Antimony porphyrins as red-light powered photocatalysts for solar fuel production from halide solutions in the presence of air.
Halide ions (X(-)) as cheap and abundant electron donors are oxidized with simulated sunlight in a photocatalytic reaction based on robust antimony porphyrins acting as red-light harvesting multielectron transfer sensitizers. Besides halogen formation (X2/X3(-)) this solar energy storing process under certain conditions also accumulates hydrogen peroxide (H2O2) as a second energy-rich compound.


Effects of aging on neural connectivity underlying selective memory for emotional scenes.
Older adults show age-related reductions in memory for neutral items within complex visual scenes, but just like young adults, older adults exhibit a memory advantage for emotional items within scenes compared with the background scene information. The present study examined young and older adults' encoding-stage effective connectivity for selective memory of emotional items versus memory for both the emotional item and its background. In a functional magnetic resonance imaging (fMRI) study, participants viewed scenes containing either positive or negative items within neutral backgrounds. Outside the scanner, participants completed a memory test for items and backgrounds. Irrespective of scene content being emotionally positive or negative, older adults had stronger positive connections among frontal regions and from frontal regions to medial temporal lobe structures than did young adults, especially when items and backgrounds were subsequently remembered. These results suggest there are differences between young and older adults' connectivity accompanying the encoding of emotional scenes. Older adults may require more frontal connectivity to encode all elements of a scene rather than just encoding the emotional item.


Mental health ubiquitous monitoring supported by social situation awareness: A systematic review.
Traditionally, the process of monitoring and evaluating social behavior related to mental health has based on self-reported information, which is limited by the subjective character of responses and various cognitive biases. Today, however, there is a growing amount of studies that have provided methods to objectively monitor social behavior through ubiquitous devices and have used this information to support mental health services. In this paper, we present a Systematic Literature Review (SLR) to identify, analyze and characterize the state of the art about the use of ubiquitous devices to monitor users' social behavior focused on mental health. For this purpose, we performed an exhaustive literature search on the six main digital libraries. A screening process was conducted on 160 peer-reviewed publications by applying suitable selection criteria to define the appropriate studies to the scope of this SLR. Next, 20 selected studies were forwarded to the data extraction phase. From an analysis of the selected studies, we recognized the types of social situations identified, the process of transforming contextual data into social situations, the use of social situation awareness to support mental health monitoring, and the methods used to evaluate proposed solutions. Additionally, we identified the main trends presented by this research area, as well as open questions and perspectives for future research. Results of this SLR showed that social situation-aware ubiquitous systems represent promising assistance tools for patients and mental health professionals. However, studies still present limitations in methodological rigor and restrictions in experiments, and solutions proposed by them have limitations to be overcome.


Afterdischarges following M waves in patients with voltage-gated potassium channels antibodies.
To explore the correlation between afterdischarges in motor nerve conduction studies and clinical motor hyperexcitability in patients with voltage-gated potassium channels (VGKC) antibodies. Six patients with positive serum antibodies to contactin-associated protein-like 2 (CASPR2) or/and leucine-rich glioma-inactivated protein 1 (LGI1) were recruited, including 5 with autoimmune encephalitis, and 1 with cramp-fasciculation syndrome. Electromyography (EMG), nerve conduction studies (NCS) and F waves were performed, and afterdischarges were assessed. One patient was followed up. Five patients had clinical evidence of peripheral motor nerve hyperexcitability (myokymia or cramp), and four of them had abnormal spontaneous firing in concentric needle electromyography (EMG). Prolonged afterdischarges following normal M waves were present in all six patients, including the two patients who had no EMG evidence of peripheral nerve hyperexcitability (PNH). Afterdischarges disappeared after treatment with intravenous immunoglobulin (IVIG). The afterdischarges in motor nerve conduction study might be a sensitive indicator of peripheral motor nerve hyperexcitability in patients with VGKC antibodies. Afterdischarges in motor nerve conduction study might be more sensitive than needle electromyography for detecting peripheral motor nerve hyperexcitability, and could disappear gradually in accordance with clinical improvement and reduction of antibodies.


Social inequalities in health anxiety: A systematic review and meta-analysis.
The association between a low socioeconomic status and poor health, also known as social inequalities in health, has been shown in numerous studies. This also applies to most mental illnesses, such as somatization or anxiety disorders. It is unclear, however, whether social inequalities are evident in health anxiety as well. Therefore, this review and meta-analysis systematically examines social inequalities in health anxiety. A systematic literature search was conducted covering all studies published until August 2021. The databases PubMED, Web of Science, PsycINFO, and PSYNDEX were screened to identify quantitative studies using validated tools to measure health anxiety and examining the association with socioeconomic status. Meta-analytic methods were applied by using random-effect models. The study quality was assessed with the Effective Public Health Practice Project Quality Assessment Tool. Overall, 37 studies were eligible for meta-analysis. Effect sizes of 27 studies using screening scales and 10 studies using diagnostic interviews to measure health anxiety were calculated separately. Meta-analyses revealed a significant association between socioeconomic status and health anxiety (r - 0.14, 95%-CI -0.18 to -0.09 (screening scales) and OR 0.63, 95%-CI 0.43 to 0.92 (diagnostic interviews)), indicating a lower risk for health anxiety in people with higher socioeconomic status. Findings suggest that the health-related disadvantage of socially deprived people is also evident regarding health anxiety. This should be considered in programs to reduce social inequalities in health. PROSPERO registration number CRD42021237550.


Complete NMR assignments of the antibacterial biflavonoid GB1 from Garcinia kola.
From the antibacterial fraction of the roots of Garcinia kola, 3'',4',4''',5,5'',7,7''-heptahydroxy-3,8''-biflavanone (GB1) was isolated as the major constituent, whose interesting conformations were studied on the basis of its 1D and 2D NMR spectra obtained at different temperatures and in different solvents. GB1 showed antibacterial activities against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) with MIC of 32 and 128 microg/ml, respectively.


Inverse supercritical fluid extraction as a sample preparation method for the analysis of the nanoparticle content in sunscreen agents.
We demonstrate the use of inverse supercritical carbon dioxide (scCO2) extraction as a novel method of sample preparation for the analysis of complex nanoparticle-containing samples, in our case a model sunscreen agent with titanium dioxide nanoparticles. The sample was prepared for analysis in a simplified process using a lab scale supercritical fluid extraction system. The residual material was easily dispersed in an aqueous solution and analyzed by Asymmetrical Flow Field-Flow Fractionation (AF4) hyphenated with UV- and Multi-Angle Light Scattering detection. The obtained results allowed an unambiguous determination of the presence of nanoparticles within the sample, with almost no background from the matrix itself, and showed that the size distribution of the nanoparticles is essentially maintained. These results are especially relevant in view of recently introduced regulatory requirements concerning the labeling of nanoparticle-containing products. The novel sample preparation method is potentially applicable to commercial sunscreens or other emulsion-based cosmetic products and has important ecological advantages over currently used sample preparation techniques involving organic solvents.


Secondary metabolites of benzo[a]pyrene: 3-hydroxy-trans-7,8-dihydro-7,8-dihydroxybenzo[a]pyrene, a biliary metabolite of 3-hydroxybenzo[a]pyrene in the rat.
Rats administered 3-hydroxybenzo[a]pyrene (50 mg/kg, i.p.), excrete via the bile metabolites which, after treatment with beta-glucuronidase and aryl sulphatase, yield, in addition to 3-hydroxybenzo[a]pyrene, 3-hydroxy-trans-7,8-dihydro-7,8-dihydroxybenzo[a]pyrene (3-OH-BP-7,8-diol) and a minor, highly labile, metabolite tentatively identified as 3,5-dihydroxybenzo[a]pyrene. These novel metabolites are readily isolated in a pure state via preparative layer chromatography. The structure of the 3-OH-BP-7,8-diol was revealed by its u.v., proton magnetic resonance and mass spectral properties. Its hydroxyl functions are in a predominantly quasi-diequatorial conformation.


Ecological assessment of a region with PCB emissions using samples of soil, vegetation and breast milk: a case study.
An ecological assessment is performed by using a data bank of air, soil, vegetation and biology data from the area of a capacitor plant in Serpukhov, Russia. Over a number of years the use of PCBs for filling capacitors has brought about environmental contamination beyond the plant's sanitary zone, in particular along a stream where vegetables and berries are grown commercially. A correlation has been found between abnormally high concentrations of PCBs in breast milk of women living in the proximity of the plant and the occurrence of diathesis in babies. A number of remedial measures have been developed in conjunction with the city's sanitary service.


Nifedipine enhances the cardioprotective effect of an angiotensin-II receptor blocker in an experimental animal model of heart failure.
This study was designed to examine the hypothesis that a calcium channel blocker nifedipine (CCB) could enhance the cardioprotective effect of an angiotensin-ll receptor blocker candesartan (ARB) in the treatment for heart failure. Isoproterenol (ISP) was injected into male rats at 300 mg/kg to produce progressive heart failure. Three months later, the rats were divided into 4 groups and treated for 4 weeks with 1) vehicle (n = 20), 2) ARB at 0.2 mg/kg/day (n = 6), 3) CCB at 10 mg/kg/day (n = 6), or 4) both drugs (n = 8). Rats injected with saline served as controls (n = 13). ISP caused severe myocardial degeneration and decreased the capillary density (D(cap)) of the left ventricular (LV) myocardium (mean +/- SD: 2,197 +/- 627 vs. 2,847 +/- 298 N/mm2 for normal controls), while increasing plasma thiobarbituric acid-reactive substances (TBARS; 3.6 +/- 1.1 vs. 1.9 +/- 0.5 nmol/ml). Although ARB therapy preserved cardiac morphology, it had little effect on D(cap) or oxidative stress. On the other hand, CCB decreased plasma TBARS and 4-hydroxy-2-nonenal protein expression in LV myocardium. Furthermore, the combination of CCB and ARB increased D(cap) and preserved the ultrastructure of LV myocardium, so this combination may be a useful option for the treatment of heart failure.


Academic Productivity of US Cardiothoracic Surgical Centers.
We investigated which metrics represent valid correlates of the academic productivity of cardiothoracic (CT) surgical centers. We collected data from 57 US academic CT surgical groups (663 surgeons), including H index and institutional role of each surgeon, ranking by US News & World Report (USNWR) and NIH funding, and designation as department versus division. Academic productivity (median H-index of each group) was significantly different across institutions (p < 0.001). Several USNWR hospital categories ("Honor Roll", "Adult Cardiology and Heart Surgery", "Adult Pulmonology", "Adult Cancer") and medical school ranking for research were associated with differences in academic productivity, while ranking by NIH funding or designation as department versus division was not. Groups with chairperson's individual H-index ≥50 were overall more productive than those with chairperson's H-index <50 (median H-index: 18 vs. 14; p = 0.005). USNWR rankings provide a good representation of academic productivity, while NIH funding ranking or designation as department versus division does not. The individual productivity of the chairperson is correlated with that of the whole group. doi: 10.1111/jocs.12773 (J Card Surg 2016;31:423-428).


Neutrophils from asthmatics exhibit diminished responsiveness to 2-chloroadenosine which is reversed by theophylline. Evidence for a cyclic-AMP-independent pathway on human neutrophils.
We have shown previously that neutrophils (PMNs) from patients with asthma have a more potent stimulated respiratory burst than normals and that their respiratory burst is significantly less suppressed with exposure to 2-chloroadenosine (2-CADO). The present studies investigated the basis of this defect in responsiveness to 2-CADO. PMNs obtained from asthmatics either not on theophylline (minus theophylline) or taking theophylline (plus theophylline) generated significantly more superoxide in response to 2 x 10(-8) M FMLP (2.08 +/- 0.36 nmol/5 x 10(5) PMNs (minus theophylline) (P less than 0.01 compared to controls) vs. 2.16 +/- 0.44 (plus theophylline) (P less than 0.01) as compared to controls (1.05 +/- 0.17 nmol). In the presence of FMLP (2 x 10(-8) M), PMNs from the minus theophylline cohort had less 2-CADO (10(-6) M)-mediated suppression of superoxide generation as compared to controls (38.3 +/- 3.8% vs. 67.1 +/- 3.8%; (P less than 0.001). The plus theophylline group exhibited suppression values similar to controls (64.5 +/- 7.2%). Theophylline, in the presence of a physiological concentration of 2-CADO (0.1 microM) accentuated the suppression of the respiratory burst in normals (74.1 +/- 5.9%, 80.1 +/- 4.9% (P less than 0.02) and 84.7 +/- 3.8% (P less than 0.02) at 0, 10, and 100 microM, respectively). PMNs from asthmatics not taking theophylline demonstrated suppression values of 46.2 +/- 6%, 53.8 +/- 6.6% (P = NS), and 63.2 +/- 7.1% (P less than 0.01), respectively. Resting PMNs from normal controls generated 0.97 +/- 0.20 pmol cAMP/10(7) cells compared to 2.83 +/- 0.75 pmol in the presence of 0.1 microM 2-CADO. The combination of 2-CADO and theophylline (10-100 microM) produced cAMP concentrations not significantly different from that observed with 2-CADO alone. These findings support the existence of a novel cAMP-independent adenosine receptor in PMNs. The specific binding of 10(-8)M 3H-labeled 2-CADO (in delta cpm) was 10,358 +/- 1502 (P less than 0.001 compared to controls), 5468 +/- 843 (NS compared to controls), and 3751 +/- 477 in the plus theophylline group, minus theophylline group, and controls, respectively. Such up-regulation of specific binding may represent the effects of theophylline as shown by the specific binding of [3H]2-CADO in PMNs from normal controls exposed to 10 microM theophylline for 30 min (6013 +/- 969) compared to unexposed PMNs (3768 +/- 656; P less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)


Analysis of the peripheral blood lymphocyte subsets in patients with bladder carcinoma.
To evaluate the immune system of patients with bladder transitional cell carcinoma (TCC) by using peripheral blood lymphocyte subsets and to further compare the relationship between these subsets with respect to tumor stage and grade (superficial versus invasive and low versus high grade). Thirty patients with superficial TCC of the bladder, 30 patients with invasive TCC of the bladder, and 30 age- and sex-matched control subjects without any malignancy or immunologic abnormality were included in this study. The peripheral blood lymphocyte subset analysis was performed in all patients before any treatment was performed. All lymphocyte subset values of patients with invasive bladder cancer, except B cell value, were significantly lower (P < 0.01) than the values of the control group. There were no significant differences between the lymphocyte subset values of patients with superficial bladder cancer and those of control subjects. The comparison of the lymphocyte subset values of the patients with superficial versus invasive bladder carcinoma revealed that in patients with invasive bladder carcinoma, the numbers of T and natural killer (NK) cells were significantly lower (P < 0.05) than those of patients with superficial bladder carcinoma. Patients with high-grade tumors had significantly fewer (P < 0.05) T and NK cells than patients with low-grade tumors. Our results indicate that analysis of mean NK and T cell values and the mean ratio of CD4+/CD8+ cells in peripheral blood might be a useful adjunct for the clinical evaluation of patients with bladder cancer.


Catalytic AgF-Initiated Intramolecular 1,3-Sulfonyl Migration of gem-Difluorovinyl Sulfonates to α,α-Difluoro-β-ketosulfones.
A 1,3-sulfonyl migration of difluorovinyl sulfonates initiated by a catalytic amount of silver fluoride is presented. α,α-Difluoro-β-ketosulfones were successfully prepared in excellent yields. This method features high chemoselectivity, good functional group tolerance, high atom economy, and mild, environmentally benign reaction conditions. Furthermore, mechanistic experiments indicate that this migration proceeds in an intermolecular pathway and the corresponding sulfinates are possible intermediates.


Task-based functional connectivity in aging: How task and connectivity methodology affect discovery of age effects.
Past studies have found that healthy aging has a significant effect on the organization and function of networks in the human brain. Many of these studies have examined how functional connectivity during one task or at rest is affected by aging; however, few studies have systematically examined how the effect of age on functional connectivity may vary as a function of choice of in-scanner task. The present study included healthy adults between the ages of 20 and 80 and examined a variety of metrics of functional connectivity during performance of 11 in-scanner tasks, falling into 4 cognitive domains: vocabulary, processing speed, fluid reasoning, and episodic memory. Functional connectivity was assessed at three levels: average correlations within and between 10 networks, system segregation (sensorimotor vs. association networks), and whole-brain graph theory metrics (global efficiency and modularity). Results showed that the effect of age on these metrics differed as a function of task-for example, age had a more consistent effect on functional connectivity metrics computed during fluid reasoning tasks; however, there was less of an effect of age on functional connectivity metrics computed during tasks of episodic memory. Further, some of these measures showed relationships with behavioral performance on the in-scanner task, with different networks playing a role in the different cognitive domains. These findings suggest that while aging may be generally associated with reductions in within- and between-network connectivity, system segregation, global efficiency, and modularity, the magnitude and presence of these effects varies by in-scanner task.


Pyroptotic cell death in the R6/2 mouse model of Huntington's disease: new insight on the inflammasome.
Mechanisms of tissue damage in Huntington's disease involve excitotoxicity, mitochondrial damage, and neuroinflammation, including microglia activation. In the present study, we investigate the role of pyroptosis process in the striatal neurons of the R6/2 mouse model of Huntington's disease. Transgenic mice were sacrificed at 4 and 13 weeks of age. After sacrifice, histological and immunohistochemical studies were performed. We found that NLRP3 and Caspase-1 were intensely expressed in 13-week-old R6/2 mice. Moreover, NLRP3 expression levels were higher in striatal spiny projection neurons and in parvalbumin interneurons, which are prone to degenerate in HD.


Primary cutaneous CD30 (Ki-1)-positive non-anaplastic B-cell lymphoma.
The CD30 (Ki-1)-positive lymphoproliferative disorders show a non-epidermotropic infiltrate of large-sized pleomorphic T cells, being extremely rare those that develop from B cells. A 51-year-old white man presented with a 8x10 cm lobulated scalp tumor that had developed during the previous 8 months. Histopathology demonstrated a proliferation of medium and large-sized non-anaplasic lymphocytes. The tumor cells (CD45+) expressed B phenotype (CD20+, CD3) and were CD30 (Ki-1) positive. The extension study revealed no extracutaneous involvement and the lesion resolved following chemotherapy. Two years later, after six cycles of chemotherapy that followed the CHOP protocol the patient remained asymptomatic. We report a patient with a diffuse lymphoma of medium and large-sized B phenotype cells on the scalp that showed the unusual characteristic of being both non-anaplastic and CD30 (Ki-1) positive.


Identification of the high affinity binding site of transforming growth factor-alpha (TGF-alpha) for the chicken epidermal growth factor (EGF) receptor using EGF/TGF-alpha chimeras.
Human epidermal growth factor (hEGF) and human transforming growth factor-alpha (hTGF-alpha) are structurally related growth factors that share relatively little sequence homology. They both exert their biological action by binding to the cell-surface EGF receptor. hEGF and hTGF-alpha bind with similar affinity to the hEGF receptor, but hEGF binds with an approximately 100-fold lower affinity to the chicken EGF receptor compared with hTGF-alpha. To map the region in hTGF-alpha that confers its ability to bind with high affinity to the chicken EGF receptor, 10 hybrids of hEGF and hTGF-alpha were constructed by exchanging domains bordered by the third, fourth, and sixth conserved cysteine residues. The activity of the expressed chimeric proteins was determined by their ability to compete with 125I-labeled mouse EGF for binding to NIH-3T3 cells transfected with the hEGF receptor. Subsequent binding competition studies of NIH-3T3 cells transfected with the chicken EGF receptor showed that chimeras carrying TGF-alpha sequences COOH-terminal of the sixth cysteine have a high affinity for this receptor, similar to hTGF-alpha. In contrast, chimeras with EGF sequences in this COOH-terminal domain have only low binding affinity, similar to hEGF. We conclude that the COOH-terminal linear region of hTGF-alpha is important for its high affinity interaction with the chicken EGF receptor.


Colibacter massiliensis gen. nov. sp. nov., a novel Gram-stain-positive anaerobic diplococcal bacterium, isolated from the human left colon.
The gut microbiota is considered to play a key role in human health. As a consequence, deciphering its microbial diversity is mandatory. A polyphasic taxonogenomic strategy based on the combination of phenotypic and genomic analyses was used to characterize a new bacterium, strain Marseille-P2911. This strain was isolated from a left colon sample of a 60-year old man who underwent a colonoscopy for an etiological investigation of iron-deficiency anemia in Marseille, France. On the basis of 16S rRNA sequence comparison, the closest phylogenetic neighbor was Anaeroglobus geminatus (94.59% 16S rRNA gene sequence similarity) within the family Veillonellaceae. Cells were anaerobic, Gram-stain-positive, non-spore-forming, catalase/oxidase negative cocci grouped in pairs. The bacterium was able to grow at 37 °C after 2 days of incubation. Strain Marseille-P2911 exhibited a genome size of 1,715,864-bp with a 50.2% G + C content, and digital DNA-DNA hybridization (dDDH) and OrthoANI values with A. geminatus of only 19.1 ± 4.5% and 74.42%, respectively. The latter value being lower than the threshold for genus delineation (80.5%), we propose the creation of the new genus Colibacter gen. nov., with strain Marseille-P2911T (=DSM 103304 = CSUR P2911) being the type strain of the new species Colibacter massiliensis gen. nov., sp. nov.


Dendritic cells license regulatory B cells to produce IL-10 and mediate suppression of antigen-specific CD8 T cells.
Regulatory B cells (Bregs) suppress and reduce autoimmune pathology. However, given the variety of Breg subsets, the role of Bregs in the pathogenesis of type 1 diabetes is still unclear. Here, we dissect this fundamental mechanism. We show that natural protection from type 1 diabetes in nonobese diabetic (NOD) mice is associated with increased numbers of IL-10-producing B cells, while development of type 1 diabetes in NOD mice occurs in animals with compromised IL-10 production by B cells. However, B cells from diabetic mice regain IL-10 function if activated by the innate immune receptor TLR4 and can suppress insulin-specific CD8 T cells in a dendritic cell (DC)-dependent, IL-10-mediated fashion. Suppression of CD8 T cells is reliant on B-cell contact with DCs. This cell contact results in deactivation of DCs, inducing a tolerogenic state, which in turn can regulate pathogenic CD8 T cells. Our findings emphasize the importance of DC-Breg interactions during the development of type 1 diabetes.


Chemotactic LTB4 metabolites produced by hepatocytes in the presence of ethanol.
Ethanol in low concentrations significantly alters the hepatocyte metabolism of the neutrophil chemotactic lipid leukotriene B4 (LTB4). Two novel metabolites of LTB4 which are encountered only when ethanol is present, retained significant biological activity. One metabolite, 3-hydroxy-LTB4 increased intracellular free calcium in the human neutrophil at concentrations as low as 3 x 10(-10) M as well as induced shape change and adherence to albumin-coated latex beads at 10 nM. The 3-hydroxy-LTB4 and 3,20-hydroxy-LTB4 metabolites were also potent chemotactic agonists with an ED50 at 3.0 and 9.0 nM, respectively. These results suggest that the presence of ethanol can substantially alter inactivation of LTB4 by the liver and may mediate neutrophil accumulation into the liver, thereby contributing to the pathogenesis of alcoholic hepatitis even when LTB4 biosynthesis occurs at some site distant to the liver.


Cryo-EM structures of Gid12-bound GID E3 reveal steric blockade as a mechanism inhibiting substrate ubiquitylation.
Protein degradation, a major eukaryotic response to cellular signals, is subject to numerous layers of regulation. In yeast, the evolutionarily conserved GID E3 ligase mediates glucose-induced degradation of fructose-1,6-bisphosphatase (Fbp1), malate dehydrogenase (Mdh2), and other gluconeogenic enzymes. "GID" is a collection of E3 ligase complexes; a core scaffold, RING-type catalytic core, and a supramolecular assembly module together with interchangeable substrate receptors select targets for ubiquitylation. However, knowledge of additional cellular factors directly regulating GID-type E3s remains rudimentary. Here, we structurally and biochemically characterize Gid12 as a modulator of the GID E3 ligase complex. Our collection of cryo-EM reconstructions shows that Gid12 forms an extensive interface sealing the substrate receptor Gid4 onto the scaffold, and remodeling the degron binding site. Gid12 also sterically blocks a recruited Fbp1 or Mdh2 from the ubiquitylation active sites. Our analysis of the role of Gid12 establishes principles that may more generally underlie E3 ligase regulation.


Noninvasive imaging of the lung NETosis by anti-Ly6G iron oxide nanoparticles.
It is challenging to visualize noninvasively the formation of neutrophil extracellular traps, known as NETosis, and therefore difficult to monitor disease progression. A desirable molecular imaging probe is the iron oxide nanoparticle (NP) that could induce reactive oxygen species. Here, we used C57BL/6 mice with pristane-induced lupus, which mimics systemic lupus erythematosus. Administration of anti-Ly6G antibody-conjugated NP allowed detection of NETosis with fluorescent molecular imaging, as evidenced by flow cytometric analysis of citrullinated histone H3 expression in lung neutrophils. This finding was consistent with NP-induced blood NETosis in a spontaneous lupus model of B6.MRL-lpr mice. A chronic assessment was performed in which the lupus mice were protected from enhanced oxidative burst by anti-Ly6G NP. This NP can migrate from the peritoneal cavity to the lungs, as visualized by magnetic particle imaging. Overall, our study provides evidence for a highly sensitive assessment of NETosis in lupus through magnetic particle imaging.


Zwitterionic dipoles as a dielectric probe for investigating head group mobility in phospholipid membranes.
For phospholipid membranes with zwitterionic head groups, the dipole can be considered as a specific label for tracing the changes in the dynamic behaviour of this region of the bilayer in its various phases. Measurements of the dielectric properties of fully hydrated 1,2-dipalmitoyl-sn-glycero-3-phosphocholine bilayers in the frequency range 1--50 MHz show a dispersion which is attributed to the motion of the phosphocholine dipoles in the plane of the bilayers. When the temperature is varied, both the permittivity and loss factor increase sharply at the pretransition (35 degrees C) and the main transition (42 degress C). The relaxation time and amplitude were also determined for this dispersion and these further reflect the structural changes occurring with temperature. The relaxation times varied between 4 ns at 30 degrees C and 2.3 ns at 50 degrees C. Due to steric hindrances a restriction in the angle of head group rotation occurs at lower temperatures but is greatly reduced above the main transition.


Enhancing needle visualization during parasagittal approach in paravertebral block for patients undergoing simple mastectomy using in-plane, multiangle ultrasound needle guidance system.
Ultrasound-guided paravertebral blocks during breast surgeries with in-plane needle approaches can be challenging due to difficult needle visualization. The purpose of this study was to assess the usefulness of using a needle guide while performing in-plane parasagittal approach paravertebral block for breast surgery. Eighty patients, American Society of Anesthesiologists physical status I-III, aged 20-40 years with breast mass scheduled for simple mastectomy surgery, were involved in prospective, controlled, randomized study, and were randomly divided by closed envelope method into two groups: Group I (n = 40): Scheduled for ultrasound-guided paravertebral block or group II (n = 40): Scheduled for ultrasound-guided paravertebral block using the needle guide. Both techniques compared as regards: (i) Needle visibility and block performance time; (ii) number of needle passes; (iii) duration of the block; (iv) doctor and patient satisfaction; and (v) incidence of complications. Needle visibility score was better in group II (2.92 ± 0.26 vs. 1.9 ± 0.44, P < 0.0001). Block performance time was shorter in group II (90.92 ± 15 vs. 128.25 ± 16s, P < 0.0001). A number of needle passes were less in group II (1.27 ± 0.45 vs. 2.2 ± 0.68, P < 0.0001). Doctor and patient satisfaction were better in group II (P = 0.015). No differences were found regarding the duration of the block and incidence of complications between groups. A needle guide can help reduce the time needed to perform a parasagittal in-plane thoracic paravertebral block, with a significant reduction in the block performance time, the number of needle passes, better needle visibility and better doctor and patient's satisfaction. However, there was no significant difference regarding the duration of the block or incidence of complications.


Spermatogenesis proceeds normally in mice without linker histone H1t.
The histone gene H1t is expressed exclusively in pachytene spermatocytes of the testis. In this report we have eliminated the single copy H1t gene by homologous recombination from the mouse genome to analyse the function of the H1t protein during spermatogenesis. Mice homozygous for the mutated H1t gene locus developed normally and showed no anatomic abnormalities until the adult stage. In addition, H1t-deficient mice were fertile and reproduced as wild-type mice. The process of spermatogenesis and the testicular morphology remained unchanged in the absence of H1t. RNase protection analysis demonstrated that H1.1, H1.2 and H1.4 histone gene expression is enhanced during spermatogenesis in H1t-deficient mice.


Quantitative fluorescence cytometric measurement of estrogen and progesterone receptors: correlation with the hormone binding assay.
We describe, here, a rapid flow cytometry technique for the detection and quantification of estrogen (ER) and progesterone (PgR) receptors in several human cell lines and in clinical samples obtained from breast cancer tumors. ER and PgR quantitation can be very useful in patients with breast cancer as their role in diagnosis and prognosis is well established. However ligand binding assays and immunohistochemical assays are difficult to measure heterogeneity in individual cells. On the other hand, flow cytometry is a convenient tool for quantification in individual cells. Flow cytometric results with breast cancer cell lines and clinical samples were compared to those obtained by quantitative biochemical ER and PgR performed by the standard dextran-coated charcoal biochemical assay. The latter assay is affected by the level of endogenous steroids. This is also the case in the routine measurement of ER/PgR in patient's tumor cells whereby estradiol molecules in patient's serum produced negative or low values in the biochemical assay. The mAbs used in our flow cytometric method bind to their specific ER or PgR independently of whether they are preoccupied by their ligands and they produce reliable results. With the use of beads calibrated in MESF (Molecules of Equivalent Soluble Fluorochrome) units, the ER and PgR can be measured on a per cell basis. The flow cytometric method showed a strong correlation with biochemical receptor assessments of either ER alpha (ER alphaDCC, r = 0.918, p = 0.073) or PgR (PgRDCC, r = 0.75, p = 0.001). This study demonstrates that ER alpha and PgR can be detected by flow cytometry on a per cell basis in intact cells, and can be quantitated reliably in terms of MESF without the limitations of competition with serum's estradiol molecules.


One-pot Synthesis and Photophysical Studies of Α-cycloamino-substituted 5-aryl-2,2'-bipyridines.
A series of α-cycloamine substituted 2,2'-bipyridines 3ae'-3ce' was obtained via the one-pot approach based on ipso-substitution of a cyano-group in 1,2,4-triazines, followed by aza-Diels-Alder reaction in good yields. Photophysical properties, including fluorosolvatochromism, were studied for 3ae'-3ce' and were compared with α-unsubstituted 2,2'-bipyridines. In addition, dipole moments differences in ground and excited states were calculated by both Lippert-Mataga equation and DFT studies and were compared to each other. The correlation between the size of cycloamine unit and the dipole moments differences value (based on Lippert-Mataga equation) was observed. In addition charge transfer indices (DCT, Λ, H and t) were calculated to demonstrate influence of molecular structure on the intramolecular charge transfer degree.


Ultrasensitive immunoradiometric assay for chorionic gonadotropin which does not cross-react with luteinizing hormone nor free beta chain of hCG and which detects hCG in blood of non-pregnant humans.
We have developed a sensitive, non-competitive, two-monoclonal antibody, sandwich-type or immunoradiometric assay for human chorionic gonadotropin (hCG) which shows no cross-reaction with the free beta chain of hCG nor with human luteinizing hormone (LH). In the assay procedure, two, highly selected monoclonal antibodies reacted in solution with hCG to be quantified. One antibody was covalently conjugated to biotin. This antibody was specific for the beta subunit of hCG, and showed no reaction with LH nor the alpha subunit. The second antibody was labelled with 125I and was specific for intact hCG and LH, showing no cross-reaction with beta hCG nor the alpha subunit. The separation system was a polystyrene ball conjugated with biotin. This ball bound via an avidin bridge the monoclonal 'sandwich' containing hCG. Counts per minute bound to the ball were directly proportional to the amount of hCG present. The assay was specific for whole hCG and showed no reaction with beta hCG, beta LH, intact LH nor the free alpha subunit. Sensitivity was adequate to detect 'hCG-like' material in all post menopausal women and, when single samples were obtained, in over 2/3 of normal men. When multiple samples were obtained, 'hCG-like' material was detectable in all eugonadal adults studied.


[Mobile geriatric rehabilitation in functionally severely impaired patients. Investigations on effectiveness].
Mobile geriatric rehabilitation is an outpatient rehabilitative treatment in which a multidisciplinary team treats elderly patients at home. This kind of treatment has been performed in rare cases in Germany but there are no data available on the effectiveness in patients with severe cognitive and functional impairment. In a retrospective study design the data of all patients who had participated in mobile geriatric rehabilitation between 1 September 2009 and 23 May 2011 were evaluated. Before treatment a comprehensive geriatric assessment was performed and after treatment an assessment of mobility and activities of daily living (ADL). After 6 months a random sample of 20 patients were contacted by telephone to check the ADL. A total of 87 patients were treated between 1 September 2009 and 23 May 2011. The median age was 83 years and 56 % of the patients lived in nursing homes. Only 24 % of the patients had MMSE scores > 23 points, 77 patients completed the treatment with an assessment and in these patients the ADL could be improved significantly (Barthel index at the beginning 36.2 points and on completion 50.9 points, p < 0.001). The Barthel index 6 months after treatment was only 1.25 points lower compared to the assessment at the end of the treatment. Significant improvement after therapy could also be demonstrated in the mobility assessment (timed up and go test, Tinetti mobility score and Esslinger transfer scale). The results of this non-randomized and non-blinded trial indicated the efficacy of mobile geriatric rehabilitation. In functionally and cognitively impaired elderly patients ADL and mobility can be improved. These effects seem to persist for at least for 6 months.


Combining multiple biomarkers differentiates between active SJIA, SJIA-MAS and EBV-HLH.
Cytokine storm syndromes are a clinically heterogeneous group of conditions resulting from a maladaptive host response to an inflammatory trigger. These syndromes lead to rapid progression of immune-mediated damage to healthy tissues resulting in life-threatening multi-system organ failure. Prompt recognition of disease and medical intervention to limit damage to healthy tissues is essential to prevent cytokine storm morbidity and mortality. However, the diagnosis of cytokine storm syndromes is challenging, given the clinical heterogeneity in disease presentations. Therefore, expeditious and readily available tests to diagnose disease and differentiate between the various types of cytokine storm syndromes are of clinical utility. The recently published work of Shimizu and colleagues brings us closer to making this a reality.


Pharmacological actions of 17 beta-oestradiol on articular cartilage chondrocytes and chondrosarcoma chondrocytes in the absence of oestrogen receptors.
(1) Pharmacological concentrations (greater than 10(-5) M) of 17 beta-oestradiol inhibited 35S-labelled proteoglycan synthesis in bovine articular cartilage explant cultures. They also inhibited 35S-labelled proteoglycan synthesis and 3H-labelled protein synthesis in cell cultures of chondrocytes from bovine articular cartilage and Swarm rat chondrosarcoma. Maximal inhibition was about 30-50%. Physiological concentrations (10(-9)-10(-8) M) of oestradiol had no effect on the synthesis of either protein or proteoglycan. (2) The inhibitory action of high concentrations of oestradiol on these biosynthetic pathways is not common to all steroids since 10(-4) M cortisol had no effect on articular chondrocyte cell cultures. 10(-4) M testosterone had a similar action to oestradiol. (3) Neither physiological nor pharmacological concentrations of 17 beta-oestradiol had any effect on 35S-labelled proteoglycan turnover in the cartilage explant system. (4) 10(-5) M oestradiol inhibited cell division in cultures of articular chondrocytes which had entered the log growth phase. 10(-7) M oestradiol had no effect on articular chondrocyte growth. (5) In male rats implanted with silastic capsules releasing 17 beta-oestradiol, increase in body weight was retarded by about 25% over a period of 6 weeks, compared to control rats. Rat chondrosarcoma grew to the same size in oestrogen-treated rats as it did in controls. (6) Oestrogen receptors could not be detected in freshly isolated bovine articular chondrocytes or in rat chondrosarcoma. (7) In conclusion, neither the mitotic rate of articular chondrocytes nor their proteoglycan metabolism is under the direct physiological control of oestradiol. Growth and biosynthetic activity of the rat chondrosarcoma chondrocytes are independent of either direct control by the hormone or control effected by oestradiol regulation of a second hormone or growth factor.


Chronically ill adults' educational needs on self-medication.
The present study, which was conducted in Jordan, examined chronically ill adults' educational needs concerning the self-administration of medications. The study utilized a cross-sectional design. Five hundred adults were recruited from primary health centers. Chronically ill adults needed education on medication benefits, odds ratio (OR)= 4.23, 95% confidence interval (95% CI) = (3.07-5.81), p = .00; side effects, OR = 2.25, 95% CI = (1.48-4.28), p = .00; and management of side effects, OR = 2.20, 95% CI = (1.35-4.10), p = .00. Nurses, physicians, and pharmacists need to collaborate to ensure that patients receive adequate education on their prescribed medications.


Role of interferon regulatory factor 1 in governing Treg depletion, Th1 polarization, inflammasome activation and antitumor efficacy of cyclophosphamide.
The antitumor effectiveness of cyclophosphamide (CTX) and other chemotherapeutics was shown to rely not only on direct cytotoxicity but also on immunogenic tumor cell death and systemic immunomodulatory mechanisms, including regulatory T cell (Treg) depletion, Th1 cell polarization, type I interferon (IFN) and proinflammatory cytokine production. IFN regulatory factor (IRF)-1 is a transcriptional regulator of IFNs and IFN-inducible genes, involved in the control of Th1 and Treg differentiation and in sterile inflammation. Aim of this study was to explore the role of IRF-1 in CTX-induced antitumor effects and related immune activities. This study shows for the first time that IRF-1 is important for the antitumor efficacy of CTX in mice. Moreover, experiments in tumor-bearing C57BL/6 mice showed that Irf1 gene expression in the spleen was transiently increased following CTX administration and correlated with the induction of Th1 cell expansion and of Il12p40 gene expression, which is the main Th1-driving cytokine. At the same time, CTX administration reduced both Foxp3 expression and Treg cell percentages. These effects were abrogated in Irf1-/- mice. Further experiments showed that the gene and/or protein expression of caspase-1, iNOS, IL-1β, IL-6 and CXCL10 and the levels of nitric oxide were modulated following CTX in an IRF-1-direct- or -indirect-dependent manner, and highlighted the importance of caspase-1 in driving the sterile inflammatory response to CTX. Our data identify IRF-1 as important for the antitumor efficacy of CTX and for the regulation of many immunomodulatory activities of CTX, such as Th1 polarization, Treg depletion and inflammation.


Regulation of IkappaB kinase-related kinases and antiviral responses by tumor suppressor CYLD.
The IkappaB kinase (IKK)-related kinases, IKKepsilon and TBK1, participate in the induction of type I interferons (IFNs) during viral infections. Deregulated activation of IKKepsilon and TBK1 also contributes to the abnormal cell survival and transformation. However, how these kinases are negatively regulated remains unclear. We show here that the tumor suppressor CYLD has an essential role in preventing aberrant activation of IKKepsilon/TBK1. CYLD deficiency causes constitutive activation of IKKepsilon/TBK1, which is associated with hyper-induction of IFNs in virus-infected cells. We further show that CYLD targets a cytoplasmic RNA sensor, RIG-I, and inhibits the ubiquitination of this IKKepsilon/TBK1 stimulator. Consistent with the requirement of ubiquitination in RIG-I function, CYLD potently inhibits RIG-I-mediated activation of the IFN-beta promoter. These findings establish CYLD as a key negative regulator of IKKepsilon/TBK1 and suggest a role for CYLD in the control of RIG-I ubiquitination.


Histological and histochemical observations on salivary microliths in ferret.
The fortuitous observation of salivary microliths in ferret was pursued in the present investigation. Major salivary glands obtained post-mortem from mature ferrets of either sex were examined with the use of histology and light microscopical histochemistry for calcium, protein, amino acids, mucosubstances and hydrolytic enzymes. Microliths were detected in most parotids, but were absent from submandibular and sublingual glands. The microliths were usually seen in lumens, and occasionally in parenchyma and interstices. They were variably stained for calcium, tryptophan, and neutral and acidic mucosubstances, similarly to acinar or ductal secretory granules. Unlike secretory granules, microliths showed autofluorescence, high levels of tyrosine and a low concentration of -SS- groups. Acid phosphatase and beta-glucuronidase reaction surrounded non-luminal microliths. The present data establish ferret as a new model for the investigation of salivary microliths and do not support the notion of microliths being almost absent from the parotid. Probably there is secretory inactivity in ferret parotid and this fosters the formation and accumulation of microliths containing calcium and disintegrated secretory material.


Tumor volume improves the long-term prediction of biochemical recurrence-free survival after radical prostatectomy for localized prostate cancer with positive surgical margins.
To develop a novel application evaluating the effect of tumor volume (TV) and percentage of high-grade tumor volume (%HGTV) on long-term biochemical recurrence-free survival rate (BCRFS) after radical prostatectomy (RP) in patients with pT2 PCa. Retrospective analysis of 903 men with pT2 PCa between 1992 and 2004 at a single European tertiary care center was performed. Cox regression models identified risk factors for BCR. A nomogram was developed to predict the BCRFS at 5, 10 and 15 years after RP. Decision curve analyses were performed to identify the net increase in cases identified by the full model. BCR-free survival rates at 5, 10 and 15 years were 94, 90 and 86 %. In Cox regression analyses, TV, %HGTV and positive surgical margin status (SM) were independent predictors of BCR. Predictive accuracies (PA) at 5, 10 and 15 years of the base model (PSA, Gleason score, SM) were 76.8 % (95 % CI 67.9-78.2 %), 70.5 % (95 % CI 64.9-75.0 %) and 68.1 % (95 % CI 60.6-73.5 %). The full model, including TV and %HGTV, achieved 76.9, 72.4 and 70.7 %. These PA differences were statistically significant at 10 and 15 years (p < 0.001). TV and %HGTV could potentially serve as valuable measures to stratify patients at high risk of BCR. The use of our nomogram should be considered to counsel patients with pT2 disease and SM and to design appropriate follow-up or treatment regimens.


What constitutes adequate surgical therapy for benign nodular goiter?
It is our hypothesis that the extent of thyroid resection for benign nodular thyroid disease (NTD) should be based on the extent of disease. Patients operated on for benign NTD from 1990 through 2007 were divided into 3 groups: those who underwent lobectomy for unilateral NTD (Group 1); near-total or total thyroidectomy for bilateral NTD (Group 2); and reoperation for NTD initially treated at other institutions (Group 3). The incidence of recurrence was determined for Groups 1 and 2 and the timing of diagnosis was compared to Group 3. Potential risk factors for recurrent disease were examined. Five hundred forty-five patients were operated on for benign NTD. Contralateral disease was excluded in Group 1 patients using ultrasound (47.7%) and/or intraoperative palpation (100%). Five (1.9%) of 260 patients in Group 1 and 1 (0.4%) of 248 patients in Group 2 developed recurrent NTD after 7 +/- 4 (median = 8) and 4 y compared to a mean 19 +/- 11 (median = 20) y for the 37 patients in Group 3 following 1 to 3 previous thyroidectomies. Recurrent disease was diagnosed by physical exam in 24 (55.8%) and imaging in 19 (44.2%) patients. Thyroid hormone was required for postsurgical hypothyroidism in 70 (26.9%) patients in Group 1. Thyroid lobectomy is optimal therapy when benign NTD is limited to 1 lobe, as evidenced by a 2% recurrence rate and maintenance of euthyroidism in 73% of patients. When NTD is bilateral, total thyroidectomy is indicated to eliminate recurrence, underscoring the importance of routine preoperative ultrasound.


Prognostic significance of plasma placental growth factor levels in renal cell cancer: an association with clinical characteristics and vascular endothelial growth factor levels.
Angiogenesis plays important roles in pathogenesis in human cancer. We assessed the relationship between clinical features of renal cell cancer (RCC) and PlGF levels in relation to VEGF. Plasma PlGF and VEGF levels were measured in patients with renal cell cancer. The levels of these angiogenetic factors were analyzed in relation to clinical parameters. PlGF and VEGF levels in patients with RCC were significantly higher than those in non-cancer controls. PlGF levels were significantly associated with histological grade and total tumor vascularity (TTV), and VEGF levels were significantly associated with T, M stage, histological grade, venous invasion and TTV. Multivariate analysis showed plasma PlGF was an independent prognostic factor. These findings suggested that plasma PlGF levels were significantly associated with the clinical features of RCC, especially prognostic significance.


Platelet-rich plasma treatment for talar cartilage repair: a systematic review and meta-analysis.
To systematically review the studies regarding to the safety, efficacy and application methods of PRP in promoting the talar cartilage repair. A systematic review was performed by searching PubMed, Web of Science, OVID and EMBASE to identify studies that compared the clinical efficacy of PRP for talar cartilage repair. Main outcome was the American Orthopedic Foot and Ankle Society (AOFAS) score for function and Visual Analog Scale (VAS) for pain was the second outcome. A total of 10 studies were included in this systematic review, including 4 randomized controlled trials, 1 controlled trial, 3 case series and 2 cohort studies. Four RCTs were analyzed using meta-analysis. For all outcomes, statistical results favored PRP group (AOFAS: MD = 7.84; 95% CI= [-0.13, 15.80], I2 = 83%, P < 0.01; VAS: MD = 1.86; 95% CI= [0.68, 3.04], I2 = 85%, P < 0.01). There were almost no reports of adverse events related to PRP intervention. Subgroup analysis showed that whether PRP was used alone or combined with other treatments could result in high heterogeneity but no more specific factors were identified to contribute to this. PRP is safe and effective for talar cartilage repair. In addition to the standardization of PRP preparation and application, it is necessary to distinguish the effects of PRP used alone or in combination with other treatments. In PRP studies, surgical treatment of talar cartilage repair remains the mainstream. The regulation of PRP in surgical applications are worth exploring. The most relative component is the mesenchymal stem cell because it is the only exposed chondrocyte precursor in the articular cavity whether it is microfracture or cell transplantation. The study was registered in the PROSPERO International prospective register of systematic reviews (CRD42022360183).


Structure of the proteolipid protein extracted from bovine central nervous system myelin with nondenaturing detergents.
As a basis for attempts to define the structures of the proteins within myelin, methods have been developed for their extraction and isolation in solutions of non-denaturing detergents. With use of solutions of deoxycholate or Triton X-100, up to 90% of the protein has been extracted from bovine CNS myelin, along with most of the phospholipid. The proteolipid protein has been purified in deoxycholate solutions by chromatography on a blue dye-ligand column, which retained all of the basic protein and 2',3'-cyclic nucleotide-3'-phosphodiesterase, and then on Sephacryl S300, which separated proteolipid protein from phospholipid and high-molecular-weight proteins. The proteolipid protein was isolated from Triton X-100 extracts of myelin by adsorption onto phosphocellulose resin, with subsequent elution by 0.5 M sodium chloride. Gel permeation chromatography was used as the final purification step. Sedimentation equilibrium experiments gave a monomer molecular weight of 134,000 +/- 8000 in deoxycholate and 145,000 +/- 17,000 in Triton X-100 solutions. On the basis of an apparent subunit molecular weight of 23,500 it was deduced that the native protein is probably hexameric. Above 0.2 gL-1 in Triton X-100 solutions and 0.5 gL-1 in deoxycholate solutions the protein aggregated. In deoxycholate solutions the protein adopts the highly helical conformation expected for an intrinsic membrane protein.


Transgenic indica rice plants harboring a synthetic cry2A* gene of Bacillus thuringiensis exhibit enhanced resistance against lepidopteran rice pests.
A novel synthetic cry2A* gene was introduced into the elite indica rice restorer line Minghui 63 by Agrobacterium-mediated transformation. A total of 102 independent transformants were obtained. Among them, 71 transformants were positive cry2A* plants according to PCR analysis. Four highly insect-resistant lines with single-copy insertion (designated as 2A-1, 2A-2, 2A-3, and 2A-4) were selected based on field assessment and Southern blot analysis in the T(1) generation. All four transgenic lines showed Mendelian segregation by seed germination on 1/2 MS medium containing Basta. Homozygous transgenic plants were selected according to germination ratio (100%) in the T2 generation. Cry2A* protein concentrations were determined in homozygous transgenic lines, their derived hybrids, and their backcross offspring. The Cry2A* protein concentrations of four homozygous transgenic lines ranged from 9.65 to 12.11 microg/g of leaf fresh weight. There was little variation in the hybrids and backcross offspring. Insect bioassays were conducted in both the laboratory and field. All four transgenic lines were significantly resistant to lepidopteran rice pests. These cry2A* transgenic lines can be used to produce insect-resistant hybrids and serve as a resistant source for the development of two-toxin Bt rice.


SPP2 Mutations Cause Autosomal Dominant Retinitis Pigmentosa.
Retinitis pigmentosa (RP) shows progressive loss of photoreceptors involved with heterogeneous genetic background. Here, by exome sequencing and linkage analysis on a Chinese family with autosomal dominant RP, we identified a putative pathogenic variant, p.Gly97Arg, in the gene SPP2, of which expression was detected in multiple tissues including retina. The p.Gly97Arg was absent in 800 ethnically matched chromosomes and 1400 in-house exome dataset, and was located in the first of the two highly conserved disulfide bonded loop of secreted phosphoprotein 2 (Spp-24) encoded by SPP2. Overexpression of p.Gly97Arg and another signal peptide mutation, p.Gly29Asp, caused cellular retention of both endogenous wild type and exogenous mutants in vitro, and primarily affected rod photoreceptors in zebrafish mimicking cardinal feature of RP. Taken together, our data indicate that the two mutations of SPP2 have dominant negative effects and cellular accumulation of Spp-24 might be particularly toxic to photoreceptors and/or retinal pigment epithelium. SPP2 has a new role in retinal degeneration.


Neurofibrillar changes in goldfish (Carassius auratus L.) brain in relation to environmental temperature.
Neurofibrillar changes occur in the brains of goldfish, Carassius auratus L., maintained at 5 degrees C for 177 days or more under a 12-hour photoperiod. In paraffin sections impregnated by the silver method of Holmes, the light microscopic appearance of the neurofibrillary change was evidenced by black rings 1 micrometer to 2 micrometer in outside diameter. A quantitative study showed the mean number of rings to be: optic tectum layer 3, 1.34 X 10(2)/mm(3), layer 5, 1.14 X 10(2)/MM(3), nucleus prerotundus 4.63 X 10(2)/mm(3), olfactory bulbs, 6.2 X 10(2)/mm3. In the brains of fish kept at 15 degrees C a few rings are found only in some olfactory bulbs and not in any other region of the brain. The brain wet weight also changes and was found to be significantly (p less than 0.01) less in 5 degrees C than in 15 degrees C fish, the mean values being 0.1588 g and 0.2091 g respectively. The significance of the observed changes is discussed. It is suggested that the smaller brain wet weight may reflect a change in the vasculature of the brain and be related to the acclimation process. It is hypothesized that the rings are a morphological expression of a physiological change caused by prolonged exposure to low temperatures; a neuronal response reflecting either early degeneration or a functional adaptation which may involve an alteration in axoplasmic transport. It is suggested that this system is a useful model for the study of relationships between neurofibrillar disturbances and neuronal function.


Pathologic endoplasmic reticulum stress induced by glucotoxic insults inhibits adipocyte differentiation and induces an inflammatory phenotype.
Adipocyte differentiation is critical in obesity. By controlling new adipocyte recruitment, adipogenesis contrasts adipocyte hypertrophy and its adverse consequences, such as insulin resistance. Contrasting data are present in literature on the effect of endoplasmic reticulum (ER) stress and subsequent unfolded protein response (UPR) on adipocyte differentiation, being reported to be either necessary or inhibitory. In this study, we sought to clarify the effect of ER stress and UPR on adipocyte differentiation. We have used two different cell lines, the widely used pre-adipocyte 3T3-L1 cells and a murine multipotent mesenchymal cell line, W20-17 cells. A strong ER stress activator, thapsigargin, and a pathologically relevant inducer of ER stress, glucosamine (GlcN), induced ER stress and UPR above those occurring in the absence of perturbation and inhibited adipocyte differentiation. Very low concentrations of 4-phenyl butyric acid (PBA, a chemical chaperone) inhibited only the overactivation of ER stress and UPR elicited by GlcN, leaving unaltered the part physiologically activated during differentiation, and reversed the inhibitory effect of GlcN on differentiation. In addition, GlcN stimulated proinflammatory cytokine release and PBA prevented these effects. An inhibitor of NF-kB also reversed the effects of GlcN on cytokine release. These results indicate that while ER stress and UPR activation is "physiologically" activated during adipocyte differentiation, the "pathologic" part of ER stress activation, secondary to a glucotoxic insult, inhibits differentiation. In addition, such a metabolic insult, causes a shift of the preadipocyte/adipocyte population towards a proinflammatory phenotype.


Upregulation of the KIAA1199 gene is associated with cellular mortality.
The microcell-mediated transfer of a normal human chromosome 3 induces replicative senescence in otherwise immortal renal cell carcinoma cells. To identify the genes involved in the chromosome 3-induced cellular mortality, we previously performed a cDNA subtraction experiment using the immortal renal cell carcinoma cells (RCC23) and the mortal counterpart with the transferred chromosome 3 (RCC23+3). We here report the cDNA cloning and characterization of one of the differentially expressed genes, which encodes KIAA1199 protein of unknown function. Northern blot and RT-PCR analyses revealed striking upregulation of KIAA1199 mRNA in mortal RCC23+3 compared with immortal RCC23. However, no significant change in KIAA1199 mRNA expression was observed during replicative aging in vitro (from early passage culture to senescent culture) of mortal human cells including RCC23+3, normal fibroblasts and prostate epithelial cells. Interestingly, an immortal fibroblast cell line and two breast cancer cell lines expressed much lower amounts of KIAA1199 mRNA than their normal counterparts. KIAA1199 mRNA is expressed in a wide range of normal human tissues, with the highest level of expression in brain. The gene is located on chromosome band 15q25, where a brain tumor suppressor gene has been mapped. These findings suggest that KIAA1199 gene may play a role in cellular mortality of normal human cells, which counters cell immortalization and carcinogenesis.


Vascular effects of oxygen-derived free radicals.
This review attempts to summarize the available data regarding the vascular actions of free oxygen radicals. Studies on blood vessels in situ and in vitro demonstrate that free oxygen radicals can evoke both vasodilation and vasoconstriction. Free oxygen radicals can modulate the tone of vascular smooth muscle by acting directly on the smooth muscle cells, and also via indirect mechanisms by changes in the production or biological activity of vasoactive mediators. The individual oxygen radicals may have different (sometimes opposite) vascular effects. Superoxide anion inactivates endothelium-derived relaxing factor and the adrenergic neurotransmitter norepinephrine. Hydrogen peroxide and the hydroxyl radical evoke vasodilation by acting directly on vascular smooth muscle and also by stimulating the synthesis/release of endothelium-derived relaxing factor. In acute arterial hypertension or experimental brain injury oxygen radicals are important mediators of vascular damage. Production of oxygen-derived free radicals by activated neutrophils may be responsible for vasodilation and increased permeability of capillary membrane during the acute inflammatory process. Free oxygen radicals also play an important role in reperfusion injury of various organs, and vascular actions of the free radicals may contribute to the damage of parenchymal tissues.


The DEXD/H-box RNA helicase RHII/Gu is a co-factor for c-Jun-activated transcription.
Tandem affinity purification (TAP) and mass spectrometric peptide sequencing showed that the DEAD-box RNA helicase RHII/Gu is a functional interaction partner of c-Jun in human cells. The N-terminal transcription activation region of, c-Jun interacts with a C-terminal domain of RHII/Gu. This interaction is stimulated by anisomycin treatment in a manner that is concurrent with, but independent of, c-Jun phosphorylation. A possible explanation for this effect is provided by the observation that RHII/Gu translocates from nucleolus to nucleoplasm upon anisomycin or UV treatment or when JNK signaling is activated by overexpression of a constitutively active form of MEKK1 kinase. Several experiments show that the RNA helicase activity of RHII/Gu supports c-Jun-mediated target gene activation: dominant-negative forms of RHII/Gu, as well as a neutralizing antibody against the enzyme, significantly interfered with c-Jun target gene activity but not with transcription in general. These findings clarify the mechanism of c-Jun-mediated transcriptional regulation, and provide evidence for an involvement of RHII/Gu in stress response and in RNA polymerase II-catalyzed transcription in mammalian cells.


Decline of melanism in two British moths: spatial, temporal and inter-specific variation.
The decline in industrial melanism over the last quarter century constitutes an exceptional case of an evolutionary change, varying in both time and space, and between species. In Biston betularia and Odontoptera bidentata, the change in melanic frequency is closely replicated at two sites 0.5 km apart. Between seven sites 50-100 km apart, there is heterogeneity in both the speed and timing of change. At sites that were heavily industrialized, the change is faster, from an initially higher frequency, and starts later than at sites which are more rural.We propose a method for estimating systematic change during sigmoid declines in melanic frequencies. This fails to show any significant change over time in selective coefficients. It is concluded that the overall pattern of change has been driven largely by events in the most polluted and industrialized parts of the country. Although migration may contribute to the estimated selective values, natural selection is the only credible explanation for the overall decline.


Sensitive analysis of amino acids with carrier-mediated single drop microextraction in-line coupled with capillary electrophoresis.
In order to analyze amino acids sensitively without derivatization, we have developed carrier-mediated single drop microextraction (SDME). Nonane-1-sulfonic acid was added to an acidic sample donor solution as a carrier to form neutral ion pair complexes with amino acids. The ion pair complexes were extracted to the organic phase, covering a drop of an aqueous basic acceptor phase hanging at the tip of a capillary, and then back-extracted to the basic acceptor phase, where both the amino acids and the carrier have negative charges and the ion pair complexes are broken. The resulting extract of enriched amino acids was injected into the capillary and analyzed by capillary electrophoresis. With 20-min SDME with agitation of the donor phase, enrichment factors of four aromatic amino acids were up to 120-fold, yielding the LOD of 70-500 nM. The linear dynamic ranges for corrected peak areas were 1-100 μM with linear correlation coefficients larger than 0.9959. With internal standardization, the intraday RSDs of migration times and corrected peak areas were 0.01-0.04% and 2.0-3.7%, respectively. The capabilities of sample cleanup including desalting and preconcentration of carrier-mediated SDME were demonstrated with the analysis of human urine after minimal pretreatment of acidification and centrifugation.


Engineering Candida tenuis Xylose reductase for improved utilization of NADH: antagonistic effects of multiple side chain replacements and performance of site-directed mutants under simulated in vivo conditions.
Six single- and multiple-site variants of Candida tenuis xylose reductase that were engineered to have side chain replacements in the coenzyme 2'-phosphate binding pocket were tested for NADPH versus NADH selectivity (R(sel)) in the presence of physiological reactant concentrations. The experimental R(sel) values agreed well with predictions from a kinetic mechanism describing mixed alternative coenzyme utilization. The Lys-274-->Arg and Arg-280-->His substitutions, which individually improved wild-type R(sel) 50- and 20-fold, respectively, had opposing structural effects when they were combined in a double mutant.


Short-course androgen deprivation therapy and the risk of death from high-risk prostate cancer in men undergoing external beam radiation therapy and brachytherapy.
We estimated the risks of prostate cancer-specific mortality (PCSM) and all-cause mortality (ACM) in men with high-risk prostate cancer (PC) undergoing external beam radiation therapy and brachytherapy with short-course androgen deprivation therapy (ADT) (median 4 months) as compared with men with more favorable-risk PC undergoing standard of care as per the National Comprehensive Cancer Network guidelines. The prospective study cohort comprised 6595 consecutively treated men with T1-4 N0M0 PC whose treatment included brachytherapy between October 16, 1997, and May 28, 2013. Fine and Gray competing risk regression and Cox regression analyses were used to assess the risks of PCSM and ACM in men with high, unfavorable intermediate, and favorable intermediate risk as compared with low-risk PC. After median followup of 7.76 years, 820 men died (12.43%): 72 of PC (8.78%). Men with favorable intermediate-risk PC did not have significantly increased PCSM risk as compared with men with low-risk PC (adjusted hazard ratio [AHR], 1.26; 95% confidence interval [CI] 0.56, 2.88; p-Value 0.58), whereas men with high-risk PC (AHR, 3.74; 95% CI 1.12, 12.53; p-Value 0.032) and unfavorable intermediate-risk PC (AHR, 3.10; 95% CI 1.43, 6.72; p-Value 0.004) did. Based on 10-year adjusted point estimates of PCSM and ACM for men with high-risk PC being 6.01% (95% CI 3.79%, 8.94%) and 21.30% (95% CI 17.45%, 25.42%), respectively, PCSM comprised 28% of ACM. In the setting of external beam radiation therapy and brachytherapy, men with high-risk PC have low absolute adjusted estimates of PCSM (~6%) during the first decade after treatment despite receiving only short-course ADT. Whether long-term ADT can lower PCSM and improve survival in these men requires additional study.


Probabilistic noninvasive prediction of wall properties of abdominal aortic aneurysms using Bayesian regression.
Multiple patient-specific parameters, such as wall thickness, wall strength, and constitutive properties, are required for the computational assessment of abdominal aortic aneurysm (AAA) rupture risk. Unfortunately, many of these quantities are not easily accessible and could only be determined by invasive procedures, rendering a computational rupture risk assessment obsolete. This study investigates two different approaches to predict these quantities using regression models in combination with a multitude of noninvasively accessible, explanatory variables. We have gathered a large dataset comprising tensile tests performed with AAA specimens and supplementary patient information based on blood analysis, the patients medical history, and geometric features of the AAAs. Using this unique database, we harness the capability of state-of-the-art Bayesian regression techniques to infer probabilistic models for multiple quantities of interest. After a brief presentation of our experimental results, we show that we can effectively reduce the predictive uncertainty in the assessment of several patient-specific parameters, most importantly in thickness and failure strength of the AAA wall. Thereby, the more elaborate Bayesian regression approach based on Gaussian processes consistently outperforms standard linear regression. Moreover, our study contains a comparison to a previously proposed model for the wall strength.


5T4 oncofetal antigen in gastric carcinoma and its clinical significance.
To evaluate the role of 5T4 antigen in gastric cancer progression and prognosis. A prospective study of 5T4 antigen expression in primary, secondary and recurrent gastric carcinoma, the relationship to selected prognostic parameters and the course of disease. Eighty six patients operated on for gastric cancer. TISSUE: One hundred and twenty two gastric tumours were studied, including 86 primary carcinomas, 32 coexisting lymph node metastases and four recurrent carcinomas. Immunohistochemistry using 5T4 monoclonal antibody on frozen sections. The 5T4 antigen was detected in 41% of primary gastric tumours including early gastric cancer. A strong relationship was found between 5T4 positivity and tumour histology. Thus, 52% of gastric carcinomas of intestinal type expressed 5T4 antigen compared with 28% of the diffuse type (P = 0.028). Among 16 sets of primary gastric carcinomas and regional lymph node metastases, coordinate 5T4 expression was seen in 14 cases; the other two showed acquisition of positivity on metastatic tumour cells (carcinomas of diffuse type). 5T4 antigen was detected more frequently in carcinomas with p53 accumulation compared with those with undetectable p53 levels (P = 0.015). The presence of 5T4 in cancer cells was correlated with poor short-term prognosis (24% vs 49% of 2 year survival for 5T4 positive and negative tumours respectively, P = 0.024). The effect on survival was evident in the p53 negative group, with patients 5T4 positive showing worse survival (28% vs 60% in 2 years). Our results suggest that the assessment of 5T4 expression in gastric carcinoma can be helpful in identifying patients with poor short-term prognosis.


Evaluating the Potential Severity of Look-Alike, Sound-Alike Drug Substitution Errors in Children.
Look-alike, sound-alike (LASA) drug name substitution errors in children may pose potentially severe consequences. Our objective was to determine the degree of potential harm pediatricians ascribe to specific ambulatory LASA drug substitution errors. We developed a unified list of LASA pairs from published sources, removing selected drugs on the basis of preparation type (eg, injectable drugs). Using a modified Delphi method over 3 rounds, 38 practicing pediatricians estimated degree of potential harm that might occur should a patient receive the delivered drug in error and the degree of potential harm that might occur from not receiving the intended drug. We identified 3550 published LASA drug pairs. A total of 1834 pairs were retained for the Delphi surveys, and 608 drug pairs were retained for round 3. Final scoring demonstrated that participants were able to identify pairs where the substitutions represented high risk of harm for receiving the delivered drug in error (eg, did not receive methylphenidate/received methadone), high risk of harm for not receiving the intended drug (eg, did not receive furosemide/received fosinopril), and pairs where the potential harm was high from not receiving the intended drug and from erroneously receiving the delivered drug (eg, did not receive albuterol/received labetalol). Pediatricians have identified LASA drug substitutions that pose a high potential risk of harm to children. These results will allow future efforts to prioritize pediatric LASA errors that can be screened prospectively in outpatient pharmacies.


Interleukin 18 and IL-18 BP response to Sars-CoV-2 virus infection.
The immune response to the SARS-CoV-2 infection is crucial to the patient outcome. IL-18 is involved in the lymphocyte response to the disease and it is well established its important role in the complex developing of the host response to viral infection. This study aims at the analysis of the concentrations of IL-18, IL-18BP, INF-γ at the onset of the SARS-CoV-2 infection. The serum levels of measured interleukins were obtained through enzyme-linked immunosorbent assay. Furthermore, the free fraction of IL-18 was numerically evaluated. The enrolled patients were divided in two severity groups according to a threshold value of 300 for the ratio of arterial partial pressure of oxygen and fraction of inspired oxygen fraction and according to the parenchymal involvement as evaluated by computerized tomography at the admittance. In the group of patients with a more severe disease, a significant increase of the IL-18, INF-γ and IL-18BP levels have been observed, whereas the free IL-18 component values were almost constant. The results confirm that, at the onset of the disease, the host response keep the inflammatory cytokines in an equilibrium and support the hypothesis to adopt the IL-18BP modulation as a possible and effective therapeutic approach.


Empowering and protecting children by enhancing knowledge, skills and well-being: A randomized trial of Learn to BE SAFE with Emmy.
Australia needs effective programs to protect children and prevent abuse, but there is little information available for policymakers or families. Using a randomized controlled trial, Learn to BE SAFE with Emmy, a school-based protection program for young children designed by Act for Kids, was evaluated to determine its effectiveness for promoting young children's knowledge and skills. Grade one children (n=245) from 15 classrooms across 5 primary schools completed assessment measures. A subset of these children received the program (n=131) or acted as a comparison group (n=114). Parents (n=72) completed questionnaires about their child's participation in the program. When compared with children who had not received the program, children who completed Learn to BE SAFE with Emmy demonstrated increased knowledge of interpersonal safety and were more likely to choose a safe response option to hypothetical unsafe scenarios 6 months after participation than at both pre- and post-intervention. Parents reported their children who participated used more safety strategies immediately and 6 months after participation compared to pre-intervention. Outcomes can assist in guiding future policies around the prevention of child abuse and protect the well-being of Australian children.


Impact of Global and Segmental Hypertrophy on Two-Dimensional Strain Derived from Three-Dimensional Echocardiography in Hypertrophic Cardiomyopathy: Comparison with Healthy Subjects.
Patients with hypertrophic cardiomyopathy (HCM) present unusual myocardial mechanics. The aim of this study was to assess the impact of hypertrophy on global and regional two-dimensional (2D) strain derived from both tomographic images (2D/2D) and volumetric image acquisition (2D/three-dimensional [3D]) in patients with HCM compared with control subjects. Comprehensive resting 2D and 3D echocardiography was performed in 40 patients with HCM and in 53 control subjects, with comparable distributions of age, gender, and left ventricular (LV) ejection fraction. LV global and segmental measurements of all 2D/2D and 2D/3D peak strain components (global and segmental longitudinal strain, global and segmental circumferential strain, global and segmental radial strain, and global and segmental area strain) and 3D indexed LV end-diastolic myocardial mass were obtained from all patients. LV wall thickness was assessed in short-axis views and classified in four quartiles (<10.5, 10.5-13.0, 13.0-16.5, and >16.5 mm). The reproducibility of 2D/3D strain was similar or greater and more consistent for all components compared with 2D/2D strain analysis. There was a significant correlation between 3D LV end-diastolic mass and all 2D/3D strain components (P < .05). Two-dimensional/3D global circumferential strain had the strongest association with 3D LV ejection fraction (r = 0.50, P = .001). For segmental deformation, patients with HCM had lower longitudinal deformation whatever the LV wall thickness, whereas circumferential function was increased in nonhypertrophied and poorly hypertrophied segments compared with control subjects. Two-dimensional/3D strain is a reliable technique to assess myocardial deformation. Myocardial mass is related to 2D/3D strain components in patients with HCM. Circumferential deformation, compared with longitudinal deformation, seems to be the main component of the maintenance of systolic function in HCM.


Glycosylation of endogenous protein(s) of the rough and smooth microsomes by a lipid sugar intermediate.
Several problems regarding the protein acceptor of the oligosaccharide from GEA (glucosylated endogenous acceptor) were investigated in the present work using rat liver microsomal subfractions. It was found that the acceptor molecule is present in rough and smooth liver microsomes. Furthermore both fractions have closely similar specific activities. The problem of whether nascent peptides must be ribosome bound for glycosylation to occur was studied. The results suggests that binding of peptides to ribosomes is not a necessary condition for the transfer of GEA oligosaccharide to protein. The increase in specific activity found after partial release of the microsomal vesicular content suggests that the acceptor protein for GEA is membrane bound. Evidence obtained in attempting to elucidate whether nascent or completed chains are glycosylated favours the later possibility.


Thermal properties of municipal solid waste components and their relative significance for heat retention, conduction, and thermal diffusion in landfills.
Significant amounts of heat can be generated during the initial stages after wastes are deposited in landfills, primarily due to decomposition of food waste. Objectives of this study are to compile, examine and compare thermal properties of municipal solid waste (MSW) components, and liquid and gas phases in MSW landfills and their thermal responses that effect temperature increases in gas and leachate. Specific thermal properties examined include thermal conductivity, thermal diffusivity, and specific heat of waste materials deposited in landfills, liquids (water), and gases present. Compilation of these properties will allow in depth thermal analyses to evaluate heat transfer dynamics in landfills with different waste compositions. Examination of thermal characteristics of MSW components indicate that heat generated during decomposition of waste components would primarily be transferred to liquid (leachate) due to formation of water and gaseous components and their high specific heats. As a result, both the leachate and gases released from a landfill during the initial stages after wastes are deposited and when some oxygen is present as an electron acceptor will be warmer. Except for the metals and construction waste, it is likely that most waste components will have a significant temperature gradient during warming up and cooling off stages due to their low thermal conductivities and low thermal diffusivities. Even when the gas phase is at higher temperatures, it will take long time for waste materials (other than food waste and metals) to come to a uniform temperature during the heat generation (primarily due to decomposition of food waste) in a landfill.


Learning in higher order Boltzmann machines using linear response.
We introduce an efficient method for learning and inference in higher order Boltzmann machines. The method is based on mean field theory with the linear response correction. We compute the correlations using the exact and the approximated method for a fully connected third order network of ten neurons. In addition, we compare the results of the exact and approximate learning algorithm. Finally we use the presented method to solve the shifter problem. We conclude that the linear response approximation gives good results as long as the couplings are not too large.


DFT simulations and microkinetic modelling of 1-pentyne hydrogenation on Cu20 model catalysts.
Adsorption and dissociation of H2 and hydrogenation of 1-pentyne on neutral and anionic Cu20 clusters have been investigated using the density functional theory and microkinetic modelling. Molecular adsorption of H2 is found to occur strictly at atop sites. The H2 dimer is activated upon adsorption, and the dissociation occurs with moderate energy barriers. The dissociated H atoms reside preferentially on 3-fold face and 2-fold edge sites. Based on these results, the reaction paths leading to the partial and total hydrogenation of 1-pentyne have been studied step-by-step. The results suggest that copper clusters can display selective activity on the hydrogenation of alkyne and alkene molecules. The hydrogenated products are more stable than the corresponding initial reactants following an energetic staircase with the number of added H atoms. Stable semi-hydrogenated intermediates are formed before the partial (1-pentene) and total (pentane) hydrogenation stages of 1-pentyne. The microkinetic model analysis shows that C5H10 is the dominant product. Increasing the reactants (C5H8/H2) ratio enhances the formation of products (C5H10 and C5H12).


Neural correlates of cognitive dysfunction after cardiac surgery.
Patients who underwent cardiac surgery and their relatives often complain on postoperative memory impairment. Most prospective neuropsychological studies also found postoperative cognitive decline early after surgery. Nevertheless, recently several reports questioned the existence of long-term brain alterations in these patient cohorts. The present review was aimed to clear up the true cardiac surgery effects on brain and cognitive functions. The reviewed data evidence that cardiac surgery interventions induce persistent localized brain ischemic lesions along with rapidly reversing global brain swelling and decreased metabolism. A range of studies showed that left temporal region was especially prone to perioperative ischemic injury, and these findings might explain persistent verbal short-term memory decline in a considerable proportion of cardiac surgery patient cohorts. Speed/time of cognitive performance is commonly decreased early after on-pump surgery either. Nevertheless, no association between psychomotor speed slowing and intraoperative embolic load was found. The rapid recovery of the latter cognitive domain might be better explained by surgery related acute global brain metabolism changes rather than ischemic injury effects. Hence, analyses of performance on separate cognitive tests rather than summarized cognitive indexes are strongly recommended for future neuropsychological studies of cardiac surgery outcomes.


Comparative transcriptome analysis of latex from rubber tree clone CATAS8-79 and PR107 reveals new cues for the regulation of latex regeneration and duration of latex flow.
Rubber tree (Hevea brasiliensis Muell. Arg.) is the primarily commercial source of natural rubber in the world. Latex regeneration and duration of latex flow after tapping are the two factors that determine rubber yield of rubber tree, and exhibit a huge variation between rubber tree clones CATAS8-79 and PR107. To dissect the molecular mechanism for the regulation of latex regeneration and duration of latex flow, we sequenced and comparatively analyzed latex of rubber tree clone CATAS8-79 and PR107 at transriptome level. More than 26 million clean reads were generated in each pool and 51,829 all-unigenes were totally assembled. A total of 6,726 unigenes with differential expression patterns were detected between CATAS8-79 and PR107. Functional analysis showed that genes related to mass of categories were differentially enriched between the two clones. Expression pattern of genes which were involved in latex regeneration and duration of latex flow upon successive tapping was analyzed by quantitative PCR. Several genes related to rubber biosynthesis, cellulose and lignin biosynthesis and rubber particle aggregation were differentially expressed between CATAS8-79 and PR107. This is the first report about probing latex regeneration and duration of latex flow by comparative transcriptome analysis. Among all the suggested factors, it is more important that the level of endogenous jasmonates, carbohydrate metabolism, hydroxymethylglutaryl-CoA reductase (HMGR) and Hevea rubber transferase (HRT) in mevalonate (MVA) parthway for latex regeneration while the level of endogenous ethylene (ETH), lignin content of laticifer cell wall, antioxidants and glucanases for the duration of latex flow. These data will provide new cues for understanding the molecular mechanism for the regulation of latex regeneration and duration of latex flow in rubber tree.


A multidisciplinary approach to evaluate the efficiency of a clean-up technology to remove mercury from water.
A microporous material denoted ETS-4 was used as the decontaminant agent to treat water with a low level of Hg contamination. The effectiveness of the treatment was evaluated by assessment of the efficiency of Hg removal and ecotoxicological responses. The results showed that under highly competitive conditions the removal of Hg ranged between 58 % and 73 % depending upon the initial Hg concentration, and that Hg removal was reflected in decreased toxicity to some organisms. The ecotoxicological data indicated that the bacterium Vibrio fischeri was the least sensitive organism tested, as no toxicity was observed in either pre- or post-treatment waters. Daphnia magna was highly sensitive to Hg. Mercury removal by ETS-4 was not sufficient to completely remove the toxicity of Hg to D. magna. However, it was effective in the complete reduction of toxicity for the green alga, Pseudokirchneriella subcapitata.


Electrophoretic transfer of proteins from fixed and stained gels.
A procedure by which sodium dodecyl sulfate gels can be fixed and stained with Coomassie blue and subsequently transferred to nitrocellulose for immunostaining is outlined. The procedure involves the complete removal of the stain followed by equilibration of the gel in sodium dodecyl sulfate running buffer. The efficiency of transfer is comparable to unfixed gels and the protein pattern of the transfer appears to be sharper, presumably due to less diffusion during the transfer process. The procedure does not affect the antigenicity of the proteins that have been examined by subsequent immunostaining. This method is particularly useful for situations in which sample size or concentration are limiting factors resulting in insufficient material for duplicate gels.


[The evaluation of apoptosis in peripheral blood lymphocytes in laryngeal cancer patients].
The laryngeal cancer is the most often cancer among others in head and neck region. It occurs mostly among 55 and 69. Its development depends on immunological state of the body. Vitality of the immunological system cells was considered due to growth, treatment sensitivity and prognosis of some neoplasms. The aim of this work were estimation and comparison the phenomenon of lymphocytes T and B apoptosis in laryngeal cancer patients treated with surgery and radiotherapy. The material were 30 patients hospitalized in The Department of Otolaryngology Medical University of Lublin. They all were treated with surgery or surgery and radiotherapy. Apoptosis was estimated on different stages of the treatment process. All samples were examined with the flow cytometry method. The control group were 21 patients hospitalized because of the suspicion of the apnea syndrome, which wasn't confirmed with polysomnographic examination. Results of this study show significantly increasing percentage of peripheral blood apoptotic B (CD19+) cells caused by surgical treatment. The results considering radiotherapy showed different influence on the phenomenon of immunological cells apoptosis, still those results weren't significant. The surgical treatment causes increased amount of apoptotic peripheral blood lymphocytes.


Association of lung cancer and airway particle concentration.
Using 15 autopsy lungs containing nonresected lung cancers, we evaluated the exogenous mineral particle content of the upper and lower lobe parenchyma and of the upper and lower lobe bronchial mucosa. The bronchial mucosa from the lobe with the cancer was found to contain about three times the median mineral particle load of the bronchial mucosa from the lobe without the cancer, a statistically significant difference. No such difference was seen for mineral content of the parenchyma. This observation suggests that mineral particle deposition or accumulation in the bronchial mucosa may be related to respiratory carcinogenesis.


Simultaneous quantitative determination of the HIV protease inhibitors amprenavir, indinavir, nelfinavir, ritonavir and saquinavir in human plasma by ion-pair high-performance liquid chromatography with ultraviolet detection.
A reversed-phase high-performance liquid chromatographic assay for the simultaneous quantitative determination of five HIV protease inhibitors (i.e. amprenavir, indinavir, nelfinavir, ritonavir, and saquinavir) in human plasma is described. Sample pretreatment consisted of solid-phase extraction prior to ion-pair, reversed-phase high-performance liquid chromatography with ultraviolet detection at 210 nm (amprenavir, indinavir and nelfinavir) and 239 nm (saquinavir and ritonavir). For amprenavir, indinavir and saquinavir the method has been validated over the range of 25 ng/ml to 25 microg/ml using a 0.6 ml sample volume. For nelfinavir and ritonavir the method has been validated over the range of 50 ng/ml to 25 microg/ml. The method proved to be accurate, with an average accuracy at four concentrations ranging from 90.6 to 109.2%, and precise, with the within-day and between-day precision ranging from 1.8 to 6.7%, and 0.7 to 7.6%, respectively. The protease inhibitors which can be quantified by using this assay proved to be stable under various conditions. This assay can readily be used in a hospital laboratory for the routine monitoring of plasma concentrations of these protease inhibitors.


The DNA bending by acetylaminofluorene residues and by apurinic sites.
We have studied the distortions induced in double-stranded oligonucleotides by covalently bound acetylaminofluorene residues and by apurinic sites. Within the acetylaminofluorene-modified oligonucleotide three base-pairs are unpaired as detected by the chemical probes chloroacetaldehyde and osmium tetroxide. These two probes reveal that the bases adjacent to the apurinic site are paired. In both the modified double-stranded oligonucleotides, the backbone on the 5' side of the modification is more reactive with 1,10-phenanthroline copper than the backbone on the 3' side. On polyacrylamide gels, the ligated multimers of acetylaminofluorene or apurinic site-modified oligonucleotides migrate slower than the multimers of the unmodified oligonucleotides. It is suggested that the acetylaminofluorene-modified guanine residues and the apurinic sites behave more as hinge joints than as the centres of directed bends.


Progress in living donor liver transplantation for biliary atresia and challenges faced: A thirty-year single institutional experience.
Biliary atresia (BA) is the most common indication for liver transplantation in the pediatric population, and living donor liver transplantation (LDLT) and deceased donor liver transplantation (DDLT) have been established as a radical treatment for BA .The aim of this study was to clarify the long term outcomes and risk factors affecting the LDLT outcomes, as well as the challenges faced. Between 1990 and 2019, 666 BA patients underwent LDLT in our institution and were enrolled in this study. Data regarding the recipient's age, anatomic findings of the biliary tree at Kasai's portoenterostomy, basic characteristics at transplantation, transplant profiles, donor characteristics, and outcomes of LDLT were analyzed. The 1-, 5-, 10-, 15-, 20-, and 25-year graft survival rates of BA patients who underwent LDLT were 88.1%, 85.4%, 81.5%, 78.9%, 76.6%, and 75.5%, respectively. The transplant era, age at transplantation, ABO incompatible transplant, and presence of pulmonary vascular complications were identified as significant risk factors for overall graft survival. When the study period was divided into the first (1990-1999) and second (2000-2019) phases and re analyzed, the outcomes of ABO-incompatible transplants and LDLT for adult BA patients remained inferior to others in the second phase. The 20-year graft survival rate in patients who underwent re transplantation in the second phase was 54.2%. The outcomes of LDLT in children are generally good, but the immunosuppression procedures need to be further improved for ABO-incompatible cases in the future. Further improvements in LDLT results for adult patients and re transplantation remain challenges to be addressed in this field, and future attempts, including revision to the organ allocation system of deceased donors, are necessary. Level III (case control study).


The hormone-binding role of 2 cysteines near the C terminus of the mouse glucocorticoid receptor.
Previous biochemical analyses with covalent affinity labels and thiol-blocking reagents suggested possible roles for one methionine residue and multiple cysteine residues in binding of steroid to the 250-amino acid hormone-binding domain at the C-terminal end of mammalian glucocorticoid receptors. To test the functional roles of these residues in the receptor's ability to bind hormone and active transcription of target genes, the mouse glucocorticoid receptor cDNA was specifically mutated to cause single amino acid substitutions for methionine 610 and for each of the 5 cysteines (at positions 628, 644, 649, 671, and 742) in the hormone-binding domain. Among these 6 residues, only mutations in cysteine 671 and cysteine 742 caused substantial reductions in function. In transient transfection assays, the concentration of dexamethasone required for half-maximal activation of a glucocorticoid-responsive reporter gene was increased by 10-40-fold by changing cysteine 742 to serine or cysteine 671 to serine or alanine. At saturating concentrations of dexamethasone, the mutant receptors activated the reporter gene to the same extent as the wild type receptor, indicating that the mutations affected only the hormone-binding function of the receptor and not its ability to bind DNA or activate transcription once the hormone was bound.