It's time to increase physical activity promotion among pregnant women in France.
The lockdown linked to COVID-19 was shown to have negative effects on healthy behaviors in the general population, prompting the implementation of adapted public health measures. However, more vulnerable populations, such as pregnant women, were not specifically taken into account. At the time of writing this study, we know little about how the COVID-19 pandemic impacted the physical activity (PA) behavior of pregnant women in France. Given the many reports in the literature about women's low level of PA throughout pregnancy and the negative effects of the COVID-19 lockdown on behaviors in the general population; however, we might assume that their sedentary behaviors increased. The current study aimed to analyze the French recommendations and PA promotion among pregnant women. A literature search was done using PubMed for the key terms "physical activity/physical activity promotion," "pregnant women/pregnancy,", and "French population/France." All the relevant studies were included to support the argument for this narrative review. Efforts to promote PA for pregnant women often seem ineffective and even unrealistic, and many women become overweight or obese during pregnancy. Health professionals need evidence-based guidelines and continuous training and skills development in order to convincingly encourage women to be more active during pregnancy and the postpartum period. Health policies should strengthen PA promotion among pregnant women with detailed evidence-based guidelines on PA during postpartum.


Oxytocin Reduces Brain Injury and Maintains Blood-Brain Barrier Integrity After Ischemic Stroke in Mice.
The present study was designed to determine the effect of different doses of oxytocin (OXT) on neuronal injury, spatial memory, blood-brain barrier (BBB) integrity and to explore possible underlying molecular mechanisms in the early stage of stroke in mice. Stroke model was generated by middle cerebral artery occlusion (MCAO) for 60 min and 24 h reperfusion in mice. OXT at doses of 1, 2, 4 and 8 IU/per mouse was administrated intranasally at the beginning of brain ischemia. Brain injury, BBB integrity, and spatial memory were evaluated by standard methods. Changes in the expression of nuclear factor-kappa B (NF-κB), and TUNEL positive cell were detected by immunohistochemistry. The levels of vascular endothelial growth factor (VEGF), aquaporin-4 (AQP4) and brain-derived neurotrophic factor (BDNF) proteins were determined by western blotting and ELISA methods. OXT at doses of 4 and 8 IU/per mouse reduced the infarct size by 42% and 52%, respectively, and improved spatial memory function (p < 0.001). OXT (8 IU/per mouse) significantly reduced brain edema, BBB disruption and upregulated the AQP4 expression (p < 0.001). Finally, OXT significantly diminished the number of apoptotic, NF-κB positive cells and enhanced the expression of BDNF and VEGF proteins in the brain tissue (p < 0.001). These findings provide important evidences that OXT significantly suppresses neuronal damage in the early stage of stroke by inhibiting apoptotic and NF-κB signaling pathway, increasing the expression of VEGF, AQP4 and BDNF proteins and reducing the BBB leakage.


The effects of 1% prednisolone acetate on pupil diameter and intraocular pressure in healthy dogs treated with 0.005% latanoprost.
Prostaglandin analogs contribute to blood-aqueous barrier breakdown and may exacerbate uveitis. As these analogs induce de novo synthesis of endogenous prostaglandins, their therapeutic, hypotensive effect could potentially be inhibited by anti-inflammatory treatment. We therefore evaluated whether topical 1% prednisolone acetate alters the effects of 0.005% latanoprost on pupil diameter (PD) and intraocular pressure (IOP). Ten healthy Labrador retriever dogs from the Israel Guide Dog Center for the Blind. Pupil diameter and IOP were measured hourly, 8 AM-4 PM, with the right and left eyes serving as control (CE) and treated (TE) eyes, respectively. Measurements were conducted during four sessions: (1) without treatment (n = 10), (2) following latanoprost treatment (n = 10) at 8 AM, (3) following prednisolone treatment (n = 7) at 8 AM, and (4) bilateral latanoprost treatment at 8 AM, prednisolone treatment in TE at 11 AM (n = 8). The different number of dogs in sessions 3 and 4 is because some dogs were matched with their new owners earlier than expected. Pupil diameters were not affected by the addition of prednisolone and, at 4 PM, were 3.82 ± 0.47 and 3.97 ± 0.36 mm in TE and CE, respectively (P = 0.175, Wilcoxon). IOPs were not affected by the addition of prednisolone and, at 4 PM, were 9.0 ± 0.8 and 9.3 ± 0.8 mm Hg in TE and CE, respectively (P = 0.339, Wilcoxon). Prednisolone did not alter latanoprost's miotic and hypotensive effects in normal dogs during this study period.


[A new criminologic procedure: study of L-cystine-aminopeptidase activity for the detection of seminal stains].
L-Cystine-aminopeptidase (CAP) enzyme cleaves L-cystine-di-beta-naphthylamide substrate to beta-naphthylamine. The final product of this process is a violet azostain. CAP enzyme determination was developed for criminological procedures. CAP shows almost 100 times greater activity in human semen than in other body fluids. Samples taken at different time intervals (1 week, 1-3-6 months, 1 year, and 5-10 years) were investigated; they were dried on cloth and filter paper and then stored at room temperature for various time periods. The enzyme was also shown to be specific in the comparative studies performed with other human secretions (nasal discharge, saliva, vaginal secretion, feces, urine, breast milk) and fruit juices (raspberry, strawberry, apple, pear, plum, tomato, etc.).


Single-cell biology meeting marks rebirth of an old science.
A report on the third Single Cell Analyses meeting, held at the Cold Spring Harbor Laboratory, New York, USA, March 6-9, 2013.


Pathologic evaluation of disc dysfunction and osseous abnormalities of the temporomandibular joint.
The arthrographic findings in 141 patients with TMJ symptoms were compared with the linear tomographic evaluation to assess the incidence and severity of associated osseous abnormalities. Thirty-nine temporomandibular joints (28%) had significant osseous abnormalities compatible with degenerative joint disease. The incidence and severity of the degenerative changes compared favorably with the arthrographic assessment of the degree of soft-tissue injury. A cause-and-effect relationship between TMJ disc damage and dysfunction and degenerative arthritis is suggested.


BDNF-AAV has protective effects on morphine-induced conditioned place preference through BDNF, TrkB, and CREB concentration changes in the VTA and NAc.
Brain-derived neurotrophic factor (BDNF) is one of the neurotrophic factors that promotes the survival and protection of neurons in many disorders. The potential protective effect of BDNF and its mechanisms on morphine addiction are unclear. In this study, morphine-induced conditioned place preference (CPP) in mice was used to show the effect of BDNF on rewarding behavior. Western blot assays were used to determine the changes caused by BDNF, for example, changes in total BDNF, tropomyosin-related kinase receptor B (TrkB), and cAMP response element binding protein (CREB) in the ventral tegmental area (VTA) and nucleus accumbens (NAc). The results showed that the BDNF-adeno-associated viral vector (BDNF-AAV) injected in the VTA, attenuated morphine-induced CPP with synergistic changes in BDNF/TrkB/CREB concentrations in the VTA and NAc in the CPP acquisition and recurrence phases; however, the attenuation was lower in the extinction phase, with different changes in molecules downstream of the BDNF.


Enhanced enzymatic activity and conformational stability of catalase in presence of tetrahedral DNA nanostructures: A biophysical and kinetic study.
The emergence of DNA nanotechnology has shown enormous potential in a vast array of applications, particularly in the medicinal and theranostics fields. Nevertheless, the knowledge on the biocompatibility between DNA nanostructures and cellular proteins is largely unknown. Herein, we report the biophysical interaction between proteins (circulatory protein bovine serum albumin, BSA, and the cellular enzyme bovine liver catalase, BLC) and tetrahedral DNA (tDNA), which are well-known nanocarriers for therapeutics. Interestingly, the secondary conformation of BSA or BLC was unaltered in the presence of tDNAs which supports the biocompatible property of tDNA. In addition, thermodynamic studies showed that the binding of tDNAs with BLC has a stable non-covalent interaction via hydrogen bond and van der Waals contact, which is indicative of a spontaneous reaction. Furthermore, the catalytic activity of BLC was increased in the presence of tDNAs after 24 h of incubation. These findings indicate that the presence of tDNA nanostructures not only ensures a steady secondary conformation of proteins, but also stabilize the intracellular proteins like BLC. Surprisingly, our investigation discovered that tDNAs have no effect on albumin proteins, either by interfering or by adhering to the extracellular proteins. These findings will aid in the design of future DNA nanostructures for biomedical applications by increasing the knowledge on the biocompatible interaction of tDNAs with biomacromolecules.


Relative population of S-form and F-form conformers of bryonolic acid and its derivatives in equilibrium in CdCl3 solutions.
Relative populations of S-form (D-E rings: boat-boat form) and F-form (D-E rings: chair-chair form) conformers, in equilibrium in CDCl3 solutions, of 20 derivatives (2-21) of bryonolic acid (D:C-friedoolean-8-en-3beta-ol-29-oic acid) (1) were calculated from NMR spectral data (J-values and chemical shifts), with the aid of molecular mechanic calculation using a MM2/CONFLEX program system. The principal deciding factor of the population ratio was found to be whether the functionality at C-29 is trigonal or tetrahedral; the S-form: F-form was 0:100-32:68 for the "trigonal" type and 48:52-100:0 for the "tetrahedral." The reliability of the results is discussed.


The Saccharomyces cerevisiae MEC1 gene, which encodes a homolog of the human ATM gene product, is required for G1 arrest following radiation treatment.
The Saccharomyces cerevisiae gene MEC1 represents a structural homolog of the human gene ATM mutated in ataxia telangiectasia patients. Like human ataxia telangiectasia cell lines, mec1 mutants are defective in G2 and S-phase cell cycle checkpoints in response to radiation treatment. Here we show an additional defect in G1 arrest following treatment with UV light or gamma rays and map a defective arrest stage at or upstream of START in the yeast cell cycle.


Sudden sensorineural hearing loss: Recovery rates according to audiometric patterns.
The aim of this study was to evaluate the different audiometric patterns in sudden sensorineural hearing loss (SSNHL), assess recovery rates based on the initial pattern and also, analyse the impact on speech discrimination scores (SDS). A retrospective, descriptive, study was completed for patients with SSNHL from January 2010 until June 2020. Outcome measures included audiometric patterns, recovery rates, improvements over time for hearing loss as well as for SDS at 14 days and 3 months follow-up. The Kruskal-Wallis test and Mann-Whitney U test were used to compare differences between the different groups. Post-hoc testing involved the Wilcoxon signed-rank test. A P<0.05 was considered statistically significant. We included 211 patients, 64.3% showed downward-sloping or flat audiometric curves. Overall, 40% of the patients had recovered 50% or more of their hearing by day 14. We observed that hearing did improve over time, and this was more common for the upward-sloping cases, with 65% recovering to at least 50% of the maximum possible recovery by 3 months follow-up. Thirty percent had concomitant vertigo and/or dizziness; these patients had worse initial speech PTAs (pure tone average) (P≤0.0001) and inferior recovery rates (P=0.0007) as compared to patients without vertigo and/or dizziness. SSNHL is still a controversial topic. Variability was observed with regards to audiometric curves, recovery rates and SDS recovery. We provide a table with recovery rates based on audiometric patterns that may help guide clinicians when explaining this condition to their patients.


The Fibrinogen-to-Albumin Ratio Is Associated with Poor Prognosis in Patients with Coronary Artery Disease: Findings from a Large Cohort.
We aimed to evaluate the association of the fibrinogen-to-albumin ratio (FAR) with the clinical outcomes of coronary artery disease (CAD). All 14,944 patients with CAD evaluated in the present study were from a prospective cohort that recruited 15,250 patients admitted in the First Affiliated Hospital of Xinjiang Medical University between December 2016 and October 2021. The all-cause mortality (ACM) and cardiac mortality (CM) were selected as the primary endpoints. The secondary endpoints were major adverse cardiovascular events (MACEs), major adverse cardiac and cerebrovascular events (MACCEs), and non-fatal myocardial infarction (NFMI). The optimal FAR cutoff value was determined by using a receiver operating characteristic (ROC) curve analysis. Using 0.1 as the cutoff value, all the patients were divided into two groups: a low-FAR group (FAR < 0.1, n = 10,076) and a high-FAR group (FAR ≥ 0.1, n = 4918). The incidence of outcomes between the two groups was compared. The high-FAR group exhibited a higher incidence of ACM (5.3% vs. 1.9%), CM (3.9% vs. 1.4%), MACEs (9.8% vs. 6.7%), MACCEs (10.4% vs. 7.6%), and NFMI (2.3% vs. 1.3%) than the low-FAR group. To adjust the confounders, multivariate Cox regression analyses showed that the risk in the high-FAR group was increased 2.182 fold in ACM (HR = 2.182, 95% CI: 1.761 ~ 2.704, P < 0.001), 2.116 fold in CM (HR = 2.116, 95% CI: 1.761 ~ 2.704, P < 0.001), 1.327 fold in MACEs (HR = 1.327, 95% CI: 1.166 ~ 1.510, P < 0.001), 1.280 fold in MACCEs (HR = 1.280, 95% CI: 1.131 ~ 1.448, P < 0.001), and 1.791 fold in NFMI (HR = 1.791, 95% CI:1.331 ~ 2.411, P < 0.001), compared to the low-FAR group. The present study suggested that the high-FAR group was an independent and powerful predictor of adverse outcomes in CAD patients.


Hydrogen Bonding Directed Reversal of 13 C NMR Chemical Shielding.
The deshielding or downfield 13 C NMR chemical shift of amide carbonyl carbon upon H-bonding is a widely observed phenomenon. This downfield shift is commonly used as a spectroscopic ruler for H-bonding. However, the very first observation of an upfield 13 C NMR of thiocarbonyl carbon in thioamides upon H-bonding encouraged us to explore the physical origin of the reversal of 13 C NMR chemical shielding. Careful NMR analysis shows that sulfur and selenium-centered H-bonds (S/SeCHBs) induce a shielding effect on the 13 CC=S(Se) while changing from amides to thioamides or selenoamides. In addition, natural chemical shielding (NCS) analysis shows that the σ11 and σ22 components of the isotropic shielding tensor (σ) have a crucial role in this unusual shielding.


IP3R activity increases propensity of RyR-mediated sparks by elevating dyadic [Ca2+].
Calcium (Ca2+) plays a critical role in the excitation contraction coupling (ECC) process that mediates the contraction of cardiomyocytes during each heartbeat. While ryanodine receptors (RyRs) are the primary Ca2+ channels responsible for generating the cell-wide Ca2+ transients during ECC, Ca2+ release, via inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs) are also reported in cardiomyocytes to elicit ECC-modulating effects. Recent studies suggest that the localization of IP3Rs at dyads grant their ability to modify the occurrence of Ca2+ sparks (elementary Ca2+ release events that constitute cell wide Ca2+ releases associated with ECC) which may underlie their modulatory influence on ECC. Here, we aim to uncover the mechanism by which dyad-localized IP3Rs influence Ca2+ spark dynamics. To this end, we developed a mathematical model of the dyad that incorporates the behaviour of IP3Rs, in addition to RyRs, to reveal the impact of their activity on local Ca2+ handling and consequent Ca2+ spark occurrence and its properties. Consistent with published experimental data, our model predicts that the propensity for Ca2+ spark formation increases in the presence of IP3R activity. Our simulations support the hypothesis that IP3Rs elevate Ca2+ in the dyad, sensitizing proximal RyRs towards activation and hence Ca2+ spark formation. The stochasticity of IP3R gating is an important aspect of this mechanism. However, dyadic IP3R activity lowers the Ca2+ available in the junctional sarcoplasmic reticulum (JSR) for release, thus resulting in Ca2+ sparks with similar durations but lower amplitudes.


Fish chemokines 14, 20 and 25: A comparative statement on computational analysis and mRNA regulation upon pathogenic infection.
In this study, we reported a molecular characterization of three CC chemokines namely, CsCC-Chem14, CsCC-Chem20 and CsCC-Chem25 which are were identified from the established cDNA library of striped murrel Channa striatus. Multiple sequence alignment of all the three chemokines revealed the presence of gene specific domains and motifs including small cytokine domain, IL8 like domain, receptor binding site and glycosaminoglycan (GAG) binding sites. Three dimensional structures of the chemokines under study showed an important facet on their anti-microbial property. Tissue specific mRNA expression showed that the CsCC-Chem14 is highly expressed in spleen, CsCC-Chem20 in liver and CsCC-Chem25 in trunk kidney. On challenge C. striatus with oomycete fungus Aphanomyces invadans, both CsCC-Chem20 and CsCC-Chem25 showed significant (P < 0.05) up-regulation compared to CsCC-Chem14. The increase in the expression levels of CsCC-Chem20 and CsCC-Chem25 due to infection showed that they are antimicrobial proteins. But considering the CsCC-Chem14 expression, it is found to be a constitutive chemokine and is involved in homeostatic function in spleen of C. striatus. C. striatus challenged with bacteria Aeromonas hydrophila also exhibited different up-regulation pattern in all the three chemokines at various time points. However, extensive studies are required to determine the functional activities of CsCC-Chem14, CsCC-Chem20 and CsCC-Chem25 in vitro and in vivo to gain more knowledge at the molecular and proteomic levels.


Enzyme cytochemical staining of individual cells with the use of a polyacrylamide carrier. Studies on the synthetizing reaction technique, the indigogenic method, the metal salt method, the post-azo-coupling technique, and the tetrazolium salt technique.
A recently developed method for the (quantitative) demonstration of glucose-6-phosphate dehydrogenase activity in individual cells with the use of a polyacrylamide carrier has been extended for other enzyme cytochemical techniques. Isolated hepatocytes have been incorporated in the matrix of a thin transparent polyacrylamide gel prior to incubation in a cytochemical medium. The techniques which have been applied are the synthetizing reaction technique for glycogen phosphorylase, the indigogenic method for nonspecific esterase, the metal salt method for glucose-6-phosphatase, the post-azo-coupling technique for acid phosphatase, and the tetrazolium salt technique for succinate and lactate dehydrogenase activities. In all cases a few major problems which occur in the cytochemistry on single cells seem to be solved. The morphology is very well preserved, the final reaction product seems to be precipitated at the expected site of enzyme activity and the coloured end-product is highly specific for the enzyme activity to be studied, as has been demonstrated well with control experiments. The conclusion is reached, therefore, that this relatively simple device can be used routinely for the optimalization of enzyme cytochemistry of single cells.


Effects of the application sequence of calcium-containing desensitising pastes during etch-and-rinse adhesive restoration.
To evaluate the effects of different application sequence of calcium-containing desensitising pastes on bonding effectiveness and tubule occlusion during etch-and-rinse (E&R) adhesive restoration. Seventy molars were sectioned parallel to the occlusal plane, polished and randomly divided into seven groups (n=10). Group 1 was etched with 35% phosphoric acid for 15s. Groups 2-4 were treated with different calcium-containing desensitisers, including an arginine-calcium carbonate (Arg-CaCO3)-containing paste, a casein phosphopeptide-amorphous calcium phosphate (CPP-ACP)-containing paste and a calcium-sodium phosphosilicate (Novamin)-containing paste, respectively. Afterward, these groups were etched with 35% phosphoric acid for 15s. Groups 5-7 were initially etched and then treated with Arg-CaCO3-, CPP-ACP- and Novamin-containing desensitisers, respectively. In each group, samples were equally distributed into two subgroups (n=5) to bond with either a two-step E&R adhesive Adper SingleBond 2 (SB) or a three-step E&R adhesive Adper ScotchBond Multi-Purpose (SBMP). The microtensile bond strengths (MTBS) were tested and fracture modes were analyzed by stereomicroscopy and field-emission scanning electron microscopy (FESEM). Eight additional dentine disks were prepared to evaluate tubule occlusion prior to bonding using FESEM. The application sequence of calcium-containing desensitising pastes did not significantly affect MTBS irrespective of two-step SB (P>0.05) or three-step SBMP E&R adhesives (P>0.05). Effective dentinal tubule occlusion was observed in the mode of etching-desensitising. Applying calcium-containing desensitisers (particularly Arg-CaCO3- and Novamin-based) after etching during E&R adhesive restoration could achieve effective tubule occlusion without affecting the bonding strength.


Lumbosacral spinal neurons in the cat that are candidates for being activated by collaterals from the spinocervical tract.
Lumbosacral spinal neurons activated via the spinocervical tract were stained by intracellular injection of horseradish peroxidase in cats anaesthetized with chloralose and paralysed with gallamine triethiodide. The neurons were activated orthodromically by single shock stimulation of the ipsilateral dorsolateral funiculus at the second to third cervical segment, but not from the rostral part of the first cervical segment. Twenty nine cells were recovered from the histological material and subsequently reconstructed from transverse sections. Sixteen cells (55%) had axons that projected ipsilaterally to the lateral funiculus and their somata were located in two regions of the spinal cord, one group in the dorsal horn (laminae IV-V) and the other in the intermediate gray matter (laminae VI-VII). The axons of 10 of these cells gave off collaterals, and in seven of them the collaterals ramified in the grey matter deep to the cell body. The axons of five cells (17%) projected medially towards the central canal, four crossing the mid line in the ventral white commissure and ascending in the contralateral ventral funiculus. Only one of these cells had an axon collateral that crossed into the contralateral dorsal horn. Of the remaining eight cells, three had no obvious long axons but had many local axon collaterals, the axons of three cells were not stained, one had an axon projecting towards the ipsilateral ventral funiculus and one was a motoneuron and its axon projected into a ventral root. A feature of the dendritic trees of many cells was their wide spread in the mediolateral and/or the dorsoventral directions, although no dendrites reached dorsally into lamina II. Twenty-two cells (76%) were excited by moving hairs and by noxious pinch, three (10%) by hair movement alone, two (7%) by noxious pinch and pressure, and for two cells (7%) no receptive field could be found. It is concluded that not only postsynaptic dorsal column neurons receive input from the spinocervical tract but also other cells in the dorsal and ventral horns and the intermediate gray matter. Possible identities for these cells are discussed.


An Opportunistic Cooperative Packet Transmission Scheme in Wireless Multi-Hop Networks.
Cooperative routing, combining cooperative communication in the physical layer and routing technology in the network layer, is one of the most widely used technologies for improving end-to-end transmission reliability and delay in the wireless multi-hop networks. However, the existing cooperative routing schemes are designed based on an optimal fixed-path routing so that the end-to-end performance is greatly restricted by the low spatial efficiency. To address this problem, in this paper an opportunistic cooperative packet transmission (OCPT) scheme is explored by combining cooperative communication and opportunistic routing. The proposed scheme divides the multi-hop route into multiple virtual multiple-input-multiple-output (MIMO) transmissions. Before each transmission, based on the idea of opportunistic routing, a cluster head (CH) is introduced to determine the multiple transmitters and multiple receivers to form a cluster. Then, the single-hop transmission distance is defined as the metric of forward progress to the destination. Each intra-cluster cooperative packet transmission is formulated as a transmit beamforming optimization problem, and an iterative optimal beamforming policy is proposed to solve the problem and maximize the single-hop transmission distance. CH organizes multiple transmitters to cooperatively transmit packets to multiple receivers with the optimized transmit beamforming vector. Finally, according to the transmission results, the cluster is updated and the new cooperative transmission is started. Iteratively, the transmission lasts until the destination has successfully received the packet. We comprehensively evaluate the OCPT scheme by comparing it with conventional routing schemes. The simulation results demonstrate that the proposed OCPT scheme is effective on shortening the end-to-end transmission delay, increasing the number of successful packet transmissions and improving the packet arrival ratio and transmission efficiency.


[Role of growth factor signaling in epithelial cell plasticity during development and in carcinogenesis].
The work described in this review addresses the multifunctional roles of growth factors and their cognate receptors in normal development and in tumorigenesis. The concept of epithelial cell plasticity is described in the context of embryonic development during which frequent remodeling occurs in epithelial tissues. The critical role of one member of the FGF family of growth factors is demonstrated in lung branching morphogenesis. Several members of this family have been shown to induce an epithelial-mesenchymal transition in a bladder carcinoma line. In vivo the same factors act in an autocrine or paracrine mode to favor tumor progression. It is suggested that an EGFR-ligand autocrine loop exerts a positive role in tumor progression of human bladder carcinoma whereas FGFR2 acts as a phenotypic tumor suppressor gene. Unexpectedly, constitutive activating mutations in FGFR3 have been uncovered in the majority of the Ta superficial tumors which progress only very rarely to the invasive stages. In contrast, in situ carcinoma, which are considered to be associated with a strong malignant potential, do not carry the FGFR3 mutations. The presence or absence of the mutations defines two distinct oncogenic pathways in bladder carcinogenesis. The studies reveal the complexities in defining the putative functions of growth factors at different times and differentiation stages during development and in tumor progression. These results emphasize the need for caution in the interpretation of studies evaluating the potential of novel anti-cancer agents and for better designs of in vitro and in vivo biological assays.


In vitro and in vivo evaluation of the mechanisms of citalopram-induced hepatotoxicity.
Even though citalopram is commonly used in psychiatry, there are several reports on its toxic effects. So, the current study was designed to elucidate the mechanisms of cytotoxic effects of in vitro and in vivo citalopram treatment on liver and the following cytolethal events. For in vitro experiments, freshly isolated rat hepatocytes were exposed to citalopram along with/without various agents. To do in vivo studies liver function enzyme assays and histological examination were performed. In the in vitro experiments, citalopram (500 µM) exposure demonstrated cell death, a marked elevation in ROS formation, mitochondrial potential collapse, lysosomal membrane leakiness, glutathione (GSH) depletion and lipid peroxidation. In vivo biochemistry panel assays for liver enzymes function (AST, ALT and GGTP) and histological examination confirmed citalopram (20 mg/kg)-induced damage. citalopram-induced oxidative stress cytotoxicity markers were significantly prevented by antioxidants, ROS scavengers, MPT pore sealing agents, endocytosis inhibitors, ATP generators and CYP inhibitors. Either enzyme induction or GSH depletion were concomitant with augmented citalopram-induced damage both in vivo and in vitro which were considerably ameliorated with antioxidants and CYP inhibitors. In conclusion, it is suggested that citalopram hepatotoxicity might be a result of oxidative hazard leading to mitochondrial/lysosomal toxic connection and disorders in biochemical markers which were supported by histomorphological studies.


Estimating the dual-enzyme kinetic parameters for Cr (VI) reduction by Shewanella oneidensis MR-1 from soil column experiments.
Hexavalent chromium (Cr (VI)) contamination of soil and groundwater is considered a major environmental concern. Bioreduction of Cr (VI) to trivalent chromium (Cr (III)) can be considered an effective technology in remediating Cr (VI) contaminated sites. Among the Cr (VI) reducing bacteria, Shewanella oneidensis MR-1 (MR-1) is relatively effective. Reduction of Cr (VI) by MR-1 is defined by the dual-enzyme kinetic model. The feasibility of bioreduction of Cr (VI) is usually evaluated by performing batch experiments which may not accurately represent the subsurface environment. The objective of this paper is to present a rational approach to determine the dual-enzyme reaction kinetics of Cr (VI) reduction by MR-1 from continuous flow soil column experiments which more accurately simulate the subsurface environment. Kinetic parameters obtained from soil column data were found to be much smaller than that obtained from batch data. The stable enzyme induced reaction rate was found to be 127th and the average deactivating enzyme induced reaction rate was 117th of the corresponding values obtained from batch reactor data as reported in literature.


The Parkinson's disease-associated genes ATP13A2 and SYT11 regulate autophagy via a common pathway.
Forms of Parkinson's disease (PD) are associated with lysosomal and autophagic dysfunction. ATP13A2, which is mutated in some types of early-onset Parkinsonism, has been suggested as a regulator of the autophagy-lysosome pathway. However, little is known about the ATP13A2 effectors and how they regulate this pathway. Here we show that ATP13A2 depletion negatively regulates another PD-associated gene (SYT11) at both transcriptional and post-translational levels. Decreased SYT11 transcription is controlled by a mechanism dependent on MYCBP2-induced ubiquitination of TSC2, which leads to mTORC1 activation and decreased TFEB-mediated transcription of SYT11, while increased protein turnover is regulated by SYT11 ubiquitination and degradation. Both mechanisms account for a decrease in the levels of SYT11, which, in turn, induces lysosomal dysfunction and impaired degradation of autophagosomes. Thus, we propose that ATP13A2 and SYT11 form a new functional network in the regulation of the autophagy-lysosome pathway, which is likely to contribute to forms of PD-associated neurodegeneration.


Designs of metal-organic framework nanoparticles for protein delivery.
Recently, there has been high interest in developing metal-organic framework (MOF) nanoparticles (NPs) for delivering therapeutic proteins, propelled mainly by the unique hierarchical porous structures of MOFs for protein encapsulation. Novel design strategies have emerged for broad therapeutic applications and clinical translations, leading to multifunctional MOF-NPs with improved biointerfacing capabilities and higher potency. This review summarizes recent MOF-NP designs specifically for protein delivery. The summary focuses on four design categories, including environment-responsive MOF-NPs for on-demand protein delivery, cell membrane-coated MOF-NPs for biomimetic protein delivery, cascade reaction-incorporated MOF-NPs for combinatorial protein delivery, and composite MOF-NPs for intelligent protein delivery. The major challenges and opportunities in using MOF-NPs for protein delivery are also discussed. Overall, this review will promote designs of MOF-NPs with unique properties to address unmet medical needs.


Effect of photoperiod on the growth of reproductive organs and on pineal N-acetyltransferase rhythm in male rats treated neonatally with testosterone propionate.
Neonatal administration of testosterone propionate (TP) sensitized male Wistar rats to photoperiodic regulation of reproduction. Testicular, seminal vesicle and prostate weights in 105-day-old TP-treated rats housed under natural photoperiodic conditions were lower during autumn and winter than during spring and summer. The reproductive organ weights in 75-day-old TP-treated rats kept under artificial lighting regimens were smaller under photoperiods shorter than 10 h per day than under photoperiods of 10 h per day or longer. In TP-treated males, pinealectomy prevented the decrease in reproductive organ weights induced by short days. The pineal involvement in the photoperiodic regulation of reproduction might be mediated by the rhythm in melatonin. Under a 8L:16D regimen, when reproductive organ weights were decreased, the period of high night N-acetyltransferase activity, which determines the period of the increased melatonin production, lasted about 1 h longer than under 12L:12D.


A randomized controlled intervention trial to relieve and prevent neck/shoulder pain.
The objective of this study is to investigate the effect of three different workplace interventions on long-term compliance, muscle strength gains, and neck/shoulder pain in office workers. A 1-yr randomized controlled intervention trial was done with three groups: specific resistance training (SRT, n = 180), all-round physical exercise (APE, n = 187), and reference intervention (REF, n = 182) with general health counseling. Physical tests were performed and questionnaires answered at pre-, mid-, and postintervention. The main outcome measures were compliance, changes in maximal muscle strength, and changes in intensity of neck/shoulder pain (scale 0-9) in those with and without pain at baseline. Regular participation was achieved by 54%, 31%, and 16% of those of the participants who answered the questionnaire in SRT (78%), APE (81%), and REF (80%), respectively, during the first half of the intervention period, and decreased to 35%, 28% and 9%, respectively, during the second half. Shoulder elevation strength increased 9-11% in SRT and APE (P < 0.0001). Participants with neck pain at baseline decreased the intensity of neck pain through SRT, from 5.0 +/- 0.2 to 3.4 +/- 0.2 (P < 0.0001), and through APE, from 5.0 +/- 0.2 to 3.6 +/- 0.2 (P < 0.001), whereas REF caused no change. For participants without shoulder pain at baseline, there was a significantly greater increase in pain over the 1-yr period in REF compared with SRT and APE (P < 0.01). Compliance was highest in SRT but generally decreased over time. SRT and APE caused increased shoulder elevation strength, were more effective than REF to decrease neck pain among those with symptoms at baseline, and prevent development of shoulder pain in those without symptoms at baseline.


The role of the computer in cardiac pacemaker technology.
To satisfy the need for even higher performance and smaller size, pacemaker design makes use of the latest available technology. This high level of technology in pacemakers requires an even higher level of technology in the follow-up system. Present methods consisting of independent programmers for each pacer model are confusing to the operator and inadequate. The solution to pacer follow-up developed at Montefiore consists of a three step program: 1) computer-assisted program which guides the personnel interactively through the follow-up; 2) computer-controlled program which enables the person to send identical commands to various programmers and let the computer program identify the various pacer models; and 3) built-in program where the testing and the follow-up are performed internally to the pacer through stored programs. The system is developed in the MIIS version of the MUMPS language but was extensively modified to make conversion to other languages easy.


Gender and ethnic differences in the health practices of the elderly poor.
Gender and ethnic differences in personal health practices were examined with a sample of 1021 Medicaid recipients aged 65 years and older. Six different health practices were examined including smoking, drinking, weight control, exercise, and social networks. Although gender differences were noted with respect to all of the health practices examined, neither men nor women were consistently favored in adhering to desired health practices. Perhaps most importantly, women were more apt than men to refrain from smoking and drinking which have been strongly linked to major health consequences. Ethnicity was also a factor in all of the health practices examined. Whereas Hispanic and Black elderly were very similar to White elderly, Asian elderly generally adhered to more desired health practices than did either Whites, Blacks, or Hispanics. The implications of these findings could benefit health promotion programs for the elderly.


Staying alive--naïve CD4(+) T cell homeostasis.
The immune system must maintain a broad repertoire of naïve T cells in order to respond to the diverse range of pathogens that it will encounter over the course of a lifetime. Although it is known that contact with IL-7 is crucial for the survival of naïve T cells, the precise intracellular signals that mediate its effects remain obscure. An article in this issue of the European Journal of Immunology has found that IL-7 requires the coordinated action of multiple pathways to maintain naïve CD4(+) T cells.


The distraction reduction fixation system and its application in spondylolisthesis.
An internal fixation apparatus--distraction reduction fixation system (DRFS) was designed to satisfy the clinical needs for spondylolisthesis. Since 1996, 53 patients were treated with DRFS. Among them, 35 had spondylolisthesis, 12 had lumbar canal stenosis accompanied with instability, 2 had vertebral tumors and 4 suffered from spinal fracture. The average age was 53.6 years old (ranged 24 approximately 72 yrs). The mean time for follow-up was 30.6 months (16 approximately 44 months). The slip rate was 0.15 +/- 0.10 before operation, and decreased to 0.09 +/- 0.07 after operation. Entire slip reposition was achieved in 19 cases (54.3%). The change in height of the intervertebral space within the fixation segments was 0.7 +/- 0.17. DRFS achieved better results for spondylolisthesis less II degree and no other adverse effects were found. Compared with other foreign and domestic techniques, it had advantages in less implants, less operation gears required and ease to utilize in operation. It was proved to be an ideal internal fixation apparatus.


Rapid prenatal diagnosis of HG-PRT deficiency using ultra-microchemical methods.
A previously developed simple ultramicromethod has been used for the rapid prenatal diagnosis of hypoxanthine-guanine phosphoribosyl transferase (HG-PRT) deficiency. The method is based on the incubation of small numbers of visually selected, lyophilized fibroblasts (in the present study five cells per incubation) with radioactive substrate in an end volume of 0.3 microliter. Fibroblasts derived from the amniotic fluid of a 15-week male fetus in a heterozygote for the X-linked Lesch-Nyhan syndrome showed a severe degree of HG-PRT deficiency. In total 50 fibroblasts were used. The diagnosis was confirmed upon termination of the pregnancy by the demonstration of HG-PRT deficiency in fetal erythrocytes and cultured skin fibroblasts.


The expression in yeast of the Escherichia coli galK gene on CYC1::galK fusion plasmids.
A set of gene fusions have been constructed between the transcriptional and translational initiation signals of the yeast CYC1 gene, encoding iso-1-cytochrome c, and the coding sequence of the Escherichia coli galK gene encoding galactokinase. These fusions are contained on plasmids which have both yeast and E. coli replication origins and selectable markers and, therefore, can be used to transform either yeast or E. coli cells. When galactokinase-deficient (gall-) yeasts were transformed with these plasmids the resulting Gal+ transformants were heterogeneous with respect to their galactokinase levels. The galactokinase levels in all were subject to glucose repression, characteristic of the transcriptional regulation of the CYC1 gene. The fusion points for representative plasmids were determined by DNA sequence analysis, and from these data, the differential expression of the galK gene could be explained. One fusion plasmid, YRpR1, which gave the highest level of galK expression, was characterized further. As an additional demonstration that galactokinase expression from the fusion was under CYC1 transcriptional control, a cis-dominant, CYC1-linked mutation known to drastically reduce CYC1 gene transcription was introduced into YRpR1 and shown to similarly effect galK expression. The galK mRNA produced from the fused gene of YCpR1, a centromere-containing derivative of YRpR1, consisted of the mRNA leader sequence plus the first four codons of the CYC1 gene, the galK coding sequence, then the remainder of the CYC1 coding sequence and the 175 nucleotide non-translated 3' sequence. As a demonstration of the usefulness of these plasmids for the selection of regulatory mutants, two mutants capable of greatly enhanced levels of galactokinase expression were isolated. Preliminary characterization of these mutations indicates that they likewise affect the expression of the chromosomal CYC1 gene.


Methylenedioxy designer drugs: mass spectrometric characterization of their glutathione conjugates by means of liquid chromatography-high-resolution mass spectrometry/mass spectrometry and studies on their glutathionyl transferase inhibition potency.
Methylenedioxy designer drugs of abuse such as 3,4-methylenedioxymethamphetamine (MDMA) can be selectively toxic to serotonergic neurons and glutathione (GSH) adducts have been implicated in its neurotoxicity. The catecholic demethylenyl metabolites of MDMA, 3,4-dihydroxymethamphetamine and 3,4-dihydroxyamphetamine, are metabolically oxidized to the corresponding ortho-quinones, which are highly reactive intermediates. These intermediates can then be conjugated with GSH preventing cellular damage. Furthermore, glutathionyl transferase (GST) activity was described to be irreversibly inhibited by the catechols dopamine, α-methyldopa and their GSH conjugates. Therefore, the aims of the present work were the detection and characterization of GSH conjugates of ten methylenedioxy drugs of abuse and their phase I metabolites as well as to assess their inhibition potency on GST activity. The substrates were incubated using human placental GST with or without preincubation by cytochrome P450 enzymes preparations. GST inhibition was tested using chlorodinitrobenzene GSH conjugation as marker reaction. GSH conjugates were analyzed and characterized using LC-high-resolution-MS/MS. For confirmation of postulated fragmentation patterns, formation of GSH conjugates of selected deuterated analogs (deuterated analogue approach, DAA) of the investigated drugs was explored. For the methylenedioxy amphetamines the following steps could be identified: conjugation of the parent compounds at position 2, 5, 6, of the demethylenyl metabolites at position 2 and 5, and of the further deaminated demethylenyl metabolites at position 2. For the β-keto-phenylalkylamine and pyrrolidinophenone, conjugation of the demethylenyl metabolites and of the deaminated demethylenyl metabolites at position 2 could be identified. The DAA allowed the differentiation of the 2 and 5/6 isomers by confirmation of the postulated mass spectral fragments. Finally, the tested drugs and phase I metabolites showed no inhibition potency on GST activity.


Energy dispersive X-ray microanalysis of zinc and calcium in organelles of insulin-producing cells of the mouse, rat, and a fish.
By means of energy dispersive X-ray microanalysis in the scanning-transmission electron microscope, spectra were obtained from quick-frozen, cryo-ultramicrotome cut, freeze-dried sections of insulin cells from a fish and a mouse. It was shown that both zinc and calcium are present in significant quantities in native islet-cell beta-granules. In the beta-granules of the rat RINm5F insuloma cells calcium, but not zinc, seemed to accumulate; the zinc contents in the secretion granules of these neoplastic beta-cells were probably below the detection limit.


AML consolidation therapy: timing matters.
Infections due to severe neutropenia are the most common therapy-associated causes of mortality in patients with acute myeloid leukemia (AML). New strategies to lessen the severity and duration of neutropenia are needed. Cytarabine is commonly used for AML consolidation therapy; we compared high- and intermediate-dose cytarabine administration on days 1, 2, and 3 (AC-123) versus days 1, 3, and 5 (AC-135) in consolidation therapy of AML. Recently, clinical trials demonstrated that high-dose AC-123 resulted in a shortened white blood cell (WBC) recovery time compared with high-dose AC-135. Our main hypothesis is that this is also the case for different cytarabine dosage, granulocyte colony-stimulating factor (G-CSF) administration, and cycle lengths. We analyzed 334 treatment schedules on virtual cohorts of digital twins. Comparison of 32,565 simulated consolidation cycles resulted in a reduction in the WBC recovery time for AC-123 in 99.6% of the considered cycles (median reduction 3.5 days) without an increase in the number of leukemic blasts (lower value in 94.2% of all cycles), compared to AC-135. Our numerical study supports the use of AC-123 plus G-CSF as standard conventional AML consolidation therapy to reduce the risk for life-threatening infectious complications.


Neurotrophin receptor activation and expression in human postmortem brain: effect of suicide.
The physiological functions of neurotrophins occur through binding to two receptors: pan75 neurotrophin receptor (p75(NTR)) and a family of tropomyosin receptor kinases (Trks A, B, and C). We recently reported that expression of neurotrophins and TrkB were reduced in brains of suicide subjects. This study examines whether expression and activation of Trk receptors and expression of p75(NTR) are altered in brain of these subjects. Expression levels of TrkA, B, C, and of p75(NTR) were measured by quantitative reverse transcription polymerase chain reaction and Western blot in prefrontal cortex (PFC) and hippocampus of suicide and normal control subjects. The activation of Trks was determined by immunoprecipitation followed by Western blotting using phosphotyrosine antibody. In hippocampus, lower mRNA levels of TrkA and TrkC were observed in suicide subjects. In the PFC, the mRNA level of TrkA was decreased, without any change in TrkC. However, the mRNA level of p75(NTR) was increased in both PFC and hippocampus. Immunolabeling studies showed similar results as observed for the mRNAs. In addition, phosphorylation of all Trks was decreased in hippocampus, but in PFC, decreased phosphorylation was noted only for TrkA and B. Increased expression ratios of p75(NTR) to Trks were also observed in PFC and hippocampus of suicide subjects. Our results suggest not only reduced functioning of Trks in brains of suicide subjects but also that increased ratios of p75(NTR) to Trks indicate possible activation of pathways that are apoptotic in nature. These findings may be crucial in the pathophysiology of suicide.


Associations Between Coronary Artery Calcification and Left Ventricular Global Longitudinal Strain and Diastolic Parameters: the ELSA-Brasil Study.
Atherosclerosis burden can be evaluated in asymptomatic patients by measuring coronary artery calcification (CAC), whereas the global longitudinal strain (GLS) and diastolic function parameters (mitral E/e' ratio, septal e', and lateral e') are used to evaluate subclinical left ventricular (LV) dysfunction. We investigated whether subjects with CAC (CAC >0 Agatston units) would present with an impairment in LV functional parameters. Among the participants of the ELSA-Brasil cohort free of clinically prevalent cardiovascular disease who performed cardiac computed tomography and echocardiography within the study protocol, we tested whether those with CAC >0 presented with worse GLS and diastolic function parameters. CAC >0 was present in 203 of the 612 included participants (33.17%; age 51.4 ± 8.6 years, 52.1% women). Absolute CAC values did not correlate with GLS (ro = 0.07, p = 0.105) but did so with E/e' (ro = 0.19, p <0.001), septal e' (ro = 0.28, p <0.001), and lateral e' (ro = 0.30, p <0.001), with stronger correlations in men. Those with CAC >0 had worse mitral E/e' ratios (7.75 ± 0.13 vs 7.01 ± 0.09; p ≤0.001), septal e' (8.25 ± 0.15 vs 9.59 ± 0.11 cm/s; p <0.001), and lateral e' (10.13 ± 0.20 vs 11.99 ± 0.14 cm/s; p ≤0.001), respectively. However, these associations were not independent of diabetes, obesity, hypertension, smoking, and low-density lipoprotein cholesterol, persisting only as significant associations of CAC >0 with mitral E/e' ratio and septal e' in men. There is an association between subclinical coronary atherosclerosis and impaired LV functional parameters. These associations are more likely attributed to the presence of common cardiovascular risk factors in the general population. However, in men, it seems to exist as an independent association.


Sodium and potassium intakes among US infants and preschool children, 2003-2010.
Data are limited on usual sodium and potassium intakes relative to age-specific recommendations and the sodium:potassium ratio in infants and preschoolers, especially among those aged <2 y, who are black or breastfed. The usual sodium intake above the Tolerable Upper Intake Levels (ULs), potassium intakes above Adequate Intakes (AIs), the sodium:potassium ratio, and sodium density (mg/kcal) among US infants and preschoolers by age group, as applicable, were estimated and compared by race-ethnicity and current breastfeeding status. Data were analyzed among 3 groups of children (aged 7-11 mo, 1-3 y, and 4-5 y) from the NHANES 2003-2010 by using measurement error models. Seventy-nine percent of children aged 1-3 y and 87% of those aged 4-5 y exceeded their sodium UL; among non-Hispanic black children, the estimates were 84% and 97%, respectively. For potassium, 97% of infants, 5% of children aged 1-3 y, and 0.4% aged 4-5 y met their AIs. Compared with non-Hispanic whites and Mexican Americans, non-Hispanic black infants and preschoolers had higher mean sodium density and sodium:potassium ratios. Currently breastfed infants and children consumed, on average, less sodium than those who were not breastfed (382 ± 53 compared with 538 ± 22 mg in those aged 7-11 mo and 1154 ± 88 compared with 1985 ± 24 mg in those aged 1-3 y, respectively), but the sodium:potassium ratio did not differ. Most US preschoolers, particularly non-Hispanic blacks, consume too much sodium, and nearly all do not consume enough potassium. Data that suggest that currently breastfed infants consume less sodium than do those who are not breastfeeding merit further investigation.


New Approaches to Critical Illness Polyneuromyopathy: High-Resolution Neuromuscular Ultrasound Characteristics and Cytokine Profiling.
Diagnosis of intensive care unit acquired weakness (ICUAW) is challenging. Pathogenesis of underlying critical illness polyneuromyopathy (CIPNM) remains incompletely understood. This exploratory study investigated whether longitudinal neuromuscular ultrasound examinations and cytokine analyses in correlation to classical clinical and electrophysiological assessment contribute to the understanding of CIPNM. Intensive care unit patients were examined every 7 days until discharge from hospital. Clinical status, nerve conduction studies, electromyography as well as ultrasound of peripheral nerves and tibial anterior muscle were performed. Cytokine levels were analyzed by a bead-based multiplex assay system. Of 248 screened patients, 35 patients were included at median of 6 days (IQR: 8) after admission to intensive care unit. Axonal damage was the main feature of CIPNM. At the peak of CIPNM (7 days after inclusion), nerve ultrasound showed cross-sectional area increase of tibial nerve as a sign of inflammatory edema as well as hypoechoic nerves as a possible sign of inflammation. Cytokine analyses showed signs of monocyte and macrophage activation at this stage. Fourteen days after inclusion, cytokines indicated systemic immune response as well as profiles associated to neovascularization and regeneration. Exploratory neuromuscular ultrasound and cytokine analyses showed signs of inflammation like macrophage and monocyte activation at the peak of CIPNM followed by a systemic immune response parallel to axonal damage. This underlines the role of both axonal damage and inflammation in pathogenesis of CIPNM.


PKC-epsilon-dependent cytosol-to-membrane translocation of pendrin in rat thyroid PC Cl3 cells.
We studied the expression and the hormonal regulation of the PDS gene product, pendrin, which is, in thyrocytes, responsible for the iodide transport out of the cell. We show that PC Cl3 cells, a fully differentiated thyroid cell line, grown without TSH and insulin, express very low level of PDS mRNA; such expression is greatly increased after stimulation with insulin or TSH. (125)I pre-loaded cells showed an (125)I efflux accelerated in chloride-containing buffer with respect to chloride-free buffer, suggesting that this efflux is chloride dependent. By immunoblotting, pendrin was found in agonists-stimulated cells, whereas it was barely detectable in un-stimulated cells. An increase in both PDS mRNA and protein was also obtained using phorbol ester PMA, or using 8-Br-cAMP and forskolin. Stimulation with insulin (1 microg/ml; 0-40 min) provoked the cytosol-to-membrane translocation of pendrin and a decrease of intracellular I(-) content in (125)I pre-loaded cells. Insulin- or PMA-treated cells also showed a cytosol-to-membrane translocation of PKC-delta and -epsilon. Inhibition of both PKC-delta and -epsilon activities by GF109203X blocked pendrin translocation, whilst the inhibition of PKA did not. The selective inhibition of PKC-delta by rottlerin did not affect the insulin-provoked translocation of pendrin whilst it was inhibited by a PKC-epsilon translocation inhibitor peptide and also by PKC-epsilon downregulation using the small interfering RNA, thus indicating that such translocation was due to PKC-epsilon activity. In conclusion, our study demonstrates that, in PC Cl3 cells, pendrin expression and localisation are regulated by insulin and influenced by a PKC-epsilon-dependent intracellular pathway.


Radiochemical analysis of 93Zr.
The zirconium isotope 93Zr is a long-lived pure beta-particle-emitting radionuclide, which is produced by nuclear fission and neutron activation of the stable isotope 92Zr. This element is a constituent of the structural components of nuclear reactor vessels. A selective liquid-liquid extraction method for radiochemical separation of Zr, based on liquid-liquid extraction with 1-(2'-thenoyl)-3,3,3-trifluoroacetone in xylene and a subsequent stripping of 93Zr by an aqueous acid solution, has been developed. The method was utilised to separate Zr from other pure beta-particle and beta-gamma emitters in different kinds of samples. Decontamination factors higher than 99% for the pure beta-particle and beta-gamma emitters and an overall chemical yield of 80% were obtained. The sensitivity of the method allows the determination of the isolated 93Zr by liquid scintillation counting and the minimum detectable activity value obtained was 0.067 Bq over a counting period of 60 min.


EEG source localization: implementing the spatio-temporal decomposition approach.
The spatio-temporal decomposition (STD) approach was used to localize the sources of simulated electroencephalograms (EEGs) to gain experience with the approach for analyzing real data. The STD approach used is similar to the multiple signal classification method (MUSIC) in that it requires the signal subspace containing the sources of interest to be isolated in the EEG measurement space. It is different from MUSIC in that it allows more general methods of spatio-temporal decomposition to be used that may be better suited to the background EEG. If the EEG data matrix is not corrupted by noise, the STD approach can be used to locate multiple dipole sources of the EEG one at a time without a priori knowledge of the number of active sources in the signal space. In addition, the common-spatial-patterns method of spatio-temporal decomposition is superior to the eigenvector decomposition for localizing activity that is ictal in nature. The STD approach appears to be able to provide a means of localizing the equivalent dipole sources of realistic brain sources and that, even under difficult noise conditions and only 2 or 3 s of available EEG, the precision of the localization can be as low as a few mm.


Signaling Pathways that Regulate Macropinocytosis in Mammalian Cells.
Macropinocytosis is an evolutionarily conserved endocytic pathway that mediates non-selective uptake of extracellular fluid in bulk. Macropinocytosis is initiated by localized polymerization of the actin cytoskeleton, which generates plasma membrane protrusions that enclose part of the environment into large endocytic vesicles. From amoebae to mammalian cells, the actin dynamics that drive macropinosome formation are regulated by a conserved set of intracellular signaling proteins including Ras superfamily GTPases and PI3-kinases. In mammalian cells, multiple upstream signaling pathways control activity of these core regulators in response to cell-extrinsic and cell-intrinsic stimuli. Growth factor signaling pathways play a central role in macropinocytosis induction. In addition, an increasing number of functionally diverse processes has been identified as macropinocytosis regulators, including several nutrient-sensing and developmental signaling pathways. Many of these signaling pathways have proto-oncogenic properties, and their dysregulation drives the high macropinocytic activity that is commonly observed in cancer cells. These regulatory principles illustrate how macropinocytosis is controlled by complex upstream inputs to exert diverse cellular functions in physiological and pathological contexts.


Prevalidation of in vitro continuous flow exposure systems as alternatives to in vivo inhalation safety evaluation experimentations: outcome from MAAPHRI-PCRD5 research program.
Diesel engine emission aerosol-induced toxicity patterns were compared using both in vitro (organotypic cultures of lung tissue) and in vivo experimentations mimicking the inhalation situation with continuous aerosol flow exposure designs. Using liquid media resuspended diesel particles, we show that toxic response pattern is influenced by the presence of tensioactive agent in the medium which alter particle-borne pollutant bioavailability. Using continuous aerosol exposure in vitro, we show that with high sulfur fuel (300ppm) in the absence of oxidation catalysis, particulate matter was the main toxic component triggering DNA damage and systemic inflammation, while a very limited oxidant stress was evidenced. In contrast, with ultra-low sulfur fuel in the presence of strong diesel oxidation catalysis, the specific role of particulate matter is no longer evidenced and the gas phase then becomes the major component triggering strong oxidant stress, increased NO(2) being the most probable trigger. In vivo, plasma tumor necrosis factor alpha (TNFalpha), lung superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) activity levels varied in agreement with in vitro observations. Diesel emission treatment with oxycat provokes a marked systemic oxidant stress. Again NO(2) proved to account for a major part of these impacts. In conclusion, similar anti-oxidant responses were observed in in vitro and in vivo experiments after diesel emission aerosol continuous flow exposures. The lung slice organotypic culture model-exposed complex aerosol appears to be a very valuable alternative to in vivo inhalation toxicology experimentations in rodents.


Identification of health care problems in a county jail.
The health care needs of prisoners often are not systematically addressed. To evaluate the ability of a questionnaire, that is administered at the time of arrest, to identify inmates' medical problems and to predict subsequent use of care, we reviewed and analyzed medical records and questionnaires for 594 individuals incarcerated in a county jail. Only 54% of subjects had a completed questionnaire filed in a medical record; the questionnaire identified only 48% of persons who eventually sought care for such major problems as drug and alcohol abuse and cardiopulmonary disease. No question screened for psychiatric problems, even though five percent of subjects were diagnosed as having major psychiatric disorders during their incarcerations. On the other hand, many items addressed problems that were far less common. Uniform screening of prison populations may be effective if conducted with greater rigor and better instruments than were employed in the institution studied. Health care for inmates might be served better by universal screening and follow-up, targeted to such important problems as drug and alcohol abuse and psychiatric disorders.


2'-5' oligo(A) synthetase levels in interferon-sensitive or -resistant lymphoid cells.
The activity of 2'-5' oligo(A) synthetase has been measured both in control and interferon-treated lymphoid cells. While control or interferon-treated Raji and Namalwa cells exhibit low activity of the 2'-5' oligo(A) synthetase, the activity of this enzyme is increased 20-fold when Daudi cells are treated with the same dose of human interferon. The same enzyme is induced by interferon in R3HR-1 cells to a lesser extent.


Cavernous Hemangioma of the Adrenal Gland: A Rare Adrenal Incidentaloma.
Cavernous hemangiomas of the adrenal gland are rare, benign, endocrinologically inactive tumors. They present as adrenal incidentalomas, mostly asymptomatic, but patients may have abdominal pain. Surgery is the mainstay of treatment.


Manulex-infra: distributional characteristics of grapheme-phoneme mappings, and infralexical and lexical units in child-directed written material.
It is well known that the statistical characteristics of a language, such as word frequency or the consistency of the relationships between orthography and phonology, influence literacy acquisition. Accordingly, linguistic databases play a central role by compiling quantitative and objective estimates about the principal variables that affect reading and writing acquisition. We describe a new set ofWeb-accessible databases of French orthography whose main characteristic is that they are based on frequency analyses of words occurring in reading books used in the elementary school grades. Quantitative estimates were made for several infralexical variables (syllable, grapheme-to-phoneme mappings, bigrams) and lexical variables (lexical neighborhood, homophony and homography). These analyses should permit quantitative descriptions of the written language in beginning readers, the manipulation and control of variables based on objective data in empirical studies, and the development of instructional methods in keeping with the distributional characteristics of the orthography.


Synthesis of new pyridines with oligocations and oxygen nucleophiles.
Nucleophilic substitution of 4-(dimethylamino)pyridine on pentachloropyridine yielded pentakis(pyridine-2,3,4,5,6-pentayl)pyridinium, tris-(3,5-dichloropyridine-2,4,6-triyl)pyridinium, or (tetrachloropyridin-4-yl)pyridinium depending on the reaction conditions. Nucleophilic substitution with water, hydroxides, and alcoholates resulted in new betaines and highly substituted pyridines. [structure: see text]


Early detection of a twin tubal pregnancy by Doppler sonography allows fertility-conserving laparoscopic surgery.
Vital twin tubal gestation is a rare complication in pregnancy. An asymptomatic gravida 1 female presented with increasing beta-HCG levels without an intrauterine gestation. Doppler sonography revealed cardiac motion of unilateral tubal twins. The patient was treated laparoscopically by partial tubectomy allowing a subsequent tubal re-anastomosis. Doppler sonography may detect vital ectopic pregnancies early enabling fertility-conserving surgery.


Relationships between body burdens of trace metals (As, Cu, Fe, Hg, Mn, Se, and Zn) and the relative body size of small tooth flounder (Pseudorhombus jenynsii).
Several studies have described strong relationships between body size and the accumulation of trace metals in animal tissues. However, few of these studies have utilized aging techniques to control for age related effects. We utilized relative body size (gy(-1)) of a model flounder species, Pseudorhombus jenynsii, in order to control for age related effects on growth and size measurements. We investigated links between relative body size, concentrations of trace metals in flounder muscle tissue, physico-chemical variables (temperature, salinity, pH, and turbidity), and levels of trace metals in the sediment. Flounder were sampled using an otter trawl net in the inner areas of eight estuaries that were either heavily modified or relatively unmodified by urbanization and industrial activity. Our results indicate that this commonly eaten fish is accumulating significant levels of some trace metals in their muscle tissue, both in relatively unmodified and heavily modified estuaries. Concentrations of Cu, Zn and Fe in muscle tissue, as well as temperature, showed a negative relationship to the relative body size of flounder. In contrast, Se and Hg in muscle showed a positive relationship to relative body size. Observed growth patterns indicate that these effects are not driven by age related differences in metabolic activity. Instead, our results suggest that differences in food supply or toxicological effects may be responsible for the observed relationships between relative body size and concentrations of Cu, Zn, and Se in muscle tissues. The use of otolith aging and growth measurement techniques represents a novel method for assessing the relationships between trace metal accumulation and the relative body size of fish in a field environment.


Burnout, Mental Health, and Workplace Discrimination in Lesbian, Gay, Bisexual, Transgender, Queer/Questioning, Intersex, and Asexual Anesthesiologists.
Increasing attention is being paid to both anesthesiologist well-being and commitments to diversity, equity, and inclusion. Sexual minorities (ie, members of the lesbian, gay, bisexual, transgender, queer/questioning, intersex, and asexual [LGBTQIA] communities) face many challenges in society and the workplace, including mental health conditions, discrimination, and increased risk for burnout. In this review, we outline the current state of mental health conditions and burnout in sexual minority individuals, discrimination and harassment faced both in society and the workplace, and steps that workplaces can take to become more inclusive and welcoming.


Differentiation of HELLP patients from healthy pregnant women by proteome analysis--on the way towards a clinical marker set.
As the pathogenesis of the HELLP-syndrome is unknown, a complex consideration regarding the changes in the plasma as the main transport medium in the body is of great benefit because it is well available and can rapidly be investigated in the clinics. Besides that, the liver which is early affected in HELLP-syndrome produces the main part of plasma proteins. For the purpose of our study plasma protein abundances from patients with HELLP-syndrome and from control individuals were determined before and after delivery. In the differential analysis using two-dimensional gel electrophoresis, six areas with variable protein spot intensities were detected. The reference gel that we developed for HELLP plasma samples integrates the changes of plasma proteins when comparing HELLP patients to healthy women prior to and after delivery. A specific plasma protein profile for the HELLP-syndrome was generated involving protein areas that contain inter-alpha-trypsin inhibitor heavy chain H4, kininogen 1, fibrinogen gamma chain, transthyretin, haptoglobins, and serum amyloid A with statistically significant expression differences when compared to controls. The most striking difference between the majority of the gels from HELLP patients and the gels from non-HELLP samples were clearly overexpressed protein spots at about 11 kDa which were identified as serum amyloid A (SAA). This differential expression was validated and quantitatively assayed by ELISA measurements against human SAA in plasma. Our results show that significant differences in SAA expressions between healthy controls and HELLP patients were obtained, that could function as markers for the HELLP-syndrome. According to our data it is possible to draw a line of separation with no overlap between the HELLP group for which SAA plasma levels were found to be above 3.51 mg/L and the non-HELLP groups in which SAA plasma levels were below 3.51 mg/L. It now is possible to clinically elucidate if the differentially expressed proteins are suited for longitudinal studies concerning both, to function as markers or perhaps even as disease predictors that might become relevant for diagnostic tests.


Nonequilibrium site distribution governs charge-transfer electroluminescence at disordered organic heterointerfaces.
The interface between electron-donating (D) and electron-accepting (A) materials in organic photovoltaic (OPV) devices is commonly probed by charge-transfer (CT) electroluminescence (EL) measurements to estimate the CT energy, which critically relates to device open-circuit voltage. It is generally assumed that during CT-EL injected charges recombine at close-to-equilibrium energies in their respective density of states (DOS). Here, we explicitly quantify that CT-EL instead originates from higher-energy DOS site distributions significantly above DOS equilibrium energies. To demonstrate this, we have developed a quantitative and experimentally calibrated model for CT-EL at organic D/A heterointerfaces, which simultaneously accounts for the charge transport physics in an energetically disordered DOS and the Franck-Condon broadening. The 0-0 CT-EL transition lineshape is numerically calculated using measured energetic disorder values as input to 3-dimensional kinetic Monte Carlo simulations. We account for vibrational CT-EL overtones by selectively measuring the dominant vibrational phonon-mode energy governing CT luminescence at the D/A interface using fluorescence line-narrowing spectroscopy. Our model numerically reproduces the measured CT-EL spectra and their bias dependence and reveals the higher-lying manifold of DOS sites responsible for CT-EL. Lowest-energy CT states are situated ∼180 to 570 meV below the 0-0 CT-EL transition, enabling photogenerated carrier thermalization to these low-lying DOS sites when the OPV device is operated as a solar cell rather than as a light-emitting diode. Nonequilibrium site distribution rationalizes the experimentally observed weak current-density dependence of CT-EL and poses fundamental questions on reciprocity relations relating light emission to photovoltaic action and regarding minimal attainable photovoltaic energy conversion losses in OPV devices.


A Promising Future for Stem-Cell-Based Therapies in Muscular Dystrophies-In Vitro and In Vivo Treatments to Boost Cellular Engraftment.
Muscular dystrophies (MD) are a group of genetic diseases that lead to skeletal muscle wasting and may affect many organs (multisystem). Unfortunately, no curative therapies are available at present for MD patients, and current treatments mainly address the symptoms. Thus, stem-cell-based therapies may present hope for improvement of life quality and expectancy. Different stem cell types lead to skeletal muscle regeneration and they have potential to be used for cellular therapies, although with several limitations. In this review, we propose a combination of genetic, biochemical, and cell culture treatments to correct pathogenic genetic alterations and to increase proliferation, dispersion, fusion, and differentiation into new or hybrid myotubes. These boosted stem cells can also be injected into pretreate recipient muscles to improve engraftment. We believe that this combination of treatments targeting the limitations of stem-cell-based therapies may result in safer and more efficient therapies for MD patients. Matricryptins have also discussed.


Probing the Link between Pancratistatin and Mitochondrial Apoptosis through Changes in the Membrane Dynamics on the Nanoscale.
Pancratistatin (PST) is a natural antiviral alkaloid that has demonstrated specificity toward cancerous cells and explicitly targets the mitochondria. PST initiates apoptosis while leaving healthy, noncancerous cells unscathed. However, the manner by which PST induces apoptosis remains elusive and impedes the advancement of PST as a natural anticancer therapeutic agent. Herein, we use neutron spin-echo (NSE) spectroscopy, molecular dynamics (MD) simulations, and supporting small angle scattering techniques to study PST's effect on membrane dynamics using biologically representative model membranes. Our data suggests that PST stiffens the inner mitochondrial membrane (IMM) by being preferentially associated with cardiolipin, which would lead to the relocation and release of cytochrome c. Second, PST has an ordering effect on the lipids and disrupts their distribution within the IMM, which would interfere with the maintenance and functionality of the active forms of proteins in the electron transport chain. These previously unreported findings implicate PST's effect on mitochondrial apoptosis.


Chromosomal aberrations in mouse lymphocytes exposed in vivo and in vitro to aliphatic epoxides.
Mouse lymphocytes in vivo or in vitro were exposed for 24 h to 4 aliphatic epoxides, glycidyl 1-naphthyl ether, glycidyl 4-nitrophenyl ether, 1-naphthyl-propylene oxide and trichloropropylene oxide (TCPO), and tested for the induction of chromosomal aberrations (CA). These epoxides were among the most genotoxic aliphatic epoxides in our previous studies. With the exception of TCPO, the test epoxides caused significant increases in CA in vivo compared to a negative control. There were concentration related increases in CA for all 4 epoxides in vitro and TCPO produced the greatest cellular toxicity and genotoxic effects towards cultured lymphocytes. The difference in the order of genotoxicity for the two test systems can be explained on the basis of a much shorter half-life for TCPO than for the other epoxides.


Transfer of the herpes simplex thymidine kinase gene from human cells to mouse cells by means of metaphase chromosomes.
Thymidine kinase (TK)-deficient human cells were infected with ultraviolet light-inactivated Herpes simplex virus type 1, and "transformed" cells that expressed Herpes TK activity were isolated. Purified metaphase chromosomes were isolated from the transformed human line and incubated with TK-deficient mouse cells. TK+ cells were selected, and it was shown that these cells were gene transferents which expressed Herpes TK activity, identical to that found in the transformed human cells. The gene transferents contained no intact human chromosomes. When removed from selective pressure, the gene transferents rapidly lost the TK+ phenotype. However, upon continued growth in nonselective medium, a subpopulation in which the TK+ phenotype had become more stabilized appeared. These results suggest that the Herpes gene for thymidine kinase has integrated into the genome of the HSV-transformed human cells and that it can be transferred to other cells by means of purified metaphase chromosomes.


Silicone T-tube for complex laryngotracheal problems.
The use of a T-tube to manage complex laryngotracheal lesions, such as tracheal stenosis, tracheomalacia and tracheal injury, has previously been reported by other surgeons in the past. However, further validation of clinical details, including operative management and postoperative care, is needed. From January 1991 to May 2000, 53 patients, including 24 with post-tracheostomy stenosis, received 55 silicone T-tubes for transient or permanent stenting of the airway. There were 20 patients for subglottic stenosis; eight for long segment tracheostensis; seven with tracheal stenosis for severe cervicomediastinal fibrosis not amenable for reconstruction; six for complex tracheal injury; four for glottic injury; two each for tracheomalacia, failed tracheal surgery and tuberculotic tracheostenosis; and one each for tracheo-esophageal fistula and necrotizing tracheitis. We retrospectively analyzed these patients. Thirty-eight out of 53 patients (71.8%) with T-tube stenting from 3 to 15 months was considered successful. Fifteen patients' operations failed due to patients' underlining diseases, previous intractable pulmonary infection, poor cognition and/or inadequate tube position. After removal of the tube, three patients (10.7%) developed partial airway obstruction with mild subglottic granulation tissue, which was resolved by carbon dioxide laser therapy. Two patients (7.1%) with prolonged tracheocutaneous fistula were conservatively treated by silver nitrate. Silicone T-tube can effectively resolve the complex laryngotracheal lesions with limited complications. Concurrent cardiopulmonary diseases and intractable infection were the two major causes for failure after the T-tube reconstruction.


Associations of Pregnancy After Bariatric Surgery with Long-Term Weight Trajectories and Birth Weight: LABS-2 Study.
This study aimed to examine whether pregnancy following bariatric surgery affects long-term maternal weight change and offspring birth weight. Using data from the Longitudinal Assessment of Bariatric Surgery (LABS)-2 study, linear regression was used to evaluate percent change in total body weight over a 5-year follow-up period among reproductive-aged women who underwent Roux-en-Y gastric bypass or laparoscopic adjustable gastric banding as well as evaluate the association of bariatric procedure type and offspring birth weight. Of 727 women with preoperative age of 36.1 (6.3) years (mean [SD]) and BMI of 46.9 (7.0) kg/m2 , 80 (11%) reported at least one pregnancy. After adjusting for covariates, percent change in total body weight was not significantly different between women who became pregnant and those who did not during a 5-year follow-up period (β = 2.02; 95% CI: -1.03 to 5.07; P = 0.19). Additionally, mean birth weight was not significantly different between mothers who underwent Roux-en-Y gastric bypass versus laparoscopic adjustable gastric banding (P = 0.99). Postoperative pregnancy did not diminish long-term weight loss in women in the LABS-2 study. The finding of comparable weight loss is relevant for providers counseling women of reproductive age on weight-loss expectations and family planning following bariatric surgery.


Meta-analysis: Exposure to Early Life Stress and Risk for Depression in Childhood and Adolescence.
Early life stress (ELS) is associated with increased risk for the development of major depressive disorder (MDD) in adulthood; however, the degree to which ELS is associated with an early onset of MDD (ie, during childhood or adolescence) is not known. In this meta-analysis, we estimated the associations between ELS and the risk for onset of MDD before age 18 years. In addition, we examined the associations between eight specific forms of ELS (ie, sexual abuse, physical abuse, poverty, physical illness/injury, death of a family member, domestic violence, natural disaster, and emotional abuse) and risk for youth-onset MDD. We conducted a systematic search in scientific databases for studies that assessed both ELS and the presence or absence of MDD before age 18 years. We identified 62 journal articles with a total of 44,066 unique participants. We assessed study quality using the Newcastle-Ottawa Scale. When heterogeneous effect sizes were detected, we tested whether demographic and/or methodological factors moderated the association between ELS and MDD. Using a random-effects meta-analysis, we found that individuals who experienced ELS were more likely to develop MDD before the age of 18 years than were individuals without a history of ELS (odds ratio = 2.50; 95% confidence interval 2.08, 3.00). Separate meta-analyses revealed a range of associations with MDD: whereas some types of ELS (eg, poverty) were not associated with MDD, other types (eg, emotional abuse) were associated more strongly with MDD than was ELS considered more broadly. These findings provide important evidence that the adverse effect of ELS on MDD risk manifests early in development, prior to adulthood, and varies by type of ELS.


Characterization of two new CTX-M-25-group extended-spectrum β-lactamase variants identified in Escherichia coli isolates from Israel.
We characterized two new CTX-M-type extended-spectrum β-lactamase (ESBL) variants in Escherichia coli isolates from stool samples of two elderly patients admitted at the Tel Aviv Sourasky Medical Center, Israel. Both patients underwent treatment with cephalosporins prior to isolation of the E. coli strains. ESBLs were detected by the double-disk synergy test and PCR-sequencing of β-lactamase genes. The bla(CTX-M) genes were cloned into the pCR-BluntII-TOPO vector in E. coli TOP10. The role of amino-acid substitutions V77A and D240G was analyzed by site-directed mutagenesis of the bla(CTX-M-94) and bla(CTX-M-100) genes and comparative characterization of the resulting E. coli recombinants. MICs of β-lactams were determined by Etest. Plasmid profiling, mating experiments, replicon typing and sequencing of bla(CTX-M) flanking regions were performed to identify the genetic background of the new CTX-M variants. The novel CTX-M β-lactamases, CTX-M-94 and -100, belonged to the CTX-M-25-group. Both variants differed from CTX-M-25 by the substitution V77A, and from CTX-M-39 by D240G. CTX-M-94 differed from all CTX-M-25-group enzymes by the substitution F119L. Glycine-240 was associated with reduced susceptibility to ceftazidime and leucine-119 with increased resistance to ceftriaxone. bla(CTX-M-94) and bla(CTX-M-100) were located within ISEcp1 transposition units inserted into ∼93 kb non-conjugative IncFI and ∼130 kb conjugative IncA/C plasmids, respectively. The plasmids carried also different class 1 integrons. This is the first report on CTX-M-94 and -100 ESBLs, novel members of the CTX-M-25-group.


Spatiotemporal three-dimensional mapping of nonlinear X waves.
The spatiotemporal intensity profile of a 100-fs wave packet at the output of a X2 crystal, tuned for mismatched second-harmonic generation, is probed via sum-frequency generation with a compressed, 20-fs pulse, revealing the appearance of an X-type wave shape.


Lineage-related susceptibility of human hemopoietic cell lines to apoptosis.
Apoptosis plays a fundamental role in shaping normal hematopoiesis. We have investigated the relationship existing between susceptibility to apoptosis and lineage commitment in hemopoietic cells. The presence and degree of apoptosis were investigated in myeloid (HL-60 and K562), T (Jurkat and MOLT-4), and B (CESS and Raji) lymphoid cell lines by using a variety of techniques-transmission electron and light microscopy, flow cytometry and DNA gel electrophoresis. The major achievement of this study is that hematopoietic cells respond to different chemical (staurosporin, tiazofurin, camptothecin) and physical (hyperthermia or hypothermia) stimuli by apoptosis in a lineage-related way. Moreover, with respect to the methods used to detect apoptosis, a strong correlation was observed between the presence of the hypodiploid peak determined by flow cytometry and the DNA laddering evaluated by gel electrophoresis, but both techniques failed to demonstrate the presence of apoptosis in some cases. We conclude that cells of different hematopoietic lineages mostly show a lineage-related behaviour in their apoptotic response to different stimuli, suggesting that the lineage commitment and the stage of differentiation can confer different sensitivities to specific apoptotic stimuli. Moreover, morphological techniques still represent the most reliable approach to detect apoptosis in hemopoietic cells.


Retinal abnormalities in universal eye screening of healthy, full-term newborn infants in Jakarta. The incidence and its risk factors: a pilot study.
To screen for ocular abnormalities in healthy full-term newborn infants using wide-field digital imaging and to analyze factors associated with the findings. A total of 1208 full-term newborn infants at a tertiary eye hospital (Cipto Mangunkusumo National Referral Hospital) and a district hospital in Jakarta (Koja Hospital) were enrolled to the study. All eligible newborns underwent fundus examination within 48 h after birth using the RetCam shuttle (Natus Medical Incorporated, USA). Retinal findings were documented and analyzed according to obstetric and neonatal risk factors. Of the 1208 newborn infants enrolled, ocular abnormalities were found in 150 infants (12.4%). Retinal hemorrhage (RH) was the most common finding (88%) in which 2.67% involved the macula, followed by chorioretinitis (4.67%). Univariate analysis showed caesarean section (C-section) (OR 0.27, 95% CI 0.18-0.41, p < 0.001) was a protective factor against RH, while prolonged labor increased the risk of developing RH (OR 1.84, 95% CI 1.24-2.72, p = 0.002). Further multivariate analysis showed similar protective association between C-section and risk of RH (OR 0.29, 95% CI 0.19-0.44, p < 0.001), while other risk factors were not. Our study showed that universal eye screening in healthy neonates is beneficial in the early diagnosis, monitoring and treatment of ocular abnormalities such as retinal hemorrhage, chorioretinitis and retinoblastoma. Retinal hemorrhage is the most common ocular abnormality and is associated with the delivery method and the duration of labor. Universal eye screening is visual-saving and life-saving for neonates with chorioretinitis, retinoblastoma as well as other abnormalities and should be mandatory in newborn screening.


Evaluation of the posterior insular cortex involvement in anxiogenic response to emotional stress in male rats: Functional topography along the rostrocaudal axis.
The insular cortex (IC) is engaged in behavioral and physiological responses to emotional stress. Control of physiological functions and behavioral responses has been reported to occur in a site-specific manner along the rostrocaudal axis of the IC. However, a functional topography of the IC regulation of anxiogenic responses caused by stress has never been evaluated. Therefore, we investigated the role of rostrocaudal subregions in the posterior IC in anxiogenic-like effect caused by exposure to acute restraint stress in male rats. For this, rats received bilateral microinjection of the non-selective synaptic inhibitor CoCl2 or vehicle into either the rostral, intermediate or caudal portions of the posterior IC before exposure to acute restraint stress. Then, behavior in the elevated plus maze (EPM) was evaluated immediately after restraint stress. The behavior of non-stressed animals in the EPM was also investigated. We observed that acute restraint stress decreased the exploration of the EPM open arms in animals treated with vehicle in all regions of the posterior IC, thus indicating an anxiogenic-like effect. The avoidance of the EPM open arms was completely inhibited in animals subjected to microinjection of CoCl2 into the intermediate posterior IC. Nevertheless, the same pharmacological treatment into either the rostral or caudal subregions of the posterior IC did not affect the restraint-evoked behavioral changes in the EPM. Taken together, these results suggest that regulation of anxiogenic-like effect to emotional stress along the rostrocaudal axis of the posterior IC might occur in a site-specific manner, indicating a role of the intermediate subregion.


Exploiting collateral sensitivity controls growth of mixed culture of sensitive and resistant cells and decreases selection for resistant cells in a cell line model.
CDK4/6 inhibitors such as ribociclib are becoming widely used targeted therapies in hormone-receptor-positive (HR+) human epidermal growth factor receptor 2-negative (HER2-) breast cancer. However, cancers can advance due to drug resistance, a problem in which tumor heterogeneity and evolution are key features. Ribociclib-resistant HR+/HER2- CAMA-1 breast cancer cells were generated through long-term ribociclib treatment. Characterization of sensitive and resistant cells were performed using RNA sequencing and whole exome sequencing. Lentiviral labeling with different fluorescent proteins enabled us to track the proliferation of sensitive and resistant cells under different treatments in a heterogeneous, 3D spheroid coculture system using imaging microscopy and flow cytometry. Transcriptional profiling of sensitive and resistant cells revealed the downregulation of the G2/M checkpoint in the resistant cells. Exploiting this acquired vulnerability; resistant cells exhibited collateral sensitivity for the Wee-1 inhibitor, adavosertib (AZD1775). The combination of ribociclib and adavosertib achieved additional antiproliferative effect exclusively in the cocultures compared to monocultures, while decreasing the selection for resistant cells. Our results suggest that optimal antiproliferative effects in heterogeneous cancers can be achieved via an integrative therapeutic approach targeting sensitive and resistant cancer cell populations within a tumor, respectively.


Author Correction: Pupil size reflects successful encoding and recall of memory in humans.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Comparison of procedures for measuring the quaternary N-glucuronides of amitriptyline and diphenhydramine in human urine with and without hydrolysis.
The activities of beta-glucuronidases from Helix pomatia, Escherichia coli and rat towards the N-glucuronides of amitriptyline and diphenhydramine were considerably lower than those towards standard substrates. The two N-glucuronides were analysed in urine samples by the following procedures: HPLC of the intact conjugate after solid-phase extraction on a cation exchanger cartridge or after direct injection of urine; HPLC of the aglycone after hydrolysis with beta-glucuronidase from H. pomatia or E. coli or after alkaline hydrolysis. Solid-phase extraction led to the highest recovery and precision, and sensitivity can be improved by extracting a larger volume of urine. On application to samples from patients under treatment with amitriptyline, the results of all procedures except alkaline hydrolysis were in good agreement. When diphenhydramine N-glucuronide was analysed in urine samples of volunteers, solid-phase extraction, hydrolysis by E. coli glucuronidase and alkaline hydrolysis resulted in similar values.


A positive 2-item Patient Health Questionnaire depression screen among hospitalized heart failure patients is associated with elevated 12-month mortality.
Given the association of depression with poorer cardiac outcomes, an American Heart Association Science Advisory has advocated routine screening of cardiac patients for depression using the 2-item Patient Health Questionnaire (PHQ-2) "at a minimum." However, the prognostic value of the PHQ-2 among HF patients is unknown. We screened hospitalized HF patients (ejection fraction [EF] <40%) that staff suspected may be depressed with the PHQ-2, and then determined vital status at up to 12-months follow-up. At baseline, PHQ-2 depression screen-positive patients (PHQ-2+; n = 371), compared with PHQ-2 screen-negative patients (PHQ-2-; n = 100), were younger (65 vs 70 years) and more likely to report New York Heart Association (NYHA) functional class III/IV than class II symptoms (67% vs. 39%) and lower levels of physical and mental health-related quality of life (all P ≤ .002); they were similar in other characteristics (65% male, 26% mean EF). At 12 months, 20% of PHQ-2+ versus 8% of PHQ-2- patients had died (P = .007) and PHQ-2 status remained associated with both all-cause (hazard ratio [HR] 3.1, 95% confidence interval [CI] 1.4-6.7; P = .003) and cardiovascular (HR 2.7, 95% CI 1.1-6.6; P = .03) mortality even after adjustment for age, gender, EF, NYHA functional class, and a variety of other covariates. Among hospitalized HF patients, a positive PHQ-2 depression screen is associated with an elevated 12-month mortality risk.


Preclinical In Vitro and In Vivo Characterization of Synaptic Vesicle 2A-Targeting Compounds Amenable to F-18 Labeling as Potential PET Radioligands for Imaging of Synapse Integrity.
Current synaptic vesicle 2A (SV2A) positron emission tomography (PET) imaging agents include the nanomolar affinity probes [11C]UCB-J and [18F]UCB-H derived from the anti-epileptic drug levitaracetam (Keppra®). An industry-utilized "de-risking" approach was used to carry out initial pharmacological characterization and to assess potential next-generation candidates amenable to F-18 radiolabeling for preliminary evaluation. Radioligand binding methods were employed in mammalian brain homogenates to determine the SV2A affinity (Kd) and maximal binding capacity (Bmax) of [3H]UCB-J. Novel leads were then screened to identify compounds minimally with comparable binding affinities with UCB-J in order to select a F-18-labeled candidate for subsequent in vivo assessment in rat. In parallel, mammalian brain tissue section autoradiography was performed to assess specific SV2A distribution. [3H]UCB-J bound with high affinity to a single population of sites in the rat brain (Kd = 2.6 ± 0.25 nM; Bmax = 810 ± 25 fmol/mg protein) and control human cortex (Kd = 2.9 ± 0.54 nM; Bmax = 10,000 ± 640 fmol/mg protein). Distribution of specific SV2A binding was shown to be homogeneous throughout the rodent brain and primarily in gray matter regions of rodent and human brain sections. Analog screening identified MNI-1038, MNI-1126/SDM-8, and SDM-2 as having comparable binding affinities with the currently available PET ligands. Subsequent [18F]MNI-1126/[18F]SDM-8 dynamic micro-PET imaging in rats revealed in vivo uptake and accumulation in the brain with favorable kinetics. Chase studies using 30 mg/kg levetiracetam confirmed that in vivo brain uptake of [18F]MNI-1126/[18F]SDM-8 was reversible. Taken together, these data suggest [18F]MNI-1126/[18F]SDM-8 (since renamed as [18F]SynVesT-1) characterized via an in vitro screening cascade provided a measurable in vivo SV2A specific signal in the rodent brain. This tracer as well as the close analog [18F]SDM-2 (since renamed as [18F]SynVesT-2) is currently undergoing further evaluation in preclinical and clinical studies.


Cytokinin modulates endocytic trafficking of PIN1 auxin efflux carrier to control plant organogenesis.
Cytokinin is an important regulator of plant growth and development. In Arabidopsis thaliana, the two-component phosphorelay mediated through a family of histidine kinases and response regulators is recognized as the principal cytokinin signal transduction mechanism activating the complex transcriptional response to control various developmental processes. Here, we identified an alternative mode of cytokinin action that uses endocytic trafficking as a means to direct plant organogenesis. This activity occurs downstream of known cytokinin receptors but through a branch of the cytokinin signaling pathway that does not involve transcriptional regulation. We show that cytokinin regulates endocytic recycling of the auxin efflux carrier PINFORMED1 (PIN1) by redirecting it for lytic degradation in vacuoles. Stimulation of the lytic PIN1 degradation is not a default effect for general downregulation of proteins from plasma membranes, but a specific mechanism to rapidly modulate the auxin distribution in cytokinin-mediated developmental processes.


Relationship between lateral angle and shape of internal acoustic canal: cautionary note for diagnosis of sex.
Lateral angle (LA) of the internal acoustic canal (IAC) at its opening at the surface of the petrous portion of the temporal bone is known in literature as a dimorphic measurement useful for sex assessment. However, its reliability is still debated. Moreover, no information is available about the possible relationship between LA and shape and size of the IAC. This study aims at assessing breadth, length and lateral angle of IAC on 100 CT scans belonging to subjects aged between 20 and 70 years, equally divided between males and females. Possible differences in the three measurements according to side and sex were assessed through two-way ANCOVA test, using three cranial measurements (distance between anterior and posterior nasal spine, upper facial height, bizygomatic breadth) as covariates (p < 0.05). Possible correlations among IAC measurements and with age were assessed through Pearson's test (p < 0.05). Results showed a significantly greater IAC breadth on the left side than on the right side; moreover, LA was significantly wider in females than in males and was positively correlated with IAC breadth in both males and females. In addition, LA was negatively correlated with age only in the female group. The study first showed that LA is strictly related to IAC morphology; moreover, it is significantly affected by masculinization of skeletal traits in females. Therefore, caution is suggested in using this measurement for sex assessment.