A practical fully automated radiosynthesis of [18F]Flurpiridaz on the module modular lab-pharmtracer without external purification.
[18F]Flurpiridaz is a promising novel cardiac PET imaging tracer formed by the radiolabeling of pyridaben derivative with fluorine-18. Clinical studies on [18F]Flurpiridaz are currently at the phase III level for the assessment of MPI. Providing high image quality thanks to its relatively long half-life, F-18 is a high-potential radionuclide for the early detection of CAD. In this study, we aimed to develop a fully automated synthesis of [18F]Flurpiridaz without further preparative HPLC purification. Precursor 6 was obtained by multi-step synthesis starting from mucochloric acid (1) as a sole product with 35% yield and identified by spectroscopic measurement. Manually cold labeling experiments were performed using the stable isotope [19F]F, and TBA-HCO3 PTC provided desirable fluorinated compound with high yield. A fully automated [18F]Flurpiridaz synthesis on the ML-PT device provided 55-65% radiochemical yield with more than 98% radiochemical purity. The final product purification method demonstrated that [18F]Flurpiridaz could be obtained without an external preparative HPLC system as a pharmaceutical quality. A novel and fascinating strategy was developed for the fully automated synthesis of [18F]Flurpiridaz (7) on ML PT. Organic synthesis of precursor 6 was achieved with a desirable yield and characterized by NMR and HR-MS. A detailed set of cold experiments were completed for optimization conditions before hot trials and TBA-HCO3 increased molar activity with a minimum amount of side products. Radiolabeling showed that our self-designed automated synthesis method enables high radiochemical yield and radiochemical purity for the production of [18F]Flurpiridaz. The desirable radiopharmaceutical quality of the product was obtained without using an additional preparative HPLC system. [18F]Flurpiridaz (7) preserved its stability within 12 h and final specifications were consistent with the acceptance criteria in Ph. Eur. regulations.


Central precocious puberty in girls: Diagnostic study and auxological response to triptorelin treatment.
There are several controversies regarding the diagnostic tests and management of central precocious puberty (CPP). The aim of this study is to present the experience acquired in a group of girls with CPP treated with triptorelin, and to analyze the auxological characteristics and diagnostic tests. An observational, retrospective study in a group of 60 girls with CPP was conducted between January 2010 and December 2017. Sociodemographic, auxological and hormonal data were recorded at diagnosis, and pelvic ultrasound and magnetic resonance imaging of the head were performed. Girls were treated with triptorelin and monitored after treatment discontinuation until menarche. At treatment start, chronological age and bone age were 7.7±0.7 and 9.7±0.8 years respectively, and growth velocity was 8.3±1.6cm/year. Target height was 161.1±5.8cm. Peak LH level after stimulation was 16.6±12.1 IU/l. Ovarian volumes were greater than 3mL in 35% of cases. MRI of the head was pathological in seven girls (11.7%). At treatment completion, chronological age and bone age were 10.3±1.1 and 11.2±0.8 years respectively, and growth velocity was 4.7±1.4cm/year. At the age of menarche (11.9±0.9 years), height was 157.5±5.7cm. Treatment of CPP with triptorelin appears to be beneficial. The possibility to block pubertal development and slow skeletal maturation allows patients to reach their target height. However, individualized auxological monitoring would be mandatory.


N2 neutrophils may participate in spontaneous recovery after transient cerebral ischemia by inhibiting ischemic neuron injury in rats.
Neutrophils have been traditionally considered as the major mediators of harmful inflammatory responses in ischemic stroke, whereas accumulating evidence indicates that neutrophils can be polarized into an N2 phenotype. Similar to M2 microglia, N2 neutrophils contribute to resolution of inflammation and may participate in neuroprotection. However, it remains unclear whether N2 neutrophils protect ischemic neurons and whether they are associated with long-term outcomes after transient cerebral ischemia in rats. The present study proved that N2 neutrophils protected against oxygen glucosedeprivation/re-oxygenation (OGD/R)-induced primary cortical neuron injury via brain-derived neurotrophic factor/tropomyosin-related kinase B (BDNF/TrkB) signaling. In addition, in vivo studies revealed that transient middle cerebral artery occlusion (tMCAO)-induced injury exhibited spontaneous recovery over time in rats. Moreover, neutrophils could infiltrate the ipsilateral brain parenchyma from the periphery after transient cerebral ischemia. Pearson's correlation analysis indicated that the proportion of N2 neutrophils in ipsilateral brain parenchyma was negatively correlated with the number of degenerating neurons, modified Neurological Severity Score (mNSS), brain water content and infarct volume, and positively correlated with the number of surviving neurons and grip strength. In summary, the present study shows that N2 neutrophils likely participate in spontaneous recovery after transient cerebral ischemia by inhibiting ischemic neuron damage in rats, which indicates that N2 neutrophils may represent promising therapeutic target for promoting recovery after ischemic stroke.


ECM remodeling regulates angiogenesis: endothelial integrins look for new ligands.
The process of angiogenesis is a dynamic one. Vascular endothelial cells are changing at the same time the extracellular matrix is being remodeled. Stupack and Cheresh discuss how remodeling of the extracellular matrix (ECM) and changes in the endothelial cell protein production and integrin expression contribute to the complex process of new blood vessel growth from an existing vascular bed.


Health professions students' perceptions of interprofessional relationships.
To make a preliminary assessment of the perceptions of health professions students about interprofessional cooperation. Health professions students (588 students from eight professions) at the Iowa Geriatric Education Center's partner institutions received a questionnaire of demographics questions and the Interdisciplinary Education Perception Scale (IEPS). The IEPS is an 18-item questionnaire that uses a six-point Likert-type scale to measure attitudes toward interprofessional cooperation on four factors: competence and autonomy, perceived need for cooperation, perception of actual cooperation, and understanding others' value. Total mean IEPS scores differed significantly among professional groups (p =.001), with physician assistant students scoring highest (most positive attitudes) and chiropractic students scoring lowest. The medical students' mean total score was significantly lower than was that of physician assistant students (p =.003) and higher than was that of chiropractic students (p =.000), but medical students' scores did not differ significantly at the alpha =.05 level from those of osteopathy, physical therapy, nursing, podiatry, or social work students. This study provides the first normative data for the IEPS for students from these eight health professions. This instrument may be valuable when designing an evaluation scheme for training programs that have interdisciplinary components, which may be increasingly prevalent in the future.


Microplastics everywhere: A review on existing methods of extraction.
Microplastics (MPs) are a ubiquitous pollutant. They take the form of fibers, fragments, microbeads, pellets, and foams, slowly choking up waterways, the atmosphere, and, eventually, the food chain. As the need for analysis and removal of these polymers takes on a role of utmost importance, it becomes imperative to know the methods by which these micro-pollutants can be extracted from the environmental matrices in which they appear. This review aims to compile the various methods of MP extraction available in literature. With general methods like density separation and the oil extraction protocol listed alongside research on process-based separation using ultrasound and fluid dynamics, there may be a way to create a standardized protocol for the mass extraction of MPs from any environmental matrix.


A promising future for tele-mental health in Oman: A qualitative exploration of clients and therapists' experiences.
Tele-mental health services can play an important role in overcoming barriers in mental health services in the Eastern Mediterranean Region. However, despite its potential, tele-mental health has not been widely adopted in Oman. This study is an exploratory investigation into the experiences of therapists and their clients in utilizing video-based tele-mental health care during the COVID-19 pandemic. A total of 19 semistructured qualitative interviews were individually conducted, it included 13 adult clients with mental health conditions who received video-based tele-mental health care and six clinical psychologists who provided video-based tele-mental health care during the COVID-19 pandemic. The clients reported favorable experiences using tele-mental health, with the primary benefits being convenience, easy accessibility to subspecialized care, reduced absenteeism from work with commuting costs, and alleviated mental health stigma. The therapists also expressed experiencing benefits from tele-mental health, such as reduced risk of intrahospital infection, reduced healthcare costs, and the achievement of work-life balance. Primary concerns were related to the lack of public tele-mental health services, lack of specified tele-mental health guidelines, shortage of trained therapists, limited access to high-speed Internet, electronic devices, privacy, and concerns toward the security of telehealth systems in general. Clients and therapists report that tele-mental health offers new opportunities to improve the quality of mental healthcare services in Oman, and that the challenges could be resolved by establishing governmental tele-mental health services along with developing tele-mental health guidelines and implementing local postgraduate clinical psychology programs in universities in Oman.


Massive endoprostheses for giant cell tumours of the distal femur: A 12-year follow-up.
We performed a retrospective analysis of twenty-five consecutive massive articulating endoprostheses that were inserted at our unit during the management of patients with Giant Cell Tumours of their distal femur. Fifteen of these implants were fixed hinge devices and ten were rotating hinge devices with HA collars (since 1993). None of these cases were revised for sepsis. There had been no cases of recurrent disease or amputation. The mean follow-up was 12 years (range = 5-18 years). The average age at time of insertion was 37 years. Young patients with fixed hinged devices developed a high incidence (33%) of aseptic loosening. They also had a significant rate of re-bushing. Results of the rotating hinge prosthesis with HA collar were much more promising. Functional scores were good after a period of 12 years despite the young age group.


Hyperinsulinaemic hypoglycaemia: genetic mechanisms, diagnosis and management.
Hyperinsulinaemic hypoglycaemia (HH) is due to the unregulated secretion of insulin from pancreatic β-cells. A rapid diagnosis and appropriate management of these patients is essential to prevent the potentially associated complications like epilepsy, cerebral palsy and neurological impairment. The molecular basis of HH involves defects in key genes (ABCC8, KCNJ11, GLUD1, GCK, HADH, SLC16A1, HNF4A and UCP2) which regulate insulin secretion. The most severe forms of HH are due to loss of function mutations in ABCC8/KCNJ11 which encode the SUR1 and KIR6.2 components respectively of the pancreatic β-cell K(ATP) channel. At a histological level there are two major forms (diffuse and focal) each with a different genetic aetiology. The diffuse form is inherited in an autosomal recessive (or dominant) manner whereas the focal form is sporadic in inheritance and is localised to a small region of the pancreas. The focal form can now be accurately localised pre-operatively using a specialised positron emission tomography scan with the isotope Fluroine-18L-3, 4-dihydroxyphenyalanine (18F-DOPA-PET). Focal lesionectomy can provide cure from the hypoglycaemia. However the diffuse form is managed medically or by near total pancreatectomy (with high risk of diabetes mellitus). Recent advances in molecular genetics, imaging with 18F-DOPA-PET/CT and novel surgical techniques have changed the clinical approach to patients with HH.


Optimization of the Use of Hospital Beds as an Example of Improving the Functioning of Hospitals in Poland on the Basis of the Provincial Clinical Hospital No. 1 in Rzeszow.
An efficient health care system combines maximum accessibility with high-quality treatments, as well as cost optimization of individual health care facilities throughout the entire system. In hospitals, the critical element is the number of beds within individual wards, which generates costs and, at the same time, affects the capacity to serve patients. The aim of this article is to discuss the restructuring and optimization of hospital bed occupancy in a healthcare facility in the Podkarpackie voivodeship. The analysis covers the years 1999-2018. In the indicated period, the analyzed healthcare institution restructured the number of beds based on a forecast of the demand for services, which resulted in positive cost effects, without limiting patients' access to diagnostic and therapeutic care. The analyzed facility took part in a common trend of optimizing cost-effectiveness and efficiency of hospital operations in Poland.


Prospective cohort analyzing risk factors for chronic kidney disease progression in children.
To identify risk factors for chronic kidney disease progression in Brazilian children and to evaluate the interactions between factors. This was a multicenter prospective cohort in São Paulo, involving 209 children with CKD stages 3-4. The study outcome included: (a) death, (b) start of kidney replacement therapy, (c) eGFR decrease >50% during the followup. Thirteen risk factors were tested using univariate regression models, followed by multivariable Cox regression models. The terms of interaction between the variables showing significant association with the outcome were then introduced to the model. After a median follow-up of 2.5 years (IQR=1.4-3.0), the outcome occurred in 44 cases (21%): 22 started dialysis, 12 had >50% eGFR decrease, seven underwent transplantation, and three died. Advanced CKD stage at onset (HR=2.16, CI=1.14-4.09), nephrotic proteinuria (HR=2.89, CI=1.49-5.62), age (HR=1.10, CI=1.01-1.17), systolic blood pressure Z score (HR=1.36, CI=1.08-1.70), and anemia (HR=2.60, CI=1.41-4.77) were associated with the outcome. An interaction between anemia and nephrotic proteinuria at V1 (HR=0.25, CI=0.06-1.00) was detected. As the first CKD cohort in the southern hemisphere, this study supports the main factors reported in developed countries with regards to CKD progression, affirming the potential role of treatments to slow CKD evolution. The detected interaction suggests that anemia may be more deleterious for CKD progression in patients without proteinuria and should be further studied.


2-(4-Methoxyphenyl)ethyl-2-Acetamido-2-deoxy-β-d-pyranoside (A Salidroside Analog) Confers Neuroprotection with a Wide Therapeutic Window by Regulating Local Glucose Metabolism in a Rat Model of Cerebral Ischemic Injury.
2-(4-Methoxyphenyl)ethyl-2-acetamido-2-deoxy-β-d-pyranoside (salidroside analog-4g, SalA-4g), has shown neuroprotective prospects for the treatment of ischemic stroke. However, the dose-response and time window study for SalA-4g, and the mechanism of SalA-4g-mediated neuroprotection remain unclear. Here, we systematically investigated the therapeutic time window and dosage of SalA-4g in permanent focal cerebral ischemia in rats. SalA-4g dose-dependently improved stroke outcome. Either pre-treatment or post-treatment of SalA-4g exhibited notable neuroprotection, and maintained for up to 6 h after ischemia onset. Moreover, significant neurological functional recovery was found after SalA-4g administration in long-term functional assays. Further studies suggested that SalA-4g ameliorated neuronal cell death, elevated local glucose metabolism and enhanced the expression level of glucose transporter 1 and 3 in the ipsilateral cortex and striatum. We suggest that data of this study are critical in exploring the clinical application prospects of SalA-4g for the treatment of ischemic stroke.


Measuring oxidative damage to DNA and its repair with the comet assay.
Single cell gel electrophoresis, or the comet assay, was devised as a sensitive method for detecting DNA strand breaks, at the level of individual cells. A simple modification, incorporating a digestion of DNA with a lesion-specific endonuclease, makes it possible to measure oxidised bases. With the inclusion of formamidopyrimidine DNA glycosylase to recognise oxidised purines, or Nth (endonuclease III) to detect oxidised pyrimidines, the comet assay has been used extensively in human biomonitoring to monitor oxidative stress, usually in peripheral blood mononuclear cells. There is evidence to suggest that the enzymic approach is more accurate than chromatographic methods, when applied to low background levels of base oxidation. However, there are potential problems of over-estimation (because the enzymes are not completely specific) or under-estimation (failure to detect lesions that are close together). Attempts have been made to improve the inter-laboratory reproducibility of the comet assay. In addition to measuring DNA damage, the assay can be used to monitor the cellular or in vitro repair of strand breaks or oxidised bases. It also has applications in assessing the antioxidant status of cells. In its various forms, the comet assay is now an invaluable tool in human biomonitoring and genotoxicity testing. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.


Polyoma virus, as a model for viral skin carcinogenesis.
Polyoma virus is presented as the simplest model of transformation by a DNA virus. The three possible ways of virus/cell interaction are recalled: lytic infection, abortive infection and malignant transformation. Nuclear and surface antigenic modifications are also recalled as well as their possible correlation with the transformation mechanism. Lastly, in "in vivo" systems, we have tried to schematize the interactions of host and virus-induced tumour.


Irradiation as a safety procedure in tissue banking.
The Central Tissue Bank in Warsaw was established in 1963 and since then ionising radiation has been routinely applied to sterilise tissue grafts. Connective tissue grafts such as bone, cartilage, tendons, sclera, pericardium, skin, acellular dermis and amnion irradiated with a dose of 35 kGy in a (60)Co source and/or with an electron beam 10 MeV accelerator are prepared in our Tissue Bank and two other multi-tissue banks operating in Poland. Over 250,000 radiation-sterilised tissue grafts have been prepared and used in hospitals throughout Poland and no infectious disease transmission or other adverse post-transplantation reactions have been reported up to today. It should be kept in mind however, that high doses of ionising radiation can evoke numerous chemical and physical changes that may affect the biological quality of tissue allografts. Therefore, interdisciplinary research has been undertaken at the Central Tissue Bank in Warsaw to establish the origin and stability of free radicals and other paramagnetic entities induced by irradiation in bone. The effects of various preservation procedures (e.g. lyophilisation, deep-freezing) and irradiation conditions (doses, temperature of irradiation) on the osteoinductive potential and mechanical properties of bone and on the degradation of collagen, a major constituent of all connective tissue grafts, have been also studied. The results of these studies indicate that radiation-induced changes can be diminished by modification of tissue preservation methods and that, to some extent, it is possible to reduce undesired radiation-induced damage to the tissue grafts.


Photogenotoxicity of folic acid.
Folic acid (FA), also named vitamin B9, is an essential cofactor for the synthesis of DNA bases and other biomolecules after bioactivation by dihydrofolate reductase (DHFR). FA is photoreactive and has been shown to generate DNA modifications when irradiated with UVA (360 nm) in the presence of DNA under cell-free conditions. To investigate the relevance of this reaction for cells and tissues, we irradiated three different cell lines (KB nasopharyngeal carcinoma cells, HaCaT keratinocytes, and a melanoma cell line) in the presence of FA and quantified cytotoxicity and DNA damage generation. The results indicate that FA is phototoxic and photogenotoxic by two different mechanisms. First, extracellular photodecomposition of FA gives rise to the generation of H2O2, which causes mostly DNA strand breaks. If this is prevented, e.g., by the presence of catalase, DNA damage generated by intracellular FA becomes evident. The damage spectrum in this case consists predominantly of oxidatively generated purine modifications sensitive to the repair glycosylase Fpg, as characteristic for type I photoreactions, and is associated with the formation of micronuclei. In KB cells, the DNA damage is strongly enhanced after pretreatment with the DHFR inhibitor methotrexate, which prevents the loss of the chromophore associated with the intracellular reduction of FA by DHFR. The results indicate that FA is photoreactive in cells and gives rise to nuclear DNA damage under irradiation.


Quantitative analysis of eyes and other optical systems in linear optics.
To show that 14-dimensional spaces of augmented point P and angle Q characteristics, matrices obtained from the ray transference, are suitable for quantitative analysis although only the latter define an inner-product space and only on it can one define distances and angles. The paper examines the nature of the spaces and their relationships to other spaces including symmetric dioptric power space. The paper makes use of linear optics, a three-dimensional generalization of Gaussian optics. Symmetric 2 × 2 dioptric power matrices F define a three-dimensional inner-product space which provides a sound basis for quantitative analysis (calculation of changes, arithmetic means, etc.) of refractive errors and thin systems. For general systems the optical character is defined by the dimensionally-heterogeneous 4 × 4 symplectic matrix S, the transference, or if explicit allowance is made for heterocentricity, the 5 × 5 augmented symplectic matrix T. Ordinary quantitative analysis cannot be performed on them because matrices of neither of these types constitute vector spaces. Suitable transformations have been proposed but because the transforms are dimensionally heterogeneous the spaces are not naturally inner-product spaces. The paper obtains 14-dimensional spaces of augmented point P and angle Q characteristics. The 14-dimensional space defined by the augmented angle characteristics Q is dimensionally homogenous and an inner-product space. A 10-dimensional subspace of the space of augmented point characteristics P is also an inner-product space. The spaces are suitable for quantitative analysis of the optical character of eyes and many other systems. Distances and angles can be defined in the inner-product spaces. The optical systems may have multiple separated astigmatic and decentred refracting elements.


Efficacy of three-dimensionally integrated exercise for scoliosis in patients with adolescent idiopathic scoliosis: study protocol for a randomized controlled trial.
Adolescent idiopathic scoliosis (AIS) is one of the most prevalent spinal deformities that may progress sharply during growth. The aim of this study will be to evaluate the efficacy of three-dimensionally integrated exercise on the Cobb angle, angle of trunk rotation, sagittal profile, and quality of life in patients with AIS. The study is designed as a randomized controlled trial. Participants include 42 patients with AIS aged 10-16 years. Randomly assigned patients will follow a 6-month treatment, either in a control group with standard care of observation following the Scoliosis Research Society criteria or in an experimental group with three-dimensionally integrated exercise for scoliosis. Blinded assessments at baseline and immediately after intervention will include the change of Cobb angle, angle of trunk rotation, sagittal index, and quality of life. If we find that the intervention is effective in improving Cobb angle, angle of trunk rotation, sagittal profile, and quality of life in patients with AIS, this trial will have a positive impact and warrant a change in clinical practice. ClinicalTrials.gov, NCT03427970 . Registered on February 9, 2018, and revised on July 24, 2018.


Potential feasibility of a novel over-the-wire microelectrode catheter for recording coronary sinus electrograms in patients with cardiac resynchronization therapy devices.
The placement of an electrode catheter into the coronary sinus (CS) is important for differentiating multiple atrial tachycardias (ATs). Based on its high selective performance in placement into small veins, inserting a novel over-the-wire 2.7Fr microcatheter (EPstar Fix AIV; Japan Lifeline) into the CS may be feasible even in patients implanted with a cardiac resynchronization therapy device.


Biventricular tissue tracking demonstrating associations between left ventricular myocardial extracellular volume, pulmonary artery pressure, and reduced right ventricular ejection fraction in patients with systemic lupus erythematosus using cardiovascular MRI.
To investigate the feasibility and prognostic value of biventricular strain analysis for patients with systemic lupus erythematosus (SLE) and relationship to left ventricular (LV) myocardial fibrosis, pulmonary hypertension (PAH), and right ventricular (RV) ejection fraction (RVEF). A total of 50 SLE patients (47 women; 34.4±12 years) and 15 controls (13 women; 32.9±8.6 years) were imaged via echocardiography and a 3 T magnetic resonance imaging (MRI). Pulmonary artery systolic pressure (sPAP) was assessed using echocardiography. Biventricular global circumferential strain (GCS), global longitudinal strain (GLS), global circumferential strain rate (GCSR), and global longitudinal strain rate (GLSR) as well as LV myocardial extracellular volume (ECV) were derived for each subject. Elevated LV ECV was significantly associated with LV GCS (beta -0.428, p=0.000), GLS (beta 0.404, p=0.000), GCSR (beta -0.350, p=0.006), GLSR (beta -0.445, p=0.000) and RV GCS (beta -0.373, p=0.000), and the presence of reduced RVEF was significantly associated with LV GCS (beta -0.338, p=0.002), GLS (beta -0.465, p=0.000) and RV GCS (beta -0.465, p=0.000). Raised sPAP was significantly associated with RV GLS (beta 0.445, p=0.000) and GCSR (beta -0.387, p=0.001). Moreover, there were significantly correlations between LV and RV strain and strain rate. Biventricular strain analysis may be a better prognostic tool for patients with SLE as it allows greater comprehensive analysis and more definitive treatment planning compared to separate assessment of LV or RV contractility.


Atrial fibrillation and stroke: the view from neurology.
In summary NVAF is an important risk factor for stroke identifying a population at a six fold increased stroke risk. When stroke occurs it tends to be large and without a preceding TIA. These patients commonly have other cardiovascular disorders that must be considered as a potential cause of brain ischemia. The appropriate management of these patients to prevent cardioembolic stroke is unknown and must be individualized pending appropriate clinical trials. Following a cardioembolic stroke anticoagulation should be considered only in patients with small or moderate sized infarcts who have no evidence of hemorrhagic infarction on a CT scan and delayed until 36-48 hours post stroke onset.


Bacterial counts in cerebrospinal fluid of children with meningitis.
Eighty-five cerebrospinal fluid (CSF) specimens from the same number of pediatric patients with meningitis were examined to determine the bacterial count and the relationship of this count to the microscopy results, the ages of the patients and the bacterial species isolated. Bacterial counts ranged from 2 x 10 to 4 x 10(9) CFU/ml CSF. Twenty-five percent of the 85 CSF specimens positive for Haemophilus influenzae type b, Neisseria meningitidis, Streptococcus pneumoniae, Escherichia coli K1 and group B streptococci had counts of 10(7) CFU/ml or higher. Children between 1 and 6 months of age had significantly higher counts (p less than 0.05) than the other age groups. The three patients who had positive CSF cultures 24 h after the start of therapy all had initial bacterial counts of 10(7) CFU/ml or higher. The detection limit for Gram stain/microscopy was 10(5) CFU/ml. No correlation was found between bacterial count and the number of polymorphonuclear leukocytes.


Five epitopes of a differentiation antigen on human inducer T cells distinguished by monoclonal antibodies.
A series of mouse monoclonal antibodies reacting with human T cells of the helper/inducer subclass, OKT4, 4A, 4B, 4C, and 4D, have been reported. Using double-fluorescent staining and complement-mediated depletion, it was shown that the antigen(s) recognized by OKT4, 4A, 4B, 4C, and 4D antibodies are present on the same cell. Using FITC-labeled OKT4, 4A, 4B, 4C and 4D, a lack of competition between the antibodies for their epitopes was shown. Immunoprecipitation of the antigen recognized by each antibody yielded a molecule of approximately 60,000-62,000 Da. Sequential precipitation with several antibodies resulted in a minimum of additional precipitated antigen following removal of the first antigen. Capping of cell surface antigen with OKT4, followed by staining with OKT4, 4A, 4B, 4C, or 4D, indicated that the epitopes for all five antibodies co-cap. A sandwich ELISA assay using OKT4 and the other antibodies showed that molecules binding to OKT4A, 4B, 4C, and 4D also bound OKT4. It can therefore be concluded that monoclonal antibodies OKT4, 4A, 4B, 4C, and 4D recognize distinct epitopes present on a molecule of approximately 60-62,000 Da on human helper/inducer T cells.


Anti-IgE-induced histamine release from human basophilic leukocytes: inhibition depends on nature and concentration of inhibitor and secretagogue.
Histamine release from human basophilic leukocytes was induced by increasing concentrations of anti-IgE in the presence of various concentrations of 2-deoxyglucose (2-DOG), the histamine H2-receptor agonist dimaprit, theophylline, or enprofylline (a new antiasthmatic xanthine derivative). The results show that the degree of inhibition produced by each agent differed with the concentration of anti-IgE used and with the nature and concentration of the inhibitor. These data indicate that great care should be used when characterizing an inhibitor of mediator release by simply giving a figure for the per cent inhibition of release observed in its presence.


The influence of muscle interstitial volume on K+-induced heart rate drive in rats.
During exercise heart rate is influenced by reflexogenic drives which are elicited by receptors situated in the interstitial space. Since the structure of interstitial tissue is complex (e.g. fixed negative charges of glycosaminoglycans), the situation in the immediate surrounding of the receptors might differ from the free fluid phases of blood or lymph in which the concentrations of stimulating substances are usually determined. Physiological variations of the interstitial structure may be due to changes in interstitial volume induced by exercise or the hydrostatic effects on body fluids. The objective of the present study was to investigate the effect of the interstitial volume on the relationship between heart rate and K+ stimuli applied through the muscle blood vessels. The calves of 12 male Wistar rats were artificially perfused and separated from the rest of the body with the sciatic nerve remaining intact. In these preparations the heart rate (HR) responses to low (4 mM) and high (8 mM) potassium concentrations were determined at different interstitial volumes. Expansion of the interstitial volume was obtained by reducing the colloid-osmotic pressure of the perfusate. The combination of intracellular oedema and mechanical limitation of total volume expansion (tapeing) was utilized to decrease the interstitial volume. When switching between the low and high potassium concentrations, significant heart rate responses could be observed only with reduced interstitial volume. It is suggested that the interstitial structure surrounding the muscular receptors modifies the relationship between heart rate response and the K+ stimuli determined in blood or lymph.


Cytoplasmic colloid inclusions in thyroid lesions: a cytomorphological study based on fine needle aspiration.
Intracytoplasmic lumens (ICL) with or without magenta material and transgressing vessels are features of Hürthle cell neoplasms (HCN). After detection of intracytoplasmic colloid inclusions (CIs) including targetoid (magenta) body-like structures in the Hürthle cells (HC) in a case of Hashimoto's thyroiditis (HT), we reviewed cases of HT, thyroid neoplasms, hyperplastic nodules (HN) and colloid goitres to determine the frequency of these structures. Further, an attempt was made to find out the significance of CIs. FNA smears of 120 HT, 101 colloid goitres, 11 HN, and 76 neoplastic goitres were examined. The presence of CIs and empty ICL were noted in epithelial cells in these lesions. An attempt was made to find out the difference between HT with and without CIs in respect of various cytomorphologic features. The groups were compared using the Fisher's exact test of probability. The CIs were present in 36 (30.0%) of HT, 26 (34.2%) of neoplastic goitres, 3 (27.3%) of HN, and 4 (4.0%) colloid goitres. As compared to colloid goitres, CIs were present in a significant higher number of cases in HN (P = 0.0202), neoplastic goitres (P < 0.0001), and HT (P < 0.0001). Among neoplasms the frequency of CIs in HCN (75.0%) was significantly higher than that of papillary thyroid carcinoma (PTC) (33.3%, P = 0.0466), and follicular neoplasm (14.3%, P = 0.0083). The CIs were more frequent in HC in HT and HCN but in follicular cells (FC) in other lesions. The HT cases with CIs differed significantly from those without CIs in respect of HC and their cellularity, cellularity of reactive lymphoid cells, extracellular colloid and empty ICL. Care should be taken not to diagnose HT cases with an excessive Hürthle cell component and CIs, and PTC cases with Hürthlization and CIs, as HCN in FNA smears. Based on review of the literature and our findings, it is suggested that the Hürthle cell metaplasia in HT is a survival response of FC and the presence of CIs in Hürthle cell may represent their limited ability to synthesize colloid.


Expression of concern: A study on the luminescence properties of gamma-ray-irradiated white light emitting Ca2Al2SiO7:Dy3+ phosphors fabricated using a combustion-assisted method.
Expression of concern for 'A study on the luminescence properties of gamma-ray-irradiated white light emitting Ca2Al2SiO7:Dy3+ phosphors fabricated using a combustion-assisted method' by Geetanjali Tiwari et al., RSC Adv., 2016, 6, 49317-49327.


An experimental assessment of macroscopic enzyme techniques for the autopsy demonstration of myocardial infarction.
Experimental cardiac infarction produced in dogs by coronary artery ligation has been used to investigate the value of macroscopic histochemical enzyme loss in the diagnosis of myocardial infarction at post-mortem. Creative phosphokinase and non-specific dehydrogenase methods gave the best results but became positive only 5-6 hr after infarction. It is concluded that the method is of limited value in autopsy practice.


Intraoperative herniation of an L5-S1 disc during microdiscectomy and transforaminal lumbar interbody fusion: a case report.
We report the progression of an intraoperative L5-S1 lumbar disc herniation that occurred during a routine microdiscectomy and transforaminal lumbar interbody fusion, which, to the best of our knowledge, has never been previously reported in the literature. The objective of this report is to bring to light the possibility of a lumbar disc herniating intraoperatively, and to demonstrate that accompanying neurologic involvement can be detected and subsequently addressed with the aid of neurophysiologic monitoring. A 36-year-old African American woman, who had previously undergone minimally invasive microdiscectomy for a right L5-S1 herniated nucleus pulposus with full recovery, presented with a large reherniation of the L5-S1 disc on the right side. During her operation, while a tap was followed into the L5 left pedicle, there was a sudden profound spasm of our patient's legs and back that lasted for the duration of 15 seconds, culminating in the loss of all somatosensory evoked potentials in our patient's lower extremities. Exploration of this previous microlaminotomy site revealed a massive disc extrusion protruding through the microlaminotomy. Immediate removal of this extruded disc material restored all somatosensory evoked potentials and our patient awoke with no neurologic deficits. An intraoperative disc herniation in the lumbar spine, though very rare, can occur and can result in neurologic compromise as evidenced by the loss of somatosensory evoked potentials. By identifying the event, it can be remedied by evaluating the disc visually, removing extruded fragments and decompressing nerve roots with recovery of somatosensory evoked potentials and normal neurologic function postoperatively. If neurophysiological monitoring shows there is a sudden loss of response, then consideration should be given to the possibility of an acute intraoperative herniation.


Surgical Management of Patellar Fractures.
The patella plays a crucial role in the extensor mechanism to increase the mechanical advantage of the quadriceps. Fractures can be classified based on displacement, comminution, and fracture pattern, which often guide treatment. Modern treatment options include internal fixation using tension bands with Kirschner wires or cannulated screws, lag screw fixation, partial patellectomy, and rarely total patellectomy. Nondisplaced, closed patellar fractures or fractures with less than 2-mm articular steps can be successfully treated conservatively. Open fractures, articular step of 2 mm or greater, and loss of knee extension are indications for surgical intervention.


The Influence of In Vitro Gastrointestinal Digestion on the Chemical Composition and Antioxidant and Enzyme Inhibitory Capacities of Carob Liqueurs Obtained with Different Elaboration Techniques.
Carob liqueur is a traditional Mediterranean alcoholic beverage obtained via a wide range of production techniques contributing to the different organoleptic attributes of the final product. The aim of this research was to evaluate the stability of the chemical composition and biological capacities (antioxidant and enzyme inhibition) under in vitro simulated gastrointestinal digestion of liqueurs prepared by flavouring the fig spirit with carob pulp by maceration, distillation, percolation, or aqueous and hydro-alcoholic infusions. For this purpose, the phenolic and furanic compositions, the total phenolic (TPC) and flavonoid (TFC) contents, antioxidant capacity (AC), and enzyme inhibitory potential against acethylcholinesterase, tyrosinase, α-glucosidase and α-amylase enzymes were evaluated. The content of gallic acid decreased after gastrointestinal digestion, while TPC, TFC, and AC significantly increased after each digestion phase. Overall, no significantly different enzyme inhibitions (p < 0.05) were observed among digested liqueurs, with moderate inhibition against acethylcholinesterase and tyrosinase (enzymes related with neurodegenerative diseases), and potent and low inhibitory capacities for α-glucosidase and α-amylase, respectively (ideal conditions employed in antidiabetic therapy). The study indicates that hydro-alcoholic infusion and maceration were the most appropriate methods to obtain liqueurs with higher values of the aforementioned parameters and safe levels of toxic furanics.


A longitudinal study assessing differences in causes of death among housed and homeless people diagnosed with HIV in San Francisco.
San Francisco has implemented several programs addressing the needs of two large vulnerable populations: people living with HIV and those who are homeless. Assessment of these programs on health outcomes is paramount for reducing preventable deaths. Individuals diagnosed with HIV/AIDS and reported to the San Francisco Department of Public Health HIV surveillance registry, ages 13 years or older, who resided in San Francisco at the time of diagnosis, and who died between January 1, 2002, and December 31, 2016 were included in this longitudinal study. The primary independent variable was housing status, dichotomized as ever homeless since diagnosed with HIV, and the dependent variables were disease-specific causes of death, as noted on the death certificate. The Cochran-Armitage test measured changes in the mortality rates over time and unadjusted and adjusted Poisson regression models measured prevalence ratios (PR) and 95% confidence intervals (CI) for causes of death. A total of 4158 deceased individuals were included in the analyses: the majority were male (87%), ages 40-59 years old at the time of death (64%), non-Hispanic White (60%), men who have sex with men (54%), had an AIDS diagnosis prior to death (87%), and San Francisco residents at the time of death (63%). Compared to those who were housed, those who were homeless were more likely to be younger at time of death, African American, have a history of injecting drugs, female or transgender, and were living below the poverty level (all p values < 0.0001). Among decedents who were SF residents at the time of death, there were declines in the proportion of deaths due to AIDS-defining conditions (p < 0.05) and increases in accidents, cardiomyopathy, heart disease, ischemic disease, non-AIDS cancers, and drug overdoses (p < 0.05). After adjustment, deaths due to mental disorders (aPR = 1.63, 95% CI 1.24, 2.14) were more likely and deaths due to non-AIDS cancers (aPR = 0.63, 95% CI 0.44, 0.89) were less likely among those experiencing homelessness. Additional efforts are needed to improve mental health services to homeless people with HIV and prevent mental-health related mortality.


Loss of IκB kinase β promotes myofibroblast transformation and senescence through activation of the ROS-TGFβ autocrine loop.
Using forward and reverse genetics and global gene expression analyses, we explored the crosstalk between the IκB kinase β (IKKβ) and the transforming growth factor β (TGFβ) signaling pathways. We show that in vitro ablation of Ikkβ in fibroblasts led to progressive ROS accumulation and TGFβ activation, and ultimately accelerated cell migration, fibroblast-myofibroblast transformation and senescence. Mechanistically, the basal IKKβ activity was required for anti-oxidant gene expression and redox homeostasis. Lacking this activity, IKKβ-null cells showed ROS accumulation and activation of stress-sensitive transcription factor AP-1/c-Jun. AP-1/c-Jun activation led to up-regulation of the Tgfβ2 promoter, which in turn further potentiated intracellular ROS through the induction of NADPH oxidase (NOX). These data suggest that by blocking the autocrine amplification of a ROS-TGFβ loop IKKβ plays a crucial role in the prevention of fibroblast-myofibroblast transformation and senescence.


Nerve supply to the inner sensory cells in a human cochlea.
A choclear apical turn taken from a 66-year-old woman showed an area with marked decrease in number of the inner sensory cells. After observation by scanning electron microscope, the same specimen was thin-sectioned tangentially to the osseous spiral lamina. Numbers of myelinated nerve fibers were counted and the thickness of the fiber was measured by computed measuring equipment. Compared to the area with inner sensory cells intact, the area without inner sensory cells showed a 70% decrease in number of myelinated fibers. These degenerated fibers might have had intimate relationship with the inner sensory cells, probably being the afferent nerve supply to the inner sensory cells.


Targeted protein degradation: mechanisms, strategies and application.
Traditional drug discovery mainly focuses on direct regulation of protein activity. The development and application of protein activity modulators, particularly inhibitors, has been the mainstream in drug development. In recent years, PROteolysis TArgeting Chimeras (PROTAC) technology has emerged as one of the most promising approaches to remove specific disease-associated proteins by exploiting cells' own destruction machinery. In addition to PROTAC, many different targeted protein degradation (TPD) strategies including, but not limited to, molecular glue, Lysosome-Targeting Chimaera (LYTAC), and Antibody-based PROTAC (AbTAC), are emerging. These technologies have not only greatly expanded the scope of TPD, but also provided fresh insights into drug discovery. Here, we summarize recent advances of major TPD technologies, discuss their potential applications, and hope to provide a prime for both biologists and chemists who are interested in this vibrant field.


Turbulent Dufour effect and Onsager-type relations.
In this paper we extend the work of Elperin et al. [Phys. Rev. Lett. 80, 69 (1998)] by showing that a turbulent Dufour-type effect is also present in chemically nonreacting gaseous admixtures. This result is used to analyze the possibility of obtaining turbulent analogs of the Onsager relations, a fundamental result for molecular crossed effects in irreversible thermodynamics.


Cation exchange--a common mechanism in the storage and release of biogenic amines stored in granules (vesicles)? I. Comparative studies on the uptake of sodium and biogenic amines by the weak cation (carboxyl) exchangers Amberlite IRC-50 and Sephadex C-50 and by biogenic (granule-enriched) materials in vitro.
Studies on the uptake and storage of sodium and biogenic amines (phenylethylamine, noradrenaline, histamine) by two weak cation-exchangers, IRC-50 and Sephadex C-50, and by biogenic granule-enriched preparations demonstrated that the synthetic and biogenic materials had several common characteristics. They showed similar concentration- and pH-dependence and fitted the same cation-exchange and receptor-binding equations. The observations were taken to support the view that the matrices of amine-storing granules have the properties of weak cation-exchangers, with carboxyls as the cation-binding groups.


Familial true hermaphroditism: paternal and maternal transmission of true hermaphroditism (46,XX) and XX maleness in the absence of Y-chromosomal sequences.
We report on 46,XX true hermaphroditism and 46,XX maleness coexisting in the same pedigree, with maternal as well as paternal transmission of the disorder. Molecular genetic analysis showed that both hermaphrodites as well as the 46,XX male were negative for Y-chromosomal sequences. Thus, this pedigree is highly informative and allows the following conclusions: first, the maternal as well as paternal transmission of the disorder allows the possibility of an autosomal dominant as well as an X-chromosomal dominant mode of inheritance; second, testicular determination in the absence of Y-specific sequences in familial 46,XX true hermaphrodites as well as in 46,XX males seems to be due to the varying expression of the same genetic defect; and third, there is incomplete penetrance of the defect.


Stereotactic localization (with computerized tomographic scanning), biopsy, and radiofrequency treatment of deep brain lesions.
In eight patients stereotactic biopsy of deep brain lesions was performed. Adequate tissue was obtained, and the information helped considerably in planning further therapy. No significant complications occurred in these patients. In three of the cases, the stereotactic coordinates were determined from the computerized tomographic (CT) scan. In one patient, after biopsy, stereotactic radiofrequency (RF) lesions in the tumor resulted in temporary improvement.


Inkjet-printed polyaniline patterns for exocytosed molecule detection from live cells.
Polyaniline (PANi) patterns on flexible substrate are fabricated for biomolecule detection from live cells. PANi patterns are prepared by inkjet printing on polyethylene terephthalate film. Subsequently, arginine-glycine-aspartate (RGD) peptide is immobilized on the PANi pattern to selectively adhere cells. Rat pheochromocytoma PC12 cells are cultured on the RGD-immobilized PANi pattern, and patterned with high selectivity and growth. Additionally, the cells show focal adhesion on the RGD-immobilized PANi pattern, which are confirmed with vinculin staining and scanning electron microscopic images. To monitor dynamic biomolecular release from PC12 cells, RGD-immobilized PANi pattern is used for a real-time electrical signal detector. RGD-immobilized PANi patterning and sensing system represents outstanding ability to translate and amplify exocytosis molecules into a detectable signal as a transducer.


Bootstrapping the energy flow in the beginning of life.
This paper suggests that the energy flow on which all living structures depend only started up slowly, the low-energy, initial phase starting up a second, slightly more energetic phase, and so on. In this way, the build up of the energy flow follows a bootstrapping process similar to that found in the development of computers, the first generation making possible the calculations necessary for constructing the second one, etc. In the biogenetic upstart of an energy flow, non-metals in the lower periods of the Periodic Table of Elements would have constituted the most primitive systems, their operation being enhanced and later supplanted by elements in the higher periods that demand more energy. This bootstrapping process would put the development of the metabolisms based on the second period elements carbon, nitrogen and oxygen at the end of the evolutionary process rather than at, or even before, the biogenetic event.


Molecular Interplay between Dormant Bone Marrow-Resident Cells (BMRCs) and CTCs in Breast Cancer.
Despite widespread knowledge that bone marrow-resident breast cancer cells (BMRCs) affect tumor progression, signaling mechanisms of BMRCs implicated in maintaining long-term dormancy have not been characterized. To overcome these hurdles, we developed a new experimental model of clinical dormancy employing patient-isolated Circulating Tumor Cells (de novo CTCs) and their injection in xenografts with subsequent tumor monitoring and CTC characterization (ex vivo CTCs). We hypothesized that significant distinctions exist between signaling pathways of bone marrow-homing vs metastasis-competent CTCs upon transplantation in xenografts. Comparative transcriptomic analyses of ex vivo vs de novo CTCs identified increased mTOR signaling-a critical pathway frequently dysregulated in breast cancer and implicated in cell survival and dormancy-with contrasting actions by its two complementary arms (mTORC2/mTORC1). Heightened mTORC2 downstream targets augmented quiescent CTCs (Ki67-/RBL2+ cells) in paired breast cancer tissues, along with high mTORC2 activity in solitary BMRCs and tissue-resident CTCs. Further, shRNA mediated the knockdown of RICTOR, an essential component of mTORC2, and augmented Ki67/PCNA biomarker expression and proliferation. Collectively, these findings suggest that the balance between mTORC1 vs mTORC2 signaling regulates CTC-associated mitotic and/or dormancy characteristics.


A consistent two-mutation model of lung cancer for different data sets of radon-exposed rats.
A two-mutation carcinogenesis model, formulated in terms of biologically motivated equations for mutation and expansion steps, has been applied in a mechanistic modelling of the lung cancer incidence in two large data sets of rats exposed to radon, both separately and jointly. Results indicate that (1) the equations employed are able to provide an accurate description of the separate data sets, (2) the parameters in the equations take on similar values for both data sets, and (3) it is possible to construct a consistent and well-fitting solution for the joint data set. It proved not to be necessary to take into account the effect of uranium ore dust, administered to part of the data or the different rat strains of the data sets. The joint solution provides a firm basis to investigate the effects of exposure, exposure rate and age at exposure on cumulative incidence, excess relative risk and excess absolute risk. For the same total exposure, cumulative incidence reaches a maximum for exposure rates between 1 and 10 WLM per day. The so-called inverse-exposure-rate effect acts for higher exposure rates. The influence of age at exposure, however, seems to be even more pronounced. Exposure at a young age leads to considerably higher incidences than exposure at a later age. Parameters derived in this study compare fairly well with those derived for uranium miners, suggesting that a consistent model description for the induction of lung cancer by radon in rats and humans may be possible.


Neuralgic amyotrophy. An update.
A century after the first description of neuralgic amyotrophy (NA), its pathophysiology remains unknown. An inflammatory (auto)immune pathophysiology is presumed, with mechanical or infectious precipitating conditions, which triggers attacks. Clinically, NA is an acute and painful unique or multiple mononeuropathy that causes palsy, amyotrophy and sensory loss in an asymmetric and patchy distribution. It involves the upper brachial plexus rather than the other parts but also may involve the cervical plexus, lumbosacral plexus and cranial nerves. The impairment can be restricted to one fascicule of one nerve, plexus or root; limited to a few ones; or extensive, involving both upper limbs. Its evolution is usually monophasic and auto-limited and never leads to generalized polyneuropathy. Electrodiagnostically, NA is characterized by severe axonal damage. The recovery is usually good after 6 months to 3 years in 80% of cases. Persistent disability is present in 20% of idiopathic NA cases and is more frequent in hereditary NA, with frequent recurrences, more frequent bilateral impairment, and more atypical distribution (cervical plexus, lumbosacral plexus or cranial nerves) than with idiopathic NA. Hereditary NA is mainly linked to a mutation in the gene of the Septin-9 protein. When the patient is seen early after disease onset, treatment with corticosteroids for 2 weeks seems to shorten the pain duration and the delayed recovery. With diagnosis during the palsy period, treatment is based on pharmacologic and non-pharmacologic therapies according to the complaints of the patient.


Reaching out to patients with alcohol and drug misuse problems.
Jeff Fernandez, Lead Alcohol and Drug Nurse, Alcohol and Drug Liaison Team, Royal Free Foundation Trust, London (email@example.com). The team was the Gold Award winner in the Gastrointestinal Nurse of the Year category in the BJN Awards 2023.


Sequential release of cytokines, lipid mediators and nitric oxide in experimental colitis.
The object of this study was to establish whether different pro- and anti-inflammatory mediators were formed in colonic tissue from experimental colitis depending on the course of the disease. Concentrations of mediators of inflammation were examined in colonic tissue in dextran induced colitis in mice. Initial inflammation was produced by 5 days treatment of 10% dextran sodium sulfate (DSS) in drinking water, followed by a further 9 day period of 2% DSS in an attempt to produce a milder chronic inflammation. The degree of inflammation was scored by a standardized macroscopic and histological examination. Initially, a 60% maximum inflammation score was observed at day 4. At this time inflammation was associated with the release of interleukin-lbeta (IL-1beta) and tumour necrosis factor-alpha (TNFalpha), whereas both prostaglandins 6kPGF(1alpha) and PGE(2) and nitric oxide (NO) markedly decreased. Then a 25% inflammation score was reached which coincided with an increased production of platelet-activating factor (PAF). No significant changes were observed in leukotriene B(4) and C(4) formation. In conclusion, pro-inflammatory cytokines IL-1beta and TNFalpha are considered to be primary mediators, whereas PAF, eicosanoids and NO may reflect secondary mediators in experimental colitis.


Malignant lymphoma in four of five siblings.
In one family four out of five siblings were affected with malignant lymphoma and HD has been diagnosed in two other relatives. HLA-haplotypes were identical for the three diseased siblings who were so studied, whereas for the unaffected one they were different.


Vectored upper cervical manipulation for chronic sleep bruxism, headache, and cervical spine pain in a child.
To discuss the management of chronic sleep bruxism in a 6-year old girl. Clinical features The patient had morning headaches and cervical spine pain. Due to abnormal tooth wear, bruxism had been previously diagnosed and was verified by observation during sleep. She also had abnormal postural and palpatory findings, indicating upper cervical joint dysfunction. Intervention and outcome Bilateral rotary cervical stretching/mobilization and a vectored high-velocity, low-amplitude adjustment were performed in the upper cervical spine, using the atlas transverse process as the contact point. There was complete relief of the chronic subjective symptoms concomitant with remission of the objective signs of joint dysfunction. Cervical, particularly upper cervical, spine muscle-joint dysfunction should be considered as a potential etiology in chronic childhood sleep bruxism.


Effect of undernutrition on developing rat cerebellum. Some electrophysiological and neuromorphological correlates.
Electrophysiological studies of Purkinje cells from the developing normal and undernourished rat cerebellar cortex were correlated with the neuromorphological studies. In undernourished animals the firing pattern of Purkinje cells was simpler as compared to complex electrical activity in normal animals. The firing frequency of Purkinje cells remained reduced in all the age-matched undernourished groups. The duration of a spike was also prolonged as compared to their normal age mates. Similarly the mean amplitude was also relatively smaller in the undernourished animals when compared with the normal litter mates. Undernourished animals exhibited typical increased stimulus thresholds, prolonged N-2 peak wave latencies, reduced number of functional components, amplitude and increased duration of mossy fiber response (MFR). The mossy fiber (MF) activity exhibited a sluggish rise in its amplitude, when stimulus intensity was increased in undernourished animals. A phenomenon of fatigue was observed with a significant reduction in the amplitude and frequency of Purkinje cell unit discharge on repeated MF stimulation in the undernourished animals. However, repeated MF stimulation provided a high frequency discharge in the normal as well as undernourished adult rats. Neuromorphological studies at light-microscopic level exhibited delayed cell migration from the external granular layer in the undernourished rats. At electron-microscopic level, intracellular morphology exhibited almost similar ultrastructural details except for a few minor alterations. A free ribosomal pool, immature development of E.R., increased incidence of lysosomes and electron-dense membrane stacks were observed in the Purkinje cells at 30 days in the undernourished animals. The results of the present study indicated that undernutrition affects the maturation of Purkinje cell regional neurocircuitry.


Preoperative Opioid Use Is Associated With Inferior Patient-Reported Outcomes Measurement Information System Scores Following Rotator Cuff Repair.
To determine the influence of preoperative opioid use on Patient-Reported Outcomes Measurement Information System (PROMIS) scores pre- and postoperatively in patients undergoing arthroscopic rotator cuff repair (RCR). A retrospective review of all RCR patients aged >18 years old was performed. PROMIS pain interference ("PROMIS PI"), upper extremity function ("PROMIS UE"), and depression ("PROMIS D") scores, were reviewed. These measures were collected at preoperative, 6-month, and 1-year postoperative time points. A prescription drug-monitoring program was queried to track opioid prescriptions. Patients were categorized as chronic users, acute users, and nonusers based on prescriptions filled. Comparison of means were carried out using analysis of variance and least squares means. Effect sizes and 95% confidence intervals were calculated. In total, 184 patients who underwent RCR were included. Preoperatively, nonusers (n = 92) had superior PROMIS UE (30.6 vs 28.9 vs 26.1; P < .05) and PI scores (61.5 vs 64.9 vs 65.3; P < .001) compared with acute users (n = 65) and chronic users (n = 27), respectively. At 6 months postoperatively; nonusers demonstrated significantly greater PROMIS UE (41.7 vs 35.6 vs. 33.5; P < .001), lower PROMIS D (41.6 vs 45.8 vs 51.1; P < .001), and lower PROMIS PI scores (50.7 vs 56.3 vs 58.1; P < .01) when compared with acute and chronic users, respectively. Nonusers had lower PROMIS PI (47.9 vs 54.3 vs 57.4; P < .0001) and PROMIS D (41.6 vs 48.3 vs 49.2; P = .0002) scores compared with acute and chronic users at 1-year postoperatively. Nonusers experienced a significantly greater magnitude of improvement in PROMIS D 6 months postoperatively compared with chronic opioid users (-5.9 vs 0.0; P < .01). Patients undergoing RCR demonstrated superior PROMIS scores pre- and postoperatively if they did not use opioids within 3 months before surgery. III, retrospective comparative trial.


TAK1 promotes BMP4/Smad1 signaling via inhibition of erk MAPK: a new link in the FGF/BMP regulatory network.
FGFs and BMPs act in concert to regulate a wide range of processes in vertebrate development. In most cases, FGFs and BMPs have opposing effects, and specific developmental outcomes arise out of a balance between the two growth factors. We and others have previously demonstrated that signaling pathways activated by FGFs and BMPs interact via inhibitory crosstalk. Here we demonstrate a role for the BMP effector TGF-β Activated Kinase 1 (TAK1) in the maintenance of Smad1 activity in Xenopus embryos, via the inhibition of erk MAPK. Up- or downregulation of TAK1 levels produces an inverse alteration in the amount of activated erk MAPK. The inhibition of erk MAPK by TAK1 is mediated by p38 and a corresponding decrease in phosphorylation of MEK. TAK1 morphant embryos show a decrease in the nuclear accumulation of Smad1. Conversely, reduction of erk MAPK activity via overexpression of MAP Kinase Phosphatase1 (MKP1) leads to an increase in nuclear Smad1. Both TAK1 morphant ectoderm and ectoderm treated with FGF show a decrease in the expression of several Smad1-inducible genes. Neural-specific gene expression is inhibited in isolated ectoderm coexpressing noggin and TAK1, suggesting that TAK1 is sufficient to inhibit neural specification. Introduction of TAK1 morpholino oligonucleotide expands the expression of organizer genes, disrupts formation of the boundary between organizer and non-organizer mesoderm, and increases the spatial range of MAPK activation in response to localized FGF. Our results indicate that inhibitory interactions between FGF and BMP4 effector pathways increase the robustness of BMP signaling via a feed-forward mechanism.


Perceptions of depression among dementia caregivers: findings from the CATIE-AD trial.
For patients with Alzheimer's disease complicated by behavioral disturbances, to use single questions about perceived depression and assess patient and caregiver accuracy in recognizing patient depression. Cross-sectional interviews from the Clinical Antipsychotic Trials of Intervention Effectiveness-Alzheimer's Disease (CATIE-AD). Patients were asked 'Do you believe you are depressed?' Caregivers were asked 'Do you believe the patient is depressed?' and 'Do you believe you (caregiver) are depressed?' These responses were compared to the patient's score on the Cornell Scale for Depression in Dementia (CSDD), and to the caregiver's score on the Beck Depression Inventory (BDI) using screening test characteristics. Two hundred fifty-seven total patient/caregiver pairs were evaluated. Patients had a mean age of 78 years, were mainly female, white, and had a mean Mini-Mental State Examination Score of 15. Caregivers were on average 63 years old, and were mostly spouses or children. 37% of patients had depression (CSDD ≥ 12). Patients recognized their own depression with a sensitivity of 0.26 (CI: 0.18, 0.37) and specificity of 0.85 (0.79, 0.90). Caregivers' accuracy in recognizing depression in the patient revealed a sensitivity of 0.65 (C.I.: 0.55, 0.75) and specificity of 0.58 (CI: 0.50, 0.66). Twelve per cent of caregivers were depressed, and caregiver burden was associated with patient depression. Clinically significant depression was common, and asking a single question to the patient about depression failed to detect most cases. Caregivers identified two-thirds of patient depression when asked this one question. Caregiver depression and perceived burden were associated with patient depression.


Elevated depressive symptoms among newer and younger healthcare workers in Japan during the COVID-19 pandemic.
Depression is a frequent outcome of long-term stress, but no studies have examined depression rates among Japanese healthcare workers fighting the COVID-19 pandemic. Therefore, we conducted a web-based interview of hospital employees to assess depression prevalence and factors. This observational cohort study was conducted from July to August, 2020, as part of a mandatory health checkup of Juntendo University Hospital employees (Tokyo, Japan). A total of 4239 participants completed a web-based questionnaire on medical history and current health status. The Center for Epidemiologic Studies Depression Scale (CES-D) was used for self-assessment, with a score of ≥16 considered to indicate depression. Among all employees, the proportion of depression was 31.3% in 2020, the highest measured in the last 10 years and substantially greater than the pre-pandemic value in 2019 (27.5%). The proportion of depression for 2020 was significantly higher in new recruits than in employees with more than 2 years of experience (47.0% vs 29.9%, respectively, P < .0001) and in new recruits in 2019 (26.4%, P < .0001). When subdivided by occupation, nurses demonstrated the highest depression rate (43.2%), followed by paramedics (35.1%) and clerks (31.6%), whereas residents (22.9%), doctors (20.4%), teaching staff (18.0%), and part-time staff (15.3%) reported lower depression rates. The positive CES-D score significantly correlated with age (P < .0001). Younger and newer employees demonstrated the highest rates of depression independent of occupation. Therefore, mental healthcare programs focusing on these vulnerable groups need to be established.


Direct projection from the nucleus of the optic tract to the medial vestibular nucleus in the cat.
The nucleus of the optic tract (NOT) serves as an important visuo-motor relay between the retina and preoculomotor structures that mediate optokinetic nystagmus. In the present study, the efferent targets of NOT were investigated using biocytin as an anterograde tracer. Following biocytin injections into NOT, labeled fibers were observed in each of the following efferent pathways: (1) those that project to the contralateral NOT via the posterior commissure; (2) those that course through the nucleus reticularis pontis oralis to terminate in the nucleus reticularis tegmenti pontis; and (3) those that descend via the medial lemniscus to the level of the medulla to terminate in the dorsolateral pontine nucleus, nucleus prepositus hypoglossi, medial vestibular nucleus and the inferior olive. Direct projections from the NOT to the medial vestibular nucleus may contribute to the residual optokinetic responses of the vestibular nucleus neurons following cerebellar or inferior olivary lesions.


Effects of sodium fluoride on uptake of T-2 mycotoxin in cultured cells.
We examined the effect of sodium fluoride on uptake of tritium-labeled T-2 toxin (molecules of toxin/cell) in Chinese hamster ovary (CHO) and African green monkey kidney (VERO) cells. Correlations were made to temperature (22 and 37 degrees C) and toxin concentration (0.001 and 0.01 microgram/ml) over time (0-180 min). As expected, toxin uptake increased in both cell types with increasing time and temperature. VERO cells exhibited significant (P less than 0.05) increases in the rate (i.e. slope) of toxin uptake under all parameters, while the rate of toxin uptake in both cell types was generally greater at 37 degrees C compared to 22 degrees C. The rate of equilibrium was affected by both temperature and sodium fluoride. At 37 degrees C toxin uptake plateaued by 30 min in the presence of sodium fluoride. At 22 degrees C the rate of toxin uptake was slower, with or without sodium fluoride present. Statistical analysis of individual time points along the curve demonstrated that sodium fluoride significantly increased cell-associated toxin at most time points. Analysis of the slopes of uptake curves from 0 to 20 min indicated significant (P less than 0.05) differences in the rates of T-2 uptake in both cell types and toxin doses in the presence of sodium fluoride. The increase in toxin uptake in the presence of sodium fluoride was not due to altered cell membrane permeability caused by sodium fluoride. This study demonstrates that sodium fluoride significantly increases cell-associated T-2 toxin and the rate of toxin uptake in two cultured cell lines.


The epidemiology of mass breast cancer screening--a plea for a valid measure of benefit.
The present paper analyses the epidemiologic effects of mass breast cancer screening. Mass mammography may possibly achieve a breast cancer mortality reduction in relative risk terms. However, this does not necessarily represent a net benefit. It is argued that the benefits and adverse effects of a screening programme must be measured in terms of absolute risks. According to this measure, the mortality reduction achieved by a mass breast screening programme is only one death per approx. 15,000 women-years. Many thousands of mammograms are needed to prevent one cancer death, and for each woman who can derive a direct benefit in terms of a prevented breast cancer death, hundreds of women have to suffer the anxiety of a positive screening mammography. Moreover, it is possible that adverse effects of breast cancer screening may contribute to mortality from other causes. Even with the assumption that screening can save lives, the net health effect of mass breast cancer screening is questionable and appears to be rather detrimental. It may be an error to recommend mass breast screening.


What do patients think about year-end resident continuity clinic handoffs? A qualitative study.
Although Internal Medicine year-end resident clinic handoffs affect numerous patients, little research has described patients' perspectives of the experience. To describe patients' perceptions of positive and negative experiences pertaining to the year-end clinic handoff; to rate patient satisfaction with aspects of the clinic handoff and identify whether or not patients could name their new physicians. Qualitative study design using semi-structured interviews. High-risk patients who underwent a year-end clinic handoff in July 2011. Three months post-handoff, telephone interviews were conducted with patients to elicit their perceptions of positive and negative experiences. An initial coding classification was developed and applied to transcripts. Patients were also asked to name their primary care physician (PCP) and rate their satisfaction with the handoff. In all, 103 telephone interviews were completed. Patient experiences regarding clinic handoffs were categorized into four themes: (1) doctor-patient relationships (i.e. difficulty building rapport); (2) clinic logistics (i.e. difficulty rescheduling appointments); (3) process of the care transition (i.e. patient unaware transition occurred); and (4) patient safety-related issues (i.e. missed tests). Only 59 % of patients could correctly name their new PCP. Patients who reported that they were informed of the clinic transition by letter or by telephone call from their new PCP were more likely to correctly name them (65 % vs. 32 % p = 0.007), report that their new doctor assumed care for them immediately (81 % [68/84] vs. 53 % [10/19], p = 0.009) and report satisfaction with communication between their old and new doctors (80 % [67/84] vs. 58 % [11/19], p = 0.04). Patients reported positive experiences such as learning more about their new physician through personal sharing, which helped them build rapport. Patients who reported being aware of the medical education mission of the clinic tended to be more understanding of the handoff process. Patients face unique challenges during year-end clinic handoffs and provide insights into areas of improvement for a patient-centered handoff.


Immunocastration of colts and immunospeying of fillies.
A series of experiments using an ovalbumin conjugated gonadotrophin releasing hormone was used to stimulate antibody production, suppress testosterone secretion and depress testicular function in yearling and 2 year old colts and fillies. In the preliminary experiment, an injectable oil-based formulation was administered to yearling colts. Testicular development and testosterone secretion were retarded for a period of approximately 28-32 weeks while antibody titres were greater than 1:1000. An implant and water-soluble vaccine (200 and 400 mg) is presently being tested in 2 year old colts. Testicular dimensions and testosterone concentrations decreased following vaccination. Preliminary results indicate that the effects of the implant are reversible and last for approximately 30 weeks. The water-soluble vaccine produced effective antibodies for 10-19 weeks. Plasma testosterone of all colts was reduced to gelding levels (0.2 ng/mL) while antibody titres were greater than 1:1000. Semen evaluation, morphometric analysis and daily sperm production data indicate that the testes recovered normal function. These experiments are continuing and more definitive data on the vaccine and its reversibility will be available at their conclusion. Ovarian activity of the treated fillies (200 and 400 mg) ceased soon after vaccination. Their progesterone concentrations have remained below 1 ng/mL for 20 weeks. The untreated control fillies have continued to cycle normally as determined by ovarian ultrasonography and progesterone concentrations.


A novel canine model for prostate cancer.
No existing animal model fully recapitulates all features of human prostate cancer. The dog is the only large mammal, besides humans, that commonly develops spontaneous prostate cancer. Canine prostate cancer features many similarities with its human counterpart. We sought to develop a canine model of prostate cancer that would more fully represent the features of human prostate cancer than existing models. The Ace-1 canine prostate cancer cell line was injected transabdominally under transrectal ultrasound (TRUS) guidance into the prostates of immunosuppressed, intact, adult male dogs. Tumor progression was monitored by TRUS imaging. Some dogs were subjected to positron emission tomography (PET) for tumor detection. Time of euthanasia was determined based on tumor size, impingement on urethra, and general well-being. Euthanasia was followed by necropsy and histopathology. Ace-1 tumor cells grew robustly in every dog injected. Tumors grew in subcapsular and parenchymal regions of the prostate. Tumor tissue could be identified using PET. Histological findings were similar to those observed in human prostate cancer. Metastases to lungs and lymph nodes were detected, predominantly in dogs with intraprostatic tumors. We have established a minimally invasive dog model of prostate cancer. This model may be valuable for studying prostate cancer progression and distant metastasis.


See no allergen, hear no allergen, speak no allergen!
Segmental allergen challenge in allergic patients with asthma reveals a previously unknown role for monocytes in the T helper 2 (TH2)-dependent inflammatory response, whereas in allergic controls without asthma, allergen unresponsiveness seems to be maintained through epithelial-myeloid cell cross-talk that prevents TH2 cell activation (see related Research Article by Alladina et al.).


Detection and characterization of Shigella species isolated from food and human stool samples in Nabeul, Tunisia, by molecular methods and culture techniques.
This study was designed to isolate Shigella spp. strains from food and stool samples by a combination of PCR and culture methods and characterize their serotypes, antibiotic resistance profiles, virulence genes and pulsed-field gel electrophoresis (PFGE) patterns to investigate possible clonal relationships amongst strains circulating. Six Shigella spp. strains were isolated from 280 food samples against 16 Shigella isolates from 236 stool samples of symptomatic patients and asymptomatic food handlers during the period from January 2007 to December 2009 in Public Health Regional Laboratory of Nabeul. The detection of ipaH, ipaBCD, ial, ShET-1 and ShET-2 was performed by a PCR technique with specific primers. The use of PCR technique improved the rate of detecting Shigella in stool samples from 6·7 to 14% and in food samples from 2·1 to 8·6%. Percentage of Shigella isolates and ipaH-specific PCR demonstrated a marked pattern of seasonality, increasing in summer and fall seasons for human and food isolates. Amongst the environmental strains, 50% of isolates were invasive. However, for the 16 clinical strains isolated, nine were found to be positive for both ial and ipaBCD gene and 11 were found to produce ShET-1 and/or ShET-2. XbaI PFGE analysis revealed the presence of a predominant clone amongst Shigella sonnei strains recovered from different sources circulating in Nabeul, Tunisia, throughout the years 2007-2009. This study demonstrated the existence of Shigella in food samples and dispersion of different virulence genes amongst these isolates, which appear to constitute an environmental source of epidemic spread. The clonal relationships amongst strains isolated from food elements and human stools indicate the incrimination of different kinds of foods as vehicle of transmission of Shigella, which are usually escaped from detection by traditional culture methods.


Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic.
Whole cell and surface proteomes were analyzed together with adhesive properties of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) grown on the emerging prebiotic raffinose, exemplifying a synbiotic. Adhesion of NCFM to mucin and intestinal HT-29 cells increased three-fold after culture with raffinose versus glucose, as also visualized by scanning electron microscopy. Comparative proteomics using 2D-DIGE showed 43 unique proteins to change in relative abundance in whole cell lysates from NCFM grown on raffinose compared to glucose. Furthermore, 14 unique proteins in 18 spots of the surface subproteome underwent changes identified by differential 2DE, including elongation factor G, thermostable pullulanase, and phosphate starvation inducible stress-related protein increasing in a range of +2.1 - +4.7 fold. By contrast five known moonlighting proteins decreased in relative abundance by up to -2.4 fold. Enzymes involved in raffinose catabolism were elevated in the whole cell proteome; α-galactosidase (+13.9 fold); sucrose phosphorylase (+5.4 fold) together with metabolic enzymes from the Leloir pathway for galactose utilization and the glycolysis; β-galactosidase (+5.7 fold); galactose (+2.9/+3.1 fold) and fructose (+2.8 fold) kinases. The insights at the molecular and cellular levels contributed to the understanding of the interplay of a synbiotic composed of NCFM and raffinose with the host.


Efficacy of endoscopic ultrasound with artificial intelligence for the diagnosis of gastrointestinal stromal tumors.
Although endoscopic ultrasound (EUS) is reported to be suitable for determining the layer from which subepithelial lesions (SELs) originate, it is difficult to distinguish gastrointestinal stromal tumor (GIST) from non-GIST using only EUS images. If artificial intelligence (AI) can be used for the diagnosis of SELs, it should provide several benefits, including objectivity, simplicity, and quickness. In this pilot study, we propose an AI diagnostic system for SELs and evaluate its efficacy. Thirty sets each of EUS images with SELs ≥ 20 mm or < 20 mm were prepared for diagnosis by an EUS diagnostic system with AI (EUS-AI) and three EUS experts. The EUS-AI and EUS experts diagnosed the SELs using solely the EUS images. The concordance rates of the EUS-AI and EUS experts' diagnoses were compared with the pathological findings of the SELs. The accuracy, sensitivity, and specificity for SELs < 20 mm were 86.3, 86.3, and 62.5%, respectively for the EUS-AI, and 73.3, 68.2, and 87.5%, respectively, for the EUS experts. In contrast, accuracy, sensitivity, and specificity for SELs ≥ 20 mm were 90.0, 91.7, and 83.3%, respectively, for the EUS-AI, and 53.3, 50.0, and 83.3%, respectively, for the EUS experts. The area under the curve for the diagnostic yield of the EUS-AI for SELs ≥ 20 mm (0.965) was significantly higher than that (0.684) of the EUS experts (P = 0.007). EUS-AI had a good diagnostic yield for SELs ≥ 20 mm. EUS-AI has potential as a good option for the diagnosis of SELs.


A Light-Triggerable Nanoparticle Library for the Controlled Release of Non-Coding RNAs.
RNA-based therapies offer a wide range of therapeutic interventions including the treatment of skin diseases; however, the strategies to efficiently deliver these biomolecules are still limited due to obstacles related to the cellular uptake and cytoplasmic delivery. Herein, we report the synthesis of a triggerable polymeric nanoparticle (NP) library composed of 160 formulations, presenting physico-chemical diversity and differential responsiveness to light. Six formulations were more efficient (up to 500 %) than commercially available lipofectamine in gene-knockdown activity. These formulations showed differential internalization by skin cells and the endosomal escape was rapid (minutes range). The NPs were effective in the release of siRNA and miRNA. Acute skin wounds treated with the top hit NP complexed with miRNA-150-5p healed faster than wounds treated with scrambled miRNA. Light-activatable NPs offer a new strategy to topically deliver non-coding RNAs.


Retroperitoneal laparoscopic nephrectomy in children: at last the gold standard?
We analyzed our experience with retroperitoneal laparoscopic nephrectomy giving special attention to the learning curve, morbidity, and feasibility in a teaching institution. Between 1996 and 2004, we performed 104 laparoscopic nephrectomies in 88 children. Only four were performed via the transperitoneal approach, the others being via the retroperitoneal approach; the files of the latter were analyzed. Mean age was 5 years (20 days-15 years). Main indications were pretransplant nephrectomy for arterial hypertension, nephrotic syndrome or uremic hemolytic syndrome (51%), non-functioning kidney secondary to obstruction, reflux or ectopic ureter (38%), and multicystic dysplastic kidney (11%). Bilateral nephrectomy was performed in 13 children. The lateral retroperitoneal approach was feasible in all cases even for those who had previous renal surgery. Conversion was not needed in any patient. No significant blood loss was observed. Mean operative time was 97 min (range 44-240) for unilateral nephrectomy; 46 nephrectomies were done in less than 90 min. In bilateral cases the mean operative time was 260 min (range 160-390). The operative time was less than 4 h in seven bilateral cases. Postoperative course was uneventful. Hospital stay was 1.9 days (range 1-3) for urological indications and 5 days (range 3-7) for patients with terminal renal disease. The procedure was initially performed by one surgeon, but was then expanded to other surgeons of the team, and safely taught to residents and fellows. The procedure is safe, the learning curve is reasonable, teaching is feasible, operating time becomes with experience closer to open surgery without morbidity, and cosmetic results are excellent. This procedure may be considered as the gold standard for nephrectomy in children.


[An unusual cause of subcutaneous postsacral swelling].
A subcutaneous sacrococcygeal located myxopapillary ependymoma is a rare presentation of this malignancy. It has a tendency to metastasize, even after a long latent period. This is a report of such a case together with proposals for treatment and follow-up.


Carbohydrate-binding specificities of five lectins that bind to O-Glycosyl-linked carbohydrate chains. Quantitative analysis by frontal-affinity chromatography.
The carbohydrate-binding specificities of lectins purified from Agaricus bisporus (ABA-I), Arachis hypogaea (PNA), Bauhinia purpurea (BPA), Glycine max (SBA), and Vicia villosa (VVA-B4) have been studied by affinity chromatography on columns of the immobilized lectins, and quantitatively analyzed by frontal affinity chromatography. These five lectins could be classified into two groups with respect to their reactivities with typical mucin-type glycopeptides, beta-D-Galp-(1----3)-alpha-D-GalpNAc-(1----3)-Ser/Thr (2) and alpha-D-GalpNAc-(1----3)-Ser/Thr (3). One group, which consists of ABA-I, PNA, and BPA, preferentially binds to 2, and the other, which consists of SBA and VVA-B4, shows higher affinity for 3 than for 2. Among the lectins tested, only ABA-I was found to bind to a sialylated glycopeptide, whic which was prepared from human erythrocyte glycophorin A and contains three three tetrasaccharide chains having the structure of alpha-NeuAc-(2----3)-beta-D-GAlp-(1----3)-NeuAC-(2----6)]-alpha-D-Galp NAc-(1----, with an association constant of 15 microM, whereas the association constants of the other four lectins for this sialylated glycopeptide were less than 3.5 mM. On the other hand, removal of the beta-D-galactopyranosyl group from a glycopeptide containing sequence 2 resulted in decreased association constants for the three lectins of the first group, especially ABA-I and PNA. The two lectins of the second group showed a high affinity for 3, but SBA preferentially interacted with oligosaccharides containing the alpha-D-GalpNAc-(1----3)-beta-D-Galp-(1----3)-D-GlapNAc sequence, prepared from a blood group A-active oligosaccharide.


A rapid Q-PCR titration protocol for adenovirus and helper-dependent adenovirus vectors that produces biologically relevant results.
Adenoviruses are employed in the study of cellular processes and as expression vectors used in gene therapy. The success and reproducibility of these studies is dependent in part on having accurate and meaningful titers of replication competent and helper-dependent adenovirus stocks, which is problematic due to the use of varied and divergent titration protocols. Physical titration methods, which quantify the total number of viral particles, are used by many, but are poor at estimating activity. Biological titration methods, such as plaque assays, are more biologically relevant, but are time consuming and not applicable to helper-dependent gene therapy vectors. To address this, a protocol was developed called "infectious genome titration" in which viral DNA is isolated from the nuclei of cells ~3 h post-infection, and then quantified by Q-PCR. This approach ensures that only biologically active virions are counted as part of the titer determination. This approach is rapid, robust, sensitive, reproducible, and applicable to all forms of adenovirus. Unlike other Q-PCR-based methods, titers determined by this protocol are well correlated with biological activity.


Hemostasis using a fully covered self-expandable metal stent for marked bleeding from the bile duct following stent removal (with videos).
We describe a case of life-threatening hemorrhage from the bile duct following stent removal. Eventually, hemostasis was achieved by tamponade using a fully covered self-expandable metal stent.


FGF21 negatively affects long-term female fertility in mice.
Obesity and associated liver disease are a growing public health concern. Pharmacological agents to treat non-alcoholic fatty liver disease are limited. FGF21, a hormone secreted by the liver and potent metabolic modulator, is a promising therapeutic target for this indication with several analogs currently in clinical development. However, concerns about a negative effect of FGF21 on female fertility have not been fully addressed. After induction of obesity, female C57BL/6N mice received a 7-day course of subcutaneously administered FGF21. Control groups received either high-fat diet (HFD) or a normal diet (ND). The mothers were then mated with lean males for 12 weeks. The estrous cycle was recorded for two weeks after breeding. The metabolic phenotype, liver steatosis and reproductive organs were assessed at sacrifice 14 weeks after treatment. A short-course treatment of FGF21 leads to weight reduction during treatment but has no long-term impact on liver steatosis. A treatment with FGF21 leads to a reduction in the number of pregnancies (0 vs 1, p = 0.019) and no viable pup was born to a mother previously treated with FGF21. The FGF21 treatment affected the number of cycles (1 vs 3, p = 0.048) and amount in diestrus (54 vs 75%, p = 0.008) 12 weeks after the treatment. Additionally, the number of corpora lutea (0.8 vs 3.0, p = 0.016), and mature follicles (0 vs 1, p = 0.037) was reduced compared to the ND group while uterine histology remained unaffected. A short-term treatment with FGF21 has a long-term effect on female fertility in mice. This represents a potential safety concern for FGF21 analogs currently in clinical development. Reproductive health outcomes should be included in upcoming clinical trials.


Neuroprotection by hypoxic preconditioning: HIF-1 and erythropoietin protect from retinal degeneration.
Hypoxic exposure of cells or organisms induces expression of a number of hypoxia responsive genes through the activation of the hypoxia-inducible factor-1 (HIF-1). One of the most prominent HIF-1 targets is erythropoietin that has beneficial effects on ischemia-related injury in the brain. Exposure to low environmental oxygen concentrations can be used as a preconditioning paradigm to protect cells or tissues against a variety of harmful conditions. Here, we summarize recent work on neuroprotection of retinal photoreceptors and ganglion cells induced by hypoxic preconditioning or by systemically elevated levels of Epo in mouse plasma.


Idiopathic Hypokalemia in Lupus Nephritis: A Newly Recognized Entity.
Various causes of hypokalemia (HK) from renal potassium wasting, including distal renal tubular acidosis (RTA), have been described in lupus nephritis (LN). We report a phenomenon of otherwise unexplained HK among a population with LN. From our population of 403 patients with LN, we identified a cohort of 20 patients with idiopathic HK, defined by serum potassium <3.5 mmol/L without any apparent explanation. This cohort is compared with 90 LN controls (CON) and ten patients with LN with distal RTA from the same population. The patients with HK had lower median serum potassium compared with CON and RTA subjects (3.26 versus 4.00 versus 3.75 mmol/L, respectively; P<0.001). The median serum bicarbonate was normal in HK and CON, but low in RTA (26.0 versus 25.0 versus 19.4 mmol/L; P<0.001). The median urine pH was abnormally high only in the RTA group (6.00 versus 6.25 versus 6.67; P=0.012). The median serum magnesium was modestly lower in HK compared with the CON and RTA groups (1.73 versus 2.00 versus 1.85 mg/dl; P=0.002). Although both HK and RTA showed a higher rate of seropositivity than CON for anti-Ro/SSA (79% and 80% versus 37%, respectively; P<0.001), only HK revealed a higher rate of seropositivity than CON for anti-RNP (84% versus 42%; P=0.003) and only RTA showed a higher rate of seropositivity than CON for anti-La/SSB (40% versus 12%; P=0.05). A syndrome of idiopathic HK was revealed in 20 out of 403 (5%) of patients within our LN population, and proved to be distinct from the RTA that occurs in LN. Furthermore, it was associated with a distinct pattern of autoantibodies. We speculate that idiopathic HK is the result of a novel target of autoimmunity in LN, affecting renal tubular potassium transport.


The labeling of stem cells by superparamagnetic iron oxide nanoparticles modified with PEG/PVP or PEG/PEI.
Poly(ethylene glycol) (PEG) and poly(vinyl pyrrolidone) (PVP) co-modified superparamagnetic iron oxide nanoparticles (SPIONs) (PEG/PVP-SPIONs), and PEG and poly(ethylene imine) (PEI) co-modified SPIONs (PEG/PEI-SPIONs) synthesized by thermal decomposition have been used as magnetic resonance imaging (MRI) contrast agents to label adipose-derived stem cells (ADSCs). Efficient cell labeling was achieved after incubation with PEG/PVP-SPIONs and PEG/PEI-SPIONs for 12h, and the MRI of labeled cells was evaluated. The cell viability tests showed the low cytotoxicity of PEG/PVP-SPIONs and PEG/PEI-SPIONs. The cellular iron content incubated with PEG/PVP-SPIONs at a concentration of 25 μg/ml was 6.96 pg/cell, the cellular iron contents incubated with PEG/PEI-SPIONs at concentrations of 12 and 25 μg/ml were 20.16, 35.4 pg/cell, respectively. The SPIONs were located predominantly in the intracellular vesicles. The cellular iron oxide uptake was significantly high after incubation with PEG/PEI-SPIONs as compared with the commercial iron oxide agents (Feridex, Feridex@PLL, Resovist and Resovist@PLL) reported. This work demonstrates that PEG/PEI-SPIONs are the competent agents for the labeling of ADSCs.


Does a physically active lifestyle attenuate the association between alcohol consumption and mortality risk? Findings from the UK biobank.
Alcohol consumption is common across Western culture, despite its associations with adverse health outcomes, including cancer and cardiovascular disease (CVD). Physical activity (PA) has beneficial effects on many alcohol related outcomes, with data suggesting PA may offset the association between alcohol consumption and mortality. This study examined the joint associations of PA and alcohol on all-cause, CVD and cancer mortality. Participants were recruited between 2006 and 2010 in the United Kingdom. Alcohol consumption was categorised based on current UK guidelines (14 units/week). PA was categorised based on the Metabolic Task Equivalent of PA as low, moderate and high. Data were analysed using Cox proportional-hazard models. The final analysis, conducted in 2019, included 297,988 adults aged ≥40. Over an average follow-up of 6.9 years, 6079 deaths were recorded, including 1219 CVD deaths and 3112 cancer deaths. We observed greater point estimates for risk of all-cause mortality among low PA individuals who consumed alcohol at the same level as active individuals. For example, low PA participants who reported alcohol consumption ≥double guidelines had a greater HR (1.55, 95% CI 1.25, 1.93) than active individuals (moderate PA HR 1.21, 95% CI 0.95, 1.54; high PA HR 1.21, 95% CI 1.00, 1.46). Considering CVD, we observed a similar trend with lower point estimates of risk of mortality among active individuals. We found some evidence that PA modified the associations of alcohol and all-cause and CVD mortality in this large population sample of British adults.


Data mining on identifying diagnosis and prognosis biomarkers in head and neck squamous carcinoma.
Head and neck squamous carcinoma (HNSC) induces high cancer-related death worldwide. The biomarker screening on diagnosis and prognosis is of great importance. This research is aimed to explore the specific diagnostic and prognostic biomarkers for HNSC through bioinformatics analysis. The mutation and dysregulation data were acquired from UCSC Xena and TCGA databases. The top ten genes with mutation frequency in HNSC were TP53 (66%), TTN (35%), FAT1 (21%), CDKN2A (20%), MUC16 (17%), CSMD3 (16%), PIK3CA (16%), NOTCH1 (16%), SYNE1 (15%), LRP1B (14%). A total of 1,060 DEGs were identified, with 396 up-regulated and 665 downregulated in HNSC patients. Patients with lower expression of ACTN2 (P = 0.039, HR = 1.3), MYH1 (P = 0.005, HR = 1.5), MYH2 (P = 0.035, HR = 1.3), MYH7 (P = 0.053, HR = 1.3), and NEB (P = 0.0043, HR = 1.5) exhibit longer overall survival time in HNSC patients. The main DEGs were further analyzed by pan-cancer expression and immune cell infiltration analyses. MYH1, MYH2, and MYH7 were dysregulated in the cancers. Compared with HNSC, their expression levels are lower in the other types of cancers. MYH1, MYH2, and MYH7 were expected to be the specific diagnostic and prognostic molecular biomarkers of HNSC. All five DEGs have a significant positive correlation with CD4+T cells and macrophages.