United States nurse practitioner students' attitudes, perceptions, and beliefs working with the uninsured.
To reduce health disparities, nurses are expected to provide compassionate and high quality care to all patients regardless of socioeconomic and insurance status. In the United States (US) nurse practitioner (NP) educators need to expose students to clinical practice settings, such as free clinics, where the vulnerable populations like the non-documented immigrants and uninsured receive care. The purpose of this mixed method study was to provide an immersion experience for (NP) students at free clinics. Then researchers evaluated the impact of a clinical immersion experience on NP students' attitudes, perceptions, and beliefs about the uninsured and determine whether the clinical experience impacted their willingness to consider working with vulnerable populations in the future. Qualitative and quantitative data suggests students challenged their own beliefs and attitudes regarding the vulnerable populations, gained insight into care provided at these free clinics, and expressed their intent to volunteer at these settings. In the era of health care reform both nationally and internationally and the need to improve primary care access globally, educational initiatives are needed to expose NP students to economically vulnerable populations. Future research needs to replicate and extend the findings of this study, focusing on teaching-learning experiences for nurse practitioners and other advanced practice nursing students.


Short energy intake is associated with muscle mass loss in older patients with type 2 diabetes: A prospective study of the KAMOGAWA-DM cohort.
Past studies reported that the intake of adequate energy is more important than protein intake; however, the relationship between energy intake and muscle mass loss remains unclear thus far. This study therefore explored the association between energy intake and muscle mass loss in people with type 2 diabetes (T2D). In this prospective cohort study, impedance body composition and a brief-type self-administered diet history questionnaire were used for analyzing body composition and habitual diet intake, respectively. Skeletal muscle mass index (SMI, kg/m2) was defined as appendicular muscle mass (kg) ÷ height-squared (m2). Rate of SMI change (%) was calculated as ([SMI at baseline - SMI at follow-up]/[follow-up duration (years) × baseline SMI (kg/m2)]) × 100, and muscle mass loss was defined as rate of SMI change ≥0.5%. Energy intake was defined as total energy intake (kcal/day) divided by ideal body weight (kg), defined as 22 × patient height-squared (m2). Among non-older and older participants, 54.8% (n = 51/93) and 58.9% (n = 116/197) experienced muscle mass loss at 16.3 (6.4) and 18.1 (7.1) months' follow-up, respectively. Logistic regression analyses showed that energy intake was associated with incident muscle mass loss after adjusting for age, sex, insulin, sodium glucose cotransporter-2 inhibitor, glucagon-like peptide-1 agonist, steroids, smoking, exercise, alcohol intake, body mass index, SMI, presence of renal failure, and protein intake (g/actual body weight/day) in the older people (odds ratio [OR] 0.94 [95% confidence interval [CI] 0.88-0.996], p = 0.037), whereas energy intake was not associated with incident muscle mass loss in the non-older people (OR 0.96 [95% CI 0.86-1.06], p = 0.423). Insufficient energy intake is associated with muscle mass loss in older people with T2D. Therefore, it is recommended to consume enough energy for older people with T2D to keep muscle mass.


Estimated benefit of increased vitamin D status in reducing the economic burden of disease in western Europe.
Vitamin D has important benefits in reducing the risk of many conditions and diseases. Those diseases for which the benefits are well supported and that have large economic effects include many types of cancer, cardiovascular diseases, diabetes mellitus, several bacterial and viral infections, and autoimmune diseases such as multiple sclerosis. Europeans generally have low serum 25-hydroxyvitamin D [25(OH)D] levels owing to the high latitudes, largely indoor living, low natural dietary sources of vitamin D such as cold-water ocean fish, and lack of effective vitamin D fortification of food in most countries. Vitamin D dose-disease response relations were estimated from observational studies and randomized controlled trials. The reduction in direct plus indirect economic burden of disease was based on increasing the mean serum 25(OH)D level to 40 ng/mL, which could be achieved by a daily intake of 2000-3000 IU of vitamin D. For 2007, the reduction is estimated at euro187,000 million/year. The estimated cost of 2000-3000 IU of vitamin D3/day along with ancillary costs such as education and testing might be about euro10,000 million/year. Sources of vitamin D could include a combination of food fortification, supplements, and natural and artificial UVB irradiation, if properly acquired. Additional randomized controlled trials are warranted to evaluate the benefits and risks of vitamin D supplementation. However, steps to increase serum 25(OH)D levels can be implemented now based on what is already known.


Two approaches to comparing hospital charges between cadaveric renal transplant patients who received orthoclone OKT 3 sterile solution or Atgam sterile solution for induction therapy.
The objectives of this study were: (1) to compare total hospital charges for a sample of cadaveric renal transplant patients categorized according to the type of induction therapy used (Orthoclone OKT3 Sterile Solution or Atgam Sterile Solution); (2) to compare specific charge categories between the two groups; and (3) to examine the relationship between charges and a set of independent variables. A retrospective review was conducted of hospital charges associated with a sample of renal transplant patients. The overall sample for this study comprised 510 patient discharges from 22 hospitals in the United States. Comparisons between the OKT3 and Atgam groups were made for total and specific charge categories using two different approaches to help control variations in charges that were not related to the type of induction therapy used. The first approach consisted of t test or chi-square comparisons between the groups for subsets of observations that had been identified in a stepwise fashion. These judgment samples were defined to remove sources of variation in charges other than those resulting from the type of induction therapy selected. The second approach used multiple linear regression analysis to help statistically control variation in charges from other sources. The results showed that higher drug charges in the Atgam group were offset by lower charges in other categories (P < 0.05). These findings suggest that hospital formulary committees should consider all relevant costs, not just drug acquisition costs, when selecting products. However, further investigation is warranted to explore differences in charges due to: (1) between-hospital variation; (2) patients' severity of illness before receiving induction therapy; and (3) differences in side-effect profiles for the two induction therapies.


[Kawasaki disease: heart disease during childhood].
The aim of this study was to determine risk factors for coronary lesions, the type of heart lesion present, and long-term outcome in Kawasaki disease. We studied 150 children, aged 3 months to 9.5 years, who met the criteria for a diagnosis of Kawasaki disease. Of the 18% who were diagnosed with heart disease, 100% had coronary artery abnormalities, 11.1% had pancarditis, and 3.7%, mitral insufficiency. The coronary abnormalities were classified according severity as follows: diffuse ectasia in 40.7%; a small-to-medium-sized solitary coronary artery aneurysm in 33.3%; numerous small-to-medium-sized aneurysms in 11.1%; giant aneurysms in 11.1%; and coronary artery stenosis in 3.7%. The presence of pancarditis was a predictor of a giant coronary artery aneurysm. Mortality was 3.7%. Coronary aneurysm was the predominant heart lesion. The risk factors for coronary aneurysm in Kawasaki disease included age less than 27 months, fever lasting more than 8 days, erythrocyte sedimentation greater than 70 mm, and pancarditis.


Comparison of the effects of temperature and water activity on growth rate of food spoilage moulds.
The influence of temperature (T) and water activity (aw) on the growth rate (mu) of seven moulds (Alternaria alternata, Aspergillus flavus, Cladosporium cladosporioides, Mucor racemosus, Penicillium chrysogenum, Rhizopus oryzae and Trichoderma harzianum) was assessed in suboptimal conditions. Firstly, the dependence of fungal growth on temperature, at aw 0.99, was modelled through an approach described previously for bacteria. A dimensionless growth rate variable: mu(dimalpha)=mu/mu (optalpha) depended on the following normalised temperature: T(dim)=(T-T(min))/(T(opt)- T(min)) according to a power function: mu(dimalpha)=[T(dim)]alpha, where alpha was an exponent to be estimated. Secondly, the same approach was used to describe the influence of aw on fungal growth, at the respective optimum temperatures for each mould. Similarly, mu(dimbeta)=mu/mu(optbeta) depended on the following normalised water activity: a(wdim)=(aw-a(wmin))/(a(wopt)-a(wmin)) according to a power function: mu(dimbeta)=[a(wdim)](beta). Results show: (i) for each mould, the alpha-value is significantly less than the beta-value, confirming that water activity has a greater influence than temperature on fungal development; (ii) the alpha-values and the beta-values depend on the mould; (iii) the alpha-value is less than 1 for the mesophilic mould A. flavus, whereas the other moulds are characterised by higher alpha-values ranging from 1.10 to 1.54; (iv) the mesophilic A. flavus exhibits a low beta-value, 1.50, compared to the hydrophilic T. harzianum, beta=2.44, while beta-values are within the range (1.71-2.37) for the other moulds.


Regulatory T-cell depletion synergizes with gp96-mediated cellular responses and antitumor activity.
Despite its potent immunostimulatory properties, vaccination with autologous tumor-derived gp96 has relatively modest antitumor effect in a range of clinical trials. Based on our previous study showing a gp96-mediated immune balance between CTL and Tregs, here we investigated possible synergy between gp96 vaccine and systemic Treg depletion on induction of antitumor T-cell immunity and the mechanisms accounting for synergistic efficacy. In gp96-peptide complex immunized BALB/c mice, anti-CD25 mAb treatment significantly increased IFN-γ-producing CD8(+) and CD4(+) T cells by about 1-2-fold in spleen and 40-50% in lymph node. A significantly higher number of peptide-specific CTL were observed under anti-CD25 mAb treatment compared with no treatment. Moreover, Treg depletion synergistically improved the anticancer activity of tumor-derived gp96 vaccine in the poorly immunogenic and highly tumorigenic B16 melanoma model in C57BL/6 J mice. While gp96 immunization alone led to the modest enhancement of CTL activities in spleen, the combination with Treg depletion dramatically increased tumor-specific CTL responses. In addition, the combination resulted in a significant increase of CD8(+) T-cell infiltration in tumor, which correlated with an enhanced inhibition of tumor growth. Our results provide evidence that targeting Tregs may provide a more efficient strategy to potentiate gp96-mediated T-cell responses and enhance the antitumor efficiency of gp96-based therapeutic vaccine.


Hyaluronan secretion into the synovial cavity of rabbit knees and comparison with albumin turnover.
1. Hyaluronan is not only a lubricant but also enhances the synovial lining's resistance to fluid outflow. This finding led to the proposal that hyaluronan (> 2 x 10(6) Da, approximately 210 nm radius) may escape across the synovial lining less freely than smaller solutes (e.g. albumin, 6.7 x 10(4) Da, 3.6 nm radius) or water. Here multiple washouts were used to measure intraarticular hyaluronan mass and secretion rate in rabbit knees, leading to an estimate of hyaluronan turnover time. Plasma albumin permeation into the joint cavity was also measured to enable comparison of turnover times between molecules of very disparate size. 2. Endogenous hyaluronan mass in the joint cavity, analysed by high performance liquid chromatography of joint washes, was 182 +/- 9.9 micrograms (mean +/- S.E.M; n = 21). Since hyaluronan concentration in synovial fluid averages 3.62 +/- 0.19 micrograms microliters-1, the endogenous synovial fluid volume was calculated to be 50 microliters (mass/concentration), about double the aspiratable volume. 3. The hyaluronan secretion rate over 4 h was 4.80 +/- 0.77 micrograms h-1 (n = 5). The rate was significantly higher in contralateral joints expanded by 2 ml Ringer solution (5.80 +/- 0.84 micrograms h-1, n = 5, P = 0.01, Student's paired t test), indicating a stretch/hydration sensitive secretory mechanism. The newly secreted chains ((2.05-2.48) x 10(6) Da) were not significantly different in length from the endogenous chains (2.95 x 10(6) Da). 4. Hyaluronan turnover time, calculated as mass/secretion rate, was 31.4-37.9 h. This is more than an order of magnitude longer than turnover time for intra-articular albumin. The latter, determined from the intra-articular albumin mass and plasma-to-cavity permeation rate was 1.8 h (95% confidence intervals 1.2-3.5 h, n = 9). The big difference in turnover times support the view that, relative to albumin and water, hyaluronan is partially sieved out and retained in the joint cavity by the synovial lining. The lining cell layer is discontinuous, so it appears that interstitial matrix itself acts as a leaky size-selective molecular filter.


An evaluation of compliance with practice guidelines on interhospital trauma transfer.
To prospectively evaluate compliance with current interhospital trauma transfer guidelines in South West Sydney, before and after an implementation programme was instituted. A scoring system was developed to assess compliance with the 11 main components of the guideline. Baseline compliance was measured during an initial 3-month period (pre), followed by an implementation programme to alert staff at referring hospitals to the presence of the guidelines. Following this, compliance was again measured over 3 months (post). Twenty-two patients were transferred during the pre-implementation phase and 35 patients during the post-phase. Overall compliance with the guidelines increased from 62 to 67%. Mean pre-hospital compliance rose from 75 to 95%, and referring hospital compliance rose from 59 to 63%. While there was an improvement in compliance with the use of the dedicated trauma hotline (86-97%), the use of a transfer checklist (41-53%), and appropriateness of transfer (95-100%), none of these reached statistical significance. Practice guidelines have been developed to optimise the process of interhospital trauma transfers. An implementation programme met with limited success in improving compliance with the guidelines. Further work is needed to ensure awareness of these guidelines, with ongoing monitoring to ensure best practice and optimal patient outcome.


How occasional backstepping can speed up a processive motor protein.
Fueled by the hydrolysis of ATP, the motor protein kinesin literally walks on two legs along the biopolymer microtubule. The number of accidental backsteps that kinesin takes appears to be much larger than what one would expect given the amount of free energy that ATP hydrolysis makes available. This indicates that backsteps are not simply the forward stepping cycle run backwards. We propose here a simple effective model that consistently includes the backstep transition. Using this model, we show how more backstepping increases the entropy of the final state, and probably also the activation state, thus reducing their free energy. This free energy reduction of the activation state (related to backstepping) speeds up the catalytic cycle of the kinesin, making both forward and backward steps more frequent. As a consequence, maximal net forward speed is achieved at nonzero backstep percentage. In addition, the optimal backstep percentage coincides with the backstep percentage measured for kinesin. This result suggests that, through natural selection, kinesin could have evolved to maximal speed.


Analyzing the effects of different signs to increase the availability of designated van-accessible parking spaces.
Often, designated van accessible parking spaces are occupied by non-ramp or lift equipped vehicles (NRLEV) with accessible parking permits. This causes inconvenience, safety concerns, and reduced availability of parking spaces for ramp or lift equipped vehicles (RLEV). The researchers conducted a single subject design study with two settings to examine a differential comparison between the standard van sign versus intervention signs (pictorial representation of a ramp-equipped van plus messages) to determine which is more effective to signal NRLEV users not to park in designated van accessible spaces. The study was composed of two phases, a multiple treatment design to compare the effects of the intervention signs during phase 1 and a reversal design to test and validate the most effective intervention effects during phase 2. The results based on observation showed that the interventions signs could moderately reduce the chances of NRLEV users parking in the designated van accessible parking space. Implications of these research findings are discussed and recommendations for changes in state and national accessible parking policies are suggested.


Reliable chiral recognition with an optoelectronic nose.
Chiral discrimination is a key problem in analytical chemistry. It is generally performed using expensive instruments or highly-specific miniaturized sensors. An electronic nose is a bio-inspired instrument capable after training of discriminating a wide variety of analytes. However, generality is achieved at the cost of specificity which makes chiral recognition a challenging task for this kind of device. Recently, a peptide-based optoelectronic nose which can board up to hundreds of different sensing materials has shown promising results, especially in terms of specificity. In line with these results, we describe here its use for chiral recognition. This challenging task requires care, especially in terms of statistical reliability and experimental confounds. For these reasons, we set up an automatic gas sampling system and recorded data over two long sessions, taking care to exclude possible confounds. Two couples of chiral molecules, namely (R) and (S) Limonene and (R) and (S) Carvone, were tested and several statistical analyses indicate the almost perfect discrimination of their two enantiomers. A method to highlight discriminative sensing materials is also proposed and shows that successful discrimination is likely achieved using just a few peptides.


Observations of neutron radiation environment during Odyssey cruise to Mars.
In April 2001, Mars Odyssey spacecraft with the High Energy Neutron Detector (HEND) onboard was launched to Mars. HEND/Odyssey was switched on measurement mode for most of transit to Mars to monitor variations of spacecraft background and solar activity. Although HEND/Odyssey was originally designed to measure Martian neutron albedo and to search for Martian subsurface water/water ice, its measurements during cruise phase to Mars are applicable to evaluate spacecraft ambient radiation background. The biological impact of the neutron component of this radiation background should be understood, as it must be taken into account in planning future human missions to Mars. We have modeled the spacecraft neutron spectral density and compared it with HEND measurements to estimate neutron dose equivalent rates during Odyssey cruise phase, which occurred during the maximum period of solar cycle 23. We find that the Odyssey ambient neutron environment during May - September 2001 yields 10.6 ± 2.0 μSv per day in the energy range from 0 to 15 MeV, and about 29 μSv per day when extrapolated to the 0-1000 MeV energy range during solar quiet time (intervals without Solar Particle Events, SPEs). We have also extrapolated HEND/Odyssey measurements to different periods of solar cycle and find that during solar minimum (maximum of GCR flux), the neutron dose equivalent rate during cruise to Mars could be as high as 52 μSv per day with the same shielding. These values are in good agreement with results reported for a similar measurement made with an instrument aboard the Mars Science Laboratory during its cruise to Mars in 2011-2012.


Norepinephrine transporter knock-out alters expression of the genes connected with antidepressant drugs action.
Norepinephrine transporter knock-out mice (NET-KO) exhibit depression-resistant phenotypes. They manifest significantly shorter immobility times in both the forced swim test and the tail suspension test. Moreover, biochemical studies have revealed the up-regulation of other monoamine transporters (dopamine and serotonin) in the brains of NET-KO mice, similar to the phenomenon observed after the chronic pharmacological blockade of norepinephrine transporter by desipramine in wild-type (WT) animals. NET-KO mice are also resistant to stress, as we demonstrated previously by measuring plasma corticosterone concentration. In the present study, we used a microdissection technique to separate target brain regions and the TaqMan Low Density Array approach to test the expression of a group of genes in the NET-KO mice compared with WT animals. A group of genes with altered expression were identified in four brain structures (frontal and cingulate cortices, dentate gyrus of hippocampus and basal-lateral amygdala) of NET-KO mice compared with WT mice. These genes are known to be altered by antidepressant drugs administration. The most interesting gene is Crh-bp, which modulates the activity of corticotrophin--releasing hormone (CRH) and several CRH-family members. Generally, genetic disturbances within noradrenergic neurons result in biological changes, such as in signal transduction and intercellular communication, and may be linked to changes in noradrenaline levels in the brains of NET-KO mice.


The microbial flora of the vagina and its relationship to bacteriuria in diabetic and non-diabetic women.
The vaginal and urinary flora have been studied in 57 diabetic and 33 non-diabetic patients. In both populations, no correlation was found between urogenital symptoms and colonisation of the vagina. There was no greater prevalence of potentially pathogenic organisms in diabetic patients, even when control was poor. However, although not statistically significant, group B streptococci were isolated more frequently from poorly controlled diabetics. Bacteriuria was not significantly more common in diabetics. Gram-negative bacilli were found in the vagina significantly more often in bacteriuric patients; the prevalence of such organisms being four-fold greater after the menopause.


Seasonality, sex-specificity and transcriptional regulation of hepatic leptin system in spotted snakehead Channa punctata.
The present study deals with sex-specific reproductive phase-dependent variation and sex steroids-induced transcriptional regulation of hepatic lep and lepr in nutritionally valuable spotted snakehead, Channa punctata. The data on seasonality reveals sex-specific variation in pattern of lep transcription where a high level was recorded during resting and postspawning quiescent phases in female while during resting and spawning phases in male. Unlike lep, lepr exhibited similar expression pattern along the reproductive phases in both the sexes. As compared to female, a three-fold higher expression of lep was detected in male during reproductively active phase only. However, no sexual dimorphism was evidenced in lepr either during active or quiescent phase. To explore the implication of sex steroids in regulation of leptin system, we correlated levels of plasma testosterone (T) and 17β-estradiol (E2) with leptin system in males as well as females. Further, criss-cross in vivo and in vitro experiments with dihydrotestosterone (DHT) and E2 were conducted in male and female spotted snakehead. The leptin system was downregulated after DHT administration in both the sexes. However, with E2, a marked decrease was evidenced in male only. The sex-wise variable response of leptin system to sex steroids was validated by in vitro experiments wherein liver fragments from male and female fish were incubated individually with both the sex steroids. In conclusion, sex steroids modulate hepatic leptin system differentially depending on sex and reproductive state of spotted snakehead.


Influence of the Topology of Poly(L-Cysteine) on the Self-Assembly, Encapsulation and Release Profile of Doxorubicin on Dual-Responsive Hybrid Polypeptides.
Τhe synthesis of a series of novel hybrid block copolypeptides based on poly(ethylene oxide) (PEO), poly(l-histidine) (PHis) and poly(l-cysteine) (PCys) is presented. The synthesis of the terpolymers was achieved through a ring-opening polymerization (ROP) of the corresponding protected N-carboxy anhydrides of Nim-Trityl-l-histidine and S-tert-butyl-l-cysteine, using an end-amine-functionalized poly(ethylene oxide) (mPEO-NH2) as macroinitiator, followed by the deprotection of the polypeptidic blocks. The topology of PCys was either the middle block, the end block or was randomly distributed along the PHis chain. These amphiphilic hybrid copolypeptides assemble in aqueous media to form micellar structures, comprised of an outer hydrophilic corona of PEO chains, and a pH- and redox-responsive hydrophobic layer based on PHis and PCys. Due to the presence of the thiol groups of PCys, a crosslinking process was achieved further stabilizing the nanoparticles (NPs) formed. Dynamic light scattering (DLS), static light scattering (SLS) and transmission electron microscopy (TEM) were utilized to obtain the structure of the NPs. Moreover, the pH and redox responsiveness in the presence of the reductive tripeptide of glutathione (GSH) was investigated at the empty as well as the loaded NPs. The ability of the synthesized polymers to mimic natural proteins was examined by Circular Dichroism (CD), while the study of zeta potential revealed the "stealth" properties of NPs. The anticancer drug doxorubicin (DOX) was efficiently encapsulated in the hydrophobic core of the nanostructures and released under pH and redox conditions that simulate the healthy and cancer tissue environment. It was found that the topology of PCys significantly altered the structure as well as the release profile of the NPs. Finally, in vitro cytotoxicity assay of the DOX-loaded NPs against three different breast cancer cell lines showed that the nanocarriers exhibited similar or slightly better activity as compared to the free drug, rendering these novel NPs very promising materials for drug delivery applications.


Biochemistry of the various fractions of sarcocysts of Sarcocystis fusiformis of buffalo (Bubalus bubalis).
A comparative biochemical study on the various fractions (cyst wall, cyst fluid and zoites) of the sarcocysts of Sarcocystis fusiformis from oesophageal muscles of naturally infected Indian water buffalo (Bubalus bubalis) was carried out. The study included analysis for glycogen, glucose, pyruvic acid, lactic acid, total lipids, phospholipids, cholesterol, triglycerides and fatty acids. The pattern and the distribution of various biochemical constituents varied in the different fractions. The cell wall had the maximum concentration of glucose and phospholipids. Of all the fractions, cell fluid showed the highest contents of pyruvate and lactate, but with a higher level of pyruvate than lactate.


Interaction of Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase with pyridoxal 5'-diphospho-5'-adenosine. Affinity labeling of Lys-21 and Lys-343.
Pyridoxal 5'-diphospho-5'-adenosine (PLP-AMP) inhibits glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides competitively with respect to glucose 6-phosphate and noncompetitively with respect to NAD+ or NADP+, with Ki = 40 microM in the NADP-linked and 34 microM in the NAD-linked reaction. Incubation of glucose-6-phosphate dehydrogenase with [3H]PLP-AMP followed by borohydride reduction shows that incorporation of 0.85 mol of PLP-AMP per mol of enzyme subunit is required for complete inactivation. Both glucose 6-phosphate and NAD+ protect against this covalent modification. The proteolysis of the modified enzyme and isolation and sequencing of the labeled peptides revealed that Lys-21 and Lys-343 are the sites of PLP-AMP interaction and that glucose 6-phosphate and NAD+ protect both lysyl residues against modification. Pyridoxal 5'-phosphate (PLP) also modifies Lys-21 and probably Lys-343. Lys-21 is part of a highly conserved region that is present in all glucose-6-phosphate dehydrogenases that have been sequenced. Lys-343 corresponds to an arginyl residue in other glucose-6-phosphate dehydrogenases and is in a region that is less homologous with those enzymes. PLP-AMP and PLP are believed to interact with L. mesenteroides glucose-6-phosphate dehydrogenase at the glucose 6-phosphate binding site. Simultaneous binding of NAD+ induces conformational changes (Kurlandsky, S. B., Hilburger, A. C., and Levy, H. R. (1988) Arch. Biochem. Biophys. 264, 93-102) that are postulated to interfere with Schiff's-base formation with PLP or PLP-AMP. One or both of the lysyl residues covalently modified by PLP or PLP-AMP may be located in regions of the enzyme undergoing the NAD(+)-induced conformational changes.


Contributions of nuclear architecture to transcriptional control.
Three parameters of nuclear structure contribute to transcriptional control. The linear representation of promoter elements provides competency for physiological responsiveness within the contexts of development as well as cycle- and phenotype-dependent regulation. Chromatin structure and nucleosome organization reduce distances between independent regulatory elements providing a basis for integrating components of transcriptional control. The nuclear matrix supports gene expression by imposing physical constraints on chromatin related to three-dimensional genomic organization. In addition, the nuclear matrix facilitates gene localization as well as the concentration and targeting of transcription factors. Several lines of evidence are presented that are consistent with involvement of multiple levels of nuclear architecture in cell growth and tissue-specific gene expression during differentiation. Growth factor and steroid hormone responsive modifications in chromatin structure, nucleosome organization, and the nuclear matrix that influence transcription of the cell cycle-regulated histone gene and the bone tissue-specific osteocalcin gene during progressive expression of the osteoblast phenotype are considered.


Association Between the Telomerase rs2736098_TT Genotype and a Lower Risk of Chronic Hepatitis B and Cirrhosis in Chinese Males.
Chronic hepatitis B (CHB) is caused by infection of hepatitis B virus (HBV) and liver cirrhosis (LC) is its most common complication. The accumulated evidence indicates a genetic context of HBV infection phenotypes. Here we determine a potential association of CHB/LC with the genetic variant of telomerase reverse transcriptase (TERT), a key player in aging including immune-senescence. The study included 227 Chinese CHB patients and 315 sex/age-matched healthy controls. TERT rs2736098 and rs2736100 genotyping was performed using pre-designed TaqMan SNP genotyping assay kits. Leukocyte telomere length (LTL) was determined using quantitative PCR. The rs2736098_CT/CC genotypes were significantly associated with risk of CHB compared to the TT one (OR 2.265, 95% CI 1.202-4.269, P=0.015). A similar association was also found in CHB patients with cirrhosis (CT/CC vs TT: OR 2.398, 95% CI 1.168-4.922, P=0.02). Further analyses showed that the rs2736098_TT genotype difference occurred between male controls and patients (P=0.008) and male CT/CC-carriers exhibited highly increased risk of CHB compared to male controls (CT+CC vs TT, OR 3.182, 95% CI 1.350-7.500, P=0.01). There was no difference in the rs2736100 variants between controls and CHB patients. LTL was not different between cases and controls. The TERT rs2736098_TT genotype is associated with a lower CHB and LC risk in Chinese males, which may have implications in CHB pathogenesis and prevention.


Cocaine toxicity in cultured rat hepatocytes.
Cocaine hydrochloride was added to primary cultures of hepatocytes isolated from naive and phenobarbital-induced (80 mg/kg i.p. for 3 d) Sprague-Dawley rats. Cocaine was cytotoxic, as measured by lactate dehydrogenase release, to cells from naive rats in concentrations of 1 mM or greater. Phenobarbital induction greatly increased the cytotoxic potency of cocaine in vitro, with nearly complete loss of cell viability at cocaine concentrations in culture as low as 0.01 mM. The addition of 10 microM SK&F-525-A to the cultures blocked cocaine cytotoxicity in cells from both naive and phenobarbital-induced rats. These results suggest that the metabolic pathways leading to cocaine hepatotoxicity identified in the mouse also exist in the rat hepatocyte.


Data management in mass spectrometry-based proteomics.
Mass spectrometry-based proteomics has made the transition from the analysis of a single or only a few proteins per experiment to a high-throughput platform for discovery/validation that can analyze several hundreds to thousands of proteins in a single experiment. This increase in analytical capability hinged on four main components: (1) innovative methodologies, (2) improved instruments, (3) ready availability of comprehensive protein sequence databases, and (4) development of sophisticated software algorithms to match fragmentation mass spectra against these databases. But as the throughput of the approach increased, so did the necessity to manage and automate the data processing workflow. Indeed, modern instruments generate tens of thousands of fragmentation spectra per hour, providing a substantial bioinformatics challenge. This chapter provides insight into the specifics of this challenge, by looking at a typical workflow, the data types and user roles involved, and a broad overview of available software solutions. Finally, the increasingly important link between a local data management system and the global, centralized dissemination of proteomics data is discussed.


Redox Role of ROS and Inflammation in Pulmonary Diseases.
Reactive oxygen species (ROS), either derived from exogenous sources or overproduced endogenously, can disrupt the body's antioxidant defenses leading to compromised redox homeostasis. The lungs are highly susceptible to ROS-mediated damage. Oxidative stress (OS) caused by this redox imbalance leads to the pathogenesis of multiple pulmonary diseases such as asthma, chronic obstructive pulmonary disease (COPD), and acute respiratory distress syndrome (ARDS). OS causes damage to important cellular components in terms of lipid peroxidation, protein oxidation, and DNA histone modification. Inflammation further enhances ROS production inducing changes in transcriptional factors which mediate cellular stress response pathways. This deviation from normal cell function contributes to the detrimental pathological characteristics often seen in pulmonary diseases. Although antioxidant therapies are feasible approaches in alleviating OS-related lung impairment, a comprehensive understanding of the updated role of ROS in pulmonary inflammation is vital for the development of optimal treatments. In this chapter, we review the major pulmonary diseases-including COPD, asthma, ARDS, COVID-19, and lung cancer-as well as their association with ROS.


Correlation between cardiac autonomic modulation in response to orthostatic stress and indicators of quality of life, physical capacity, and physical activity in healthy individuals.
Increased heart rate variability (HRV) at rest is frequently associated to maximal oxygen uptake (VO2max), physical activity, and markers of quality of life (QoL). However, the HRV has not been observed during physical exercise or orthostatic (ORT) challenge. This study investigated the associations of HRV changes (ΔHRV) from rest at supine (SUP) to ORT positions with (VO2max), physical activity level, and QoL in young adults. Cardiac autonomic modulation was assessed by spectral analysis of R-R time series measured from SUP to ORT positions in 15 healthy volunteers (26 ± 7 years). Questionnaires were applied for evaluation of QoL (SF-36 score), to estimate (VO2max), and to quantify physical activity (Baecke Sport Score). All HRV indices at SUP, but not ORT, strongly correlated to QoL, estimated (VO2max), and physical activity. The ΔHRV from SUP to ORT showed significant correlations with all questionnaire scores (r = 0.52-0.61 for low frequency and r = -0.61 to -0.65 for high frequency, p ≤ 0.05). Higher vagal activity at rest and greater changes in adrenergic and parasympathetic modulation from SUP to ORT were detected in the volunteers exhibiting higher scores of QoL, estimated (VO2max), and physical activity. Taken together, the level of neural adaptations from resting SUP position to active standing, and physical activity and QoL questionnaires seem to be a simple approach to understand the physiological and lifestyle adaptations to exercise that may be applied to a large sample of subjects in almost any sports facilities at a low cost.


Clinicopathological Review of Micropapillary Urothelial Carcinoma.
This review will discuss micropapillary urothelial carcinoma with respect to biology, histopathologic characteristics, genetic and molecular features, diagnosis, clinical management, and future directions of research. Recent consensus opinion study showed only moderate interobserver reproducibility in the diagnostic criteria. The most reproducible criteria with the highest consensus were multiple nests in the same lacunar spaces. There are recent reports of high rates of intratumoral heterogeneity of ERBB2 amplification within tumor containing both micropapillary and classic urothelial components. Micropapillary urothelial carcinoma is a well-documented highly aggressive variant of urothelial carcinoma with proven worse outcomes. Accurate recognition and reporting of this pattern is critical for optimal management. Newer therapeutic strategies related to the molecular and genetic findings seen in MPUC remain to be explored further.


APP96-110 Elicits Neuroprotective Effects Following Ischemic Insult in Animal Models.
Competitive amyloidogenic pathways play an important role in many neurological diseases such as the onset of various degenerative diseases and ischemic stroke. Overexpression of amyloid precursor protein (APP) and amyloid-beta is modulated via the amyloidogenic pathway, which plays a crucial role in neuroinflammation. During ischemic conditions, the activity of the anti-inflammatory non-amyloidogenic pathway decreases, thus increasing the activity of amyloidogenic pathway. The soluble alpha form of APP (sAPPα), formed via the non-amyloidogenic pathway, exhibits neuroprotective effects against neurological diseases. sAPPα is thought to have a modulatory effect on several cell survival pathways, including its ability to inhibit the phosphoinositide 3-kinases (PI3K) pathway, thereby inhibiting the inflammatory response. The APP derivative, APP96-110, could act as a functional substitute for native sAPPα. Herein, we investigated whether APP96-110 has neuroprotective effects against neuroinflammation and damage following cerebral ischemic stroke. Treatment with diluted APP96-110 (0.005 mg/kg) in mice after 30 min of transient middle cerebral artery occlusion (tMCAO) showed improved motor function and reduced expression of the inflammatory marker CD86. APP96-110 decreased the infarct size and induced an anti-inflammatory response by inhibiting the PI3K pathway. These results suggest that the treatment of APP96-110 is efficacious in reducing neuroinflammation and infarct size in ischemic stroke.


Groundwater vulnerability assessment combining the drastic and dyna-clue model in the Argentine pampas.
Vulnerability assessment is considered an effective tool in establishing monitoring networks required for controlling potential pollution. The aim of this work is to propose a new integrated methodology to assess actual and forecasted groundwater vulnerability by including land-use change impact on groundwater quality. Land-use changes were simulated by applying a spatial dynamics model in a scenario of agricultural expansion. Groundwater vulnerability methodology DRASTIC-P, was modifyed by adding a land-use parameter in order to assess groundwater vulnerability within a future scenario. This new groundwater vulnerability methodology shows the areas where agricultural activities increase the potential level of groundwater vulnerability to pollution. The Dulce Creek Basin was the study case proposed for the application of this methodology. The study revealed that the area with Very High vulnerability would increase 20% by the year 2020 in the Dulce Creek Basin. This result can be explained by analyzing the land-use map simulated by the Dyna-CLUE model for the year 2020, which shows that the areas with increments in crop and pasture coincide with the area defined by the Very High aquifer vulnerability category in the year 2020. Through scenario analysis, land-use change models can help to identify medium or long term critical locations in the face of environmental change.


Elevated WBP2 Expression in HER2-positive Breast Cancers Correlates with Sensitivity to Trastuzumab-based Neoadjuvant Therapy: A Retrospective and Multicentric Study.
Trastuzumab-based chemotherapy has shown remarkable clinical benefits for patients with HER2-positive breast cancer. However, treatment regimens involving trastuzumab had little or no effect for a subset of patients. Preliminary studies revealed WW-binding protein 2 (WBP2), an oncogenic transcription coactivator, to be coamplified with HER2 in 36% of HER2-positive breast cancers. We hypothesize that WBP2 regulates and correlates with the response of HER2-positive breast cancer to trastuzumab. The coexpression of WBP2 and HER2 in breast tumors was validated using IHC. The role and mechanism of WBP2 in regulating breast cancer response to trastuzumab was elucidated using in vitro, patient-derived xenograft and murine xenograft models. A multicenter retrospective study involving 143 patients given neoadjuvant trastuzumab-based chemotherapy was conducted to determine whether WBP2 expression correlates with pathologic complete response (pCR). Elevated expression of WBP2 significantly enhanced breast cancer's response to trastuzumab by augmenting trastuzumab-induced HER2 downregulation and cell-cycle arrest via inhibition of cyclin D expression. High level of WBP2 correlated with better pCR (67.19%) compared with low WBP2 level (26.58%). The highest response was observed in subgroups of patients with high WBP2-expressing tumors also aged below 50 years (77.78%) or were premenopausal in status (73.33%). Retrospectively, WBP2 demonstrated sensitivity of 80% to 81% and specificity of 76.5% to 80% in discriminating between patients showing pCR and non-pCR. WBP2 expression correlates with the response of HER2-positive breast cancer to trastuzumab-based neoadjuvant chemotherapy.


Variants of Concern Are Overrepresented Among Postvaccination Breakthrough Infections of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in Washington State.
Across 20 vaccine breakthrough cases detected at our institution, all 20 (100%) infections were due to variants of concern (VOCs) and had a median Ct of 20.2 (IQR, 17.1-23.3). When compared with 5174 contemporaneous samples sequenced in our laboratory, VOCs were significantly enriched among breakthrough infections (P < .05).


Circulating CD33 and its clinical value in acute leukemia.
CD33 is a cell surface antigen for committed myelomonocytic lineage. We explored the potential of detecting CD33 as cell-free circulating protein in patients with leukemia. We developed a quantitative bead-based immunoflow cytometry assay to measure cell-free circulating CD33 (cCD33) levels in the plasma of patients with acute leukemia, and correlated these results with corresponding clinical behavior. We measured cCD33 levels in the plasma of 48 healthy subjects and in patients with acute myelogenous leukemia (n = 98), acute lymphoblastic leukemia (n = 46), myelodysplastic syndrome (MDS) (n = 50), and myeloproliferative disorder (n = 49). Patients with acute myeloid leukemia and myeloproliferative disorders had significantly higher concentrations of cCD33 than the other patient groups and normal individuals (p = 0.0001), and among these groups, MDS patients displayed the lowest cCD33 levels (p = 0.02). Circulating CD33 values correlated positively with the CD33(+) blast cell counts in these patients. While there was no correlation between cCD33 levels and survival in acute myelogenous leukemia and MDS, higher cCD33 plasma concentrations did correlate with shorter survival in acute lymphoblastic leukemia (p = 0.03), and with shorter complete remission duration in acute myelogenous leukemia (p = 0.04) and MDS (p = 0.03). Circulating CD33 can be detected in the plasma from patients with leukemias, and cCD33 levels may have clinical implication, e.g., predictive and prognostic value, in these patients.


Policy stress and social support: Mental health impacts for Latinx Adults in the Southeast United States.
Latinx, and especially Mexican and Central American, adults in the United States (U.S.) have been detrimentally impacted by policies under the Trump administration that instilled continual stress associated with immigration enforcement. Both native- and foreign-born adults are affected when either they or loved ones are deported, resulting in family disruption. Though research has established a connection between Trump-era exclusionary immigration policies and mental health status of Latinx community members, data are notably lacking for Mexicans and Central Americans in the southeastern U.S., where enforcement activity is particularly high. To inform service provision and policymaking, the current study was designed to investigate stress associated with policies under the Trump administration on two mental health outcomes (depression and anxiety) among Mexican and Central American adults in the southeastern U.S. Additionally, the mediating effect of social support on the relationship was examined. A sample of 264 Mexican and Central American adults in the southeast was surveyed via an online platform. Using structural equation modeling (SEM), we employed a two-step procedure to first test a measurement model to ensure that the latent variables were explained by the indicators. In the next step, we estimated a structural model for hypothesized direct and indirect relationships between Trump-era policy stress, social support, and two outcome variables: depression and anxiety. Findings suggest that Trump-era policy stress significantly and positively predicted depression and anxiety symptoms. In addition, policy stress predicted lower levels of social support, which in turn, predicted more symptoms of both depression and anxiety. Findings can inform service provision and policymaking for a vulnerable community that underutilizes mental health services.


Bis(2-amino-1,3-benzothia-zol-3-ium) tetra-chloridozincate(II).
The asymmetric unit of the title compound, (C(7)H(7)N(2)S)(2)[ZnCl(4)], contains a network of 2-amino-benzothia-zolium cations and tetra-hedral [ZnCl(4)](2-) anions. The crystal packing is influenced by cation-to-anion N-H⋯Cl and C-H⋯Cl hydrogen bonds. The [ZnCl(4)](2-) anions have a distorded tetra-hedral geometry. Inter-molecular π-π stacking inter-actions are present between neighboring benzene rings, thia-zole and benzene rings and neighboring thia-zole rings [centroid-centroid distances = 3.711 (2), 3.554 (1), 3.536 (2) and 3.572 (1) Å].


Chapter 15 Development of a Nationwide Network for Ovarian Tissue Cryopreservation.
Ovarian tissue cryopreservation is gaining much interest since the publication of the first live birth after retransplantation of frozen-thawed tissue in 2004 (Donnez et al., Lancet 364:1405-1410, 2004). In contrast to cryopreservation of gametes and embryos, ovarian tissue freezing is a complex requiring a proper approach in order to make this a viable option for fertility preservation of cancer patients. Due to the need in terms of laboratory space, equipment, personnel, and adequate logistics, an ovarian tissue cryobank is most economic if managed as a centralized service unit that interacts with numerous clinics covering the surgical part. Transportation of ovarian tissue under appropriate conditions from the surgical unit to the cryobank for subsequent preparation and freezing has been shown to have no impact on cryo-survival (Schmidt et al., Hum Reprod 18:2654-2659, 2003; Isachenko et al., Fertil Steril 91:1556-1559, 2009). Several children have been born after retransplantation of such tissue that was derived from the cryobank in Bonn, Germany (Homepage FertiPROTEKT. http://www.fertiprotekt.de ). This cryobank is one of the largest in the world with more than 1300 tissue samples that were frozen from 2003 until today. It is integrated in the network FertiPROTEKT (Homepage FertiPROTEKT. http://www.fertiprotekt.de ) and is served by 108 surgical centers that are located all over Germany. The concept of this cryobank is a blueprint for success and has recently been used for another regionally centralized cryobank in Beijing, China. In this chapter the most important topics that need to be considered while creating a centralized cryobank within a national or regional network are highlighted.


Hydrochemical characterization and groundwater quality in Cauvery deltaic fluvial plains of Southern India.
Groundwater sources are drastically changing in their quantity and quality depending on local and regional level natural and anthropogenic factors, influencing their suitability for drinking and irrigation purposes. The objective of this study is to characterize the hydrochemistry and assess the groundwater quality in the fluvial deltaic plains of Cauvery river basin, Tamil Nadu, India. A total of 50 georeferenced groundwater samples were collected across Needamangalam block of Thiruvarur district and analyzed for major ions and hydrochemical processes. The results showed an ionic sequence of Cl- > Na+ > HCO3- > Mg2+ > Ca2+ > CO32- > SO42- > K+ based on their relative proportions. The scatter diagram indicated that groundwater chemistry was mostly influenced by weathering dominance followed by evaporation and silicate weathering. The dominant hydro-chemical facies were Na+-Cl--HCO3- type, Na+-Mg2+-Cl--HCO3- type, Na+-Cl--HCO3--CO32- type and Na+-Mg2+-Cl--HCO3--CO32- type influenced by the ion-exchange reaction. Most of the groundwater samples are suitable for drinking and irrigation except few with higher Na+ and Cl- content caused by the mixing of salt from fluvio-marine sources or agriculture return flow. The high sodium content in irrigation water may affect the soil hydraulic and nutrient properties in the long run.


Surgical Anatomy of Vertebral Artery in Relation to Atlantoaxial Instrumentation: A Cadaveric Study.
With the advent of pedicle screws and advanced instrumentation techniques, internal fixation and stabilization of upper cervical vertebrae are possible in fractures of an axis. However, the proximity of vertebral arteries (VAs) poses a unique challenge to surgeons during these procedures and can result in profound physical impairment to patients. Cadaveric studies contributing to fine anatomical details necessitate conducting such studies. After receiving due ethical permission, this descriptive cross-sectional study was carried out on 10 cadavers in the department of Anatomy, All India Institute of Medical Science (AIIMS) Rishikesh. Twenty VAs were dissected along their course, and measurements of parameters related to the axis and atlas vertebra were noted. The length of the pre-osseous segment related to the axis (VAX-1) on the right and left sides were from 3.8 to 14.5 mm (7.48±3.88 mm) and 4.46 to 10.5mm (6.94±2.01mm) respectively. The length of the osseous segment related to the axis (VAX-2) on the right side and left sides were from 6.82 to 31 mm (17.9±7.84mm) and 7.35 to 20 mm (15.6±4.53). The osseous segment of the VA related to the axis (VAX-2) shows genu (bend), which extends to a variable distance towards the midline. The mean distance of VA genu from the midline of the axis vertebral body on the right and left sides was 15.6mm and 17.5 mm, respectively. The percentage of superior articular facet (SAF) surface area of the axis occupied by the VA was 25-50% in nine and 50-75% in 11 cadavers, reflecting incomplete occupancy. The study suggests that for instrumentation of the axis vertebra in the midline, the minimum distance between the genu of both sides of VA segments, related to an osseous segment of the axis (VAX-2) and medial extent of the VA groove of the atlas, should be considered as a safe zone to minimize inadvertent VA injury. During atlantoaxial fixation through a posterior approach in interarticular, pars, and pedicle screws, the surgical anatomy of the VA in relation to the osseous segment of the VA within the transverse process of the axis should be kept in mind to avoid inadvertent VA injury.


Super-Linear-Threshold-Switching Selector with Multiple Jar-Shaped Cu-Filaments in the Amorphous Ge3 Se7 Resistive Switching Layer in a Cross-Point Synaptic Memristor Array.
The learning and inference efficiencies of an artificial neural network represented by a cross-point synaptic memristor array can be achieved using a selector, with high selectivity (Ion /Ioff ) and sufficient death region, stacked vertically on a synaptic memristor. This can prevent a sneak current in the memristor array. A selector with multiple jar-shaped conductive Cu filaments in the resistive switching layer is precisely fabricated by designing the Cu ion concentration depth profile of the CuGeSe layer as a filament source, TiN diffusion barrier layer, and Ge3 Se7 switching layer. The selector performs super-linear-threshold-switching with a selectivity of > 107 , death region of -0.70-0.65 V, holding time of 300 ns, switching speed of 25 ns, and endurance cycle of > 106 . In addition, the mechanism of switching is proven by the formation of conductive Cu filaments between the CuGeSe and Ge3 Se7 layers under a positive bias on the top Pt electrode and an automatic rupture of the filaments after the holding time. Particularly, a spiking deep neural network using the designed one-selector-one-memory cross-point array improves the Modified National Institute of Standards and Technology classification accuracy by ≈3.8% by eliminating the sneak current in the cross-point array during the inference process.


The pre-clinical behavioural pharmacology of 3,4-methylenedioxymethamphetamine (MDMA).
3,4-Methylenedioxymethamphetamine (MDMA) is a relatively novel drug of abuse and as such little is currently known of its behavioural pharmacology. This review aims to examine whether MDMA represents a novel class of abused drug. MDMA is known as a selective serotonergic neurotoxin in a variety of animal species but acutely it is a potent releaser and/or reuptake inhibitor of presynaptic serotonin, dopamine, noradrenaline, and acetylcholine. Interaction of these effects contributes to its behavioural pharmacology, in particular its effects on body temperature. Drug discrimination studies indicate that MDMA and related drugs produce unique interoceptive effects which have led to their classification as entactogens. This is supported by results from other behavioural paradigms although there is evidence for dose dependency of MDMA-specific effects. MDMA also produces conditioned place preference but is not a potent reinforcer in self-administration studies. These unique behavioural effects probably underlie its current popularity. MDMA is found in the street drug ecstasy but it may not be appropriate to equate the two as other drugs are routinely found in ecstasy tablets


[Hydrogen sulphide stimuli shorten exhalation during sleep as well as in wakefulness].
In the attentive waking state, pleasant odours often prolong inhalation while unpleasant odours often shorten the exhalation. It should be checked whether this induced breathing pattern is maintained even during sleep. 23 healthy normosmic adults were examined by polysomnography for one night and randomized pulsed either with H2S, phenylethyl alcohol (PEA) or CO2 via a flow olfactometer. The determination of the sleep stages was based on the standard polysomnography curves. From the breathing curves (respiratory nasal pressure breathing, thoracic or abdominal belt breathing) a low-artefact curve was selected and seven breaths were measured from their breathing signal around the trigger impulse. The duration of inspiration (DIN) and the duration of the expiration (DEX) were determined from the six prestimulatory breaths and from breath with the stimulus onset and thus defined a reaction index (RI) indicating whether there is a significant change in shortening or prolonging DIN and/or DEX. These different RI were corrected by the spontaneous baseline RI and analysed according to the type of stimulus and the sleep stages. Reaction indices with reductions of DEX were larger than spontaneous RI and RI for PEA and H2S stimulation. In the case of CO2 irritation, RI with reductions of DIN and DEX were larger than with olfactory stimulation. All RI were equal in their amount between sleep stages. In REM, the difference between olfactory RI with shortened DEX between H2S and PEA stimulation was greatest. When stimulated with an unpleasant smell in sleep, the respiratory reaction pattern from the awake state is preserved. RI with shortened DEX can be considered as an imperfect arousal.


Non-coding RNA in SARS-CoV-2: Progress toward therapeutic significance.
The recently developed pathogenic virus, SARS-CoV-2, was found in the Hubei Province, China. Giving rise to a broad spectrum of symptoms, SARS-CoV-2 rapidly spread across the globe, causing multi-systemic and dangerous complications, with death in extreme cases. Thereby, the number of research cases increases every day on preventing infection and treating its resulting damage. Accumulating evidence suggests noncoding RNAs (ncRNAs) are necessary for modifying virus infection and antiviral immune reaction, along with biological processes regulating SARS-CoV-2 and subsequent disease states. Therefore, understanding these mechanisms might provide a further understanding of the pathogenesis and feasible therapy alternatives against SARS-CoV2. Consequently, the molecular biology of SARS-CoV-2, ncRNA's role in its infection, and various RNA therapy tactics against the virus have been presented in this review section.


Positional nystagmus and vertigo due to a solitary brachium conjunctivum plaque.
The authors describe two patients suffering from demyelinating central nervous system disease who developed intense vertigo and downbeat nystagmus upon tilting their heads relative to gravity. Brain MRI revealed in both cases a single, small active lesion in the right brachium conjunctivum. The disruption of otolithic signals carried in brachium conjunctivum fibres connecting the fastigial nucleus with the vestibular nuclei is thought to be causatively involved, in agreement with a recently formulated model simulating central positional nystagmus. Insufficient otolithic information results in erroneous adjustment of the Listing's plane in off-vertical head positions, thus producing nystagmic eye movements.


Two-dimensional upper airway normative values in children aged 7 to 17 years.
To develop two-dimensional (2D) upper airway normative values in pediatric patients and correlate upper airway measurements with age. Cone beam computed tomography scans of 3738 patients were examined, and a sample of 61 patients, divided into two age groups (7-11 and 12-17 years), was used to reconstruct lateral cephalograms. The mean adenoid-nasopharynx (A/N) ratio was 0.45 and 0.44 for age groups 1 and 2, respectively. The mean PNS-ad1 and PNS-ad2 were 24 and 18.7 mm, respectively, for age group 1, and 26.7 and 21.2 mm, respectively, for age group 2. Additional normative values for the hyoid bone, airway space, uvula, and tongue are presented. The highest correlation with age was the upper airway length (r = 0.557, p < 0.001). Although cephalometric radiographs remain 2D reconstructions of three-dimensional structures, they have significant value and can aid clinicians in the screening process for pediatric sleep apnea.


Focal complex formation in adult cardiomyocytes is accompanied by the activation of beta3 integrin and c-Src.
In pressure-overloaded myocardium, our recent study demonstrated cytoskeletal assembly of c-Src and other signaling proteins which was partially mimicked in vitro using adult feline cardiomyocytes embedded in three-dimensional (3D) collagen matrix and stimulated with an integrin-binding Arg-Gly-Asp (RGD) peptide. In the present study, we improved this model further to activate c-Src and obtain a full assembly of the focal adhesion complex (FAC), and characterized c-Src localization and integrin subtype(s) involved. RGD dose response experiments revealed that c-Src activation occurs subsequent to its cytoskeletal recruitment and is accompanied by p130Cas cytoskeletal binding and focal adhesion kinase (FAK) Tyr925 phosphorylation. When cardiomyocytes expressing hexahistidine-tagged c-Src via adenoviral gene delivery were used for RGD stimulation, the expressed c-Src exhibited relocation: (i) biochemical analysis revealed c-Src movement from the detergent-soluble to the -insoluble cytoskeletal fraction and (ii) confocal microscopic analysis showed c-Src movement from a nuclear/perinuclear to a sarcolemmal region. RGD treatment also caused sarcolemmal co-localization of FAK and vinculin. Characterization of integrin subtypes revealed that beta3, but not beta1, integrin plays a predominant role: (i) expression of cytoplasmic domain of beta1A integrin did not affect the RGD-stimulated FAC formation and (ii) both pressure-overloaded myocardium and RGD-stimulated cardiomyocytes exhibited phosphorylation of beta3 integrin at Tyr773/785 sites but not beta1 integrin at Thr788/789 sites. Together these data indicate that RGD treatment in cardiomyocytes causes beta3 integrin activation and c-Src sarcolemmal localization, that subsequent c-Src activation is accompanied by p130Cas binding and FAK Tyr925 phosphorylation, and that these events might be crucial for growth and remodeling of hypertrophying adult cardiomyocytes.


Oncological transformation in vitro of hepatic progenitor cell lines isolated from adult mice.
Colorectal cancer cells can transfer the oncogene KRAS to distant cells, predisposing them to malignant transformation (Genometastasis Theory). This process could contribute to liver metastasis; besides, hepatic progenitor cells (HPCs) have been found to be involved in liver malignant neoplasms. The objective of this study is to determine if mouse HPCs-Oval cells (OCs)-are susceptible to incorporate Kras GAT (G12D) mutation from mouse colorectal cancer cell line CT26.WT and if OCs with the incorporated mutation behave like malignant cells. To achieve this, three lines of OCs in different conditions were exposed to CT26.WT cells through transwell co-culture for a week. The presence of KrasG12D and capacity to form tumors were analyzed in treated samples by droplet digital PCR and colony-forming assays, respectively. The results showed that the KrasG12D mutation was detected in hepatic culture conditions of undifferentiated OCs and these cells were capable of forming tumors in vitro. Therefore, OCs are susceptible to malignant transformation by horizontal transfer of DNA with KrasG12D mutation in an undifferentiated condition associated with the liver microenvironment. This study contributes to a new step in the understanding of the colorectal metastatic process.


The Role of Hypoxia and the Immune System in Tumor Radioresistance.
Radiotherapy is given to a majority of patients with cancer, and remains one of the most (cost)effective treatment options available [...].


[Identification of medical Dendrobium herbs by ISSR marker].
A molecular biology method was studied to identify medical Dendrobium and to provide a method for quality control of these plants. ISSR primer was screened through ISSR-PCR reaction according to its gene resolving power, and digital barcodes were established for identification. Screening 2 primers which Rp value above 8, This primers can identify medical Dendrobium from 6 kinds 8 groups. ISSR molecular maker technology is useful for identifying species and habitats of medical Dendrobium plants.


Favorable immune phenotype predicts successful implantation and pregnancy.
Immune markers that may predict IVF failure and successful implantation and pregnancy were studied. Favorable immune parameters were selected based on 90% of data of women who got pregnant and had uneventful pregnancy course and outcome in present IVF cycle. Immune phenotype and NK cell activity of peripheral blood of 123 women with multiple IVF failure were studied by flow cytometry. Some parameters that were out of favorable borders (elevated expression of CD56, CD158a in T lymphocytes, decreased levels of CD4 T lymphocytes, up-regulated expression of HLA DR in CD8+ T cells and NK cells, elevated number of NK cells and increased NK cytotoxicity, increased and decreased expression of CD158a and CD8 in NK cells) were considered to be immune deviations (ID) potentially predictive for IVF failure. In women with 0-1 ID implantation rate (IR) was 50.9% (27/53), with two ID - 42.8% (12/28), with three and more ID - 21.4% (9/42). IR in group with three ID was lower than in group with 0-1 ID (p<0.01, OR=3.8, CI: 1.52-9.48) and in group with two ID (p<0.05). Live birth rate (LBR) in women with 0-1 ID was 33.9%, with two ID - 28.5%, with three and more ID - 9.5%. LBR in group with three ID was lower than in group with 0-1 ID (p<0.01, OR=4.8, CI: 1.52-15.8) and in group with two ID (p<0.05). The absence or single ID seems to be more favorable for successful IVF program. Combination of ID may predict implantation failure to a greater degree than isolated ID. Multiple immune deviations form unfavorable "immune phenotype" for implantation and pregnancy development.


Surface analysis of pure and complex mucin coatings on a real-type substrate using individual and combined mBCA, ELLA, and ELISA.
In the past, we introduced the idea of using mucin coatings to improve biomaterials performance. Here, we evaluate non-radioactive methods for the analysis of pure and human host protein-containing (complex) mucin coatings on a real-type substrate (Thermanox). A common protein quantification assay (mBCA) was combined with mass-calibrated, enzyme-amplified assays based on lectin (ELLA) and antibody (ELISA) recognition, to determine the total and specific amounts of surface-associated proteins. Model studies showed the mBCA assay to be of limited use at low mass loads, and steric effects to influence the ELLA at high surface layer densities. Non-specific responses due to substrate interaction were low for the ELLA and ELISAs. Cross-reactions were observed during ELLA analysis of analytes sharing high degree of O-glycosylation. Combined mBCA-ELLA-ELISA analysis suggested that mucin desorption was low upon protein addition and that low concentrations of ELISA-determined protein for the complex coatings could be explained in terms of low accessibility of proteins to the bulk environment. Specifically, a methodology is presented for the determination of the fraction of surface-exposed, presumed bioactive proteins in a complex mucin coating. Finally, X-ray photoelectron spectroscopy and infrared reflectance spectroscopy combined with multivariate data analysis were proven useful in the evaluation of mucin-based coatings.


Inhibition of gastrulation in Xenopus embryos by an antibody against a cathepsin L-like protease.
An antibody against cathepsin L-like protease (AACLP) was injected into one cell of 2-celled Xenopus embryos. The blastopores of AACLP-injected embryos either did not invaginate or failed to complete invagination. As a result of this failure to complete gastrulation, the body axes could not form normally and tail bud stage embryos were bent dorsally. Embryos injected with a control antibody (CA) developed normally through the tadpole stage. Mesodermal induction was not inhibited in embryos exhibiting this AACLP-induced gastrulation defect, but the mesodermal structure of these embryos was organized incorrectly due to the defective gastrulation during the early stages.


Oct4 protein remains in trophectoderm until late stages of mouse blastocyst development.
Oct-4, the marker of pluripotent cells, is crucial for murine preimplantation development. During the formation of the blastocyst Oct-4 is downregulated in the trophectoderm (TE) and its expression becomes restricted to the inner cell mass (ICM). In order to determine the exact timing of the disappearance of Oct-4 protein from TE we analyzed the localization and level of Oct-4 at different stages of blastocyst development. The presence of Oct-4 protein was determined by immunohistochemistry using confocal microscopy. We found that the downregulation of Oct-4 protein in TE of mouse blastocysts progresses gradually during development, and Oct-4 protein persists in some of the TE cells at least until the expanded blastocyst (120-140 cells) stage. Our findings indicate that the switching-off of the Oct-4 expression is not necessary for the trophectoderm formation. The complete elimination of Oct-4 protein from TE occurs at the period of blastocyst implantation, when lack of Oct-4 is required for the proper functioning of the trophectoderm.


Dosing in obesity: a simple solution to a big problem.
The global epidemic of obesity has led to an increased prevalence of chronic diseases and need for pharmacological intervention. However, little is known about the influence of obesity on the drug exposure profile, resulting in few clear dosing guidelines for the obese. Here we present a semi-mechanistic model for lean body weight (LBW) that we believe is sufficiently robust to quantify the influence of body composition on drug clearance, and is therefore an ideal metric for adjusting chronic dosing in the obese.


Discriminating between virilizing ovary tumors and ovary hyperthecosis in postmenopausal women: clinical data, hormonal profiles and image studies.
The presence of virilizing signs associated with high serum androgen levels in postmenopausal women is rare. Virilizing ovarian tumors (VOTs) and ovarian stromal hyperthecosis (OH) are the most common etiologies in virilized postmenopausal women. The differential diagnosis between these two conditions is often difficult. To evaluate the contribution of clinical features, hormonal profiles and radiological studies to the differential diagnosis of VOT and OH. A retrospective study. A tertiary center. Clinical data, hormonal status (T, E2, LH and FSH), pelvic images (transvaginal sonography and MRI) and anatomopathology were reviewed. Thirty-four postmenopausal women with a diagnosis of VOT (13 women) and OH (21 women) were evaluated retrospectively. Clinical signs of hyperandrogenism were more prevalent in the VOT group than the OH group. Although the VOT group showed higher T and E2 levels and lower gonadotropin levels than the OH group, a great overlap occurred among the hormone levels. A pelvic MRI provided an accurate differentiation of these two conditions. In this group of patients, the main features contributing to the differential diagnosis of VOT and OH were serum levels of testosterone and gonadotropins and the presence of an ovarian nodule identified on the MRI. Although the association of clinical, hormonal and radiological features contributes to the differential diagnosis of these two conditions, histopathological analysis remains the gold standard for the diagnosis of ovarian hyperandrogenism in postmenopausal women.


Cutting edge: cross-presentation of cell-associated antigens to MHC class I molecule is regulated by a major transcription factor for heat shock proteins.
The ability for the professional APC to cross-present Ag to MHC class I from parenchymal cells is essential for priming as well as tolerance of CD8+ T cells against intracellular Ags. Since cross-presentations of non-cell-associated free Ags are inefficient, the roles of molecular chaperones or heat shock proteins (HSPs) in chaperoning Ags to APCs have been postulated. We herein genetically addressed this hypothesis using mice that were defective of heat shock factor 1 (Hsf1), a major transcription factor for HSPs. Hsf1(-/-) mice have a decreased expression of several HSPs including HSP90 and HSP70. Using multiple Ag systems, we demonstrated that cross-priming of Ag-specific CD8+ T cells was inefficient when Ag expression was restricted to Hsf1(-/-) non-APCs. Our study provides the first genetic evidence for the roles of Hsf1 in regulating cross-presentation of MHC class I-associated Ags.


Staff Self-Efficacy and Staff Outcome Expectations for Function-Focused Care: Rasch Testing.
The purpose of this study was to test the reliability and validity for two new scales. We used baseline data from a worksite health promotion project with 98 long-term care staff and Pearson correlations and Rasch analysis testing. For Staff Self-Efficacy for Function Focused Care and Staff Outcome Expectations for Function Focused Care item, reliability was .80 and .83, respectively. All items except one had acceptable INFIT and OUTFIT mean square statistics. Both new scales were significantly correlated (r = .342, p = .007), but neither of the scales were correlated with participants' baseline exercise levels (r = .014, p = .918; r = .092, p = .454, respectively). There was some evidence of validity and internal consistency for both scales. Suggestions are provided to improve the measures for future use.


Renal stone disease.
Renal stone disease may be seen as a clinical symptom of an underlying pathological process predisposing to crystallization within the renal tract. Renal stones may be comprised of calcium salts, uric acid, cystine and various other insoluble complexes. Nephrolithiasis may be the manifestation of rare single gene disorders or part of more common idiopathic renal stone-forming diseases. Molecular genetics has allowed significant progress to be made in our understanding of certain stone-forming conditions. The molecular defect underlying single gene disorders often contributes to a significant metabolic risk factor for stone formation. In contrast, idiopathic renal stone formation relates to the interplay of environmental, dietary and genetic factors, with hypercalciuria being the most commonly found metabolic risk factor. Candidate genes for idiopathic stone formers have been identified using numerous approaches, some of which are outlined here. Despite this, the genetic basis underlying familial hypercalciuria and calcium stone formation remains elusive. The molecular basis of other metabolic risk factors such as hyperuricosuria, hyperoxaluria and hypocitraturia is being unraveled and is allowing new insights into renal stone pathogenesis. The discovery of both rare and common molecular defects leading to renal stones will hopefully increase our understanding of the disease pathogenesis. Such knowledge will allow screening for genetic defects and the use of specific drug therapies in order to prevent renal stone formation.


Hippocampal formation alterations differently contribute to autobiographic memory deficits in mild cognitive impairment and Alzheimer's disease.
Autobiographical memory (AM) is part of declarative memory and includes both semantic and episodic aspects. AM deficits are among the major complaints of patients with Alzheimer's disease (AD) even in early or preclinical stages. Previous MRI studies in AD patients have showed that deficits in semantic and episodic AM are associated with hippocampal alterations. However, the question which specific hippocampal subfields and adjacent extrahippocampal structures contribute to deficits of AM in individuals with mild cognitive impairment (MCI) and AD patients has not been investigated so far. Hundred and seven participants (38 AD patients, 38 MCI individuals and 31 healthy controls [HC]) underwent MRI at 3 Tesla. AM was assessed with a semi-structured interview (E-AGI). FreeSurfer 5.3 was used for hippocampal parcellation. Semantic and episodic AM scores were related to the volume of 5 hippocampal subfields and cortical thickness in the parahippocampal and entorhinal cortex. Both semantic and episodic AM deficits were associated with bilateral hippocampal alterations. These associations referred mainly to CA1, CA2-3, presubiculum, and subiculum atrophy. Episodic, but not semantic AM loss was associated with cortical thickness reduction of the bilateral parahippocampal and enthorinal cortex. In MCI individuals, episodic, but not semantic AM deficits were associated with alterations of the CA1, presubiculum and subiculum. Our findings support the crucial role of CA1, presubiculum, and subiculum in episodic memory. The present results implicate that in MCI individuals, semantic and episodic AM deficits are subserved by distinct neuronal systems.


Dietary intake of deuterium oxide decreases cochlear metabolism and oxidative stress levels in a mouse model of age-related hearing loss.
Age-related hearing loss (ARHL) is the most prevalent sensory disorder in the elderly. Currently, no treatment can effectively prevent or reverse ARHL. Aging auditory organs are often accompanied by exacerbated oxidative stress and metabolic deterioration. Here, we report the effect of deuterated oxygen (D2O), also known as "heavy water", mouse models of ARHL. Supplementing the normal mouse diet with 10% D2O from 4 to 9 weeks of age lowered hearing thresholds at selected frequencies in treated mice compared to untreated control group. Oxidative stress levels were significantly reduced and in the cochlear duct of treated vs. untreated mice. Through metabolic flux analysis, we found that D2O mainly slowed down catabolic reactions, and may delay metabolic deterioration related to aging to a certain extent. Experiments confirmed that the Nrf2/HO-1/glutathione axis was down-regulated in treated mice. Thus, D2O supplementation can hinder ARHL progression in mouse models by slowing the pace of metabolism and reducing endogenous oxidative stress production in the cochlea. These findings open new avenues for protecting the cochlea from oxidative stress and regulating metabolism to prevent ARHL.


Pooled CRISPR-activation screening coupled with single-cell RNA-seq in mouse embryonic stem cells.
CRISPR/Cas9 screens are a powerful approach to identify key regulators of biological processes. By combining pooled CRISPR/Cas9 screening with single-cell RNA-sequencing readout, individual perturbations can be assessed in parallel both comprehensively and at scale. Importantly, this allows gene function and regulation to be interrogated at a cellular level in an unbiased manner. Here, we present a protocol to perform pooled CRISPR-activation screens in mouse embryonic stem cells using 10× Genomics scRNA-seq as a readout. For complete information on the generation and use of this protocol, please refer to Alda-Catalinas et al. (2020).


Comparison of surveillance and control activities of infection control nurses and infection control laboratorians in United States hospitals, 1976-1977.
To study the impact of the professional background of infection control personnel, we compared the characteristics and activities of 107 infection control nurses (ICNs) with those of 13 infection control laboratorians (ICLs), all in hospitals with 300 beds or more. Although the two groups performed similarly in many respects. ICNs spent more time teaching, whereas ICLs spent more time and appeared more proficient in investigating outbreaks. Staff nurses at hospitals with ICNs found the infection control person more visible on the wards and more available for discussing infection control matters. ICNs appeared less hesitant to speak up to personnel not following correct handwashing techniques. ICNs and ICLs appear to offer different skills that should be considered when filling different infection control positions.


Why most patients with hepatitis delta virus infection are seronegative for hepatitis B e antigen. A prospective controlled study.
A prospective age/sex matched control and follow-up study was conducted to explore the reason(s) for the association of hepatitis delta virus (HDV) infection with hepatitis B e antigen (HBeAg) negative hepatitis B surface antigen (HBsAg) carrier state. Over a 3-year period, a total of 110 patients (104 males and six females) with acute HDV superinfection were documented in our unit. Twenty-four (21.8%) of them were HBeAg positive at the onset of acute HDV infection. In the control study, 110 age- and sex-matched asymptomatic HBsAg carriers with normal serum transaminase, as well as 110 age- and sex-matched patients with chronic type B hepatitis were randomly selected from the computer files of the same 3-year period of entry. The prevalence of serum HBeAg in patients with HDV infection was similar to that of asymptomatic HBsAg carriers (20.9%), but significantly lower than that of the patients with chronic type B hepatitis (72.7%). In a follow-up study of 16 HBeAg-positive patients with HDV infection, eight (50%) cleared HBeAg and three (18.8%) seroconverted to anti-HBe within 3 months. The HBeAg clearance rate was significantly higher than for chronic type B hepatitis and asymptomatic carriers (p less than 0.01). The results suggest that the low prevalence of serum HBeAg in HDV infection simply reflects the HBeAg/anti-HBe status of the asymptomatic HBsAg carriers in the population under study. Also in some patients HDV superinfection may itself suppress HBV and thus clear HBeAg.


The SARS-CoV-2 Exerts a Distinctive Strategy for Interacting with the ACE2 Human Receptor.
The COVID-19 disease has plagued over 200 countries with over three million cases and has resulted in over 200,000 deaths within 3 months. To gain insight into the high infection rate of the SARS-CoV-2 virus, we compare the interaction between the human ACE2 receptor and the SARS-CoV-2 spike protein with that of other pathogenic coronaviruses using molecular dynamics simulations. SARS-CoV, SARS-CoV-2, and HCoV-NL63 recognize ACE2 as the natural receptor but present a distinct binding interface to ACE2 and a different network of residue-residue contacts. SARS-CoV and SARS-CoV-2 have comparable binding affinities achieved by balancing energetics and dynamics. The SARS-CoV-2-ACE2 complex contains a higher number of contacts, a larger interface area, and decreased interface residue fluctuations relative to the SARS-CoV-ACE2 complex. These findings expose an exceptional evolutionary exploration exerted by coronaviruses toward host recognition. We postulate that the versatility of cell receptor binding strategies has immediate implications for therapeutic strategies.


Alcohol Screening During US Primary Care Visits, 2014-2016.
Unhealthy alcohol use is a significant health issue for the US population. The US Preventive Services Task Force (USPSTF) recommends screening adults 18 years or older for unhealthy alcohol use during primary care visits. To evaluate alcohol screening among ambulatory visits made by US adult primary care patients and identify characteristics predictive of alcohol screening. A series of cross-sectional analysis of the National Ambulatory Medical Care Survey (NAMCS) data collected from 2014 to 2016 was used to examine US primary care providers' use of alcohol screening questionnaires and delivery of counseling on alcohol use. A total of 19,213 visits made by patients aged 18 years or older to a US primary care physician trained in family medicine or internal medicine. Administration of a validated alcohol screening questionnaire and counseling/education on alcohol use. Variation in alcohol screening by patient demographic characteristics, reason for office visit, length of office visit, chronic medical conditions, evaluation by assigned primary care physician, new patient to practice, and region. Alcohol screening with a validated questionnaire occurred during 2.6% (95% Cl: 0.9%, 4.3%) of visits. Alcohol counseling, provided either by the physician or by referral, was documented in 0.8% (95% Cl: 0.3%, 1.3%) of visits. Screening was significantly more likely if patients were seen by their assigned primary care physician (adjOR 4.38 (95% Cl: 1.41, 13.61)), a new patient to the practice (adjOR 4.18 (95% Cl: 2.30, 7.79)), or had several chronic medical conditions (adjOR 3.40 (95% Cl: 1.48, 7.78)). Patients' sex, race/ethnicity, age group, or length of appointment time was not associated with screening for unhealthy alcohol use. Screening for unhealthy alcohol use using a validated questionnaire is uncommonly performed during US primary care visits. Interventions or incentives may be needed to increase uptake of USPSTF alcohol screening recommendations.


Simulating homeostatic, allostatic and goal-directed forms of interoceptive control using active inference.
The adaptive regulation of bodily and interoceptive parameters, such as body temperature, thirst and hunger is a central problem for any biological organism. Here, we present a series of simulations using the framework of active inference to formally characterize interoceptive control and some of its dysfunctions. We start from the premise that the goal of interoceptive control is to minimize a discrepancy between expected and actual interoceptive sensations (i.e., a prediction error or free energy). Importantly, living organisms can achieve this goal by using various forms of interoceptive control: homeostatic, allostatic and goal-directed. We provide a computationally-guided analysis of these different forms of interoceptive control, by showing that they correspond to distinct generative models within Active inference. We discuss how these generative models can support empirical research through enabling fine-grained predictions about physiological and brain signals that may accompany both adaptive and maladaptive interoceptive control.


Abnormal resting-state function within language network and its improvement among post-stroke aphasia.
Several studies with resting-state magnetic resonance imaging (rs-fMRI) have examined functional impairments and plasticity within language network in patients with post-stroke aphasia (PSA). However, there is still ubiquitous inconsistency across these studies, partly due to restricted to very small sample size and the absence of validation with follow-up data. In the current study, we aimed at providing relatively strong evidence to support functional impairments and its reorganization in PSA. Here, the amplitude of low frequency fluctuations (ALFF) and functional connectivity were used to assess functional alterations of PSA with moderate sample size at baseline (thirty-five PSA patients and thirty-five healthy controls). Functional abnormalities at baseline were observed whether improved among sixteen follow-up patients. Compared with controls, PSA at baseline presented decreased ALFF in the left inferior frontal gyrus (IFG) and decreased functional connectivity of the left IFG with the bilateral supplementary motor area (SMA) and right superior temporal gyrus (STG). The decreased ALFF in IFG, decreased IFG-SMA and IFG-STG connectivity were enhanced among follow-up patients and was synchronized with language-performance improvement. Our results revealed reduced intrinsic neural activity and inter-connections within language network in PSA, which would be normalized synchronously as the improvement of language performance.


[Medical prohibition of pain therapy in cancer patients - an incredible case report].
Case presentation of a 59-year old female patient, who has been treated for her cancer by a physician of internal medicine with peroxodisulphuric acid. For this therapy to be successful - according to the physician - all drugs especially the analgetics had to be stopped. Along with this case presentation the statement by the German Medical Association is shown and legal assessment is discussed.


[Physiological and psychophysiological indices dynamics in operators with three-shift working day in different working conditions].
It was shown that intense operator work in pulp and paper industry (bleachers and chlorinators) with changes in shift time (day, evening, night) causes fatigue. The fatigue in operators was indicated by increasing number of errors in sensomotor coordination test and coordination index, and among chlorinators it was accompanied by deterioration of state of health. Manifestations of 24 hours' rhythm of physiological functions were detected by changes in body temperature (among bleachers), heart rate, blood pressure and sensomotor coordination indices (all operators). Harmful chemical substances (chlorine and chlorine dioxide) in the air negatively influence psychophysiological status among clorinators.


Oxidant-mediated AA release from astrocytes involves cPLA(2) and iPLA(2).
Excessive generation of reactive oxygen species (ROS) in the central nervous system (CNS) is a leading cause of neuronal injury. Despite yet unknown mechanisms, oxidant compounds such as H(2)O(2) have been shown to stimulate the release of arachidonic acid (AA) in a number of cell systems. In this study, H(2)O(2) and menadione, a compound known to release H(2)O(2) intracellularly, were used to examine the phospholipases A(2) (PLA(2)) responsible for AA release from primary murine astrocytes. Both H(2)O(2) and menadione dose-dependently stimulated AA release, and the release mediated by H(2)O(2) was completely inhibited by catalase. H(2)O(2) also stimulated phosphorylation of extracellular signal-regulated kinases (ERK1/2) and cytosolic phospholipase A(2) (cPLA(2)). However, complete inhibition of cPLA(2) phosphorylation by U0126, an inhibitor for mitogen-activated protein kinase kinase (MEK) and GF109203x, a nonselective PKC inhibitor preferring the conventional and novel isoforms, only reduced H(2)O(2)-stimulated AA release by 50%. MAFP, a selective, active, site-directed, irreversible inhibitor of both cPLA(2) and the Ca(2+)-independent iPLA(2), nearly completely inhibited H(2)O(2)-mediated AA release; but, HELSS, a potent irreversible inhibitor of iPLA(2), only inhibited H(2)O(2)-mediated AA release by 40%. Along with the observation that H(2)O(2)-mediated AA release was only partially inhibited upon chelating intracellular Ca(2+) by BAPTA, these results indicate the involvement of both cPLA(2) and iPLA(2) in H(2)O(2)-mediated AA release in murine astrocytes.


Mechanistic interaction of ciprofloxacin on zeolite modified seaweed (Sargassum crassifolium) derived biochar: Kinetics, isotherm and thermodynamics.
Modification of biochar for efficient removal of antibiotics from water could be a valuable approach in the environmental applications. In this study, a brown seaweed (Sargassum crassifolium) was pyrolyzed at 500 °C and the obtained biochar (SWBC) was modified with zeolite through the slurry method maintaining the ratio at 1:5 (zeolite: biochar) (SWBC-Z). Batch adsorption experiments were conducted to evaluate the adsorption tendency of SWBC and SWBC-Z for the removal of ciprofloxacin (CPX) from water via pH edge, kinetics, isotherm and thermodynamic experiments. The highest adsorption was in the pH range of 6.5-8, supported by the electrostatic attractions and hydrogen bonding with zwitterionic CPX. Experimental kinetics data was well-fitted to the pseudo-second-order and Elovich models (R2 of 0.992 and 0.976, respectively), while the Langmuir and Freundlich isotherm models best described the isotherm data (R2 of 0.954 and 0.976, respectively). The maximum adsorption capacity of 93.65 mg g-1 was recorded for the SWBC-Z. The models predicted chemisorption and physisorption interactions on the heterogenous biochar surface. Well-defined peaks of silanol groups in the FTIR spectrum of SWBC-Z and its electron microscopy confirmed the incorporation of zeolite minerals. Post adsorption FTIR analysis elucidated the changes in the surface functional groups of the SWBC-Z. Thermodynamic data revealed spontaneous and exothermic reaction between CPX and both the biochars. It was concluded that modification of pristine biochar with zeolite imparted greater surface area and additional active sites, which subsequently enhanced the overall CPX adsorption by the SWBC-Z.


Advances in activity-based diagnostics for infectious disease and microbiome health.
In the human body, pathogens and the endogenous microbiome produce enzymes that aid in replication and survival. The activity from these enzymes as well as energy-dependent transport processes can be used as functional biomarkers for pathogen identification, antimicrobial treatment monitoring, and surveillance of microbiome health. To produce visual and/or quantifiable readouts from this activity, concepts from chemical biology and nanomedicine have been utilized to develop signal-producing probes for patient samples or for direct administration in vivo. In the context of infection, activity-based diagnostics offer several potential advantages over current diagnostics including the ability to differentiate between active infection and sterile inflammation, which is made possible by targeting microbial enzymes with orthogonal activity to that of the host. In this review, we discuss new developments in the making of activity-based infection diagnostics and the beginnings of microbiome activity-based diagnostics.


Metabolomic profiling in a Hedgehog Interacting Protein (Hhip) murine model of chronic obstructive pulmonary disease.
Genetic variants annotated to the hedgehog interacting protein (HHIP) are robustly associated with chronic obstructive pulmonary disease (COPD). Hhip haploinsufficiency in mice leads to increased susceptibility towards the development of emphysema following exposure to chronic cigarette smoke (CS). To explore the molecular pathways which contribute to increased susceptibility, we performed metabolomic profiling using high performance liquid chromatography tandem mass spectroscopy (LC/MS-MS) on plasma, urine, and lung tissue of Hhip +/- heterozygotes and wild type (Hhip +/+) C57/BL6 mice exposed to either room-air or CS for six months. Univariate comparisons between groups were made with a combined fold change ≥2 and Student's t-test p-value < 0.05 to denote significance; associations with mean alveolar chord length (MACL), a quantitative measure of emphysema, and gene-by-environment interactions were examined using empiric Bayes-mediated linear models. Decreased urinary excretion of cotinine despite comparable plasma levels was observed in Hhip +/- heterozygotes; a strong gene-by-smoking association was also observed. Correlations between MACL and markers of oxidative stress such as urinary methionine sulfoxide were observed in Hhip +/- but not in Hhip +/+ mice. Metabolite set enrichment analyses suggest reduced antioxidant capacity and alterations in macronutrient metabolism contribute to increased susceptibility to chronic CS-induced oxidative stress in Hhip haploinsufficiency states.


Molecular Imaging with Single-Walled Carbon Nanotubes.
Nanoparticle-based molecular imaging has emerged as an interdisciplinary field which involves physics, chemistry, engineering, biology, and medicine. Single-walled carbon nanotubes (SWCNTs) have unique properties which make them suitable for applications in a variety of imaging modalities, such as magnetic resonance, near-infrared fluorescence, Raman spectroscopy, photoacoustic tomography, and radionuclide-based imaging. In this review, we will summarize the current state-of-the-art of SWCNTs in molecular imaging applications. Multifunctionality is the key advantage of nanoparticles over traditional approaches. Targeting ligands, imaging labels, therapeutic drugs, and many other agents can all be integrated into the nanoparticle to allow for targeted molecular imaging and molecular therapy by encompassing many biological and biophysical barriers. A multifunctional, SWCNT-based nanoplatform holds great potential for clinical applications in the future.


Upper airway obstruction caused by bilateral giant tonsilloliths.
Tonsilloliths are rare dystrophic calcifications caused by chronic inflammation of the tonsils. They are usually small and occur on one side. Herein, we report a case involving a 75-year-old man presenting odynophagia and progressive dyspnea for days who was found by computed tomography image to have bilateral giant tonsilloliths. Hyperdensity lesions were found over the tonsillar fossa on both sides. Tonsillectomy was performed leading to immediate relief of symptoms. A review of relevant literature revealed that most patients with tonsilloliths are asymptomatic and need only conservative treatment. Severe symptoms such as dyspnea are extremely rare. Although tonsillolith can be easily diagnosed by computed tomography, otolaryngologists should be careful to differentiate this entity.


Pharmacological characterization of second generation FXR agonists as effective EphA2 antagonists: A successful application of target hopping approach.
It is well demonstrated the key role of Eph-ephrin system, specifically of EphA2 receptor, in supporting tumor growth, invasion, metastasis and neovascularization. We previously identified FXR agonists as eligible antagonists of Eph-ephrin system. Herein we characterize new commercially available FXR (Farnesoid X Receptor) agonists as potential Eph ligands including Cilofexor, Nidufexor, Tropifexor, Turofexorate isopropyl and Vonafexor. Our exploration based on molecular modelling investigations and binding assays shows that Cilofexor binds specifically and reversibly to EphA2 receptor with a Ki value in the low micromolar range. Furthermore, Cilofexor interferes with the phosphorylation of EphA2 and the cell retraction and rounding in PC3 prostate cancer cells, both events depending on EphA2 activation. In conclusion, we can confirm that target hopping can be a successful approach to discover new moiety of protein-protein inhibitors.