Cytokine dysregulation associated with exam stress in healthy medical students.
The mechanisms of stress-related immune alterations have not been fully elucidated. Cell-mediated immune responses as well as antibody and certain cytokines are reported as being suppressed during times of high stress. However, the role of suppression vs dysregulation has not been established in human stress models. The effect of exam stress on regulatory cytokines in 16 healthy medical students was assessed by measuring type-1 (IFN-gamma) and type-2 (IL-10) cytokines from 72-h PHA/PMA-stimulated PBMC 4 weeks before and 48 h after exams. Results demonstrated decreased IFN-gamma accompanied by increased IL-10 during exam stress that resulted in a decreased IFN-gamma:IL-10 ratio. There was a significant correlation between the cytokine response to PHA/PMA and number and subjective adjustment to daily hassles. Additionally, students who reported greater levels of loneliness also reported greater numbers of and poorer subjective adjustment to hassles. The differences were consistent in both males and females but did not correlate with AM cortisol levels. Additionally, when individuals were grouped into high vs low preexam hassle levels, the type-1/type-2 shift in the IFN-gamma:IL-10 ratio occurred in the low hassles group only. These data suggest that psychologically stressful situations shift type-1/type-2 cytokine balance toward type-2 and result in an immune dysregulation rather than overall immunosuppression. This may partially explain the increased incidence of type-2-mediated conditions such as increased viral infections, latent viral expression, allergic/asthmatic reactions, and autoimmunity reported during periods of high stress.


Structure and Scm3-mediated assembly of budding yeast centromeric nucleosomes.
Much controversy exists regarding the structural organization of the yeast centromeric nucleosome and the role of the nonhistone protein, Scm3, in its assembly and architecture. Here we show that the substitution of H3 with its centromeric variant Cse4 results in octameric nucleosomes that organize DNA in a left-handed superhelix. We demonstrate by single-molecule approaches, micrococcal nuclease digestion and small-angle X-ray scattering that Cse4-nucleosomes exhibit an open conformation with weakly bound terminal DNA segments. The Cse4-octamer does not preferentially form nucleosomes on its cognate centromeric DNA. We show that Scm3 functions as a Cse4-specific nucleosome assembly factor, and that the resulting octameric nucleosomes do not contain Scm3 as a stably bound component. Taken together, our data provide insights into the assembly and structural features of the budding yeast centromeric nucleosome.


Reproductive performance of genetically engineered mice housed in different housing systems.
The genetically engineered mice require special husbandry care and are mainly housed in Individually Ventilated Cage (IVC) systems and Static Micro Isolator Cages (SMIC) to minimize the risk for spreading undesirable microorganisms. However, the static micro isolation cage housing like SMIC are being replaced with IVC systems in many facilities due to a number of benefits like a higher density housing in limited space, better protection from biohazards and allergens and decreased work load due to decreased frequency of cage changing required in this system. The purpose of this study was to examine the reproductive performance of genetically engineered mice housed in individually ventilated cages (IVC) and Static Micro Isolator Cages (SMIC). When the B6C3-Tg (APPswe, PSEN1dE9) 85Dbo/Mmjax transgenic mice were housed in these two housing systems, the number of litters per dam, number of pups born per dam and number of pups weaned per dam were found to be slightly higher in the IVC as compared to the SMIC but the difference was not significant (P<0.05). In case of Growth Associated Protein 43 (GAP-43) knockout mice, the number of litters born per dam and the number of pups born per dam were marginally higher in the IVC as compared to those housed in SMIC but the difference was not significant (P<0.05). Only the number of pups weaned per dam were found to be significantly higher as compared to those housed in the SMIC system at P<0.05.


Within-Day Energy Balance and Metabolic Suppression in Male Collegiate Soccer Players.
Metabolic suppression due to relative energy deficiency can cause various physiological impairments in athletes. The purpose of this study was to evaluate within-day energy balance (WDEB) and the ratio between measured and predicted resting energy expenditure (REEratio) and to investigate the relationships between the markers of metabolic suppression. Ten male collegiate soccer players completed a 7-day food diary, physical activity, and heart rate records during the training and rest days. Energy intake (EI) and energy expenditure (EE) were analyzed to evaluate WDEB components. Body composition was measured using dual-energy X-ray absorptiometry (DXA), and blood sampling was conducted for hormonal analysis. The REE was measured using the Douglas bag method and predicted using the DXA-predicted method to calculate the REEratio. Participants were categorized into the normal (REEratio ≥ 0.94, n = 5) and suppressed (REEratio < 0.94, n = 5) groups. There were no group differences in the components of WDEB, except diet-induced thermogenesis (DIT), but EI was significantly higher in the normal group than in the suppressed group (7-day total: 3660 ± 347 vs. 3024 ± 491 kcal/day, p = 0.046 and rest days: 3772 ± 463 vs. 2796 ± 800 kcal/day, p = 0.046). Analysis of hormonal markers of metabolic suppression only showed a significant positive association between insulin-like growth factor-1 (IGF-1) and REEratio (r = 0.771, p = 0.009). The relationships between metabolic suppression and the markers of energy deficiency were inconclusive. There are possible associations of insufficient EI and IGF-1 levels with metabolic suppression, and further study is required to understand energy deficiency in male soccer players.


Congenital nevi less than or equal to 10 cm as precursors to melanoma. 52 cases, a review, and a new conception.
Fifty-two congenital melanocytic nevi (CMN) as precursors to melanoma or severe melanocytic dysplasia were reviewed macroscopically and microscopically. Forty-eight "small" CMN measured less than 10 cm in diameter. Histologically, only five reached to the lower third of the dermis or subcutis ("deep type"); the remaining 47 were limited to the upper two thirds of the corium ("superficial type"). There were 47 invasive melanomas, two in situ melanomas, and three severe focal melanocytic dysplasias. All melanomas were of "epidermal" origin and primarily of the superficial spreading type. The age at diagnosis ranged from 18 to 79 years. Prepubertal melanomas were not observed. Melanoma may also arise on small CMN. The most frequent origin of the melanomas in small CMN and the preferred age at manifestation seem to be different from that for giant nevi.


Quantifying the effects of elastic collisions and non-covalent binding on glutamate receptor trafficking in the post-synaptic density.
One mechanism of information storage in neurons is believed to be determined by the strength of synaptic contacts. The strength of an excitatory synapse is partially due to the concentration of a particular type of ionotropic glutamate receptor (AMPAR) in the post-synaptic density (PSD). AMPAR concentration in the PSD has to be plastic, to allow the storage of new memories; but it also has to be stable to preserve important information. Although much is known about the molecular identity of synapses, the biophysical mechanisms by which AMPAR can enter, leave and remain in the synapse are unclear. We used Monte Carlo simulations to determine the influence of PSD structure and activity in maintaining homeostatic concentrations of AMPARs in the synapse. We found that, the high concentration and excluded volume caused by PSD molecules result in molecular crowding. Diffusion of AMPAR in the PSD under such conditions is anomalous. Anomalous diffusion of AMPAR results in retention of these receptors inside the PSD for periods ranging from minutes to several hours in the absence of strong binding of receptors to PSD molecules. Trapping of receptors in the PSD by crowding effects was very sensitive to the concentration of PSD molecules, showing a switch-like behavior for retention of receptors. Non-covalent binding of AMPAR to anchored PSD molecules allowed the synapse to become well-mixed, resulting in normal diffusion of AMPAR. Binding also allowed the exchange of receptors in and out of the PSD. We propose that molecular crowding is an important biophysical mechanism to maintain homeostatic synaptic concentrations of AMPARs in the PSD without the need of energetically expensive biochemical reactions. In this context, binding of AMPAR with PSD molecules could collaborate with crowding to maintain synaptic homeostasis but could also allow synaptic plasticity by increasing the exchange of these receptors with the surrounding extra-synaptic membrane.


Detection of ricin contamination in ground beef by electrochemiluminescence immunosorbent assay.
Ricin is a highly toxic protein present in the seeds of Ricinus communis (castor), grown principally as a source of high quality industrial lubricant and as an ornamental. Because ricin has been used for intentional poisoning in the past and could be used to contaminate food, there is a need for analytical methodology to detect ricin in food matrices. A monoclonal antibody-based method was developed for detecting and quantifying ricin in ground beef, a complex, fatty matrix. The limit of detection was 0.5 ng/g for the electrochemiluminescence (ECL) method and 1.5 ng/g for enzyme-linked immunosorbent assay (ELISA). The detection of nanogram per gram quantities of ricin spiked into retail samples of ground beef provides approximately 10,000-fold greater sensitivity than required to detect a toxic dose of ricin (>1 mg) in a 100 g sample.


Wideband holographic recorder.
This paper summarizes the design considerations and developmental status of a wideband holographic system breadboard designed to prove the feasibility of recording and reproducing digital data at 400 Mbits/sec and 40 Mbits/sec, respectively. The following critical system elements were investigated: multiplexer-demultiplexer electronics, linear data composer, polygonal mirror scanner, holographic recording techniques, and data readout devices. The feasibility of recording digital data in multichannel holographic form at high densities has been demonstrated on a subsystem basis.


Accounting for temporal and individual variation in the estimation of Von Bertalanffy growth curves.
Growth and growth limitation are important indicators of density dependence and environmental limitation of populations. Estimating individual growth trajectories is therefore an important aspect of understanding and predicting the life history and dynamics of a population. Variation in individual growth trajectories arises due to variation in the environmental factors limiting individual growth. This environmental limitation can vary over time, between cohorts and between individuals within a cohort. For a complete and accurate understanding of individual growth in a population, it is important to include all these sources of variation. So far, statistical models only accounted for a subset of these factors or required an extensive growth history of individuals. Here, we present a novel model describing the growth curves of cohorts in a population. This model is derived from a stochastic form of the Von Bertalanffy growth equation describing individual growth. The model is specifically tailored for use on length-at-age data in which the growth trajectory of an individual is unknown and every individual is only measured once. The presented method can also be used if growth limitation differs strongly between age or length classes. We demonstrate the use of the model for length-at-age data of North Sea plaice (Pleuronectes platessa) from the last 30 years. Fitting this model to length-at-age data can provide new insights in the dynamics of the environmental factors limiting individual growth and provides a useful tool for ecological research and management.


Serum C-reactive protein in Nigerians with type 2 diabetes mellitus.
C-reactive protein is an acute-phase proteins, produce in the liver, its release is stimulated by cytokines (interleukin 6 and tumour necrosis factor alpha). Elevated level of it is a risk factor for coronary heart disease. Baseline levels of C-reactive protein in apparently healthy men and women predict long-term risk of a first myocardial infarction. Diabetics are at increased risk for coronary heart disease, data from the Framingham Study showed a two- to three-fold elevation in the risk of clinically evident atherosclerotic disease in patients with type II diabetes compared to those without diabetes. However, but data regarding CRP in Nigerian diabetic is lacking. A cross-sectional study conducted among patients attending out patient clinic of the Obafemi Awolowo University Teaching Hospitals complex OAUTHC) Ile Ife, Osun State south western Nigeria. Measurement of C-reactive protein was based on the principle of solid phase enzyme-linked immunosorbent assay (ELISA). A total of 125 consecutive subjects were recruited comprising 75 patients with type II diabetes mellitus with or without hypertension and 50 apparently healthy age-and-sex comparable controls. There was a significant difference between the mean systolic and diastolic blood pressures of the patients and controls. The fasting blood glucose and C-reactive protein were significantly higher in diabetics compared to controls. there was a positive and significant correlation between FBG and CRP in both patients and controls. This study showed that diabetics have significantly higher serum C-reactive protein compared to the apparently controls. Also there was a positive and significant correlation between C-reactive protein and fasting blood glucose among both patients and controls.


Dietary abomasal impaction in a herd of dairy replacement heifers.
An episode of dietary abomasal impaction in a herd of dairy replacement heifers was found to be caused by excess almond shells in the ration. Clinical signs and necropsy findings led to the diagnosis. Removal of the almond shells and increasing the energy and digestibility of the ration corrected the problem. Factors contributing to the dietary impactions included advanced stages of pregnancy, high energy demands of growing heifers, and cold weather. Dietary abomasal impactions are not common in dairy cattle because of the high-quality ration a dairy cow generally receives. Replacement heifers in advanced stages of pregnancy have nutritional requirements similar to those of dairy cows, yet are often nutritionally neglected. The clinical findings in this report may help make veterinarians aware of the possibility of dietary abomasal impactions in dairy replacement heifers fed low-quality feeds.


Osteopathic postdoctoral training institutions.
The Osteopathic Postdoctoral Training Institution (OPTI) program was established by the American Osteopathic Association (AOA) in 1995 as a support and accreditation structure for osteopathic graduate medical education. Since 1999, all AOA-approved postdoctoral training programs are required to take place in OPTIs. In a continuing effort to further define standards for quality and to promote program development, the AOA's 2007 Annual OPTI Workshop focused on recommended changes to the OPTI standards document. In addition, an OPTI Inspectors Workshop was held in January 2008 to increase interrater reliability in determining whether individual OPTIs have met specific criteria.


The Parasol Protocol: An Implementation Science Study of HIV Continuum of Care Interventions for Gay Men and Transgender Women in Burma/Myanmar.
Efforts to improve HIV diagnosis and antiretroviral therapy (ART) initiation among people living with HIV and reduce onward transmission of HIV rely on innovative interventions along multiple steps of the HIV care continuum. These innovative methods are particularly important for key populations, including men who have sex with men (MSM) and transgender women (TW). The HIV epidemic in Myanmar is concentrated among key populations, and national efforts now focus on reducing stigma and improving engagement of MSM and TW in HIV prevention and care. This study aims to test the use of several innovations to address losses in the HIV care continuum: (1) use of respondent-driven sampling (RDS) to reach and engage MSM and TW in HIV testing, (2) HIV self-testing (HIVST) to increase HIV testing uptake and aid early diagnosis of infection, (3) community-based CD4 point-of-care (POC) technology to rapidly stage HIV disease for those who are HIV infected, and (4) peer navigation support to increase successful health system navigation for HIV-infected MSM and TW in need of ART or HIV engagement in care. To assess the effect of HIVST, we will implement a randomized trial in which MSM and TW adults in the greater Yangon metropolitan area who are HIV uninfected will be recruited via RDS (N=366). Participants will complete a baseline socio-behavioral survey and will be randomized to standard, voluntary counseling and testing (VCT) or to HIVST. Biologic specimens will be collected during this baseline visit for confirmatory testing using dried blood spots. Participants will be asked to return to the study office to complete a second study visit in which they will report their HIV test result and answer questions on the acceptability of the assigned testing method. Aim 1 participants with confirmed HIV infection and who are not engaged in care (N=49) will be offered direct enrollment into Aims 2 and 3, which include immediate CD4 POC and the option for peer navigation, respectively. Aims 2 and 3 participants will be prospectively followed for 12 months with data collection including interviewer-administered sociobehavioral survey, CD4 POC, and viral load testing occurring biannually. Participants who accept peer navigation will be compared to those who decline peer navigation. Analyses will estimate the impact of CD4 POC on engagement in care and the impact of peer navigation on ART adherence and viral load. Formative qualitative research was conducted in June and September 2015 and led to further refinement of recruitment methods, HIVST instructions and counseling, and peer navigation methods. Aim 1 recruitment began in November 2015 with subsequent enrollment into Aims 2 and 3 and is currently ongoing. These innovative interventions may resolve gaps in the HIV care continuum among MSM and TW and future implementation may aid in curbing the HIV epidemic among MSM and TW in Myanmar.


Factors regulating apoptosis and homeostasis of CD4+ CD25(high) FOXP3+ regulatory T cells are new therapeutic targets.
CD4+ CD25(high) Foxp3+ regulatory T cells (Tregs), as an active mechanism of immune suppression, have been targeted due to their tremendous therapeutic potentials to prevent autoimmune diseases, transplant rejection, and to inhibit progression of tumors and chronic viral diseases. In last twelve years, substantial molecular differences between homeostasis of Tregs and that of other subsets of T cells and some factors specific in regulation of Treg survival have been characterized. In this overview we focus on panoramic reviewing of 91 factors, pathways and drugs, both well-characterized and newly defined, regarding the survival and homeostasis of Tregs in the following sections: 2: Tregs, an essential mechanism of immune tolerance; 3: nTregs, aTregs and other regulatory T cells; 4: co-stimulation receptor signaling; 5: innate immunity and Toll-like receptor (TLR) signaling; 6: effects of cytokines and hormones; 7: transcription factors in regulation of Tregs; 8: Treg intracellular signaling pathways; 9: drugs and potential therapeutics; and 10: Treg cell survival and death. We recently reported that removal of Tregs via a Bax-dependent apoptotic pathway significantly enhances anti-self antigen immune responses, which demonstrated for the first time the proof of principle that apoptosis of Tregs is a new therapeutic target. Therefore, continued characterization of Treg apoptosis and homeostasis pathways would further improve our understanding in regulation of immune responses by cytokines, hormones and drugs, and would also lead to development of new therapeutics.


Identification of supplier induced demand in the health care sector.
This paper explores the issues and pitfalls encountered when attempting to test empirically the hypothesis that physician, hospital, or any other input supply level induces increasing demand for health services in the strict sense of demand shift and, through that, increased demand for the input in question. Evidence is presented which suggests that an empirical test of the supplier induced demand (SID) hypothesis of the type traditionally performed may not in fact be feasible with cross-sectional aggregate data such as is usually used.


Topographic and tomographic properties of forme fruste keratoconus corneas.
To investigate the efficacy of topography and tomography indices combined in discriminant functions to detect mild ectatic corneas. The authors retrospectively reviewed the data of 143 eyes separated into three groups by the Corneal Navigator OPD scanning system (Nidek, Gamagori, Japan): normal (N; LASIK surgery with a 2-year follow-up; n = 72), forme fruste keratoconus (N topography with contralateral KC; FFKC; n = 40), and KC (n = 31). Topography and tomography indices, corneal thickness spatial profile (CTSP), and anterior and posterior curvature spatial profiles were obtained with the Orbscan IIz (Bausch & Lomb Surgical, Rochester, NY). The percentage of thickness increase (PTI) from the thinnest point to the periphery, the percentage of variation of anterior (PVAK), and posterior curvature were calculated and compared by Kruskal-Wallis test. The usefulness of these data to discriminate among the three groups was assessed by receiver operating characteristic (ROC) curve analysis. Posterior elevation of the thinnest point (TP), all positions of CTSP, PTI for all distances from the TP, and PVAK from a 5- to 7-mm distance from the TP were significantly different in the FFKC compared with the N group. The discriminant functions between the FFKC and the N groups and between the KC and the N groups reached an area under the ROC curve of 0.98 and 0.99, respectively. PTI indices and maximum posterior central elevation were the most important contributors to the discriminant function. Indices generated from corneal thickness and curvature measurements over the entire cornea centered on the TP can identify very mild forms of ectasia undetected by a Placido-based neural network program.


The role of lines and corners of geometric figures in recognition performance.
A relative role of lines and corners of images of outline geometric figures in recognition performance was studied psychophysically. Probability of correct response to the shape of the whole figure (control) and figures with lines or corners masked to a different extent was compared. Increase in the extent of masking resulted in a drop of recognition performance that was significantly lower for figures without corners, than for figures without part of their lines. The whole 3D figures were recognized better than 2D ones, whereas the opposite relations were observed under conditions of masking. Significant gender difference in a recognition performance was found: men recognize entire and partly masked figures better than women. Possible mechanisms of relatively better recognition of figures with corners than with lines are discussed in connection with finding of high sensitivity of many neurons in the primary visual cortex to line crossing and branching.


[Alcohol consumption and environmental influence among school children].
The objective of this study was to know the attitude towards alcohol and its consumption among schoolchildren. We also studied the influence of the immediate environment. Participants were a random sample of sixth and eight grade schoolchildren during the 1995 school year from our rural area (n = 14,537). Data were collected from a confidential and anonymous survey about attitude and alcohol consumption. We found that 63% of children had tried alcohol occasionally. Forty-seven percent of them drank on a regular basis. Furthermore, 8% of this group consumed beer daily. Among frequent drinkers, 20.4% had gotten drunk at some time and this was more frequent among boys (p < 0.01) and older students. Forty percent of children did not think alcohol was a drug. Regarding family environment, 37% of fathers consumed alcohol daily in contrast to 4% of mothers. Logistic-regression analyses showed an independent association between alcohol consumption and smoking habits. A great number of schoolchildren and consumed alcohol occasionally. Boys and older students had gotten drunk more frequently than girls. Moreover, they believe that alcohol is not a drug and cannot perceive it as a harmful substance.


Hemophilus influenzae pharyngitis and cellulitis in adults.
Hemophilus influenzae infections in adults are becoming more common but are often unsuspected in this age group by the primary care physician. Two case reports illustrate pharyngitis, and pharyngitis associated with cellulitis of the neck, in which H influenzae was cultured from the blood. The throat and skin are only two of the many sites for H influenzae infections in adults. As no physical signs are pathognomonic for this organism, its possible role should influence the choice of antibiotics while awaiting culture results. Newer cephalosporins, especially cefamandole and cefoxitin, appear promising in the treatment of these infections.


Examining the association between implementation and outcomes : state-wide scale-up of school-wide positive behavior intervention and supports.
Although there is an established literature supporting the efficacy of a variety of prevention programs, there has been less empirical work on the translation of such research to everyday practice or when scaled-up state-wide. There is a considerable need for more research on factors that enhance implementation of programs and optimize outcomes, particularly in school settings. The current paper examines how the implementation fidelity of an increasingly popular and widely disseminated prevention model called, School-wide Positive Behavioral Interventions and Supports (SW-PBIS), relates to student outcomes within the context of a state-wide scale-up effort. Data come from a scale-up effort of SW-PBIS in Maryland; the sample included 421 elementary and middle schools trained in SW-PBIS. SW-PBIS fidelity, as measured by one of three fidelity measures, was found to be associated with higher math achievement, higher reading achievement, and lower truancy. School contextual factors were related to implementation levels and outcomes. Implications for scale-up efforts of behavioral and mental health interventions and measurement considerations are discussed.


Purification and characterization of the polycation-stimulated protein phosphatase catalytic subunit from porcine renal cortex.
The predominant form of phosphorylase phosphatase activity in porcine renal cortical extracts was a polycation-stimulated protein phosphatase. This activity was present in extracts in a high-molecular-weight form which could be converted to a free catalytic subunit by treatment with ethanol, urea, or freezing and thawing in the presence of beta-mercaptoethanol. The catalytic subunit of the polycation-stimulated phosphatase was purified by chromatography on DEAE-Sephacel, heparin-Sepharose, and Sephadex G-75. The phosphatase appeared to be homogeneous on SDS-polyacrylamide gel electrophoresis. The enzyme had an apparent Mr of 35 000 on gel filtration and SDS-polyacrylamide gel electrophoresis. The purified phosphatase could be stimulated by histone H1, protamine, poly(D-lysine), poly(L-lysine) or polybrene utilizing phosphorylase a as the substrate. It preferentially dephosphorylated the alpha-subunit of phosphorylase kinase. The phosphatase was highly sensitive to inhibition by ATP. These results suggest that the renal polycation-stimulated phosphatase catalytic subunit is very similar to or identical with the skeletal muscle phosphatase form which has been previously designated phosphatase-2Ac.


Temperature mediated variation of DNA secondary structure in (A.T) clusters; evidence by use of the oligopeptide netropsin as a structural probe.
The titration viscometric investigation of the multi-mode interaction of netropsin (Nt) with (A.T) clusters of NaDNA12 and NH4DNA10 has been extended to different temperatures. The position of two boundaries on the r-scale (r= [Nt]bound/[DNA-P]) with increasing temperature steadily (rI/II) or more abruptly (rO/I) shifts to lower values. For the most (A.T) rich Nt-binding sites of modes (O), (I) and (II) this observation suggests the existence of an equilibrium between different DNA secondary structures with a different translation per base pair. The mode specific changes delta L1Nt of DNA contour length as induced by one Nt molecule proved to be almost independent of temperature. Concomitant stiffening effects increase with decreasing temperature, contrary to initial expectation. Conformational variability of (A.T) clusters may represent an essential feature in specific or selective DNA-protein interaction.


Inactivation of Fam20b in the neural crest-derived mesenchyme of mouse causes multiple craniofacial defects.
The glycosaminoglycan (GAG) chains attached to the core proteins of proteoglycans exert multiple roles, such as enriching signal molecules and regulating the binding of ligands to the corresponding receptors. A newly identified kinase - family with sequence similarity 20 member B (FAM20B) - is essential for the formation of GAG chains. The FAM20B protein phosphorylates the initial xylose on the side chain of a serine residue in the protein. Although the GAG chains of proteoglycans are believed to be indispensable during craniofacial development, there are few reports on their exact functions in craniofacial organogenesis. In this study, by mating Wnt1-cre mice with Fam20b-floxed mice (Fam20bflox/flox), we created Wnt1-Cre;Fam20bflox/flox mice in which Fam20b is ablated in the neural crest-derived mesenchyme. The Wnt1-Cre;Fam20bflox/flox mice died immediately after birth because of complete cleft palates. In addition to cleft palate, Wnt1-Cre;Fam20bflox/flox mice also manifested tongue elevation, micrognathia, microcephaly, suture widening, and reduced mineralization in the calvaria, facial bones, and temporomandibular joint. These findings indicate that the proteoglycans formed through the catalysis of FAM20B are essential for the morphogenesis and mineralization of the craniofacial complex.


The sequence of the ribosomal 16S RNA from Proteus vulgaris. Sequence comparison with E. coli 16S RNA and its use in secondary model building.
The complete nucleotide sequence of the 16S RNA from Proteus vulgaris has been determined. The molecule (1544 nucleotides) shows 93% homology with the sequence of E. coli 16S RNA. Six methylated nucleotides have been localized in positions homologous to those observed in the E. coli RNA molecule. Both E. coli and P. vulgaris 16S RNA chains can be folded up into a common secondary structure scheme. Comparative sequence analysis of the two molecules has provided a valuable contribution to 16S RNA secondary structure model building.


Frameshift suppression by thyA mutants of Escherichia coli K-12.
We have extended our previous study on the suppression of frameshift mutants by Escherichia coli thyA mutants by assaying suppression of 15 rIIB frameshift mutants of bacteriophage T4 on one of our suppressing thyA mutant strains. The majority of insertion mutants were suppressible, whereas none of the deletion mutants tested was suppressible. Frameshift suppression could be inhibited by adding thymidine to the assay medium, but was not affected by the presence of a restrictive rpsL mutation in the host strain. We suggest that the frameshift suppression event occurs at a nonsense codon generated by the frameshift mutation.


PEEP: its use in young patients with apparently normal lungs.
PEEP has been advocated for use in patients with the acute respiratory distress syndrome characterized by a marked reduction in FRC. It has not been advocated for patients with apparently normal lungs requiring prolonged mechanical ventilation. We have done inert gas washouts on five young patients comparing no PEEP to 10 cm H2O PEEP. Four of the five showed a significant improvement in PaO2, a decrease in AaDO2 and Qs/Qt, the fifth a slight improvement on PEEP. Arterial washouts were significantly faster on PEEP in all patients suggesting a decrease in intrapulmonary shunting. Airway washouts in the initial phase were faster off PEEP suggesting an increase in lung volume and FRC with PEEP. Later portions of the curve were faster on PEEP suggesting improved distribution of ventilation. These findings suggest that PEEP might be of significant value in treatment of patients requiring prolonged mechanical ventilation for reasons other than lung abnormalities.


Low-wage maternal employment and outcomes for children: a study.
Despite the importance of anticipating how children may be affected by policies that move mothers off welfare and into employment, as the article by Zaslow and Emig in this journal issue points out, few research studies have addressed this critical policy question. To help fill that gap, this article presents the results of a new study using national survey data to examine child outcomes among families that had previously received welfare. About half the families studied had mothers who remained at home, the others were working at varying wage levels. The findings reported here echo themes discussed in the two preceding articles. Maternal employment does not appear to undermine children's social or cognitive development from ages 5 to 14, and it may yield advantages. Children whose mothers earned more than $5.00 per hour, particularly, had somewhat better outcomes than others. The authors emphasize, however, that background characteristics specific to the mothers who chose employment contributed to these positive outcomes. The authors add that it would be risky to apply these generalizations based on these findings to families forced into employment by welfare reform.


What cognitive processes are "sluggish" in sluggish cognitive tempo?
Sluggish cognitive tempo refers to a constellation of symptoms that include slowed behavior/thinking, reduced alertness, and getting lost in one's thoughts. Despite the moniker "sluggish cognitive tempo," the evidence is mixed regarding the extent to which it is associated globally with slowed (sluggish) mental (cognitive) information processing speed (tempo). A well-characterized clinical sample of 132 children ages 8-13 years (M = 10.34, SD = 1.51; 47 girls; 67% White/non-Hispanic) were administered multiple, counterbalanced neurocognitive tests and assessed for sluggish cognitive tempo symptoms via multiple-informant reports. Bayesian linear regressions revealed significant evidence against associations between sluggish cognitive tempo and computationally modeled processing speed (BF01 > 3.70), and significant evidence for associations with slower working memory manipulation speed. These findings were consistent across parent and teacher models, with and without control for attention-deficit/hyperactivity disorder inattentive symptoms and IQ. There was also significant evidence linking faster inhibition speed with higher parent-reported sluggish cognitive tempo symptoms. These findings provide strong evidence against characterizing children with sluggish cognitive tempo symptoms as possessing a globally sluggish cognitive tempo. Instead, these symptoms appear to be related, to a significant extent, to executive dysfunction characterized by working memory systems that are too slow and inhibition systems that are too fast. Behaviorally, these findings suggest that requiring extra time to rearrange the active contents of working memory delays responding, whereas an overactive inhibition system likely terminates thoughts too quickly and therefore prevents intended behaviors from starting or completing, thereby giving the appearance that children are absent-minded or failing to act when expected. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Association between serum gamma-glutamyltransferase and early-onset coronary artery disease: a retrospective case-control study.
Serum gamma-glutamyltransferase (GGT) activity has been proposed as a promising predictor of atherosclerosis-related complications and a prognostic marker for cardiovascular diseases. The objective of this study was to investigate the potential correlation between serum levels of GGT and early-onset coronary artery disease (EOCAD). A retrospective, hospital-based case-control study was conducted, which included 860 patients with EOCAD and gender- and age-matched controls. Serum levels of GGT were measured using the reference measurement procedure on an automatic biochemistry analyser. The serum GGT levels of patients with EOCAD (34.90 ± 31.44 U/L) were significantly higher than those of the control group (21.57 ± 16.44 U/L, p < .001). Elevated serum levels of GGT were found to be an independent risk factor for EOCAD, with an odds ratio (OR) of 1.021 (95% confidence interval (CI): 1.014-1.029). Additionally, for every quartile increase in serum GGT levels, the risk of developing EOCAD increased by 1.6-fold. Moreover, serum GGT levels were significantly associated with disease severity, with lower GGT levels observed in patients without significant vascular disease (31.74 ± 24.06 U/L) compared to those with two-vessel disease (33.06 ± 25.00 U/L, p = .002) and three-vessel disease (37.75 ± 36.76 U/L, p = .001). The results of this study suggest that elevated serum GGT levels are associated with the development of EOCAD, and GGT may be implicated in the pathogenesis of the disease. Further large-scale prospective studies are needed to explore the potential relationship between serum GGT levels and the dynamic development of EOCAD.


Chemotaxis of 3T3 and SV3T3 cells to fibronectin is mediated through the cell-attachment site in fibronectin and a fibronectin cell surface receptor.
Fibronectin (FN) is a multidomain extracellular matrix protein that induces attachment and chemotactic migration of fibroblastic cells. In this study we analyzed the molecular determinants involved in the FN-induced chemotactic migration of normal and SV40-transformed 3T3 cells. Two different monoclonal antibodies to the cell-binding site of FN blocked chemotaxis to a 140-kD FN fragment (Ca 140) containing the cell-binding domain. A monoclonal antibody to a determinant distant from the cell-binding site did not affect chemotaxis. A synthetic tetrapeptide, RGDS, which represents the major cell-attachment sequence, was able to compete with FN and the Ca 140 fragment in chemotaxis assays, but this peptide itself had no significant chemotactic activity. A larger peptide encompassing this sequence, GRGDSP, was chemotactic, while the peptide GRGESP, where a glutamic acid residue was substituted for aspartic acid, was inactive. Chemotactic migration could be prevented in a dose-dependent manner by a rabbit polyclonal antiserum to a 140-kD cell surface FN receptor. This antibody was more effective on normal than on transformed 3T3 cells. Neither the anti-FN receptor antiserum nor a monoclonal antibody to the cell-binding site of FN blocked migration induced by another potent chemoattractant, platelet-derived growth factor. These data indicate that FN-induced chemotaxis of 3T3 and SV3T3 cells is mediated via the RGDS cell-attachment site of FN and the 140-kD cell surface FN receptor. The interaction is specific and can be altered by transformation.


Cardiac remodeling in erythropoietin-transgenic mice.
Transgenic (tg) mice with chronic overexpression of the human erythropoietin gene are characterized by an increased hematocrit of about 0.80 in adulthood. This is accompanied by cardiac dysfunction and premature death. The aim of this study was to examine whether this cardiac dysfunction was accompanied by hypertrophy of the heart with remodeling of the extracellular matrix (ECM). 3-months-old wild type (wt) and tg mice without cardiac hypertrophy were compared with the respective 7-months-old mice. The mRNA of brain natriuretic peptide (BNP), of the matrix metalloproteinases (MMP)-2, -8, -9, -13, of the tissue inhibitor of metalloproteinase (TIMP)-1, -2, -3, -4 and of collagen I and III was detected by ribonuclease protection assay. The activity of MMPs was measured by zymography. There was hypertrophy of both ventricles in 7-months-old tg mice, which was accompanied by elevated mRNA expression of BNP. MMP-2 activity was increased and MMP-9 activity was decreased in the left ventricle (LV) of 3-months-old tg mice. This was accompanied by elevated TIMP-4 expression, followed by a shift of collagen mRNA expression from type III to type I in this ventricle. The shift to collagen I in the heart of tg mice might be associated with a stiffer ventricle resulting in diastolic dysfunction. This may be responsible for a relative and intermittent LV- and right ventricle (RV)-insufficiency which was likely to have occurred as evidenced by the elevation of lung and liver weight with hemorrhage and interstitial fibrosis after 7 months.


Phosphorylation of importin-α1 by CDK1-cyclin B1 controls mitotic spindle assembly.
Importin-α serves as an adaptor linking importin-β to proteins carrying a nuclear localization sequence (NLS). During interphase, this interaction enables nuclear protein import, while in mitosis it regulates spindle assembly factors (SAFs) and controls microtubule nucleation, stabilization and spindle function. Here, we show that human importin-α1 is regulated during the cell cycle and is phosphorylated at two sites (threonine 9 and serine 62) during mitosis by the major mitotic protein kinase CDK1-cyclin B. Mutational analysis indicates that the mitotic phosphorylation of importin-α1 inhibits its binding to importin-β and promotes the release of TPX2 and KIFC1, which are then targeted like importin-β to the spindle. Loss of importin-α1 or expression of a non-phosphorylated mutant of importin-α1 results in the formation of shortened spindles with reduced microtubule density and induces a prolonged metaphase, whereas phosphorylation-mimicking mutants are functional in mitosis. We propose that phosphorylation of importin-α1 is a general mechanism for the spatial and temporal control of mitotic spindle assembly by CDK1-cyclin B1 that acts through the release of SAFs such as TPX2 and KIFC1 from inhibitory complexes that restrict spindle assembly.


The four mechanisms of dental resorption initiation.
The aim of this study is to present a classification with a clinical application for root resorption, so that diagnosis will be more objective and immediately linked to the source of the problem, leading the clinician to automatically develop the likely treatment plan with a precise prognosis. With this purpose, we suggest putting together all diagnosed dental resorptions into one of these four criteria: 1) Root resorption caused by cementoblast cell death, with preservation of the Malassez epithelial rests. 2) Root resorption by cementoblasts and Malassez epithelial rests death. 3) Dental resorption by odontoblasts cell death with preservation of pulp vitality. 4) Dental resorption by direct exposure of dentin to gingival connective tissue at the cementoenamel junction gaps.


Planar-chiral ferrocenylphosphine-borane complexes featuring agostic-type B-HE (E = Hg, Sn) interactions.
The synthesis of ferrocenylphosphine-borane adducts 1,2-fc(E)(PPh2·BH3) (E = SnR2R', HgX; 1,2-fc = 1,2-ferrocenediyl) that are substituted with organotin or organomercury Lewis acid moieties in ortho-position is presented. Several compounds that feature two ferrocenylphosphine-borane moieties bridged by Sn or Hg are also introduced. The products are fully characterized by multinuclear NMR spectroscopy, high-resolution MALDI-TOF mass spectrometry and elemental analysis. The attachment of the Lewis acid substituent to the same Cp ring of the ferrocene results in planar-chirality and the close proximity between the boron hydride group and the Lewis acid is expected to allow for agostic-type B-HE (E = Sn, Hg) interactions. Structural investigations by X-ray diffraction reveal a short B-HSn contact of 2.755(4) Å for 1,2-fc(SnMe2Cl)(PPh2·BH3), which is only the second reported example of such a short agostic-type contact involving a coordinatively saturated tin(iv) center. In contrast, for the tetraorganotin derivatives 1,2-fc(SnMe3)(PPh2·BH3) and [1,2-fc(PPh2)](μ-SnMe2)[1,2-fc(PPh2·BH3)], in which the Lewis acidity of the tin atom is weaker than in 1,2-fc(SnMe2Cl)(PPh2·BH3), the B-HSn distances are much longer but still within the sum of the van der Waals radii of Sn and H (∑vdW = 3.27 Å). The chloromercury-substituted ferrocenylphosphine-borane 1,2-fc(HgCl)(PPh2·BH3) shows a similarly short B-HHg contact of 2.615(5) Å (∑vdW = 3.15 Å). Inspection of the extended structure of 1,2-fc(HgCl)(PPh2·BH3) reveals that the Lewis acidic mercury atom is also involved in intermolecular HgCl interactions with a neighboring molecule. An analysis of 31P and 11B NMR data reveals a correlation between the chemical shifts and the Lewis acidity of the adjacent organotin/mercury substituent. Structure optimization of 1,2-fc(SnMe3)(PPh2·BH3) and 1,2-fc(SnMe2Cl)(PPh2·BH3) by density functional theory (DFT) indicates B-HSn contacts respectively of 3.129 and 2.631 Å that are close to the experimental values. Natural bond orbitals (NBO) and atom in molecules (AIM) analyses reveals a B-H→Sn donor-acceptor interaction energy of 5.46 kcal mol-1 and a B-HSn bond path for the chlorodimethyltin-substituted derivative with a modest electron density ρ(r) of 0.0082 a.u. and a positive Laplacian at the bond critical point.


Wideband-tunable nanotube Q-switched low threshold erbium doped fiber laser.
An all-fiber passively wideband-tunable nanotube Q-switched low threshold erbium doped fiber laser is presented. With a low insertion loss carbon nanotube based saturable absorber, C- and L-band tunable Q-switched lasers are achieved by tuning the transmission wavelength of one C-band tunable filter and the other L-band one, respectively. The threshold of the Q-switched operation is only 12.8 mW. The self-mode-locking effect on the Q-switching can be effectively reduced by introducing a spatial hole burning effect with two optical circulators. The tunable wavelength range of the Q-switching laser can be changed by inserting a variable optical attenuator in the laser cavity to tune the gain spectrum.


An EOG-Based Human-Machine Interface for Wheelchair Control.
Nonmanual human-machine interfaces (HMIs) have been studied for wheelchair control with the aim of helping severely paralyzed individuals regain some mobility. The challenge is to rapidly, accurately, and sufficiently produce control commands, such as left and right turns, forward and backward motions, acceleration, deceleration, and stopping. In this paper, a novel electrooculogram (EOG) based HMI is proposed for wheelchair control. A total of 13 flashing buttons, each of which corresponds to a command, are presented in the graphical user interface. These buttons flash on a one-by-one manner in a predefined sequence. The user can select a button by blinking in sync with its flashes. The algorithm detects the eye blinks from a channel of vertical EOG data and determines the user's target button based on the synchronization between the detected blinks and the button's flashes. For healthy subjects/patients with spinal cord injuries, the proposed HMI achieved an average accuracy of 96.7% / 91.7% and a response time of 3.53 s/3.67 s with 0 false positive rates (FPRs). Using one channel of vertical EOG signals associated with eye blinks, the proposed HMI can accurately provide sufficient commands with a satisfactory response time. The proposed HMI provides a novel nonmanual approach for severely paralyzed individuals to control a wheelchair. Compared with a newly established EOG-based HMI, the proposed HMI can generate more commands with higher accuracy, lower FPR, and fewer electrodes.


To B(enign) or Not to B: functionalisation of variant in a mild form of argininosuccinate lyase deficiency identified through newborn screening.
Argininosuccinate lyase (ASL) deficiency is an autosomal recessive disorder of the urea cycle with a diverse spectrum of clinical presentation that is detectable in newborn screening. We report an 8-year-old girl with ASL deficiency who was detected through newborn screening and was confirmed using biochemical and functional assay. She is compound heterozygous for a likely pathogenic variant NM_000048.4(ASL):c.283C>T (p.Arg95Cys) and a likely benign variant NM_000048.4(ASL): c.1319T>C (p.Leu440Pro). Functional characterisation of the likely benign genetic variant in ASL was performed. Genomic sequencing was performed on the index patient presenting with non-specific symptoms of poor feeding and lethargy and shown to have increased serum and urine argininosuccinic acid. Functional assay using HEK293T cell model was performed. ASL enzymatic activity was reduced for Leu440Pro. This study highlights the role of functional testing of a variant that may appear benign in a patient with a phenotype consistent with ASL deficiency, and reclassifies NM_000048.4(ASL): c.1319T>C (p.Leu440Pro) variant as likely pathogenic.


B-type natriuretic peptide prevents acute hypertrophic responses in the diabetic rat heart: importance of cyclic GMP.
Stimulation of cardiomyocyte guanosine 3',5'-cyclic monophosphate (cyclic GMP) via endothelial-derived nitric oxide (NO) is an important mechanism by which bradykinin and ACE inhibitors prevent hypertrophy. Endothelial NO dysfunction and cardiac hypertrophy are morbid features of diabetes not entirely prevented by ACE inhibitors. In cardiomyocyte/endothelial cell cocultures, bradykinin efficacy is abolished by high-glucose-induced endothelial NO dysfunction. We now demonstrate that antihypertrophic actions of natriuretic peptides, which stimulate cyclic GMP independently of NO, are preserved in cardiomyocytes despite high-glucose-induced endothelial dysfunction. Further, streptozotocin-induced diabetes significantly impairs the effectiveness of acute antihypertrophic strategies in isolated rat hearts. In hearts from citrate-treated control rats, angiotensin II-stimulated [(3)H]phenylalanine incorporation and atrial natriuretic peptide and beta-myosin heavy chain mRNA expression were prevented by B-type natriuretic peptide (BNP), bradykinin, the ACE inhibitor ramiprilat, and the neutral endopeptidase inhibitor candoxatrilat. These antihypertrophic effects were accompanied by increased left ventricular cyclic GMP. In age-matched diabetic hearts, the antihypertrophic and cyclic GMP stimulatory actions of bradykinin, ramiprilat, and candoxatrilat were absent. However, the blunting of hypertrophic markers and accompanying increases in cyclic GMP stimulated by BNP were preserved in diabetes. Thus BNP, which increases cyclic GMP independently of NO, is an important approach to prevent growth in the diabetic myocardium, where endothelium-dependent mechanisms are compromised.


Beta-globin Gene Mutations in Turkish Children with Beta-Thalassemia: Results from a Single Center Study.
The beta thalassemias are common genetic disorders in Turkey and in this retrospective study our aim was to evaluate β-globin chain mutations and the phenotypic severity of β-thalassemia patients followed-up in our hospital, a tertiary center which serves patients from all regions of Turkey. 106 pediatric patients were analysed for β-globin gene mutations by using DNA analysis. Patients were classified as having β-thalassemia major or β-thalassemia intermedia based on age at diagnosis, transfusion frequency and lowest hemoglobin concentration in between transfusions. There were 106 patients (52.8% female and 47.2% male) with a mean age of 11.2±5 years (1.6 - 22.3 years). Eighty-four (79.2%) patients had β-thalassemia major, whereas the remaining 22 patients (20.8%) were identified as having β-thalassemia intermedia. Overall, 18 different mutations were detected on 212 alleles. The most frequently encountered mutation was IVS I.110 (G>A) (35.3%), followed by Codon 8 del-AA (10.4%), IVS II.1 (G>A) (8%), IVS I.1 (G>A) (7.5%), Codon 39 (C>T) (7.1%) and Codon 5 (-CT) (6.6%), which made up 79.4% of observed mutations. According to present results, IVS I.110 (G>AA) was the most frequent mutation observed in this study, as in other results from Turkey. Evaluation of β-thalassemia mutations in 106 patients with 212 alleles, revealed the presence of homozygous mutation in 85 patients (80.2%) and compound heterozygous mutation in 21 patients (19.8%). The mutations detected in patients with homozygous mutation were IVS I.110 (G>A) (38.8%), Codon 8 del -AA (11.8%), IVS II.1 (G>A) (8.2%) and IVS I.1 (G>A) (8.2%). Observed mutations in the compound heterozygotes were Codon 39 (C>T)/Codon 41-42 (-CTTT) (14.3%), IVS I.110 (G>A)/Codon 39(C>T) (14.3%), IVS I.110 (G>A)/Codon 44(-C) (14.3%), and IVS II.745 (C>G)/5'UTR + 22 (G>A) (9.5%). Our hospital is a tertiary referral center that provides care to patients from all over the country, and thus the distribution of mutations observed in the current study is significant in term of representing that of the country as a whole.


Bimodal affect recognition based on autoregressive hidden Markov models from physiological signals.
Affect provides contextual information about the emotional state of a person as he/she communicates in both verbal and/or non-verbal forms. While human's are great at determining the emotional state of people while they communicate in person, it is challenging and still largely an unsolved problem to computationally determine the emotional state of a person. Emotional states of a person manifest in the physiological biosignals such as electrocardiogram (ECG) and electrodermal activity (EDA) because these signals are impacted by the peripheral nervous system of the body, and the peripheral nervous system is strongly coupled with the mental state of the person. In this paper, we present a method to accurately recognize six emotions using ECG and EDA signals and applying autoregressive hidden Markov models (AR-HMMs) and heart rate variability analysis on these signals. The six emotions include happiness, sadness, surprise, fear, anger, and disgust. We evaluated our method on a comprehensive new dataset collected from 30 participants. Our results show that our proposed method achieves an average accuracy of 88.6% in distinguishing across the 6 emotions. The key technical depth of the paper is in the use of the AR-HMMs to model the EDA signal and the use of LDA to enable accurate emotion recognition without requiring a large number of training samples. Unlike other studies, we have taken a hierarchical approach to classify emotions, where we first categorize the emotion as either positive or negative and then identify the exact emotion.


Motif-specific sampling of phosphoproteomes.
Phosphoproteomics, the targeted study of a subfraction of the proteome which is modified by phosphorylation, has become an indispensable tool to study cell signaling dynamics. We described a methodology that linked phosphoproteome and proteome analysis based on Ba2+ binding properties of amino acids. This technology selected motif-specific phosphopeptides independent of the system under analysis. MudPIT (Multidimensional Identification Technology) identified 1037 precipitated phosphopeptides from as little as 250 microg of proteins. To extend coverage of the phosphoproteome, we sampled the nuclear extract of HeLa cells with three values of Ba2+ ions molarity. The presence of more than 70% of identified phosphoproteins was further substantiated by their nonmodified peptides. Upon isoproterenol stimulation of HEK cells, we identified an increasing number of phosphoproteins from MAPK cascades and AKAP signaling hubs. We quantified changes in both protein and phosphorylation levels of 197 phosphoproteins including a critical kinase, MAPK1. Integration of differential phosphorylation of MAPK1 with knowledge bases constructed modules that correlated well with its role as node in cross-talk of canonical pathways.


A NOVEL IMAGE-SPECIFIC TRANSFER APPROACH FOR PROSTATE SEGMENTATION IN MR IMAGES.
Prostate segmentation in Magnetic Resonance (MR) Images is a significant yet challenging task for prostate cancer treatment. Most of the existing works attempted to design a global classifier for all MR images, which neglect the discrepancy of images across different patients. To this end, we propose a novel transfer approach for prostate segmentation in MR images. Firstly, an image-specific classifier is built for each training image. Secondly, a pair of dictionaries and a mapping matrix are jointly obtained by a novel Semi-Coupled Dictionary Transfer Learning (SCDTL). Finally, the classifiers on the source domain could be selectively transferred to the target domain (i.e. testing images) by the dictionaries and the mapping matrix. The evaluation demonstrates that our approach has a competitive performance compared with the state-of-the-art transfer learning methods. Moreover, the proposed transfer approach outperforms the conventional deep neural network based method.


NK Cell and Ig Interplay in Defense against Herpes Simplex Virus Type 1: Epistatic Interaction of CD16A and IgG1 Allotypes of Variable Affinities Modulates Antibody-Dependent Cellular Cytotoxicity and Susceptibility to Clinical Reactivation.
HSV-1 latently infects most humans, causing a variable clinical picture that depends, in part, on host genetic factors. Both IgG and its cellular FcRs, CD16A and CD32A-C (encoded by FCGR3A and FCGR2A-C, respectively, on chromosome 1), display polymorphisms that could affect their defensive function. Of potential relevance are a FCGR3A dimorphism resulting in CD16A-valine/phenylalanine-158 allotypes with different IgG affinity, variations conditioning NK cell expression of CD32B or CD32C, and IgG1 H chain (IGHG1) and kappa-chain (IGKC) polymorphisms determining allotypes designated G1m and Km. In this study, we assessed the contribution of Ig genetic variations and their interaction with FcR polymorphism to HSV-1 susceptibility, as well as their impact on NK cell-mediated Ab-dependent cellular cytotoxicity (ADCC). Our results show an epistatic interaction between IGHG1 and FCGR3A such that the higher affinity CD16A-158V/V genotype associates with an asymptomatic course of HSV-1 infection only in homozygotes for G1m3. Furthermore, CD16A-158V and G1m3 allotypes enhanced ADCC against opsonized HSV-1-infected fibroblasts. Conversely, Km allotypes and CD32B or CD32C expression on NK cells did not significantly influence HSV-1 susceptibility or ADCC. NK cells degranulating against immune serum-opsonized HSV-1-infected fibroblasts had heterogeneous phenotypes. Yet, enhanced ADCC was observed among NK cells showing a differentiated, memory-like phenotype (NKG2C(bright)NKG2A(-)CD57(+)FcRγ(-)), which expand in response to human CMV. These results extend our knowledge on the importance of immunogenetic polymorphisms and NK cell-Ab interplay in the host response against HSV-1 and point to the relevance of interactions between immune responses elicited during chronic coinfection by multiple herpesviruses.


The increase of miR-27a affects the role of cisplatin on proliferation and migration capacities of liver cancer cells.
To study the effects of chronic virus-mediated micro ribonucleic acid miR-27a on proliferation and migration capacities of liver cancer cells. A total of 60 patients with primary liver cancer from January 2015 to December 2016 were selected as observation group, 60 patients with chronic liver disease were selected as control group, and another 60 healthy subjects who received physical examination during the same period were selected as healthy group. All patients received serum miRNA detection. The different expressions of serum miRNAs in healthy people, patients with chronic liver disease, and patients with liver cancer were analyzed. The correlations of miR-27a with growth and proliferation of liver cancer MCC-7721 cells were studied. The levels of miR-27a and miR-664b in subjects in control group and healthy group were significantly lower than those in observation group, but the expression levels of miR-30a were statistically elevated (p<0.05). The expression of serum miR-27a in liver cancer patients with higher tumor-node-metastasis (TNM) staging (stage III-IV, number of tumors >1, tumor size >5 cm, and vascular invasion) was significantly higher than those in patients with lower TNM staging (stage I-II, number of tumors =1, tumor size ≤5 cm, and no vascular invasion) (p<0.05). At 48 h and 72 h after transfection, the proliferation of liver cancer MCC-7721 cells was significantly enhanced compared to that in non-transfection group (p<0.05). The level of miR-27a was significantly upregulated in cisplatin-resistant liver cancer A549/CDDP cells compared with that in parental A549 cells. MiR-27a regulated epithelial-mesenchymal transition (EMT) and cisplatin resistance in vitro, while modulated the in vivo response of liver cancer cells to cisplatin. Further studies identified the Raf kinase inhibitor protein (RKIP) as a direct and functional target of miR-27a. The knockdown of RKIP by RNAi showed a similar effect to ectopic miR-27a expression, whereas the over-expression of RKIP weakened the function of miR-27a in liver cancer cells. MiR-27a participated in the proliferation and migration capacities of liver cancer cells and influenced the effect of cisplatin via targeting RKIP. The high expression of miR-27a is closely related to the malignant degree of liver cancer, which provides guidance for the diagnosis, targeted therapy, and prognostic evaluation of liver cancer.


Characterization of the avian GLUT1 glucose transporter: differential regulation of GLUT1 and GLUT3 in chicken embryo fibroblasts.
Vertebrate cells that are transformed by oncogenes such as v-src or are stimulated by mitogens have increased rates of glucose uptake. In rodent cells, the mechanisms whereby glucose transport is up-regulated are well understood. Stimulation of glucose transport involves an elevation in mRNA encoding the GLUT1 glucose transporter that is controlled at the levels of both transcription and mRNA stability. Cloning and sequencing of chicken GLUT1 cDNA showed that it shares 95% amino acid sequence similarity to mammalian GLUT1s. Nevertheless, unlike mammalian GLUT1 mRNA, it was not induced by v-src, serum addition, or treatment with the tumor promoter 12-O-tetradecanoylphorbol 13-acetate in chicken embryo fibroblasts. Rather, the induction of glucose transport in chicken embryo fibroblasts by v-src, serum, and 12-O-tetradecanoylphorbol 13-acetate was associated with induction of GLUT3 mRNA level and GLUT3 transcription. Rat fibroblasts were also found to express both GLUT1 and GLUT3 isoforms, but v-src induced GLUT1 and not GLUT3. This suggests that animal cells require both a basal and an upregulatable glucose transporter and that these functions have been subsumed by different GLUT isoforms in avian and mammalian cells.


Parental experiences of homeschooling during the COVID-19 pandemic: differences between seven European countries and between children with and without mental health conditions.
The aim of the present study was to examine parental experiences of homeschooling during the COVID-19 pandemic in families with or without a child with a mental health condition across Europe. The study included 6720 parents recruited through schools, patient organizations and social media platforms (2002 parents with a child with a mental health condition and 4718 without) from seven European countries: the UK (n = 508), Sweden (n = 1436), Spain (n = 1491), Belgium (n = 508), the Netherlands (n = 324), Germany (n = 1662) and Italy (n = 794). Many parents reported negative effects of homeschooling for themselves and their child, and many found homeschooling to be of poor quality, with insufficient support from schools. In most countries, contact with teachers was limited, leaving parents with primary responsibility for managing homeschooling. Parents also reported increased levels of stress, worry, social isolation, and domestic conflict. A small number of parents reported increased parental alcohol/drug use. Some differences were found between countries and some negative experiences were more common in families with a child with a mental health condition. However, differences between countries and between families with and without a mental health condition were generally small, indicating that many parents across countries reported negative experiences. Some parents also reported positive experiences of homeschooling. The adverse effects of homeschooling will likely have a long-term impact and contribute to increased inequalities. Given that school closures may be less effective than other interventions, policymakers need to carefully consider the negative consequences of homeschooling during additional waves of the COVID-19 pandemic and future pandemics.


[Computer modeling of ESR spectra of the plasma in patients with Down's syndrome].
ESR technique at 77 degrees K was used for studying the blood plasma ESR signals of patients with Down syndrome and of healthy people. It was observed that the first exhibited a tendency towards a decrease of ESR signal with g = 4.3 and increase of the ratio of ceruplasmin (g = 2.05) and transferrin (g = 4.3) ESR signal amplitude. A computer simulation has been carried out by means of special mathematical program of experimental ESR spectra of the blood plasma.


12-month trajectories of depressive symptoms among nurses-Contribution of personality, job characteristics, coping, and burnout.
Job related factors have been associated with higher risk for developing depression, but past studies lacked full consideration of individual factors such as personality and coping. We sought to evaluate associations of personality, coping, job characteristics, and burnout with 12-month trajectories of depressive symptoms among nursing workers. Cohort of nursing workers (N = 281) in a private hospital system, with baseline assessments of personality, job characteristics, and coping. Burnout and depression were measured at baseline and during monthly follow-ups. Linear mixed modeling was used to examine contributions to between- and within-individual variation in monthly depressive symptoms. Personality trait of negative affectivity accounted for 36% of between-individual variation in depressive symptoms over 12 months, while job characteristics and coping explained an additional 5% and 8% of this variation, respectively. Exhaustion dimension of burnout was associated with between-individual variation in depressive symptoms (fixed effect β coefficient 2.44, p < 0.001), but not with within-individual variation in symptoms. Disengagement dimension of burnout was not associated with between-individual variation in depressive symptoms, but contributed to within-individual variation in depressive symptoms over time (fixed effect β coefficient 0.52, p = 0.01). Participants were nursing workers within a single hospital system. Participants who were excluded due to missing baseline data were more likely of non-white race, which may also limit the generalizability of our results. We used latent variables to represent certain job and coping characteristics, which may make our results less comparable with other studies examining the role of these factors in work-associated depression. Future interventions to prevent depression in healthcare workers should consider multiple job and individual factors. Potential components include strategies to manage negative affectivity and reduce avoidant coping, such as cognitive reframing and mindfulness-based techniques, and organizational approaches to address burnout through augmentation of job resources.


Suppressing glucose uptake and acetic acid production increases membrane protein overexpression in Escherichia coli.
The production of integral membrane spanning proteins (IMP's) constitutes a bottleneck in pharmaceutical development. It was long considered that the state-of-the-art was to produce the proteins as inclusion bodies using a powerful induction system. However, the quality of the protein was compromised and the production of a soluble protein that is incorporated into the membrane from which it is extracted is now considered to be a better method. Earlier research has indicated that a slower rate of protein synthesis might overcome the tendency to form inclusion bodies. We here suggest the use of a set of E. coli mutants characterized by a slower rate of growth and protein synthesis as a tool for increasing the amount of soluble protein in high- throughput protein production processes. A set of five IMP's was chosen which were expressed in three mutants and the corresponding WT cell (control). The mutations led to three different substrate uptake rates, two of which were considerably slower than that of the wild type. Using the mutants, we were able to express three out of the five membrane proteins. Most successful was the mutant growing at 50% of the wild type growth rate. A further effect of a low growth rate is a low acetic acid formation, and we believe that this is a possible reason for the better production. This hypothesis was further supported by expression from the BL21(DE3) strain, using the same plasmid. This strain grows at a high growth rate but nevertheless yields only small amounts of acetic acid. This strain was also able to express three out of the five IMP's, although at lower quantities. The use of mutants that reduce the specific substrate uptake rate seems to be a versatile tool for overcoming some of the difficulties in the production of integral membrane spanning proteins. A set of strains with mutations in the glucose uptake system and with a lower acetic acid formation were able to produce three out of five membrane proteins that it was not possible to produce with the corresponding wild type.


Subjective neuronal coding of reward: temporal value discounting and risk.
A key question in the neurobiology of reward relates to the nature of coding. Rewards are objects that are advantageous or necessary for the survival of individuals in a variety of environmental situations. Thus reward appears to depend on the individual and its environment. The question arises whether neuronal systems in humans and monkeys code reward in subjective terms, objective terms or both. The present review addresses this issue by dealing with two important reward processes, namely the individual discounting of reward value across temporal delays, and the processing of information about risky rewards that depends on individual risk attitudes. The subjective value of rewards decreases with the temporal distance to the reward. In experiments using neurophysiology and brain imaging, dopamine neurons and striatal systems discount reward value across temporal delays of a few seconds, despite unchanged objective reward value, suggesting subjective value coding. The subjective values of risky outcomes depend on the risk attitude of individual decision makers; these values decrease for risk-avoiders and increase for risk-seekers. The signal for risk and the signal for the value of risky reward covary with individual risk attitudes in regions of the human prefrontal cortex, suggesting subjective rather than objective coding of risk and risky value. These data demonstrate that important parameters of reward are coded in a subjective manner in key reward structures of the brain. However, these data do not rule out that other neurons or brain structures may code reward according to its objective value and risk.


Cucurbitacin delta 23-reductase from the fruit of Cucurbita maxima var. Green Hubbard. Physicochemical and fluorescence properties and enzyme-ligand interactions.
Cucurbitacin delta 23-reductase from Cucurbita maxima var. Green Hubbard fruit displays an apparent Mr of 32,000, a Stokes radius of 263 nm and a diffusion coefficient of 8.93 X 10(-7) cm2 X s-1. The enzyme appears to possess a homogeneous dimeric quaternary structure with a subunit Mr of 15,000. Two tryptophan and fourteen tyrosine residues per dimer were found. Emission spectral properties of the enzyme and fluorescence quenching by iodide indicate the tryptophan residues to be buried within the protein molecule. In the pH range 5-7, where no conformational changes were detected, protonation of a sterically related ionizable group with a pK of approx. 6.0 markedly influenced the fluorescence of the tryptophan residues. Protein fluorescence quenching was employed to determine the dissociation constants for binding of NADPH (Kd 17 microM), NADP+ (Kd 30 microM) and elaterinide (Kd 227 microM). Fluorescence energy transfer between the tryptophan residues and enzyme-bound NADPH was observed.


[Informative value of complex electromyographic and immunologic examinations of patients with diabetic neuropathies].
As many as 43 patients suffering from diabetes mellitus associated with the clinical manifestations of diabetic neuropathy underwent ++clinico-electromyographic and immunologic examinations. The control group comprised 30 practically healthy persons of the same age. The electromyographic++ examination included measurements of the rate of wave spreading in motor fibers of the limb nerves in the distal and proximal parts (in accordance with the F-wave), a study of H-reflex from the musculus soleus of the leg together with M-response of hand and foot muscles. The immunologic examination consisted in measurements of different class immunoglobulins in the blood serum. Based on the electromyographic findings, 3 groups of patients were distinguished: group I included patients with primary injury to the myelin membrane of the nerves; group II was made up of patients with primary injury to the axon, and group III of patients with concomitant injury to the nervous membranes and axons. There was a significant elevation of the concentration of anti-myelin activity possessing IgM in the blood serum of patients belonging to groups I and III, which attests to the involvement of the immunopathological mechanism into myelopathy development.


Environmental risk factors for respiratory infection and wheeze in young children: A multicentre birth cohort study.
Respiratory infections and wheeze have a considerable impact on the health of young children and consume significant healthcare resources. We aimed to evaluate the effect of environmental factors on respiratory infections and symptoms in early childhood. Environmental risk factors including: daycare attendance; breastfeeding; siblings; damp within the home; environmental tobacco smoke (ETS); child's bedroom flooring; animal exposure; road traffic density around child's home; and solid fuel pollution within home were assessed in children recruited to the GO-CHILD multicentre prospective birth cohort study. Follow-up information on respiratory infections (bronchiolitis, pneumonia, otitis media and cold or flu), wheeze and cough symptoms, healthcare utilisation and medication prescription was collected by postal questionnaires at 12 and 24 months. Log binomial and ordered logistic regression models were fitted to the data. Follow-up was obtained on 1344 children. Daycare was associated with increased odds of pneumonia (odds ratio [OR] = 2.39, 95% confidence interval [CI]: 1.04-5.49), bronchiolitis (OR = 1.40, 1.02-1.90), otitis media (OR = 1.68, 1.32-2.14) and emergency department attendance for wheeze (RR = 1.81, 1.17-2.80). Breastfeeding beyond 6 months was associated with a reduced odds of bronchiolitis (OR = 0.55, 0.39-0.77) and otitis media (OR = 0.75, 0.59-0.99). Siblings at home was associated with an increased odds of bronchiolitis (OR = 1.65, 1.18-2.32) and risk of reliever inhaler prescription (RR = 1.37, 1.02-1.85). Visible damp was associated with an increased odds of wheeze (OR = 1.85, 1.11-3.19), and risk of reliever inhaler (RR = 1.73, 1.04-2.89) and inhaled corticosteroid prescription (RR = 2.61, 1.03-6.59). ETS exposure was associated with an increased odds of primary care attendance for cough or wheeze (OR = 1.52, 1.11-2.08). Dense traffic around the child's home was associated with an increased odds of bronchiolitis (OR = 1.32, 1.08-2.29). Environmental factors likely influence the wide variation in infection frequency and symptoms observed in early childhood. Larger population studies are necessary to further inform and guide public health policy to decrease the burden of respiratory infections and wheeze in young children.


The impact of Medicaid peer support utilization on cost.
Peer support programs have proliferated over the past decade, building on recovery oriented programming, yet relationships between peer support services and the costs to public programs have not been well described in literature. The purpose of this study is to fill gaps in the literature related to peer support programs and cost: lack of comparison groups, small sample sizes, and the availability of research examining utilization of Medicaid mental health services. The study employed a retrospective design with treatment and comparison groups created from three administrative databases. Three ordinary least squares regression models were constructed to predict crisis stabilization cost, psychiatric hospitalization cost, and total Medicaid cost while controlling for other factors. The Chronic Illness and Disability Payment System + Rx was used to control for illness severity. Peer support was associated with $5,991 higher total Medicaid cost (p < .01). Peer support was also associated with higher crisis stabilization cost and lower psychiatric hospitalization cost, but the relationships were not statistically significant. Peer support was associated with $2,100 higher prescription drug cost (p < .01), $5,116 higher professional services cost (p < .01), and $1,225 lower facility cost (p < .01). While the implementation of Medicaid financed peer support programs may not result in savings from reductions of costly crisis stabilizations and psychiatric hospitalizations, it does support the principles of self-direction and recovery from severe mental illness. State policy makers must weigh the potential higher cost associated with peer support programs with efforts to redesign the delivery of mental health services.


Structural and functional characterization of the mitochondrial complex IV assembly factor Coa6.
Assembly factors play key roles in the biogenesis of many multi-subunit protein complexes regulating their stability, activity, and the incorporation of essential cofactors. The human assembly factor Coa6 participates in the biogenesis of the CuA site in complex IV (cytochrome c oxidase, COX). Patients with mutations in Coa6 suffer from mitochondrial disease due to complex IV deficiency. Here, we present the crystal structures of human Coa6 and the pathogenic W59CCoa6-mutant protein. These structures show that Coa6 has a 3-helical bundle structure, with the first 2 helices tethered by disulfide bonds, one of which likely provides the copper-binding site. Disulfide-mediated oligomerization of the W59CCoa6 protein provides a structural explanation for the loss-of-function mutation.


An attempt to reproduce 'mal seco' in horses by feeding them Festuca argentina.
'Mal seco' is an almost invariably fatal disease of horses in Argentina and Chile, which resembles grass sickness, a dysautonomia of horses in Europe. The aetiology of mal seco remains unknown. An attempt to reproduce the disease was made by feeding horses with Festuca argentina, a plant considered to be toxic to animals and which was consistently found in the diet of nine horses suffering from mal seco. Three horses were fed with F argentina ad libitum for 28 days. The plant was infected with an endophytic fungus, whose morphological characteristics were in agreement with descriptions of Acremonium chlamydosporioides. No clinical abnormalities were observed in two of the horses, but one died on the fifth day of the trial after becoming incoordinated, unsteady and ataxic in the fore- and hindlimbs. No gross changes were observed post mortem in any of the horses, with the exception of a small number of Fasciola hepatica in the liver of the horse which died, and a moderate number of Gasterophilus species in the stomach of all three horses. No histopathological changes were observed in any of the organs examined, including several autonomic ganglia, brain including most brain stem nuclei, spinal cord, liver, kidney, stomach and small and large intestine. The results of this study suggest that F argentina is either not implicated in the aetiology of mal seco or produces its effects only when they are triggered by other unknown factors.


Using measured stocks of biomass and litter carbon to constrain modelled estimates of sequestration of soil organic carbon under contrasting mixed-species environmental plantings.
Reforestation of agricultural land with mixed-species environmental plantings of native trees and shrubs contributes to abatement of greenhouse gas emissions through sequestration of carbon, and to landscape remediation and biodiversity enhancement. Although accumulation of carbon in biomass is relatively well understood, less is known about associated changes in soil organic carbon (SOC) following different types of reforestation. Direct measurement of SOC may not be cost effective where rates of SOC sequestration are relatively small and/or highly spatially-variable, thereby requiring intensive sampling. Hence, our objective was to develop a verified modelling approach for determining changes in SOC to facilitate the inclusion of SOC in the carbon accounts of reforestation projects. We measured carbon stocks of biomass, litter and SOC (0-30cm) in 125 environmental plantings (often paired to adjacent agricultural sites), representing sites of varying productivity across the Australian continent. After constraining a carbon accounting model to observed measures of growth, allocation of biomass, and rates of litterfall and litter decomposition, the model was calibrated to maximise the efficiency of prediction of SOC and its fractions. Uncertainties in both measured and modelled results meant that efficiencies of prediction of SOC across the 125 contrasting plantings were only moderate, at 39-68%. Data-informed modelling nonetheless improved confidence in outputs from scenario analyses, confirming that: (i) reforestation on agricultural land highly depleted in SOC (i.e. previously under cropping) had the highest capacity to sequester SOC, particularly where rainfall was relatively high (>600mmyear-1), and; (ii) decreased planting width and increased stand density and the proportion of eucalypts enhanced rates of SOC sequestration. These results improve confidence in predictions of SOC following environmental reforestation under varying conditions. The calibrated model will be a useful tool for informing land managers and policy makers seeking to understand the dynamics of SOC following such reforestation.


Synthesis and characterization of the crystal and magnetic structures and properties of the hydroxyfluorides Fe(OH)F and Co(OH)F.
The title compounds were synthesized by a hydrothermal route from a 1:1 molar ratio of lithium fluoride and transition-metal acetate in an excess of water. The crystal structures were determined using a combination of powder and/or single-crystal X-ray and neutron powder diffraction (NPD) measurements. The magnetic structure and properties of Co(OH)F were characterized by magnetic susceptibility and low-temperature NPD measurements. M(OH)F (M = Fe and Co) crystallizes with structures related to diaspore-type α-AlOOH, with the Pnma space group, Z = 4, a = 10.471(3) Å, b = 3.2059(10) Å, and c = 4.6977(14) Å and a = 10.2753(3) Å, b = 3.11813(7) Å, and c = 4.68437(14) Å for the iron and cobalt phases, respectively. The structures consist of double chains of edge-sharing M(F,O)6 octahedra running along the b axis. These infinite chains share corners and give rise to channels. The protons are located in the channels and form O-H···F bent hydrogen bonds. The magnetic susceptibility indicates an antiferromagnetic ordering at ∼40 K, and the NPD measurements at 3 K show that the ferromagnetic rutile-type chains with spins parallel to the short b axis are antiferromagnetically coupled to each other, similarly to the magnetic structure of goethite α-FeOOH.


Esophageal motor disorders in terms of high-resolution esophageal pressure topography: what has changed?
The concept of high-resolution manometry (HRM) is to use sufficient pressure sensors such that intraluminal pressure can be monitored as a continuum along luminal length much as time is viewed as a continuum in conventional manometry. When HRM is coupled with pressure topography plots, pressure amplitude is transformed into spectral colors with isobaric conditions indicated by same-colored regions on the display. Together, these technologies are called high-resolution esophageal pressure topography (HREPT). HREPT has several advantages compared with conventional manometry, the technology that it was designed to replace. (i) The contractility of the entire esophagus can be viewed simultaneously in a uniform format, (ii) standardized objective metrics can be systematically applied for interpretation, and (iii) topographic patterns of contractility are more easily recognized and have greater reproducibility than with conventional manometry. Compared with conventional manometry, HREPT has improved sensitivity for detecting achalasia, largely due to the objectivity and accuracy with which it identifies impaired esophagogastric junction (EGJ) relaxation. In addition, it has led to the subcategorization of achalasia into three clinically relevant subtypes based on the contractile function of the esophageal body: classic achalasia, achalasia with esophageal compression, and spastic achalasia. Headway has also been made in understanding hypercontractile conditions, including diffuse esophageal spasm and a newly described entity, spastic nutcracker. Ultimately, clinical experience will be the judge, but it seems likely that HREPT data, along with its well-defined functional implications, will improve the clinical management of esophageal motility disorders.


[Breast cancer: can axillary dissection be restricted to Berg stages 1 and 2? Study of 895 cases].
895 axillary clearances were carried out in 3 years between 1986 and 1988 on 878 patients with primary breast cancer. Two procedures were used: either direct total axillary clearance (in stages 1, 2 and 3 of Berg) or inferior axillary clearance (stages 1 and 2 of Berg). This was carried out in association with frozen section diagnosis of the lymph node followed by clearance in Berg stage 3 if the diagnosis on frozen section was positive. The authors found a higher frequency of late complications, particularly of arm lymphoedema in patients who had complete axillary clearance than in patients who only had an inferior axillary clearance. The survival rate actuarilly calculated was not significantly different in the two groups of patients. The carrying out of total axillary clearance was "beneficial" for only 3 patients who had negative inferior axillary clearance and positive sub-clavicular clearance. Finally the frozen section diagnosis of the state of the axillary nodes shown to be correct (specificity -99% and sensitivity -76%) but it was difficult to carry out routinely.


Understanding behavioral and physiological phenotypes of stress and anxiety in zebrafish.
The zebrafish (Danio rerio) is emerging as a promising model organism for experimental studies of stress and anxiety. Here we further validate zebrafish models of stress by analyzing how environmental and pharmacological manipulations affect their behavioral and physiological phenotypes. Experimental manipulations included exposure to alarm pheromone, chronic exposure to fluoxetine, acute exposure to caffeine, as well as acute and chronic exposure to ethanol. Acute (but not chronic) alarm pheromone and acute caffeine produced robust anxiogenic effects, including reduced exploration, increased erratic movements and freezing behavior in zebrafish tested in the novel tank diving test. In contrast, ethanol and fluoxetine had robust anxiolytic effects, including increased exploration and reduced erratic movements. The behavior of several zebrafish strains was also quantified to ascertain differences in their behavioral profiles, revealing high-anxiety (leopard, albino) and low-anxiety (wild type) strains. We also used LocoScan (CleverSys Inc.) video-tracking tool to quantify anxiety-related behaviors in zebrafish, and dissect anxiety-related phenotypes from locomotor activity. Finally, we developed a simple and effective method of measuring zebrafish physiological stress responses (based on a human salivary cortisol assay), and showed that alterations in whole-body cortisol levels in zebrafish parallel behavioral indices of anxiety. Collectively, our results confirm zebrafish as a valid, reliable, and high-throughput model of stress and affective disorders.


Analysis of methylarginine metabolism in the cardiovascular system identifies the lung as a major source of ADMA.
Protein arginine methylation is catalyzed by a family of enzymes called protein arginine methyltransferases (PRMTs). Three forms of methylarginine have been identified in eukaryotes: monomethylarginine (l-NMMA), asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA), all characterized by methylation of one or both guanidine nitrogen atoms of arginine. l-NMMA and ADMA, but not SDMA, are competitive inhibitors of all nitric oxide synthase isoforms. SDMA is eliminated almost entirely by renal excretion, whereas l-NMMA and ADMA are further metabolized by dimethylarginine dimethylaminohydrolase (DDAH). To explore the interplay between methylarginine synthesis and degradation in vivo, we determined PRMT expression and DDAH activity in mouse lung, heart, liver, and kidney homogenates. In addition, we employed HPLC-based quantification of protein-incorporated and free methylarginine, combined with immunoblotting for the assessment of tissue-specific patterns of arginine methylation. The salient findings of the present investigation can be summarized as follows: 1) pulmonary expression of type I PRMTs was correlated with enhanced protein arginine methylation; 2) pulmonary ADMA degradation was undertaken by DDAH1; 3) bronchoalveolar lavage fluid and serum exhibited almost identical ADMA/SDMA ratios, and 4) kidney and liver provide complementary routes for clearance and metabolic conversion of circulating ADMA. Together, these observations suggest that methylarginine metabolism by the pulmonary system significantly contributes to circulating ADMA and SDMA levels.


Ovarian mucinous cystadenocarcinoma as a cause of Zollinger-Ellison syndrome: report of a case and review of the literature.
Gastrin producing ovarian tumor is a rare cause of the Zollinger-Ellison syndrome. We report the case of an ovarian carcinoma showing increased plasma gastrin concentration. A 60-yr-old woman presented with epigastric pain and diarrhea. Physical examination showed a large mass in the lower abdomen. Computed tomography revealed a large multilocular ovarian cyst. Upper gastrointestinal endoscopy examination showed multiple ulcerations of the stomach and duodenum. The plasma gastrin level was 1500 pg/ml. No tumors were found in the pancreas and duodenum, and salpingo-oophorectomy was performed. Histologic examination revealed a mucinous tumor of borderline malignant potential. Immunoperoxidase studies for gastrin showed many gastrin-producing cells within the epithelium of the tumor. There have been 11 cases (including our patient) of gastrin-producing ovarian tumor reported in the literature. We review here the relevant literature. Although ovarian gastrinoma is extremely rare, it should be considered as a possible cause of the Zollinger-Ellison syndrome in women.


Respirometric evaluation of biodegradation characteristics of dairy wastewater for organic carbon removal.
This study evaluates the biodegradation kinetics of an integrated dairy wastewater, with the main purpose of defining the experimental basis for modelling of the activated sludge process. Besides conventional characterization, the experiments involved detailed chemical oxygen demand (COD) fractionation and assessment of major kinetic and stoichiometric coefficients, by using respirometric methods. A multi-component model based on the endogenous decay concept was used for the kinetic interpretation. The results of conventional analyses and respirometric evaluations together with the assessment of residual components showed that the organic carbon content of the dairy wastewater was mostly soluble and biodegradable. The soluble, slowly biodegradable COD was the major COD fraction, representing around 50% of the total COD. Model calibration of the oxygen uptake rate profiles were consistent and revealed the existence of dual hydrolysis kinetics for soluble and particulate COD components. The hydrolysis rate associated with the main COD component--the soluble, slowly biodegradable COD fraction--was found to be 1.2 d(-1), which is quite low and underlines the role of this COD fraction as the rate-limiting factor for effluent quality. Simulation of process efficiency by the adopted model, calibrated with the experimentally determined parameters, indicated that effective control of the biodegradation of the soluble biodegradable COD components could be done by selection of appropriate values for the sludge age and hydraulic retention time. In this way, the total effluent soluble COD level could be lowered to 30-40 mg L(-1) range, in conformity with effluent limitations.


Cry1Ab toxin production of MON 810 transgenic maize.
Levels of Cry1Ab toxin were detected in genetically modified maize of genetic event MON 810 against near isogenic maize as negative control by two commercial immunoassays. The immunoassays were characterized for their cross-reactivity (CR) between Cry1Ab protoxin and activated toxin, and were compared with each other for toxin detection in a reference plant sample. Cry1Ab toxin levels, corrected for active toxin content using the CR values obtained, were monitored in maize DK-440 BTY through the entire vegetation period. The toxin concentration was found to show a rapid rise in the leaves to 17.15 +/- 1.66 microg/g by the end of the fifth week of cultivation, followed by a gradual decline to 9.61 +/- 2.07 microg/g by the 16th week and a slight increase again to 13.51 +/- 1.96 microg/g during the last 2 weeks due to partial desiccation. Similar but lesser fluctuation of toxin levels was seen in the roots between 5.32 +/- 0.49 microg/g at the less differentiated V1 stage and 2.25 +/- 0.30 microg/g during plant development. In contrast, Cry1Ab toxin levels appeared to be stably 1.36 +/- 0.45, 4.98 +/- 0.31, 0.47 +/- 0.03, and 0.83 +/- 0.15 microg/g in the stem, anther wall, pollen, and grain, respectively. Toxin concentrations produced at the VT-R4 phenological stages under actual cultivation conditions were compared with each other in three different years within an 8-year period.


Expanding the RNA-Guided Endonuclease Toolkit for Mouse Genome Editing.
The RNA-guided endonuclease CRISPR-Cas system from Streptococcus pyogenes (SpCas9) is widely used for generating genetically modified mice via zygotic microinjection. Although SpCas9 is a potent mutagen, it requires an NGG proto-spacer adjacent motif (PAM) at the target site, restricting sequence targetability. Here, we show that RNA-guided endonucleases that utilize a range of alternative PAM sequences can edit the mouse genome at the neurog3 (Ngn3) locus: SpCas9 VQR (NGAN PAM), SpCas9 VRER (NGCG), AsCas12a (TTTN), SaCas9 (NNGRRT), and SaCas9 KKH (NNNRRT). Additional experiments targeting tyrosinase and frizzled3 with SaCas9 KKH and its parent protein demonstrated that these endonucleases generated mutations in up to 100% of embryos across three loci. Remarkably, in contrast to wild-type SpCas9, these endonucleases frequently generated mutant embryos that retain unmodified alleles in both template-free and HDR-repair experiments. Our findings broaden PAM recognition options for mouse genome editing and identify SaCas9/SaCas9 KKH as useful alternatives when targeting genes with null lethal phenotypes.