontinuation and intestinal surgery were significantly less frequent with TDM. TDM requested to investigate loss of response resulted in change in patient management.


Comparison of injection site pain and injection site reactions in relapsing-remitting multiple sclerosis patients treated with interferon beta-1a or 1b.
This prospective, multicentre, international, observational, cohort study compared injection site pain (ISP) and injection site reactions (ISRS) between interferon beta-1b (IFNB-1b; Betaferon) 250 microg subcutaneously every other day and interferon beta-1a (IFNB-1a; Rebif) 44 microg subcutaneously three times weekly in patients with relapsing-remitting MS. Patients started treatment within 3 months before recruitment and were on full dose of therapy at inclusion. Patients self-injected IFNB and self-assessed ISP for 15 consecutive injections immediately, 30 and 60 min after injection, using a visual analogue scale diary. Study staff assessed ISRS. Of 445 patients (valid cases), approximately 90% used autoinjectors. More patients were pain-free at all timepoints with IFNB-1b than with IFNB-1a (eg, 30 min: 42.6% versus 19.7%; P<0.0001). The mean proportion of pain-free injections was greater for IFNB-1b (eg, 30 min: 79.0%) than for IFNB-1a (53.3%; P<0.0001). The proportion of patients without ISRS was greater for IFNB-1b (second visit 51.8% versus 33.8%; P<0.0001). Compared with IFNB-1a, more IFNB-1b patients either had no pain or their ISP had no influence on treatment satisfaction (76.9% versus 64.1%; P=0.006). The impact on tolerability and patient acceptability of any new IFNB product formulations would, however, have to be evaluated in comparative studies.


Phylogenetic, antigenic and biological characterization of pigeon paramyxovirus type 1 circulating in China.
For many years, ND has been one of the most important infectious pigeon diseases in China. In recent years, a high mortality has been observed in ND-infected pigeons in China. Mortality is from 40% to 80% or 100% in some cases. The full-length genomes of four pigeon paramyxovirus type 1 (PPMV-1) strains, which were isolated from infected pigeons in China in 2012 and 2013, were sequenced and analyzed to determine the phylogenetic characteristics of PPMV-1 circulating in pigeons of China in recent years. Furthermore, cross hemagglutination inhibition and cross virus neutralization assays, as well as animal experiments were conducted to determine the antigenicity and pathogenicity of those viruses. Proteolytic cleavage sites (residues 112-117) of the F proteins were identified as the typical virulence motif, 112RRQKR↓F117 for all four PPMV-1 strains investigated. Phylogenetic analysis based on sequences of complete genomes and F gene revealed that the four PPMV-1 isolates and most of recent isolates in China were highly homologous to European isolates from 1998 to 2011. All those isolates were clustered in one clade of genotype VI NDV, termed as subgroup 4bii f. The R value was calculated based on cross hemagglutination inhibition and cross virus neutralization results, and confirmed antigenic difference of the PPMV-1 strains isolated in 2013 from the LaSota vaccine strain. Several mutations were identified in the surface glycoproteins F and HN, which probably gave rise to those antigenic differences. Our result suggested that the PPMV-1 strain prevailing in China in the last decade diverged from a common ancestor and was presumably transmitted from Europe. PPMV-1 isolates displayed obvious antigenic differences from vaccine strain LaSota. Even though PPMV-1 did not cause high mortality in experimental pigeons, the infected pigeons were exhibiting viral shedding for 3 weeks after infection, suggesting PPMV-1 is a potential threat to NDV control worldwide.


Preoperative Osteopenia Is Associated with Significantly Shorter Survival in Patients with Perihilar Cholangiocarcinoma.
Background: Osteopenia is defined as low bone mineral density (BMD) and has been shown to be associated with outcomes of patients with various cancers. The association between osteopenia and perihilar cholangiocarcinoma is unknown. The aim of this study was to evaluate osteopenia as a prognostic factor in patients with perihilar cholangiocarcinoma. Methods: A total of 58 patients who underwent surgery for perihilar cholangiocarcinoma were retrospectively analyzed. The BMD at the 11th thoracic vertebra was measured using computed tomography scan within one month of surgery. Patients with a BMD < 160 HU were considered to have osteopenia and b BMD ≥ 160 did not have osteopenia. The log-rank test was performed for survival using the Kaplan−Meier method. After adjusting for confounding factors, overall survival was assessed by Cox′s proportional-hazards model. Results: The osteopenia group had 27 (47%) more females than the non-osteopenia group (p = 0.036). Median survival in the osteopenia group was 37 months and in the non-osteopenia group was 61 months (p = 0.034). In multivariable analysis, osteopenia was a significant independent risk factor associated with overall survival in patients with perihilar cholangiocarcinoma (hazard ratio 3.54, 95% confidence interval 1.09−11.54, p = 0.036), along with primary tumor stage. Conclusions: Osteopenia is associated with significantly shorter survival in patients with perihilar cholangiocarcinoma.


Exploring radiogrammetry beyond the second metacarpal: Using the third, fourth, and fifth metacarpals to quantify cortical bone.
Traditional metacarpal radiogrammetry, a method for quantifying cortical bone in metacarpals to identify bone loss, typically relies on the presence of an unaltered or undamaged second metacarpal. This study compares the cortical indices of the second to the third, fourth, and fifth metacarpals to test if an additional metacarpal can be used as substitute when the second metacarpal is not available for study. Hand and wrist radiographs from the Burlington Growth Study, belonging to 56 individuals (28 females; 28 males) between 18 and 20 years old, were included in this study. Cortical indices were calculated for metacarpals two through five. Cortical index differences were statistically compared by sex, and the second metacarpal cortical indices were correlated with those of the third, fourth, and fifth metacarpals. The third, fourth, and fifth metacarpal cortical indices were all significantly correlated with the second metacarpal cortical indices for both females and males (p < .05). Cortical indices of the second metacarpal were most strongly correlated with those of the third metacarpal (females r = .644, p < .001; males r = .643, p < .001). The results of this study indicate that the third, fourth, or fifth metacarpal may serve as substitutes for cortical index analyses when the second metacarpal is unavailable or unsuitable for analysis. While the second metacarpal should remain the primary choice in radiogrammetry analyses, the third metacarpal is the most suitable alternative for quantitative analyses of cortical bone.


Vanadate down-regulates cell surface insulin and growth hormone receptors and inhibits insulin receptor degradation in cultured human lymphocytes.
Insulin is able to down-regulate its specific cell surface receptor in cultured human lymphocytes. The effect of vanadate, a known insulinomimetic agent, was examined to determine whether it could mimic insulin to down-regulate the insulin receptor. Exposure of cultured human lymphocytes (IM-9) to vanadate (0-200 microM) resulted in a time- and dose-dependent decrease in cell surface insulin receptors to 60% of control, while insulin (100 nM) down-regulated to 40%. The vanadate effect, in contrast to the rapid effect of insulin, was slow to develop (4-6 h). Surface receptor recovery after 18 h exposure was rapid after vanadate removal (20 min), but it required hours after insulin suggesting the presence of an intracellular (cryptic) pool of receptors after vanadate treatment. Insulin binding to Triton X-100-solubilized whole cells after 18 h treatment revealed that total cell receptors had decreased to 50% of control after insulin but increased to 120 and 189% of control after 100 and 200 microM vanadate, respectively. Furthermore, vanadate inhibited the insulin-mediated loss of total cell receptors from 50 to 28%. Removal of cell surface receptors by trypsin before cell solubilization revealed that 100 microM vanadate increased insulin binding to 321% of control indicating an accumulation of intracellular receptors. Labeling of cell surface proteins with Na125I and lactoperoxidase followed by immunoprecipitation of solubilized receptors with anti-receptor antibody after incubation for various times up to 20 h and quantitation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that, while insulin shortened t1/2 from 7.3 to 5.3 h, vanadate prolonged receptor t1/2 to 14 h. No effect of vanadate was detected on insulin receptor tyrosine kinase activity with up to 4 h incubation at the vanadate concentrations used in this study. Furthermore, human growth hormone surface receptors were similarly down-regulated by vanadate. We conclude that 1) vanadate has an apparent insulin-like effect to down-regulate cell surface insulin receptors in cultured human lymphocytes; 2) in contrast to insulin-induced down-regulation which is associated with receptor degradation vanadate causes an accumulation of intracellular (cryptic) receptors and inhibits insulin receptor degradation; and 3) these effects of vanadate may be exerted on other cell surface receptors.


Papillary cystic neoplasm of the pancreas: a rare presentation.
Papillary cystic neoplasm is a rare tumor and is usually found in young female patients. Here we describe a rare case of PCN in 16 year old girl, which was presented with abdominal pain in last 3 years. Preoperative diagnosis was suggested by ECHO and CT of the abdomen. The sonographic examination of the abdomen showed a tumor measuring 6.5 x 5.5cm in diameter in epigastrium. It was excised totally. Histological description confirmed the diagnosis of PCN. The patient is currently in good health without signs of relapse 3 years after surgery. Our patient is one of the very few adolescent patients with PCN who was treated successfully with surgery. ECHO and CT of the abdomen suggested the diagnosis which was confirmed by histopatological examination. The diagnosis of PCN is suspected by ECHO and CT of the abdomen and confirmed by specific histologic features. Prognosis is good and tumor should be excised whenever detected.


Symbiotic interactions between arbuscular mycorrhizal (AM) fungi and male papaya plants: its status, role and implications.
Experiments were conducted to study the arbuscular mycorrhizal (AM) status and its role in P-uptake through assay of root phosphatases activities in four varieties of male Carica papaya L. viz. CO-1, CO-2, Honey Dew and Washington during flowering stages. In the present study, mean total root colonization of AM fungi recorded peak increase in flowering stage-II while mean root phosphatase (acid and alkaline) activities recorded peak increase in flowering stage-I. Unlike root colonization and root phosphatase activities, spore density did not exhibit any definite patterns and recorded a narrow range of fluctuation during different flowering stages of male C. papaya. The study brought out the fact that root colonization and spore density of AM fungi along with root phosphatase activities varied significantly within the four varieties of male C. papaya plants during each flowering stage. The study also recorded consistently higher acid root phosphatase activity than alkaline root phosphatase activity under P-deficient, acidic soil conditions during all flowering stages of male C. papaya plants. Studies revealed that the root colonization of AM fungi influenced root phosphatase activities (acid and alkaline) positively and significantly during all flowering stages of male C. papaya plants. A total of twelve species of AM fungi belonging to five genera viz. Acaulospora, Dentiscutata, Gigaspora, Glomus, and Racocetra were recovered from the rhizosphere of male C. papaya plants.


Quantifying microplastics in fjords along the Western Antarctic Peninsula.
Microplastics are ubiquitous around the world. Microplastics have been documented around the Southern Ocean, in coastal sediments and in Antarctic marine organisms, however microplastics data for Antarctic waters remain scarce. Microplastics concentrations were characterized from fjord habitats on the Western Antarctic Peninsula where most glaciers are rapidly retreating. Water samples were collected from 2017 to 2020 from surface and benthos, vacuum-filtered, quantified to determine the classification of microplastic, color, and size. Micro-FTIR spectrophotometry was utilized to confirm chemical composition. Comparisons over time and location were made for average microplastic per liter. Despite the new emergent youth and remoteness of these habitats, it was determined that all fjord habitats had microplastics present each year sampled and increased from 2017 to 2020 in each fjord. Despite physical 'barriers' such as the Antarctic Circumpolar Current (and particularly its strongest jet, the Polar Front), microplastics are clearly present and increasing in even recent habitats.


The rise times of miniature endplate currents suggest that acetylcholine may be released over a period of time.
Models of miniature endplate currents predict 20-80% rise times of 100 microseconds or less. These predictions are substantially less than most of the rise times recorded in the literature. New measurements were made of rise times at the frog neuromuscular junction using extracellular recording. The mean 20-80% rise time was 250 microseconds. Rise times were variable; at 20 degrees C, 95% of them fell in a range from 140 to 460 microseconds. The most questionable assumption in the models is that the acetylcholine (ACh) is released instantaneously. Modifying the model, so that ACh diffuses from the vesicle through a pore, lengthens the rise time to observed levels. It has been proposed that ACh is released from the vesicle in exchange for Na+. However, the rise times of miniature endplate currents recorded in solutions in which the Na+ is replaced by sucrose are in the normal range. The Q10 for the rise of miniature endplate currents is approximately 2, which is consistent with the models and with temperature effects on pore formation in mast cells.


Bromine-bridged Dy2 single-molecule magnet: magnetic anisotropy driven by cis/trans stereoisomers.
We report the first bromine-bridged dinuclear [Dy(Cy3PO)2(μ-Br)(Br)2]2·2C7H8 single-molecule magnet with an effective energy barrier of 684 K and magnetic hysteresis below 3 K. The asymmetric DyIII centres present two unique stereoisomeric octahedral coordination environments depending on the cis/trans disposition of the Cy3PO ligands, leading to the orthogonality of the easy magnetic axes that annihilates the dipolar interactions.


Repeated Oral Exposure to N ε-Carboxymethyllysine, a Maillard Reaction Product, Alleviates Gut Microbiota Dysbiosis in Colitic Mice.
Diet is suggested to participate in the etiology of inflammatory bowel diseases (IBD). Repeated exposure to Maillard reaction products (MRPs), molecules resulting from reduction reactions between amino acids and sugars during food heating, has been reported to be either potentially detrimental or beneficial to health. The aim of this study is to determine the effect of repeated oral ingestion of N ε-carboxymethyllysine (CML), an advanced MRP, on the onset of two models of experimental IBD and on the gut microbiota composition of mice. Mice received either saline (control) or N ε-carboxymethyllysine daily for 21 days. For the last week of treatment, each group was split into subgroups, receiving dextran sulfate sodium salt (DSS) or trinitrobenzenesulfonic acid (TNBS) to induce colitis. Intensity of inflammation was quantified, and cecal microbiota characterized by bacterial 16S ribosomal RNA (rRNA) amplicon sequencing. Daily oral administration of N ε-carboxymethyllysine did not induce intestinal inflammation and had limited impact on gut microbiota composition (Bacteroidaceae increase, Lachnospiraceae decrease). DSS and TNBS administration resulted in expected moderate experimental colitis with a shift of Bacteroidetes/Firmicutes ratio and a significant Proteobacteria increase but with distinct profiles: different Proteobacteria taxa for DSS, but mainly Enterobacteriaceae for TNBS. While N ε-carboxymethyllysine exposure failed to prevent the inflammatory response, it allowed maintenance of healthy gut microbiota profiles in mice treated with DSS (but not TNBS). Repeated oral exposure to CML limits dysbiosis in experimental colitis. IBD patients may modulate their microbiota profile by regulating the level and type of dietary MRP consumption.


Comparison of three commercial one-dose porcine circovirus type 2 (PCV2) vaccines on PCV2 shedding in semen from experimentally infected boars.
This study compared the effects of 3 different types of commercial PCV2 vaccines on PCV2 virus shedding in the semen from infected boars. Twenty-five non-PCV2 viremic and seronegative boars were randomly divided into five groups: three vaccinated and challenged groups, a non-vaccinated and challenged group, and a negative control group. The number of genomic copies of PCV2 in serum and semen samples was significantly decreased in vaccinated and challenged boars compared to non-vaccinated and challenged boars from 14 to 70 days post-inoculation (dpi). The number of PCV2 genomic copy in the semen correlated with the number of PCV2b genomic copy in the blood in vaccinated and challenged boars (r(2)=0.894-0.926, P<0.01), and non-vaccinated and challenged boars (r(2)=0.903, P<0.01). The vaccination protocol reduced the amount of PCV2 DNA shed in the semen. However, there was a significantly different amount of PCV2 DNA shed in semen among the 3 vaccinated and challenged boar groups.


Calcium entry activators: distinct sites of dihydropyridine and aminopyridine action.
The dihydropyridine compound BAY K 8644, a putative calcium entry activator, inhibits binding of the dihydropyridine calcium entry blocker, [3H]nitrendipine, to rat skeletal muscle membranes. In contrast, aminopyridine compounds also believed to stimulate calcium flux do not interact with [3H]nitrendipine binding sites in skeletal muscle or brain. These findings suggest that calcium entry activators, like calcium entry blockers, affect voltage-dependent calcium channels by diverse mechanisms.


EMPoWARed: Edmonton pediatric warfarin self-management study.
Patient self-management (PSM) in adults is safer and more cost effective than conventional management. Warfarin is a narrow therapeutic index drug with individual patient response to changes and frequently a long-term therapy. Children and their families are proposed to be able to effectively manage their child's warfarin therapy. Increased health related quality of life is highly associated with effective therapy in patients with chronic conditions. The aim of this study is to evaluate the safety and efficacy of PSM over time including HRQOL and variables that may influence PFU success at PSM. Children and their family units (PFUs) current performing patient self-testing/monitoring for ≥ 3 months were enrolled in this cohort study. PFUs participated in comprehensive education on warfarin testing and management followed by an apprenticeship. Socio-demographic, clinical, and laboratory data were collected to evaluate safety and efficacy and health related quality of life. Outcomes were compared between the first 6 months on PSM (phase 1) and the last 6 months data collected on PSM (phase 2). Forty-two patients performed PSM for a median of 2.7 years (range: 1.1-6.2 years). Time in therapeutic range was 90% and 92.9% (p=0.30) in phases 1 and 2 respectively. All measures were strongly associated with improved heath related quality of life. PFUs socio-demographic status did not influence success at PSM. All PFUs maintained warfarin knowledge and INR testing competency. Warfarin dosing decision errors median 0 (range: 0-5, p=0.73) and a median 0 (range 0-4, p=0.55) per patient in phases 1 and 2 respectively. There were no adverse hemorrhagic or thrombotic events. Empowering PFUs to self-manage warfarin results in increased knowledge and understanding of their health condition, improved commitment to their health care and adherence to medication regimens and is demonstrated to be sustainable over time.


Histology-based simulations of ultrasound imaging: methodology.
Simulations of ultrasound (US) images based on histology may shed light on the process by which microscopic tissue features translate to a US image and may enable predictions of feature detectability as a function of US system parameters. This technical note describes how whole-slide hematoxylin and eosin-stained histology images can be used to generate maps of fractional change in bulk modulus, whose convolution with the impulse response of the US system yields simulated US images. The method is illustrated by two canine mastocytoma histology images, one with and the other without signs of intra-operative hemorrhaging. Quantitative comparisons of the envelope statistics with corresponding clinical US images provide preliminary validation of the method.


Ultrastructural demonstration of a glycoproteinic surface coat in allergenic pollen grains by combined cetylpyridinium chloride precipitation and silver proteinate staining.
In allergenic birch pollen grains, highly watersoluble surface substances were precipitated by the cationic detergent cetylpyridinium chloride (CPC) during aqueous fixation. After processing the pollen for electron microscopy, ultrathin sections of pollen grains were subjected to the periodic acid - thiocarbohydrazide - silver proteinate (PA-TCH-SP) procedure according to Thiery (1967) for the detection of vicinal glycol groups. It was found that the material precipitated by CPC on the surface and within the exine cavities of the pollen wall strongly reacted with the PA-TCH-SP reagent thus indicating the presence of polysaccharides on the surface of birch pollen grains. In samples which had not been treated with the cationic detergent, PA-TCH-SP reactivity was reduced to thin linings on the surface and within the exine cavities. In both cases the exine proper did not stain whereas the intine showed moderate staining. Within the aperture region of the intine, PA-TCH-SP reactivity is preferably associated with fibrillar or reticular structures. The results are discussed with special reference to biochemical findings on allergenic birch pollen proteins.


Molecular phylogeny and missense mutations at envelope proteins across coronaviruses.
Envelope (E) protein is one of the structural viroporins (76-109 amino acids long) present in the coronavirus. Sixteen sequentially different E-proteins were observed from a total of 4917 available complete SARS-CoV-2 genomes as on 18th June 2020 in the NCBI database. The missense mutations over the envelope protein across various coronaviruses of the β-genus were analyzed to know the immediate parental origin of the envelope protein of SARS-CoV-2. The evolutionary origin is also endorsed by the phylogenetic analysis of the envelope proteins comparing sequence homology as well as amino acid conservations.


The response of the rabbit subsynovial connective tissue to a stress-relaxation test.
The subsynovial connective tissue (SSCT) in the carpal tunnel may play a role in the etiology of carpal tunnel syndrome (CTS), yet the material properties of the SSCT remain unclear. Thus, we investigated the mechanical response of the SSCT in a rabbit model. Twenty-four rabbit cadaver paws were used for mechanical testing; two paws were used for scanning electron microscopy (SEM) imaging. After testing normal tendon excursion, the divided third digit flexor digitorum superficialis (FDS) tendon was pulled to displacements of 2, 3.5, 5, or 8 mm, maintained at that position until force decay, and then the process was repeated. Normal excursion of the FDS averaged 4.8 mm. The ratio of the second peak force to the first peak force in the 2 mm group was 0.98 (SD = 0.16), which was significantly higher than the other groups (3.5 mm: 0.74, 5 mm, 0.63, and 8 mm: 0.59; p < 0.05). SEM showed ruptured fibrils in the displaced specimen. The declining force ratio with displacements >2 mm suggests damage to the SSCT within the physiological tendon excursion. These data may be useful in understanding SSCT mechanics in CTS, which is associated with SSCT fibrosis.


Utility of transesophageal echocardiography in identification of thrombogenic milieu in patients with atrial fibrillation (an ACUTE ancillary study).
The ACUTE trial randomly assigned patients who had atrial fibrillation (AF) of >2 days' duration to a transesophageal echocardiographically guided or a conventional strategy before cardioversion. In the 571 patients who underwent transesophageal echocardiography (TEE) in the ACUTE trial, we assessed the relative predictive value of baseline data derived by history, transthoracic echocardiography, and TEE for prediction of thrombus and adjudicated embolism (thromboembolism) as a composite end point. TEE was performed at 70 centers in 571 patients, 549 in the transesophageal echocardiographically guided group and 22 crossovers in the conventional group. Six patients (1.1%) who had embolism and 79 (13.8%) who had thrombi were identified in this group. Thrombus was completely resolved in 76.5% of patients who had repeat transesophageal echocardiographic procedures after 31.7 +/- 7.5 days of anticoagulation. For patients who had embolic events, none had a transesophageal echocardiographically identified thrombus; 5 of 6 (83.3%) had >/=1 transesophageal echocardiographic risk factors (including spontaneous echocardiographic contrast, aortic atheroma, patent foramen ovale, atrial septal aneurysm, mitral valve strands), and 4 of 6 (66.66%) had subtherapeutic anticoagulation or no anticoagulation. Clinical, transthoracic echocardiographic, and transesophageal echocardiographic risk factors contributed significantly to the prediction of composite thrombus/embolism. However, transesophageal echocardiographic thromboembolic risk factors were the strongest predictors of thromboembolism and provided statistically significant incremental value (chi-square 38.0, p <0.001) for identification of risk. Thus, in addition to thrombus identification, TEE has significant incremental value in the identification of patients who had high thromboembolic risk. In conclusion, this study supports the role of TEE and anticoagulation monitoring in patients who have atrial fibrillation and is useful for identifying thromboembolic risk factors.


Enhanced anaerobic degradation of nitrogen heterocyclic compounds with methanol, sodium citrate, chlorella, spirulina, and carboxymethylcellulose as co-metabolic substances.
The aim of the study was to explore the feasibility of methanol, sodium citrate, chlorella, spirulina, and carboxymethylcellulose (CMC) as co-metabolic substances in strengthening the anaerobic degradation of selected nitrogen heterocyclic compounds (NHCs). Chlorella, spirulina, and CMC as co-metabolic substances were first introduced into the enhanced anaerobic treatment of refractory compounds. With the addition of 300 μg/L sodium citrate, chlorella, spirulina, and CMC, reactor 3, reactor 4, reactor 5, and reactor 6 had higher degradation ratios than reactor 2 with methanol as co-metabolic substance. The addition of sodium citrate, chlorella, spirulina, and CMC increased the number of bacterial sequences, promoted the richness and diversity of the bacterial community structure, and enriched the functional genera (Levilinea and Longilinea) responsible for the degradation of quinoline and indole.


HLA and nasopharyngeal carcinoma in Chinese--a further study.
This paper reports the results of 8 years of HLA typing of newly diagnosed nasopharyngeal carcinoma (NPC) patients among Chinese in Singapore, and compares the HLA profile of NPC cases with that of the normal Chinese population. Earlier results had indicated an association with A2 and BW46, but the heterogeneity of the earlier case series, which contained a proportion of long-term survivors, obscured other aspects of the relationship. In this paper, the risk associated with A2 (relative risk = 1.5) and BW46 (relative risk = 1.9) is confirmed, showing a much higher relative risk in the presence of both antigens, and a risk is also demonstrated for B17 (relative risk = 2.1). Relative risks of less than unity are observed for AII (relative risk = 0.5) and B13 (relative risk = 0.5). Data from family studies indicate the importance of the haplotype A2.BW46 in determining risk for NPC.


[New growth hormone secretagogues: from physiology to the first therapeutic applications].
Growth hormone releasing peptides (GHRP) are synthetic hexapeptides that physiologically stimulate GH release through two different pathways: 1) central and 2) direct action on somatotropic cells. Animal experiments and first clinical trials show that synthetic GHRP and synthetic analogues could be useful substitutes to recombinant GH in the treatment of GH deficiency, and in pathological conditions which may benefit from amplification of the GH-IGF I axis activity.


Association between severity of illness and mortality from nosocomial infection.
For the years 1987 through 1992, a study was undertaken to analyze nosocomial infection mortality data and to stratify risk according to severity of underlying illness to compare with published data from the Centers for Disease Control and Prevention. Nosocomial infections that contributed to or caused death were identified. In addition, during 1990 through 1992, severity of illness was determined as the subjective estimate of the risk of death or lack of risk of death during the current hospital admission before the onset of the nosocomial infection. These groups were named +SIC and -SIC, respectively. It was determined that the data from death certificates in cases of known nosocomial infection were not sufficient to determine whether nosocomial infection contributed to or caused death. There was a 24% increase in cases of nosocomial infection contributing to or causing death when a physician reviewed deaths in patients with nosocomial infections who did not have a diagnosis of nosocomial infection listed on the death certificate. The rates for nosocomial infections contributing to or causing death are as follows: nosocomial pneumonia, 20%; and bloodstream infections, 19%. In patients who died and had severity of illness determination, there was a statistically significant difference in the rates of nosocomial infections contributing to or causing death between -SIC and +SIC groups for both nosocomial pneumonia and bloodstream infections. The rates for bloodstream infections were as follows: -SIC, 5%; and +SIC, 21%. For nosocomial pneumonia, the rates were as follows: -SIC, 13%; and +SIC, 23%. In published reports from the Centers for Disease Control and Prevention, a rate of 13% is given for nosocomial pneumonia and bloodstream infections contributing to or causing death; however, there is no stratification for severity of illness in these reports. The presence of life-threatening illness before the onset of nosocomial pneumonia or bloodstream infection accounts for most deaths among our patients. For valid comparisons, mortality outcome data for nosocomial infections should be stratified for risk according to severity of underlying illness.


Crocidolite asbestos suppresses the differentiation of HL-60 cells induced by DMSO.
This study was undertaken to determine if the biological function of inducers for cell differentiation is affected by asbestos fibers, which are sometimes deposited in human tissues. Protein kinase C activity, c-myc protein expression and cell surface CR3 expression were used as the markers of cell differentiation. The function of dimethylsulfoxide (DMSO), an inducer of cell differentiation, was suppressed by the co-culturing of crocidolite asbestos, because DMSO reacted with the hydroxyl radical released after the stimulation with crocidolite and spent itself. Superoxide dismutase (SOD) inhibited the effect of crocidolite, reacting rapidly with .O2- before the secondary release of .OH. Asbestos fibers deposited in tissues may inhibit the function of inducers which stimulate immature cells to differentiate, because such inducers frequently are also radical scavengers.


Evaluation of the Metabolic Relationship between Cows and Calves by Monitoring Calf Health and Cow Automatic Milking System and Metabolic Parameters.
With this study, we investigated the relationship between a cow's and calf's metabolic state, and its effect on health status. To achieve this, 20 calves of primiparous and 20 calves of multiparous cows were selected. The calves were monitored for 30 days and scored for signs of disease, as described in McQuirk (2008); according to score, they were divided into healthier calves; the Low calf score group (LCS, 5-8), Medium calf score group (MCS, 9-12) and High calf score group (HCS, 14-17); or calves most prone to disease. Their mothers were monitored for the same period with a Lely Astronaut 3 herd management system (Lely, Maassluis, The Netherlands) for rumination time, milk yield, milk fat, protein, lactose concentrations and milk fat to protein ratio. Both cows and calves were sampled for blood, and concentrations of glucose with β-hydroxybutyrate were registered. The results indicate that primiparous cows had a 16% higher blood glucose concentration (3.03 mmol/L SE = 0.093) compared with multiparous cows (2.61 mmol/L, SE = 0.102) (p < 0.01), but no difference in calf glucose was recorded. Β-hydroxybutyrate levels did not differ significantly between cows and calves by parity group. Rumination time was longest in the HCS group at 550.79 min/d. and was 16% longer compared with the LCS group (461.94 min/d.; p < 0.001) and 8% longer compared with the MCS group (505.56 min/d.; p < 0.001). The MCS group rumination time mean was statistically significantly higher compared with the LCS group by 8% (p < 0.001). Milk yield was also highest in the HCS group (44.8 kg/d.): 19% higher compared with the MCS group (36.31 kg/d., p < 0.001) and 13% higher than the LCS group (38.83 kg/d., p < 0.001). There was also a significant difference between the MCS and LCS groups of 6% (p < 0.001). The HCS group had the highest milk fat concentration (4.47%): it was 4% higher compared with the LCS group (4.28%, p < 0.001) and 5% higher than the MCS group (4.25%, p < 0.001). Milk fat to protein ratio was highest in the HCS group (1.21) and was 7% higher than in the MCS group (1.12, p < 0.001) and 8% higher than in the LCS group (1.11, p < 0.001). The LCS group was determined to have the highest concentration of milk lactose (4.66%). It was 1% higher compared with the MCS group (4.62%, p < 0.001) and 1.07% higher than the HCS group (4.61%, p < 0.001). We can conclude that parity did not affect calf health status and that cows of the HCS group showed symptoms of negative energy balance expressed through higher milk yield, higher milk fat concentration and higher milk fat to protein ratio, with lower milk lactose concentration. Further and more thorough research is needed to evaluate the relationship between pregnant cows and calves.


Comparison of genetic backgrounds of methicillin-resistant and -susceptible Staphylococcus aureus isolates from Portuguese hospitals and the community.
In order to understand the origins of the dominant methicillin-resistant Staphylococcus aureus (MRSA) clones in Portuguese hospitals, we compared the genetic backgrounds of nosocomial MRSA with methicillin-susceptible S. aureus (MSSA) isolates from the same hospitals (n=155) and from the community (n=157) where they were located. Pulsed-field gel electrophoresis, spa typing, multilocus sequence typing, and agr type analysis revealed that the genetic backgrounds correspondent to the dominant MRSA clones in Portuguese hospitals during the last 15 years (Iberian ST 247, Brazilian ST 239, and EMRSA-15 ST 22) were scarcely or not found among the present MSSA collection. The four major MSSA clones encountered (A-ST 30, B-ST 34, C-ST 5, and H-ST 45) correspond, or are very similar, to the background of other international MRSA pandemic clones, i.e., EMRSA-16, New York/Japan, Pediatric, and Berlin clones. However, with the exception of the Pediatric clone, none of these MRSA clones has been detected in Portugal. Our findings suggest the three major MRSA clones identified in Portuguese hospitals have not originated from the introduction of SCCmec into dominant MSSA backgrounds present in the Portuguese nosocomial or community environment but were probably imported from abroad. In contrast, the MRSA Pediatric clone might have originated in our country by the acquisition of SCCmec type IV into MSSA clone C. Furthermore, we provide evidence that the introduction of SCCmec into sensitive clones is most likely a relatively infrequent event that seems to depend not exclusively on the presence of a successful MSSA lineage.


Race differences in longitudinal associations between adolescent personal and peer marijuana use and adulthood sexually transmitted infection risk.
To assess whether adolescent marijuana exposure represents a modifiable predictor of risk of sexually transmitted infections as adults, we used nationally representative, longitudinal data from Waves I (1994-1995, adolescence) and III (2001-2002, adulthood) of the National Longitudinal Study of Adolescent Health (N = 10,738) to examine racial and gender differences in associations between adolescent marijuana use, current use, and peer use and adulthood multiple partnerships, self-reported sexually transmitted infections, and biologically confirmed sexually transmitted infections. The risk of sexually transmitted infections as adults was predicted by adolescent marijuana use in all groups except Black women and by peer marijuana use among Black men. Adolescents who use or have friends who use marijuana constitute priority populations for sexually transmitted infection prevention.


A study of mapping usual care and unmet need for vocational rehabilitation and psychological support following major trauma in five health districts in the UK.
To identify where and how trauma survivors' rehabilitation needs are met after trauma, to map rehabilitation across five UK major trauma networks, and to compare with recommended pathways. Qualitative study (interviews, focus groups, workshops) using soft-systems methodology to map usual care across trauma networks and explore service gaps. Publicly available documents were consulted. CATWOE (Customers, Actors, Transformation, Worldview, Owners, Environment) was used as an analytic framework to explore the relationship between stakeholders in the pathway. Five major trauma networks across the UK. 106 key rehabilitation stakeholders (service providers, trauma survivors) were recruited to interviews (n = 46), focus groups (n = 4 groups, 17 participants) and workshops (n = 5 workshops, 43 participants). None. Mapping of rehabilitation pathways identified several issues: (1) lack of vocational/psychological support particularly for musculoskeletal injuries; (2) inconsistent service provision in areas located further from major trauma centres; (3) lack of communication between acute and community care; (4) long waiting lists (up to 12 months) for community rehabilitation; (5) most well-established pathways were neurologically focused. The trauma rehabilitation pathway is complex and varies across the UK with few, if any patients following the recommended pathway. Services have developed piecemeal to address specific issues, but rarely meet the needs of individuals with multiple impairments post-trauma, with a lack of vocational rehabilitation and psychological support for this population.


Chemoproteomic target deconvolution reveals Histone Deacetylases as targets of (R)-lipoic acid.
Lipoic acid is an essential enzyme cofactor in central metabolic pathways. Due to its claimed antioxidant properties, racemic (R/S)-lipoic acid is used as a food supplement but is also investigated as a pharmaceutical in over 180 clinical trials covering a broad range of diseases. Moreover, (R/S)-lipoic acid is an approved drug for the treatment of diabetic neuropathy. However, its mechanism of action remains elusive. Here, we performed chemoproteomics-aided target deconvolution of lipoic acid and its active close analog lipoamide. We find that histone deacetylases HDAC1, HDAC2, HDAC3, HDAC6, HDAC8, and HDAC10 are molecular targets of the reduced form of lipoic acid and lipoamide. Importantly, only the naturally occurring (R)-enantiomer inhibits HDACs at physiologically relevant concentrations and leads to hyperacetylation of HDAC substrates. The inhibition of HDACs by (R)-lipoic acid and lipoamide explain why both compounds prevent stress granule formation in cells and may also provide a molecular rationale for many other phenotypic effects elicited by lipoic acid.


More than Meets the Eye: Untargeted Metabolomics and Lipidomics Reveal Complex Pathways Spurred by Activation of Acid Resistance Mechanisms in Escherichia coli.
Escherichia coli is a ubiquitous group of bacteria that can be either commensal gut microbes or enterohemorrhagic food-borne pathogens. Regardless, both forms must survive acidic environments in the stomach and intestines to reach and colonize the gut, a process that partially relies on amino acid-dependent acid resistance (AR) mechanisms and modifications to membrane phospholipids. However, only the basic tenets of these mechanisms have been elucidated. In this paper, we aim to conduct a full-scale metabolic and lipidomic characterization of E. coli's adaptations to acid stress. We hypothesized that the use of untargeted metabolomics and lipidomics would reveal mechanisms downstream of AR processes that provide novel contributions to acid stress survival. We detected significant differences in the extracellular metabolome and the lipidome induced by amino acid supplementation (glutamine, arginine, or lysine) and contextualized these results using real-time quantitative polymerase chain reaction (RT-qPCR). We additionally identified several metabolic pathways as well as a significant alteration in phospholipid synthetic pathways induced by differential amino acid supplementation. These results demonstrate that AR may extend beyond canonical mechanisms to a coordinated metabolic phenotype. Future studies may benefit from our analysis to further elucidate distinct targets for prebiotic supplements to cultivate commensal strains or therapies to combat pathogenic ones.


Occurrence of moniliformin, deoxynivalenol, and zearalenone in durum wheat (Triticum durum Desf.).
Forty-eight durum wheat samples from 5 locations in Austria were examined forFusarium infection andFusarium toxin content.F.gramlnearum andF.avenaceum were by far the prevailingFusarium species In durum wheat kernels, followed byEpoae, F.culmorum, andF.equlsetl. Ion-paired HPLC analyses of the samples showed moniliformin contents of kernels up to 0.88 mg/kg. All moniliformin contaminated samples also contained high levels of deoxynivalenol (up to 8.2 mg/kg) and lower levels of zearalenone (<0.33 mg/kg). The levels of zearalenone in naturally contaminated durum wheat samples did not correspond to the high yields of zearalenone found in cultures of the fusaria isolated from the durum wheat kernels. These conflicting results as well as some toxicologlcal aspects of the carry over ofFusarium toxins from durum wheat kernels into pasta are discussed.


CYP71D8L is a key regulator involved in growth and stress responses by mediating gibberellin homeostasis in rice.
Phytohormones are crucial in the regulation of plant growth and development, and in responses to adverse environments. Multiple cytochrome P450 monooxygenases (CYP450s) are involved in the biosynthesis and catabolism of phytohormones. Here, we report that a CYP450 member of the CYP71 clan in rice, OsCYP71D8L, participates in the control of multiple agronomic traits and abiotic stress responses by affecting gibberellin (GA) and cytokinin (CK) homeostasis. The gain-of-function mutant cyp71d8l and transgenic plants overexpressing CYP71D8L (CYP71D8L-OE) display similar phenotypes compared to the wild-type (WT), including dwarfed plants, reduced panicle length, reduced grain number per panicle, and decreased levels of endogenous GAs. Moreover, the dwarfed plant trait and the less-developed roots of CYP71D8L-OE and cyp71d8l seedlings could be rescued by application of GA3 or the CK biosynthetic inhibitor lovastatin, and exacerbated by application of the synthetic CK 6-BA. Importantly, CYP71D8L-OE and cyp71d8l seedlings maintained high chlorophyll contents and low levels of reactive oxygen species, and showed enhanced tolerance to drought and salt stress compared with the WT. Thus, our results suggest that OsCYP71D8L plays important roles in regulating rice growth and stress responses by coordinating the homeostasis of GAs and CKs, and it may therefore be a useful target for engineering stress-tolerant rice varieties.


Stand-Alone or Combinatorial Effects of Grafting and Microbial and Non-Microbial Derived Compounds on Vigour, Yield and Nutritive and Functional Quality of Greenhouse Eggplant.
The current research investigated the effects of endophytic fungi such as Trichoderma atroviride (Ta) or Ascophyllum nodosum seaweed extract (An) and their combination on growth, yield, nutritive and functional features, and mineral profile of 'Birgah' F1 eggplant either ungrafted, self-grafted or grafted onto the Solanum torvum rootstock. Eggplant exposed to An or An+Ta had a significant increase in root collar diameter 50 days after transplanting (RCD50), total yield (TY), marketable yield (MY), ascorbic acid (AA) content, Mg, Cu, and Zn concentration, and a reduction in glycoalkaloids (GLY) compared with the control. Furthermore, grafted plants had a higher TY, MY, number of marketable fruits (NMF), RCD50, AA, Cu, and Zn and a lower SSC, GLY, and Mg than the ungrafted plants. The combination of grafting and An+Ta significantly improved mean weight of marketable fruits (MF), plant height 50 days after transplanting (PH50), number of leaves 50 days after transplanting (NL50), fruit dry matter (FDM), chlorogenic acid (ClA), proteins, and K and Fe concentration. This combination also produced fruits of high premium quality as evidenced by the higher AA and ClA concentration, the lower GLY concentration, and an overall improved mineral profile.