Remdesivir Strongly Binds to Both RNA-Dependent RNA Polymerase and Main Protease of SARS-CoV-2: Evidence from Molecular Simulations.
The outbreak of a new coronavirus SARS-CoV-2 (severe acute respiratory syndrome-coronavirus 2) has caused a global COVID-19 (coronavirus disease 2019) pandemic, resulting in millions of infections and thousands of deaths around the world. There is currently no drug or vaccine for COVID-19, but it has been revealed that some commercially available drugs are promising, at least for treating symptoms. Among them, remdesivir, which can block the activity of RNA-dependent RNA polymerase (RdRp) in old SARS-CoV and MERS-CoV viruses, has been prescribed to COVID-19 patients in many countries. A recent experiment showed that remdesivir binds to SARS-CoV-2 with an inhibition constant of μM, but the exact target has not been reported. In this work, combining molecular docking, steered molecular dynamics, and umbrella sampling, we examined its binding affinity to two targets including the main protease (Mpro), also known as 3C-like protease, and RdRp. We showed that remdesivir binds to Mpro slightly weaker than to RdRp, and the corresponding inhibition constants, consistent with the experiment, fall to the μM range. The binding mechanisms of remdesivir to two targets differ in that the electrostatic interaction is the main force in stabilizing the RdRp-remdesivir complex, while the van der Waals interaction dominates in the Mpro-remdesivir case. Our result indicates that remdesivir can target not only RdRp but also Mpro, which can be invoked to explain why this drug is effective in treating COVID-19. We have identified residues of the target protein that make the most important contribution to binding affinity, and this information is useful for drug development for this disease.


The biology of spermatogenesis: the past, present and future.
The physiological function of spermatogenesis in Caenorhabditis elegans, Drosophila melanogaster and mammals is to produce spermatozoa (1n, haploid) that contain only half of the genetic material of spermatogonia (2n, diploid). This half number of chromosomes from a spermatozoon will then be reconstituted to become a diploid cell upon fertilization with an egg, which is also haploid. Thus, genetic information from two parental individuals can be passed onto their offspring. Spermatogenesis takes place in the seminiferous epithelium of the seminiferous tubule, the functional unit of the mammalian testis. In mammals, particularly in rodents, the fascinating morphological changes that occur during spermatogenesis involving cellular differentiation and transformation, mitosis, meiosis, germ cell movement, spermiogenesis and spermiation have been well documented from the 1950s through the 1980s. During this time, however, the regulation of, as well as the biochemical and molecular mechanisms underlying these diverse cellular events occurring throughout spermatogenesis, have remained largely unexplored. In the past two decades, important advancements have been made using new biochemical, cell and molecular biology techniques to understand how different genes, proteins and signalling pathways regulate various aspects of spermatogenesis. These include studies on the differentiation of spermatogonia from gonocytes; regulation of spermatogonial stem cells; regulation of spermatogonial mitosis; regulation of meiosis, spermiogenesis and spermiation; role of hormones (e.g. oestrogens, androgens) in spermatogenesis; transcriptional regulation of spermatogenesis; regulation of apoptosis; cell-cell interactions; and the biology of junction dynamics during spermatogenesis. The impact of environmental toxicants on spermatogenesis has also become an urgent issue in the field in light of declining fertility levels in males. Many of these studies have helped investigators to understand important similarities, differences and evolutionary relationships between C. elegans, D. melanogaster and mammals relating to spermatogenesis. In this Special Issue of the Philosophical Transactions of the Royal Society B: Biological Sciences, we have covered many of these areas, and in this Introduction, we highlight the topic of spermatogenesis by examining its past, present and future.


Glycosyl Cations versus Allylic Cations in Spontaneous and Enzymatic Hydrolysis.
Enzymatic prenyl and glycosyl transfer are seemingly unrelated reactions that yield molecules and protein modifications with disparate biological functions. However, both reactions employ diphosphate-activated donors and each proceed via cationic species: allylic cations and oxocarbenium ions, respectively. In this study, we explore the relationship between these processes by preparing valienyl ethers to serve as glycoside mimics that are capable of allylic rather than oxocarbenium cation stabilization. Rate constants for spontaneous hydrolysis of aryl glycosides and their analogous valienyl ethers were found to be almost identical, as were the corresponding activation enthalpies and entropies. This close similarity extended to the associated secondary kinetic isotope effects (KIEs), indicating very similar transition state stabilities and structures. Screening a library of over 100 β-glucosidases identified a number of enzymes that catalyze hydrolysis of these valienyl ethers with kcat values up to 20 s-1. Detailed analysis of one such enzyme showed that ether hydrolysis occurs via the analogous mechanisms found for glycosides, and through a very similar transition state. This suggests that the generally lower rates of enzymatic cleavage of the cyclitol ethers reflects evolutionary specialization of these enzymes toward glycosides rather than inherent reactivity differences.


Matricectomy utilizing negative galvanic current.
An established technique for matricectomy procedures has been described with the effects of negative galvanic current on tissue being elucidated. An ongoing prospective study is in progress to verify previous claims. This procedure appears to offer distinct advantages over conventional procedures and now appropriate instrumentation is readily available, being manufactured specifically for this technique.


Characterization of two human lung adenocarcinoma cell lines by reciprocal chromosome painting.
Lung cancer is a leading cause of cancer death worldwide. Some lung cancer patients correlate with a gas of radon besides smoking. To search for common chromosomal aberrations in lung cancer cell lines established from patients induced by different factors, a combined approach of chromosome sorting, forward and reverse chromosome painting was used to characterize karyotypes of two lung adenocarcinoma cell lines: A549 and GLC-82 with the latter line derived from a patient who has suffered long-term exposure to environmental radon gas pollution. The chromosome painting results revealed that complex chromosomal rearrangements occurred in these two lung adenocarcinoma cell lines. Thirteen and twenty-four abnormal chromosomes were identified in A549 and GLC-82 cell lines, respectively. Almost half of abnormal chromosomes in these two cell lines were formed by non-reciprocal translocations, the others were derived from deletions and duplication/or amplification in some chromosomal regions. Furthermore, two apparently common breakpoints, HSA8q24 and 12q14 were found in these two lung cancer cell lines.


BOSC 2023, the 24th annual Bioinformatics Open Source Conference.
The 24th annual Bioinformatics Open Source Conference ( BOSC 2023) was part of the 2023i conference on Intelligent Systems for Molecular Biology and the European Conference on Computational Biology (ISMB/ECCB 2023). Launched in 2000 and held yearly since, BOSC is the premier meeting covering open-source bioinformatics and open science. Like ISMB 2022, the 2023 meeting was a hybrid conference, with the in-person component hosted in Lyon, France. ISMB/ECCB attracted a near-record number of attendees, with over 2100 in person and about 900 more online. Approximately 200 people participated in BOSC sessions. In addition to 43 talks and 49 posters, BOSC featured two keynotes: Sara El-Gebali, who spoke about "A New Odyssey: Pioneering the Future of Scientific Progress Through Open Collaboration", and Joseph Yracheta, who spoke about "The Dissonance between Scientific Altruism & Capitalist Extraction: The Zero Trust and Federated Data Sovereignty Solution." Once again, a joint session brought together BOSC and the Bio-Ontologies COSI. The conference ended with a panel on Open and Ethical Data Sharing. As in prior years, BOSC was preceded by a CollaborationFest, a collaborative work event that brought together about 40 participants interested in synergistically combining ideas, shaping project plans, developing software, and more.


Systematic review of the effects of pertussis vaccines in children.
To assess the efficacy and safety of whole-cell and acellular pertussis vaccines administered to children singly or within diphtheria, tetanus and pertussis (DTP) vaccines. We searched the Cochrane Library, MEDLINE, EMBASE, Biological Abstracts and Science Citation Index to December 2001. Specialised websites and bibliographies of retrieved articles and reviews were assessed. Vaccine manufacturers and investigators were contacted for additional data. We included randomised and cohort studies comparing efficacy and/or safety of pertussis vaccines with placebo, DT, no intervention or each other. We included 52 studies (49 randomised controlled trials (RCTs), 3 cohort studies). All tested whole-cell and acellular vaccines were significantly more effective than placebo against pertussis. Absolute efficacy of whole-cell DTP varied from 37 to 92%. One- and two-component acellular vaccines had lower absolute efficacy (67-70%), than vaccines with >/=3 components (80-84%). Whole-cell vaccines were associated with significantly higher incidences of swelling and induration (odds ratio (OR) 11.67, 95% confidence interval (CI) 8.83-15.44), fever (OR for fever >39 degrees C 3.36, 95% CI 2.06-5.49) and crying for >2h (OR 4.72, 95% CI 2.94-7.59) than placebo or DT. Differences in incidence of hypotonic hyporesponsive episodes (HHE) and convulsions were not statistically significant. Acellular pertussis vaccines did not cause a higher incidence of local signs, fever, convulsions, HHE or prolonged crying than placebo or DT. All tested pertussis vaccines were efficacious. Whole-cell vaccines show variable efficacy, making interpretation of direct comparisons unreliable. Acellular vaccines with >/=3 antigenic components showed higher efficacy than one- and two-component vaccines. The adverse event profile of acellular vaccines was similar to that of placebo and considerably better than that of whole-cell vaccines.


Current Understanding of the Health Effects of Electromagnetic Fields.
There has been an exponential increase in the use of electronic devices over the past few decades. This has led to increased exposure to electromagnetic fields (EMF). Electric fields result from differences in voltage, whereas magnetic fields result from the flow of electric current. Higher-frequency waves of EMF have more energy than lower-frequency waves, and thus generally tend to be more harmful. An EMF activates cellular stress response and also causes breaks in DNA strands. There are many methodological barriers to effectively measuring the associations of EMF and childhood cancers. The consensus from multiple studies is that there is no causal role of extremely low-frequency EMFs in childhood cancers, including brain cancer. A recent study showed a link between EMF radiation and the development of malignant tumors in rats. In light of that study, the American Academy of Pediatrics set out new recommendations to decrease the adverse effects of cellphone exposure on children. [Pediatr Ann. 2017;46(4):e172-e174.].


PICK1/calcineurin suppress ASIC1-mediated Ca2+ entry in rat pulmonary arterial smooth muscle cells.
Acid-sensing ion channel 1 (ASIC1) contributes to Ca(2+) influx and contraction in pulmonary arterial smooth muscle cells (PASMC). ASIC1 binds the PDZ (PSD-95/Dlg/ZO-1) domain of the protein interacting with C kinase 1 (PICK1), and this interaction is important for the subcellular localization and/or activity of ASIC1. Therefore, we first hypothesized that PICK1 facilitates ASIC1-dependent Ca(2+) influx in PASMC by promoting plasma membrane localization. Using Duolink to determine protein-protein interactions and a biotinylation assay to assess membrane localization, we demonstrated that the PICK1 PDZ domain inhibitor FSC231 diminished the colocalization of PICK1 and ASIC1 but did not limit ASIC1 plasma membrane localization. Although stimulation of store-operated Ca(2+) entry (SOCE) greatly enhanced colocalization between ASIC1 and PICK1, both FSC231 and shRNA knockdown of PICK1 largely augmented SOCE. These data suggest PICK1 imparts a basal inhibitory effect on ASIC1 Ca(2+) entry in PASMC and led to an alternative hypothesis that PICK1 facilitates the interaction between ASIC1 and negative intracellular modulators, namely PKC and/or the calcium-calmodulin-activated phosphatase calcineurin. FSC231 limited PKC-mediated inhibition of SOCE, supporting a potential role for PICK1 in this response. Additionally, we found PICK1 inhibits ASIC1-mediated SOCE through an effect of calcineurin to dephosphorylate the channel. Furthermore, it appears PICK1/calcineurin-mediated regulation of SOCE opposes PKA phosphorylation and activation of ASIC1. Together our data suggest PKA and PICK1/calcineurin differentially regulate ASIC1-mediated SOCE and these modulatory complexes are important in determining downstream Ca(2+) signaling.


Re-evaluating neonatal-age models for ungulates: does model choice affect survival estimates?
New-hoof growth is regarded as the most reliable metric for predicting age of newborn ungulates, but variation in estimated age among hoof-growth equations that have been developed may affect estimates of survival in staggered-entry models. We used known-age newborns to evaluate variation in age estimates among existing hoof-growth equations and to determine the consequences of that variation on survival estimates. During 2001-2009, we captured and radiocollared 174 newborn (≤24-hrs old) ungulates: 76 white-tailed deer (Odocoileus virginianus) in Minnesota and South Dakota, 61 mule deer (O. hemionus) in California, and 37 pronghorn (Antilocapra americana) in South Dakota. Estimated age of known-age newborns differed among hoof-growth models and varied by >15 days for white-tailed deer, >20 days for mule deer, and >10 days for pronghorn. Accuracy (i.e., the proportion of neonates assigned to the correct age) in aging newborns using published equations ranged from 0.0% to 39.4% in white-tailed deer, 0.0% to 3.3% in mule deer, and was 0.0% for pronghorns. Results of survival modeling indicated that variability in estimates of age-at-capture affected short-term estimates of survival (i.e., 30 days) for white-tailed deer and mule deer, and survival estimates over a longer time frame (i.e., 120 days) for mule deer. Conversely, survival estimates for pronghorn were not affected by estimates of age. Our analyses indicate that modeling survival in daily intervals is too fine a temporal scale when age-at-capture is unknown given the potential inaccuracies among equations used to estimate age of neonates. Instead, weekly survival intervals are more appropriate because most models accurately predicted ages within 1 week of the known age. Variation among results of neonatal-age models on short- and long-term estimates of survival for known-age young emphasizes the importance of selecting an appropriate hoof-growth equation and appropriately defining intervals (i.e., weekly versus daily) for estimating survival.


The parsley plastocyanin-turnip cytochrome f complex: a structurally distorted but kinetically functional acidic patch.
In general, inter-protein electron transfer proceeds via the formation of transient complexes. The initial stage of the interaction between plastocyanin (PCu) and cytochrome f (cyt f ) from plants is mediated by complementary electrostatics. Given the diffuse nature of its acidic patch, parsley PCu is an atypical example of a plant PCu. The interaction of this PCu with turnip cyt f was investigated by stopped-flow kinetics, NMR spectroscopy and protein-docking simulations. We show that, despite the altered acidic patch, parsley PCu is as efficient as spinach PCu in accepting electrons from cyt f, over the physiological range of ionic strength. At high ionic strength, the rate constant for the reaction of cyt f with parsley PCu is twice that of the spinach protein. This difference in reactivity is attributed to variations in the hydrophobic patch of parsley PCu. The results of NMR studies and protein-docking simulations indicate that parsley PCu and its spinach analogue adopt different orientations in their complexes with cyt f.


The effectiveness of tacrolimus and minidose methotrexate in the prevention of acute graft-versus-host disease following allogeneic hematopoietic stem cell transplantation in children: a single-center study in Korea.
Knowledge of the roles of tacrolimus and minidose methotrexate (MTX) in the prevention of acute graft-versus-host disease (aGVHD) in pediatric allogeneic hematopoietic stem cell transplantation (HSCT) is limited. We retrospectively evaluated the engraftment status, incidence of aGVHD and chronic GVHD (cGVHD), and toxicities of tacrolimus and minidose MTX in aGVHD prophylaxis in children undergoing allogeneic HSCT. Seventeen children, who underwent allogeneic HSCT and received tacrolimus and minidose MTX as GVHD prophylaxis from March 2003 to February 2011, were reviewed retrospectively. All the patients received tacrolimus since the day before transplantation at a dose of 0.03 mg/kg/day and MTX at a dose of 5 mg/m(2) on days 1, 3, 6, and 11. Of the 17 patients, 9 received human leukocyte antigen (HLA)-matched related donor transplants, and 8 received HLA-matched, or partially mismatched unrelated donor transplants. The median time for follow-up was 55 months. The incidence of aGVHD in the related and unrelated donor groups was 22.2% and 42.9%, respectively. cGVHD was not observed. To maintain therapeutic blood levels of tacrolimus, the younger group (<8 years of age) required an increased mean dose compared to the older group (≥8 years) (P=0.0075). The adverse events commonly associated with tacrolimus included hypomagnesemia (88%), nephrotoxicity (23%), and hyperglycemia (23%). Tacrolimus and minidose MTX were well tolerated and effective in GVHD prophylaxis in pediatric patients undergoing allogeneic HSCT. Children <8 years of age undergoing HSCT required increased doses of tacrolimus to achieve therapeutic levels.


Observation of dissociative quasi-free electron attachment to nucleoside via excited anion radical in solution.
Damage to DNA via dissociative electron attachment has been well-studied in both the gas and condensed phases; however, understanding this process in bulk solution at a fundamental level is still a challenge. Here, we use a picosecond pulse of a high energy electron beam to generate electrons in liquid diethylene glycol and observe the electron attachment dynamics to ribothymidine at different stages of electron relaxation. Our transient spectroscopic results reveal that the quasi-free electron with energy near the conduction band effectively attaches to ribothymidine leading to a new absorbing species that is characterized in the UV-visible region. This species exhibits a nearly concentration-independent decay with a time constant of ~350 ps. From time-resolved studies under different conditions, combined with data analysis and theoretical calculations, we assign this intermediate to an excited anion radical that undergoes N1-C1' glycosidic bond dissociation rather than relaxation to its ground state.


Role of electrical stimulation added to conventional therapy in patients with idiopathic facial (Bell) palsy.
The aim of this study was to determine the efficacy of electrical stimulation when added to conventional physical therapy with regard to clinical and neurophysiologic changes in patients with Bell palsy. This was a randomized controlled trial. Sixty patients diagnosed with Bell palsy (39 right sided, 21 left sided) were included in the study. Patients were randomly divided into two therapy groups. Group 1 received physical therapy applying hot pack, facial expression exercises, and massage to the facial muscles, whereas group 2 received electrical stimulation treatment in addition to the physical therapy, 5 days per week for a period of 3 wks. Patients were evaluated clinically and electrophysiologically before treatment (at the fourth week of the palsy) and again 3 mos later. Outcome measures included the House-Brackmann scale and Facial Disability Index scores, as well as facial nerve latencies and amplitudes of compound muscle action potentials derived from the frontalis and orbicularis oris muscles. Twenty-nine men (48.3%) and 31 women (51.7%) with Bell palsy were included in the study. In group 1, 16 (57.1%) patients had no axonal degeneration and 12 (42.9%) had axonal degeneration, compared with 17 (53.1%) and 15 (46.9%) patients in group 2, respectively. The baseline House-Brackmann and Facial Disability Index scores were similar between the groups. At 3 mos after onset, the Facial Disability Index scores were improved similarly in both groups. The classification of patients according to House-Brackmann scale revealed greater improvement in group 2 than in group 1. The mean motor nerve latencies and compound muscle action potential amplitudes of both facial muscles were statistically shorter in group 2, whereas only the mean motor latency of the frontalis muscle decreased in group 1. The addition of 3 wks of daily electrical stimulation shortly after facial palsy onset (4 wks), improved functional facial movements and electrophysiologic outcome measures at the 3-mo follow-up in patients with Bell palsy. Further research focused on determining the most effective dosage and length of intervention with electrical stimulation is warranted.


Contributions of the parietal cortex to increased efficiency of planning-based action selection.
Response selection is foundational to adaptive behavior, and considerable attention has been devoted to investigating this behavior under conditions in which the mapping between stimuli and responses is fixed. Results from prior studies implicate the left supramarginal gyrus (SMg), premotor and prefrontal cortices, as well as the cerebellum in this essential function. Yet, many goal-directed motor behaviors have multiple solutions with flexible mappings between stimuli and responses whose solutions are believed to involve prospective planning. Studies of selection under conditions of flexible mappings also reveal involvement of the left SMg, as well as bilateral premotor, superior parietal cortex (SPL) and pre-supplementary motor (pre-SMA) cortices, along with the cerebellum. This evidence is, however, limited by exclusive reliance on tasks that involve selection in the absence of overt action execution and without complete control of possible confounding effects related to differences in stimulus and response processing demands. Here, we address this limitation through use of a novel fMRI repetition suppression (FMRI-RS) paradigm. In our prime-probe design, participants select and overtly pantomime manual object rotation actions when the relationship between stimuli and responses is either flexible (experimental condition) or fixed (control condition). When trials were repeated in prime-probe pairs of the experimental condition, we detected improvements in performance accompanied by a significant suppression of blood oxygen-level dependent (BOLD) responses in: left SMg extending into and along the length of the intraparietal sulcus (IPS), right IPS, bilateral caudal superior parietal lobule (cSPL), dorsal premotor cortex (dPMC), pre-SMA, and in the lateral cerebellum. Further, region-of-interest analyses revealed interaction effects of fMRI-RS in the experimental versus control condition within left SMg and cerebellum, as well as in bilateral caudal SPL. These efficiency effects cannot be attributed to the repetition of stimulus or response processing, but instead are planning-specific and generally consistent with earlier findings from conventional fMRI investigations. We conclude that repetition-related increases in the efficiency of planning-based selection appears to be associated with parieto-cerebellar networks.


Accuracy of physical examination in the detection of arteriovenous graft stenosis.
Physical examination has recently been demonstrated to detect vascular access stenosis in patients with arteriovenous fistulae. However, its accuracy in the identification of stenoses when compared with the gold standard (angiography) in patients with arteriovenous grafts has not been studied in a systematic fashion. We conducted a prospective study to examine the accuracy of physical examination in the detection of stenotic lesions when compared with angiography. Forty-three consecutive cases referred for an arteriovenous graft dysfunction were included in this analysis. Preprocedure physical examination was performed. The findings of the examination and diagnosis were recorded and secured in a sealed envelope. Angiography from the feeding artery to the right atrium was performed. The images were reviewed by an independent interventionalist with expertise in endovascular dialysis access procedures and the diagnosis was rendered. The reviewer was blinded to the physical examination. Cohen's Kappa was used as a measurement of the level of agreement beyond chance between the diagnosis made by physical examination and angiography. There was a strong agreement between the physical examination and the angiography in the diagnosis of vein-graft anastomotic stenosis (kappa = 0.52). The sensitivity and specificity for this lesion was 57% and 89%, respectively. There was a moderate agreement beyond chance regarding the diagnosis of intragraft (kappa = 0.43) and inflow stenoses (kappa = 0.40). The sensitivity and specificity for the intragraft and inflow stenosis was 100%, 73% and 33%, 73%; respectively. The findings of this study demonstrate that physical examination can assist in the detection and localization of stenoses in arteriovenous grafts.


Spectrum of diseases for house staff education on a VA general medicine inpatient service.
A significant portion of internal medicine residency training in the United States today occurs in general medicine sections of Veterans Administration hospitals. The authors studied the demographic, diagnostic, and prognostic characteristics of the patients treated by house staff members rotating through the general medical wards of the Houston, Texas, Veterans Administration (VA) Medical Center. In 2,131 admissions over 13 months, the most frequent primary causes of admissions included congestive heart failure, pneumonia, exacerbation of chronic obstructive lung disease, and malignancy. In 85 percent of the admissions, two or more chronic diseases were present. In 53 percent of admissions, the patients were deemed to be moderately or severely ill on admission. The data indicate that the residents' experience is representative of problems encountered by practicing internists and that VA hospitals make a significant contribution to internal medicine training and thus to the provision of health care for the nation.


Two-dimensional ternary pentagonal BCX (X = P, As, and Sb): promising photocatalyst semiconductors for water splitting with strong piezoelectricity.
Seeking high-performance energy conversion materials is one of the most important issues in designing 2D materials. In the framework of density functional theory, we propose a series of ternary monolayers, penta-BCX (X = P, As, and Sb), and systematically investigate their structural stability, mechanical, piezoelectric, and photocatalytic properties. All three materials are semiconductors with a bandgap ranging from 2.56 eV to 3.24 eV, so they could be promising catalysts for the photolysis of water. Penta-BCX exhibits significant piezoelectric properties attributed to their non-centrosymmetric structure and low in-plane Young's modulus, which are expected to efficiently drive photocatalytic water decomposition. Moreover, the bandgap, band edge position, and light absorption of penta-BCX can be modulated by tensile or compressive strain to enhance their photocatalytic performance in the visible light and ultraviolet regions.


Construction of a tagging system for subcellular localization of proteins encoded by open reading frames.
We have previously characterized a monoclonal antibody (SC1D7) that is directed to maltose-binding protein (MBP) of Escherichia coli and other closely related enteric bacteria. SC1D7 does not cross-react with proteins in eucaryotes and appears to be a highly specific tool in immunochemical analyses. To better map the epitope, we took advantage of an available plasmid, pMAL-c2, that encodes the E. coli MBP-coding sequence and constructed plasmids to express MBP fragments. A construct containing the N-terminal portion of MBP does not react with SC1D7, whereas a second construct expressing glutathione S-transferase fused with the C-terminal half of MBP does react with SC1D7. To precisely define the epitope, random peptides displayed on M13 were used to react with SC1D7. Sequences of reactive peptides were aligned, and a consensus sequence of XDXRIPX was deduced. This sequence matches MBP with an amino acid stretch of KDPRIAA. To consolidate the mapping result, a sequence encoding this epitope was inserted into an expression vector and the resulting recombinant protein did react with SC1D7. Thereafter, this epitope was incorporated into a eucaryotic expression plasmid containing a previously defined hepatitis delta virus epitope for protein tagging. This two-epitope-tagging vector is useful in various molecular analyses. We demonstrate its usage for localization of a bacterial virulence factor in host cells. This vector should be applicable for high-throughput characterization of new open reading frames found in genome sequencing.


Local immunotherapy for chronic respiratory allergic pathology with allergen aerosols. II. Treatment of bronchial asthma.
Efficacy of hyposensitization of patients suffering from different forms of bronchial asthma caused by bacterial, house dust and epidermal allergens was studied. Allergens were delivered to the mucous membrane of bronchi with the help of aerosol inhalers on a schedule worked out by the author. The results of treatment with this method were compared to the results in the group of patients treated by the routine method of intradermal or subcutaneous injections of allergens. There were demonstrated advantages of the method of projecting allergens directly on to the shock organ, which proved more effective and less reactogenic.


Coming Out of the Closet, Also on the News? A Longitudinal Content Analysis of Patterns in Visibility, Tone and Framing of LGBTs on Television News (1986-2017).
This study documents the results of a longitudinal content analysis of television news about LGBT people in terms of visibility, active representation, tone and framing in Flanders (1986-2017). While attention for LGBT issues has increased over time, LGBTs are not more likely to be visually represented or granted a voice. Gay men are more often actively represented than lesbians and transgender people. News remains negatively biased, although news stories in which LGBT people are depicted as the cause of negativity have become less prevalent. Patterns in framing have shifted: Deviance and abnormality frames have decreased in favor of a rise in equal rights and victim frames. Patterns in tone and framing were similar for gay men, lesbians and transgender people. Results suggest that journalists have shifted from problematizing homosexuality to problematizing homophobia. Implications of news as a source of mass-mediated contact to promote tolerance toward LGBT people are discussed.


Room-temperature cobalt-catalyzed arylation of aromatic acids: overriding the ortho-selectivity via the oxidative assembly of carboxylate and aryl titanate reagents using oxygen.
A room temperature phosphine or NHC ligand-free cobalt-catalyzed arylation of (hetero)aromatic acids has been developed. It involves an oxidative cross-coupling between carboxylate and aryl titanate reagents using oxygen as an oxidant, and the arylation at the position ortho, meta and para to the carboxylic acid group could all be achieved. As application, various (hetero)aromatic acids including xenalipin, tafamidis and the key intermediate for a cardioprotective compound have been efficiently synthesized.


Characterization of bone-marrow-derived rat mesenchymal stem cells depending on donor age.
It is generally accepted that autologous transfers, as non-immunogenic, constitute the safest approach in cellular transplantations. However, this attitude is often associated with the need for isolation and extracorporeal propagation of cells derived from aged patients. Thus the knowledge about relationship between aging and the properties of MSCs (mesenchymal stem cells) is crucial in developing new clinical strategies. The aim of this study was to perform complex comparison of MSC derived from young and aged individuals, which included phenotype, proliferating rate, osteogenic and adipogenic potential and secretory activity. Evaluated populations were isolated from bone marrow of 3-month-old and 24-month-old rats. There was no significant difference in membrane antigen expression and PDT (population doubling time). Additionally, the adipogenic and osteogenic potential did not vary between studied populations. The reaction of MSCs to either mitogen [bFGF (basic fibroblas t growth factor)] or oxidative stress (H2O2) in vitro displayed a very similar pattern in both analysed populations. There was no difference in TGFβ1 (transforming growth factor β1) and VEGF (vascular endothelial growth factor) secretion measured by ELISA test and gene expression evaluated by real-time PCR. However, the expression of the gene for IL-1α (interleukin-1α) was 8-fold lower in oMSC (MSC isolated from old rats). These results indicate that aging individuals can be considered as candidates for autologous transplantation of bone-marrow-derived MSCs.


Subaortic obstruction after the use of an intracardiac baffle to tunnel the left ventricle to the aorta.
Postoperative hemodynamic studies in five patients document subaortic obstruction after surgical repair utilizing an intracardiac baffle to establish continuity between the left ventricle and the aorta. Four of the patients had a Rastelli procedure for D-transposition of the great arteries with a ventricular septal defect and pulmonary stenosis; one patient had repair of double outlet right ventricle with a ventricular septal defect and pulmonary stenosis. The left ventricular outflow was shown to be a long narrow tunnel by angiography in four of five patients and by echocardiography in one patient. Resting aortic peak systolic pressure gradient ranged from 10 to 42 mm Hg (mean 24). The obstruction was localized to the proximal end of the left ventricule to aorta tunnel (i.e., at the site of ventricular septal defect) in five patients. One patient with a gradient of 42 mm Hg has angina and decreased exercise tolerance. Subaortic obstruction is a newly described sequelae after the Rastelli procedure for transposition or repair of double outlet right ventricle. The obstruction may be hemodynamically significant and should be searched for at postoperative cardiac catheterization.


Primary reconstruction for spinal infections.
Primary reconstruction using bone grafts and instrumentation for spinal infections remains controversial. Between 1991 and 1993, 27 infections of the spinal column were treated at the Department of Neurosurgery of the University of Florida. Of the 27 cases 20 (six cervical, eight thoracic, and six lumbar spine) required surgical debridement and spinal reconstruction to maximize eradication of the infection and maintenance of spinal alignment. All of the cervical and lumbar cases were caused by bacterial infections, and two of eight thoracic cases were caused by tuberculous infections. Spinal arthrodesis was performed in all cases: interbody grafts were used in 18 procedures and posterolateral onlay grafts in 14. Interbody grafts were autologous in 10 cases (six rib and four iliac crest) and allograft in eight (six fibular and two humerus). All of the posterolateral onlay grafts were autologous (three rib and 11 iliac crest). Spinal instrumentation was used in 15 cases: four with Caspar plates and 11 with posterior segmental fixation (five hook/rod constructs and six screw/rod constructs). Seventeen of 20 patients achieved improved clinical status postoperatively and 18 of 20 showed radiographic evidence of bone fusion. Antibiotic drugs were administered parenterally for an average of 6 weeks followed by a 3-month course of oral antibiotic medications. Tuberculous infections were treated for 1 year with antibiotic therapy. The average follow-up period was 37 months from surgery and 31 months after completion of treatment with antibiotic drugs. The authors conclude that primary arthrodesis and instrumentation can be performed in acute spinal infections; however, successful management depends on aggressive debridement of infectious foci and prolonged treatment with parenteral antibiotic drugs.


Is MCI really just early dementia? A systematic review of conversion studies.
Older people commonly present with memory loss although on assessment are not found to have a full dementia complex. Previous studies have suggested however that people with subjective and objective cognitive loss are at higher risk of dementia. We aimed to determine from the literature the rate of conversion from mild cognitive impairment to dementia. Systematic review of MedLine, PsychLit and EmBase. We identified 19 longitudinal studies published between 1991 and 2001 that addressed conversion of mild cognitive impairment to dementia. Overall the rate of conversion was 10% but with large differences between studies. The single biggest variable accounting for between study heterogeneity was source of subjects, with self-selected clinic attenders having the highest conversion rate. The most important factor accounting for heterogeneity within studies was cognitive testing, with poor performance predicting conversion with a high degree of accuracy. These data strongly support the notion that subjective and objective evidence of cognitive decline is not normal and predicts conversion to dementia. The more stringent the measures of both variables the better the prediction of conversion. Mild cognitive impairment, appropriately diagnosed, is a good measure with which to select subjects for disease modification studies.


Advanced REACH Tool: development and application of the substance emission potential modifying factor.
The Advanced REACH Tool (ART) is an exposure assessment tool that combines mechanistically modelled inhalation exposure estimates with available exposure data using a Bayesian approach. The mechanistic model is based on nine independent principal modifying factors (MF). One of these MF is the substance emission potential, which addresses the intrinsic substance properties as determinants of the emission from a source. This paper describes the current knowledge and evidence on intrinsic characteristics of solids and liquids that determine the potential for their release into workplace air. The principal factor determining the release of aerosols from handling or processing powdered, granular, or pelletized materials is the dustiness of the material, as well as the weight fraction of the substance of interest in the powder and the moisture content. The partial vapour pressure is the main intrinsic factor determining the substance emission potential for emission of vapours. For generation of mist, the substance emission potential is determined by the viscosity of the liquid as well as the weight fraction of the substance of interest in the liquid. Within ART release of vapours is considered for substances with a partial vapour pressure at the process temperature of 10 Pa or more, while mist formation is considered for substances with a vapour pressure ≤ 10 Pa. Relative multipliers are assigned for most of the intrinsic factors, with the exception of the weight fraction and the vapour pressure, which is applied as a continuous variable in the estimation of the substance emission potential. Currently, estimation of substance emission potential is not available for fumes, fibres, and gases. The substance emission potential takes account of the latest thinking on emissions of dusts, mists, and vapours and in our view provides a good balance between theory and pragmatism. Expanding the knowledge base on substance emission potential will improve the predictive power of occupational exposure models and thereby the accuracy and precision of the exposure estimates.


Effect of beef long-storage under different temperatures and vacuum-packaging conditions on meat quality, oxidation processes and microbial growth.
The global beef market demands the meat industry to ensure product quality and safety in markets that are often very distant. The present study aimed to evaluate the effects of chilled (CH, 120 d) and chilled-then-frozen (CHF, 28 d + 92 d) storage conditions of beef vacuum packaged (VP) and vacuum packaged with antimicrobial (VPAM) on meat quality, oxidative status and microbial loads. Treatments resulted from the combination of storage condition and packaging type: VP + CH, VP + CHF, VPAM + CH and VPAM + CHF. Warner-Bratzler shear force values decreased in all treatments after 28 d of chilling. Except for VP + CH, L* values (lightness) of meat color did not differ in each treatment as the storage time increased. Meat from VP + CH had greater a* values than CHF treatments on day 120 of storage. A consumer panel did not detect differences in tenderness, flavor and overall liking between VP and VPAM beef, but they preferred CHF steaks rather than CH beef. TBARS values did not differ between VP and VPAM and between CH and CHF at any time during the storage period. At the end of storage time, all treatments except VP + CHF presented a greater concentration of thiols than at 48 h post-mortem. On day 120 of storage, VP + CH had greater catalase enzyme activity than CHF treatments while VP + CH and VP + CHF showed a greater superoxide dismutase activity than VPAM + CHF. Storage condition (CH or CHF) had a greater impact on microbial counts than the type of packaging. Freezing meat after an ageing period represents a suitable strategy to extend beef storage life without a detrimental impact on its quality. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


[Lipid lowering therapy: new pathophysiologic aspects and clinical implications].
Lipid-lowering therapy reduces cardiovascular morbidity and mortality. Although atherosclerotic plaques regress under lipid-lowering therapy, only minor changes in arteriosclerotic plaques are angiographically detectable. These changes cannot fully explain all the benefits of lipid-lowering therapy. Additional factors independent of macroscopically detectable alterations in plaque size may contribute to the clinical effects of lipid-lowering therapy. Qualitative changes in the composition of atherosclerotic plaques may reduce the risk of subsequent plaque rupture. Furthermore, hypercholesterolemia is associated with endothelial dysfunction, i.e., impaired endothelium-mediated vasodilation, induction of adhesion molecules, and altered expression of growth factors. Lipid lowering seems to reverse endothelial dysfunction. Also, lipid-lowering therapy may reduce peripheral arterial resistance and hyperinsulinemia, and may improve erythrocyte deformability. These effects of lipid-lowering therapy may contribute to the clinical benefits. In particular, the concept of endothelial dysfunction as a first, reversible manifestation of target organ damage deserves further attention in the diagnosis and therapy of cardiovascular disease.


Post-prostatectomy incontinence: a guideline of guidelines.
To provide a comprehensive review of guidelines from various professional organisations on the work-up and management of post-prostatectomy Incontinence (PPI). The following guidelines were included in this review: European Association of Urology (EAU 2023), American Urological Association/Society of Urodynamics, Female Pelvic Medicine and Urogenital Reconstruction (AUA/SUFU 2019), International Consultation on Incontinence (ICI, 2018), the Canadian Urological Association (CUA, 2012) and the Urological Society of India (USI, 2018). In general, the guidelines concur regarding the significance of conducting a comprehensive history and physical examination for patients with post-prostatectomy incontinence (PPI). However, there are variations among the guidelines concerning the recommended additional investigations. In cases of troublesome PPI, male slings are typically recommended for mild to moderate urinary incontinence (UI), while artificial urinary sphincters are preferred for moderate to severe UI, although the precise definition of this severity remains unclear. The guidelines provided by AUA/SUFU and the ICI have offered suggestions for managing complications or persistent/recurrent UI post-surgery, though some differences can be observed within these recommendations as well. This is a first of its kind review encompassing Guidelines on PPI spanning over a decade. Although guidelines share overarching principles, nuanced variations persist, posing challenges for clinicians. This compilation consolidates and highlights both the similarities and differences among guidelines, providing a comprehensive overview of PPI diagnosis and management for practitioners. It is our expectation that as more evidence emerges in this and other areas of PPI management, the guidelines will converge and address crucial patient-centric aspects.


The micronutrient levels in the third trimester of pregnancy and assessment of the neonatal outcome: a pilot study.
During pregnancy, an adequate intake of vitamins and minerals is recommended, to prevent the occurrence of adverse effects in the mother and the foetus. In our study, we aimed to study the levels of the micronutrients like iron, zinc and copper in the third trimester of pregnancy and to assess the neonatal outcome in them. Fifty pregnant women who were aged 20-30 years, who had completed 24 weeks of gestation, who were on regular antenatal checkups, were included. The collected venous blood samples were used for the estimation of haemoglobin, serum ferritin, zinc and copper. They were followed up till their deliveries and the neonatal outcomes were noted. The gestational ages, weight of the babies, the lengths of the babies and their head circumferences and any complications which had occurred during and after the deliveries, were noted in the proforma. The serum ferritin levels did not significantly correlate with the other study parameters. The zinc levels decreased with an increase in the parity (p<0.05). The copper levels decreased with an increase in the BMI (p<0.05). With an increase in haemoglobin, there was an increase in the levels of zinc and ferritin (p<0.05). With an increase in the parity, there was a decrease in the neonatal birth weight. Our study clearly brought out a correlation between the microminerals such as iron, zinc and copper during late pregnancies. An improvement in the iron status brings out a positive effect on the ferritin and zinc levels, thus indicating better outcomes of the pregnancies.


Model Selection in Historical Research Using Approximate Bayesian Computation.
Computational models are increasingly being used to study historical dynamics. This new trend, which could be named Model-Based History, makes use of recently published datasets and innovative quantitative methods to improve our understanding of past societies based on their written sources. The extensive use of formal models allows historians to re-evaluate hypotheses formulated decades ago and still subject to debate due to the lack of an adequate quantitative framework. The initiative has the potential to transform the discipline if it solves the challenges posed by the study of historical dynamics. These difficulties are based on the complexities of modelling social interaction, and the methodological issues raised by the evaluation of formal models against data with low sample size, high variance and strong fragmentation. This work examines an alternate approach to this evaluation based on a Bayesian-inspired model selection method. The validity of the classical Lanchester's laws of combat is examined against a dataset comprising over a thousand battles spanning 300 years. Four variations of the basic equations are discussed, including the three most common formulations (linear, squared, and logarithmic) and a new variant introducing fatigue. Approximate Bayesian Computation is then used to infer both parameter values and model selection via Bayes Factors. Results indicate decisive evidence favouring the new fatigue model. The interpretation of both parameter estimations and model selection provides new insights into the factors guiding the evolution of warfare. At a methodological level, the case study shows how model selection methods can be used to guide historical research through the comparison between existing hypotheses and empirical evidence.


Bone-chip system to monitor osteogenic differentiation using optical imaging.
Human organoids and organ-on-chip systems to predict human responses to new therapies and for the understanding of disease mechanisms are being more commonly used in translational research. We have developed a bone-chip system to study osteogenic differentiation in vitro, coupled with optical imaging approach which provides the opportunity of monitoring cell survival, proliferation and differentiation in vitro without the need to terminate the culture. We used the mesenchymal stem cell (MSC) line over-expressing bone morphogenetic protein-2 (BMP-2), under Tet-Off system, and luciferase reporter gene under constitutive promoter. Cells were seeded on chips and supplemented with osteogenic medium. Flow of media was started 24 h later, while static cultures were performed using media reservoirs. Cells grown on the bone-chips under constant flow of media showed enhanced survival/proliferation, comparing to the cells grown in static conditions; luciferase reporter gene expression and activity, reflecting the cell survival and proliferation, was quantified using bioluminescence imaging and a significant advantage to the flow system was observed. In addition, the flow had positive effect on osteogenic differentiation, when compared with static cultures. Quantitative fluorescent imaging, performed using the osteogenic extra-cellular matrix-targeted probes, showed higher osteogenic differentiation of the cells under the flow conditions. Gene expression analysis of osteogenic markers confirmed the osteogenic differentiation of the MSC-BMP2 cells. Immunofluorescent staining performed against the Osteocalcin, Col1, and BSP markers illustrated robust osteogenic differentiation in the flow culture and lessened differentiation in the static culture. To sum, the bone-chip allows monitoring cell survival, proliferation, and osteogenic differentiation using optical imaging.


Radiopacity of endodontic sealers: development of a new method for direct measurement.
Radiopacity is an essential attribute of endodontic filling materials. It is important to see clearly the root canal filling to detect its presence, extent, and apparent condensation. National and international standards require a minimal radiopacity equivalent to 3 or 4 mm of aluminum, yet some products made by reputed companies do not meet this requirement. The result may be unjustified downgrading of clinical cases. The purpose of this investigation was 2-fold: to develop a novel and easily reproducible technique for assessing the radiopacity of endodontic sealers and to apply it to measure the opacity of 21 current cements poured in 6- x 1-mm wells. As in the currently recommended technique, the standardized samples are radiographed alongside an aluminum stepwedge but instead of using an optical densitometer to measure the density and then calculate the radiopacity, the image is digitized. The gray pixel value of the test material is compared with the stepwedge, using computer software, to find the corresponding step. The opacity of the samples ranged from 1.6 mm to more than 11 mm of aluminum. The technique can be advanced to a dry, chemical-less variant by using radiation captors, as for periapical X-rays, instead of film.


Prospectively randomized clinical trial of three intensive chemotherapy regimens for the treatment of advanced unfavorable histology non-Hodgkin's lymphoma.
Three hundred thirty-two eligible patients with advanced (Ann Arbor stage III or IV) non-Hodgkin's lymphoma of aggressive histologic subtype (Rappaport classification diffuse histiocytic [DH], diffuse poorly differentiated lymphocytic [DPDL], diffuse mixed [DM], or diffuse undifferentiated [DU]) were randomly assigned to receive induction chemotherapy with one of three intensive regimens in a clinical trial conducted by the Eastern Cooperative Oncology Group (ECOG) between 1978 and 1983. Chemotherapy regimens consisted of cyclophosphamide, vincristine, prednisone, and doxorubicin (Adriamycin; Adria Laboratories, Columbus, OH) (COPA) administered in 3-week cycles; cyclophosphamide plus doxorubicin plus prednisone beginning day 1, with vincristine plus bleomycin day 15 of each 3-week cycle (COPA + Bleo); or cyclophosphamide plus doxorubicin plus procarbazine beginning day 1, and bleomycin plus vincristine plus prednisone beginning day 15 of each 4-week cycle (CAP-BOP). The median patient follow-up from study entry for patients still alive is 5 years. The three regimens were not significantly different with respect to complete response (CR) rates (43% to 46%), time to progression of malignant disease (median, 1.0 to 1.7 years), or survival (5-year survival, 34% to 45%), although duration of complete remission appeared to be shorter in patients receiving COPA (P = .03). COPA + Bleo and CAP-BOP were significantly more toxic than the COPA regimen. This study did not demonstrate any substantial therapeutic advantage associated with the addition of a fifth or sixth chemotherapy drug, or with treatment administered on a more frequent administration schedule, compared with the COPA regimen in this population of patients with advanced diffuse non-Hodgkin's lymphoma. The relatively small proportion of long-term disease-free survivors treated with COPA underscores the need for prospective clinical trials of new and more effective treatments for patients with these potentially curable tumors.


Inhibition of the mTORC pathway in the antiphospholipid syndrome.
Although thrombosis is considered the cardinal feature of the antiphospholipid syndrome, chronic vascular lesions are common, particularly in patients with life-threatening complications. In patients who require transplantation, vascular lesions often recur. The molecular pathways involved in the vasculopathy of the antiphospholipid syndrome are unknown, and adequate therapies are lacking. We used double immunostaining to evaluate pathway activation in the mammalian target of rapamycin complex (mTORC) and the nature of cell proliferation in the vessels of patients with primary or secondary antiphospholipid syndrome nephropathy. We also evaluated autopsy specimens from persons who had catastrophic antiphospholipid syndrome. The molecular pathways through which antiphospholipid antibodies modulate the mTORC pathway were evaluated in vitro, and potential pharmacologic inhibitors were also tested in vitro. Finally, we studied the effect of sirolimus in kidney-transplant recipients with the antiphospholipid syndrome. The vascular endothelium of proliferating intrarenal vessels from patients with antiphospholipid syndrome nephropathy showed indications of activation of the mTORC pathway. In cultured vascular endothelial cells, IgG antibodies from patients with the antiphospholipid syndrome stimulated mTORC through the phosphatidylinositol 3-kinase (PI3K)-AKT pathway. Patients with antiphospholipid syndrome nephropathy who required transplantation and were receiving sirolimus had no recurrence of vascular lesions and had decreased vascular proliferation on biopsy as compared with patients with antiphospholipid antibodies who were not receiving sirolimus. Among 10 patients treated with sirolimus, 7 (70%) had a functioning renal allograft 144 months after transplantation versus 3 of 27 untreated patients (11%). Activation of mTORC was also found in the vessels of autopsy specimens from patients with catastrophic antiphospholipid syndrome. Our results suggest that the mTORC pathway is involved in the vascular lesions associated with the antiphospholipid syndrome. (Funded by INSERM and others.).


Activation of a nicotinic acetylcholine receptor.
We studied activation of the nicotinic acetylcholine (ACh) receptor on cells of a mouse clonal muscle cell line (BC3H1). We analyzed single-channel currents through outside-out patches elicited with various concentrations of acetylcholine (ACh), carbamylcholine (Carb) and suberyldicholine (Sub). Our goal is to determine a likely reaction scheme for receptor activation by agonist and to determine values of rate constants for transitions in that scheme. Over a wide range of agonist concentrations the open-time duration histograms are not described by single exponential functions, but are well-described by the sum of two exponentials, a brief-duration and a long-duration component. At high concentration, channel openings occur in groups and these groups contain an excess number of brief openings. We conclude that there are two open states of the ACh receptor with different mean open times and that a single receptor may open to either open state. The concentration dependence of the numbers of brief and long openings indicates that brief openings do not result from the opening of channels of receptors which have only one agonist molecule bound to them. Closed-time duration histograms exhibit a major brief component at low concentrations. We have used the method proposed by Colquhoun and Sakmann (1981) to analyze these brief closings and to extract estimates for the rates of channel opening (beta) and agonist dissociation (k-2). We find that this estimate of beta does not predict our closed-time histograms at high agonist concentration (ACh: 30-300 microM; Carb: 300-1,000 microM). We conclude that brief closings at low agonist concentrations do not result solely from transitions between the doubly-liganded open and the doubly-liganded closed states. Instead, we postulate the existence of a second closed-channel state coupled to the open state.


Recurrent vitreous haemorrhage and epidural haematoma in a child with hypofibrinogenaemia.
A 14-month-old male infant was brought by parents for redness of the right eye of 18 days duration. Exam and B-scan ultrasonography revealed total hyphema, dense vitreous haemorrhage and lens subluxation in the right eye while CT disclosed right small epidural haematoma. The left eye had neither retinal haemorrhage nor disc oedema. There was no sign of shaken baby syndrome. Fibrinogen level in the blood was very low. The parents are first-degree cousins with two family members having hypofibrinogenaemia. Vitreous haemorrhage recurred after surgical intervention resulting in phthisis and loss of vision. Hypofibrinogenaemia needs to be included in the differential diagnosis of ocular haemorrhage and vision loss.


Calcium/calmodulin-dependent phosphorylation of tumor protein D52 on serine residue 136 may be mediated by CAMK2delta6.
Tumor protein D52 is expressed at relatively high levels in cells within the gastrointestinal tract that undergo classical exocytosis and is overexpressed in several cancers. Current evidence supports a role for D52 in the regulation of vesicular trafficking. D52 function(s) are regulated by calcium-dependent phosphorylation; however, the intracellular mechanisms that mediate this process are not well characterized. The goal of this study was to identify the calcium-dependent phosphorylation site(s) in D52 and to characterize the protein kinase(s) that mediate this phosphorylation. Using mass spectrometry and site-directed mutagenesis, we identified a single amino acid residue, S(136), that undergoes increased phosphorylation upon elevation of intracellular Ca(2+) concentration. A phosphospecific antibody (pS(136)) was produced and used to characterize D52 kinase activity in gastric mucosal, colonic T84, and HEK293 cells. By using D52 as a substrate, a protein kinase with a molecular weight (M(r)) of approximately 50 kDa was identified with "in gel" assays. This kinase comigrated with rat brain calcium/calmodulin-dependent protein kinase (CAMK2)alpha cross-reacted with pan-specific CAMK2 antibodies as well as with anti-active CAMK2 (pT(286/287)) antibody when activated. Carbachol-stimulated phosphorylation of S(136) was inhibited by the CAMK2 inhibitor KN93 (IC(50) 38 microM) and by the calmodulin antagonist W7 (IC(50) 3.3 nM). A previously uncharacterized CAMK2 isoform, CAMK2delta6, which has the same domain structure and M(r) as CAM2alpha, was identified in gastric mucosa by RT-PCR. The cloned, expressed protein comigrated with D52 kinase and colocalized with D52 protein in T84 and HEK293 cells. These findings support a role for CAMK2delta6 in the mediation of D52 phosphorylation.


Pericardial patch tracheoplasty for severe tracheal stenosis in children: intermediate results.
Between 1982 and 1990, 15 children have undergone pericardial patch tracheoplasty, 13 for complete tracheal rings and 2 for acquired tracheal stenosis. Eleven had complete tracheal rings from the thoracic inlet or the cricoid to the carina. Diagnosis was by bronchoscopy (15), computed tomography (10), and magnetic resonance imaging (3). As first described by Idriss in 1984, the surgical technique uses a median sternotomy approach, extracorporeal circulation, bronchoscopic guidance, and the use of pericardium as a tracheal patch. There was one perioperative death of mediastinitis (operative survival, 93%). There were two late deaths caused by complications of tracheostomy (1) and direct laryngoscopy and bronchoscopy (DLB) (1). Follow-up is complete in 12 intermediate-term survivors and ranges from 0.4 to 8.2 years (mean, 4.2 +/- 2.9 years). Five children required multiple DLBs with dilation and excision of granulation tissue; one of these patients underwent tracheoplasty revision 5 months after the initial procedure for residual tracheal rings, and four of these children have had tracheostomies, two temporary and two currently (4 months, 7 years). Seven children had widely patent tracheas observed at follow-up DLB with little or no residual stenosis after tracheoplasty. All survivors are currently asymptomatic, except for the two children with tracheostomies. Pericardial patch tracheoplasty offers effective therapy with relatively low operative mortality for infants and children with severe long segment tracheal stenosis. On intermediate-term follow-up, nearly all survivors are essentially free of symptoms of residual tracheal stenosis.


Diel investments in metabolite production and consumption in a model microbial system.
Organic carbon transfer between surface ocean photosynthetic and heterotrophic microbes is a central but poorly understood process in the global carbon cycle. In a model community in which diatom extracellular release of organic molecules sustained growth of a co-cultured bacterium, we determined quantitative changes in the diatom endometabolome and the bacterial uptake transcriptome over two diel cycles. Of the nuclear magnetic resonance (NMR) peaks in the diatom endometabolites, 38% had diel patterns with noon or mid-afternoon maxima; the remaining either increased (36%) or decreased (26%) through time. Of the genes in the bacterial uptake transcriptome, 94% had a diel pattern with a noon maximum; the remaining decreased over time (6%). Eight diatom endometabolites identified with high confidence were matched to the bacterial genes mediating their utilization. Modeling of these coupled inventories with only diffusion-based phytoplankton extracellular release could not reproduce all the patterns. Addition of active release mechanisms for physiological balance and bacterial recognition significantly improved model performance. Estimates of phytoplankton extracellular release range from only a few percent to nearly half of annual net primary production. Improved understanding of the factors that influence metabolite release and consumption by surface ocean microbes will better constrain this globally significant carbon flux.


Probing the role of CXC motif in chemokine CXCL8 for high affinity binding and activation of CXCR1 and CXCR2 receptors.
All chemokines share a common structural scaffold that mediate a remarkable variety of functions from immune surveillance to organogenesis. Chemokines are classified as CXC or CC on the basis of conserved cysteines, and the two subclasses bind distinct sets of GPCR class of receptors and also have markedly different quaternary structures, suggesting that the CXC/CC motif plays a prominent role in both structure and function. For both classes, receptor activation involves interactions between chemokine N-loop and receptor N-domain residues (Site-I), and between chemokine N-terminal and receptor extracellular/transmembrane residues (Site-II). We engineered a CC variant (labeled as CC-CXCL8) of the chemokine CXCL8 by deleting residue X (CXC → CC), and found its structure is essentially similar to WT. In stark contrast, CC-CXCL8 bound poorly to its cognate receptors CXCR1 and CXCR2 (K(i) > 1 μm). Further, CC-CXCL8 failed to mobilize Ca(2+) in CXCR2-expressing HL-60 cells or recruit neutrophils in a mouse lung model. However, most interestingly, CC-CXCL8 mobilizes Ca(2+) in neutrophils and in CXCR1-expressing HL-60 cells. Compared with the WT, CC-CXCL8 binds CXCR1 N-domain with only ∼5-fold lower affinity indicating that the weak binding to intact CXCR1 must be due to its weak binding at Site-II. Nevertheless, this level of binding is sufficient for receptor activation indicating that affinity and activity are separable functions. We propose that the CXC motif functions as a conformational switch that couples Site-I and Site-II interactions for both receptors, and that this coupling is critical for high affinity binding but differentially regulates activation.


Arginine-rhamnosylation as new strategy to activate translation elongation factor P.
Ribosome stalling at polyproline stretches is common and fundamental. In bacteria, translation elongation factor P (EF-P) rescues such stalled ribosomes, but only when it is post-translationally activated. In Escherichia coli, activation of EF-P is achieved by (R)-β-lysinylation and hydroxylation of a conserved lysine. Here we have unveiled a markedly different modification strategy in which a conserved arginine of EF-P is rhamnosylated by a glycosyltransferase (EarP) using dTDP-L-rhamnose as a substrate. This is to our knowledge the first report of N-linked protein glycosylation on arginine in bacteria and the first example in which a glycosylated side chain of a translation elongation factor is essential for function. Arginine-rhamnosylation of EF-P also occurs in clinically relevant bacteria such as Pseudomonas aeruginosa. We demonstrate that the modification is needed to develop pathogenicity, making EarP and dTDP-L-rhamnose-biosynthesizing enzymes ideal targets for antibiotic development.


A randomized, controlled trial of mindfulness-based stress reduction in HIV infection.
Evidence links depression and stress to more rapid progression of HIV-1 disease. We conducted a randomized controlled trial to test whether an intervention aimed at improving stress management and emotion regulation, mindfulness-based stress reduction (MBSR), would improve immunological (i.e. CD4+ T-cell counts) and psychological outcomes in persons with HIV-1 infection. We randomly assigned participants with HIV-1 infection and CD4 T-cell counts >350 cells/μl who were not on antiretroviral therapy in a 1:1 ratio to either an MBSR group (n = 89) or an HIV disease self-management skills group (n = 88). The study was conducted at the University of California at San Francisco. We assessed immunologic (CD4, c-reactive protein, IL-6, and d-dimer) and psychological measures (Beck Depression Inventory for depression, modified Differential Emotions Scale for positive and negative affect, Perceived stress-scale, and mindfulness) at 3, 6 and 12 months after initiation of the intervention; we used multiple imputation to address missing values. We observed statistically significant improvements from baseline to 3-months within the MBSR group in depression, positive and negative affect, perceived stress, and mindfulness; between group differences in change were significantly greater in the MBSR group only for positive affect (per item difference on DES-positive 0.25, 95% CI 0.049, 0.44, p = .015). By 12 months the between group difference in positive affect was not statistically significant, although both groups had trends toward improvements compared to baseline in several psychological outcomes that were maintained at 12-months; these improvements were only statistically significant for depression and negative affect in the MBSR group and perceived stress for the control group. The groups did not differ significantly on rates of antiretroviral therapy initiation (MBSR = 39%, control = 29%, p = .22). After 12 months, the mean decrease in CD4+ T-cell count was 49.6 cells/μl in participants in the MBSR arm, compared to 54.2 cells/μl in the control group, a difference of 4.6 cells favoring the MBSR group (95% CI, -44.6, 53.7, p = .85). The between group differences in other immunologic-related outcomes (c-reactive protein, IL-6, HIV-1 viral load, and d-dimer) were not statistically significant at any time point. MBSR improved positive affect more than an active control arm in the 3 months following the start of the intervention. However, this difference was not maintained over the 12-month follow-up and there were no significant differences in immunologic outcomes between intervention groups. These results emphasize the need for further carefully designed research if we are to translate evidence linking psychological states to immunological outcomes into evidence-based clinical practices.


[Data on the study of the epidemiology of rotavirus infection in Leningrad].
The results obtained in the study of rotavirus infection in Leningrad in 1984-1987 are presented. Enzyme immunoassay techniques were used for the examination of 4,715 children aged 0-14 years and 1,162 adults with diagnosed acute enteral infection of unknown etiology, as well as the control group of 556 of healthy children aged 0-6 years and 77 healthy adults. The rotavirus antigen was detected in 1.210 sick children (25.7%) and 133 sick adults (11.4%), as well as in 6 healthy children (1.1%), but not detected in healthy adults. The following epidemiological regularities of rotavirus infection were established: the highest sick rate among children aged 0-2 years and a low level of rotavirus carriership among healthy persons; the seasonal character of rotavirus infection, its outbreaks occurring in winter and the epidemic periods varying in their character and duration in different years; the prevailing role of rotaviruses in the development of winter rises of acute enteral infection of unknown etiology in the city among children aged 0-2 years. The problem of the respiratory syndrome in rotavirus diarrhea is discussed. The rotavirus antigen was detected in 39 out of 144 children (27%) with diagnosed acute respiratory virus diseases and in 4 out of 99 nasopharyngeal washings (4%) from diarrhea patients. Adenoviruses were shown to play an insignificant role in the etiology of diarrhea (10% of cases).


Colony defense by africanized and European honey bees.
Africanized and European honey bee (Apis mellifera) populations showed quantitative differences in colony defensive behavior. Africanized bees responded faster and in much larger numbers than European honey bees and produced 8.2 and 5.9 times as many stings during two different experiments. Times to react to alarming stimuli were negatively correlated with the number of bees responding and to the total number of stings. The number of bees responding was significantly correlated to the total number of stings only for the Africanized population.


Examining the Association of Social Needs with Future Health Care Utilization in an Older Adult Population: Which Needs Are Most Important?
Abstract Social needs, such as social isolation and food insecurity, are important individual-level social determinants of health, especially for adults ages 65 years and older. These needs may be associated with future health care utilization, but this research area has not been studied extensively. The objective of this study was to examine the independent association of 5 individual social needs with future (1) emergency department (ED) visits and (2) hospital admissions. This observational study included 9649 Kaiser Permanente Northwest (KPNW) Medicare members who completed the Medicare Total Health Assessment (MTHA) quality improvement survey between August 17, 2020 and January 31, 2022. The 5 social needs assessed by the MTHA, defined as binary measures (yes/no), included (1) financial strain, (2) food insecurity, (3) housing instability, (4) social isolation, and (5) transportation needs. ED utilization (yes/no) and hospitalization (yes/no), the current study outcome measures, were measured in the 12 months after MTHA assessment. In multivariable analyses, 3 of the 5 social needs were significantly associated with higher ED utilization: financial strain (odds ratio [OR] = 1.40, 95% confidence interval [CI] = 1.11-1.76, P < 0.05), housing instability (OR = 1.43, 95% CI = 1.02-1.99, P < 0.05), and social isolation (OR = 1.19, 95% CI = 1.05-1.34, P < 0.05), and 1, financial strain, was significantly associated with hospital admissions (OR = 1.66, 95% CI = 1.23-2.23, P < 0.05). The study results identified which social needs are most strongly associated with future ED utilization and hospital admissions. Further research is needed to better understand whether addressing social needs is associated with improved patient-level health outcomes over time.


Sequence information for the splicing of human pre-mRNA identified by support vector machine classification.
Vertebrate pre-mRNA transcripts contain many sequences that resemble splice sites on the basis of agreement to the consensus,yet these more numerous false splice sites are usually completely ignored by the cellular splicing machinery. Even at the level of exon definition,pseudo exons defined by such false splices sites outnumber real exons by an order of magnitude. We used a support vector machine to discover sequence information that could be used to distinguish real exons from pseudo exons. This machine learning tool led to the definition of potential branch points,an extended polypyrimidine tract,and C-rich and TG-rich motifs in a region limited to 50 nt upstream of constitutively spliced exons. C-rich sequences were also found in a region extending to 80 nt downstream of exons,along with G-triplet motifs. In addition,it was shown that combinations of three bases within the splice donor consensus sequence were more effective than consensus values in distinguishing real from pseudo splice sites; two-way base combinations were optimal for distinguishing 3' splice sites. These data also suggest that interactions between two or more of these elements may contribute to exon recognition,and provide candidate sequences for assessment as intronic splicing enhancers.


UV-Induced Wnt7a in the Human Skin Microenvironment Specifies the Fate of Neural Crest-Like Cells via Suppression of Notch.
Multipotent stem cells with neural crest-like properties have been identified in the dermis of human skin. These neural crest stem cell (NCSC)-like cells display self-renewal capacity and differentiate into neural crest derivatives, including epidermal pigment-producing melanocytes. NCSC-like cells share many properties with aggressive melanoma cells, such as high migratory capabilities and expression of the neural crest markers. However, little is known about which intrinsic or extrinsic signals determine the proliferation or differentiation of these neural crest-like stem cells. Here we show that, in NCSC-like cells, Notch signaling is highly activated, similar to melanoma cells. Inhibition of Notch signaling reduced the proliferation of NCSC-like cells, induced cell death, and downregulated noncanonical Wnt5a, suggesting that the Notch pathway contributes to the maintenance and motility of these stem cells. In three-dimensional skin reconstructs, canonical Wnt signaling promoted the differentiation of NCSC-like cells into melanocytes. This differentiation was triggered by the endogenous Notch inhibitor Numb, which is upregulated in the stem cells by Wnt7a derived from UV-irradiated keratinocytes. Together, these data reveal a cross talk between the two conserved developmental pathways in postnatal human skin, and highlight the role of the skin microenvironment in specifying the fate of stem cells.


Clinical Features for the Diagnosis of Pediatric Urinary Tract Infections: Systematic Review and Meta-Analysis.
Accurate diagnosis of urinary tract infection in children is essential because children left untreated can experience permanent renal injury. We aimed to assess the diagnostic value of clinical features of pediatric urinary tract infection. We performed a systematic review and meta-analysis of diagnostic test accuracy studies in ambulatory care. We searched the PubMed, Embase, Web of Science, Cumulative Index to Nursing and Allied Health Literature, Cochrane Central Register of Controlled Trials, Health Technology Assessment, and Database of Abstracts of Reviews of Effects databases from inception to January 27, 2020 for studies reporting 2 × 2 diagnostic accuracy data for clinical features compared with urine culture in children aged <18 years. For each clinical feature, we calculated likelihood ratios and posttest probabilities of urinary tract infection. To estimate summary parameters, we conducted a bivariate random effects meta-analysis and hierarchical summary receiver operating characteristic analysis. A total of 35 studies (N = 78,427 patients) of moderate to high quality were included, providing information on 58 clinical features and 6 prediction rules. Only circumcision (negative likelihood ratio [LR-] 0.24; 95% CI, 0.08-0.72; n = 8), stridor (LR- 0.20; 95% CI, 0.05-0.81; n = 1), and diaper rash (LR- 0.13; 95% CI, 0.02-0.92; n = 1) were useful for ruling out urinary tract infection. Body temperature or fever duration showed limited diagnostic value (area under the receiver operating characteristic curve 0.61; 95% CI, 0.47-0.73; n = 16). The Diagnosis of Urinary Tract Infection in Young Children score, Gorelick Scale score, and UTIcalc (https://uticalc.pitt.edu) might be useful to identify children eligible for urine sampling. Few clinical signs and symptoms are useful for diagnosing or ruling out urinary tract infection in children. Clinical prediction rules might be more accurate; however, they should be validated externally. Physicians should not restrict urine sampling to children with unexplained fever or other features suggestive of urinary tract infection.


[Humoral and cellular immune phenomena in an acute viral hepatitis (author's transl)].
During the course of acute viral hepatitis A and B, several humoral and cellular immune phenomena have been observed, part of which is predominantly or even exclusively associated with hepatitis B: 1. Relative and absolute counts for T-lymphocytes depressed and for "null" -cells elevated; 2. mild elevation of serum globulin levels; 3. IgM augmentation occurring fastly, pronounced, and long lasting in typical cases; 4. IgG augmentation occurring later, less pronounced, and for a shorter period in typical cases; 5. autoantibodies to smooth muscles and mitochondria in low titers in some patients; 6. specific antibodies to "e" -antigen (early) and HB-Ag (later in the course) in part of the cases with hepatitis B; 7. immune complexes including HB-Ag, IgG and probably IgM (and IgA) as well as complement in some cases; 8. depressed levels of the fourth component of complement and - in cases complicated by "allergic" symptoms - of C3, C4, and total complement; 9. occurrence of activated lymphocytes ("virocytes") in peripheral blood; 10. enhanced spontaneous lymphocytic DNA-synthesis; 11. enchanced phytohaemagglutinin stimulation of lymphocytes; 12. mild lymphocyte proliferation to HB-Ag in part of the acute and convalescent cases of hepatitis B; 13. production of migration inhibition factor to liver specific protein (and HB-Ag) or lymphocytes in different percentages of patients with hepatitis B. Origin, diagnostic and prognostic importance, as well as pathogenetic revelance of the described immune phenomena are discussed.


Marginal ridge fracture resistance, microleakage and pulpal response to glass ionomer/glass cermet partial tunnel restorations.
Sixty sound premolars which were to be extracted for orthodontic treatment purposes were restored either with glass ionomer cement or glass cermet cements after partial tunnel preparation, and prior to the extraction after a time interval of 30 and 60 days respectively. The teeth were then subjected to marginal ridge fracture resistance, microleakage study using dye penetration and histological evaluation of the pulpal response to these materials. Both the materials exhibited increase in marginal ridge fracture resistance at 60 days, with minimal degree of microleakage and were biologically compatible with the dental pulp.


Gene array and real time PCR analysis of the adrenal sensitivity to adrenocorticotropic hormone in pig.
Variability in hypothalamic-pituitary-adrenal (HPA) axis activity has been shown to be influenced by genetic factors and related to great metabolic differences such as obesity. The aim of this study was to investigate molecular bases of genetic variability of the adrenal sensitivity to ACTH, a major source of variability, in Meishan (MS) and Large White (LW) pigs, MS being reported to exhibit higher basal cortisol levels, response to ACTH and fatness than LW. A pig cDNA microarray was used to identify changes in gene expression in basal conditions and in response to ACTH stimulation. Genotype and/or ACTH affected the expression of 211 genes related to transcription, cell growth/maintenance, signal transduction, cell structure/adhesion/extra cellular matrix and protein kinase/phosphatase activity. No change in the expression of known key regulator proteins of the ACTH signaling pathway or of steroidogenic enzymes was found. However, Mdh2, Sdha, Suclg2, genes involved in the tricarboxylic acid (TCA) pathway, were over-expressed in MS pigs. Higher TCA cycle activity in MS than in LW may thus result in higher steroidogenic activity and thus explain the typically higher cortisol levels in MS compared to LW. Moreover, up-regulation of Star and Ldlr genes in MS and/or in response to ACTH suggest that differences in the adrenal function between MS and LW may also involve mechanisms requisite for cholesterol supply to steroidogenesis. The present study provides new potential candidate genes to explain genetic variations in the adrenal sensitivity to ACTH and better understand relationship between HPA axis activity and obesity.


Chikungunya vaccine candidate is highly attenuated and protects nonhuman primates against telemetrically monitored disease following a single dose.
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes major epidemics of rash, fever, and debilitating arthritis. Currently, there are no vaccines or antivirals available for prevention or treatment. We therefore generated 2 live-attenuated vaccine candidates based on the insertion of a picornavirus internal ribosome entry site (IRES) sequence into the genome of CHIKV. Vaccination of cynomolgus macaques with a single dose of either vaccine produced no signs of disease but was highly immunogenic. After challenge with a subcutaneous inoculation of wild-type CHIKV, both vaccine candidates prevented the development of detectable viremia. Protected animals also exhibited no significant changes in core body temperature or cardiovascular rhythm, whereas sham-vaccinated animals showed hyperthermia, followed by sustained hypothermia, as well as significant changes in heart rate. These CHIKV/IRES vaccine candidates appear to be safe and efficacious, supporting their strong potential as human vaccines to protect against CHIKV infection and reduce transmission and further spread.


Misoprostol, an anti-ulcer agent and PGE2 receptor agonist, protects against cerebral ischemia.
Induction of COX-2 activity in cerebral ischemia results in increased neuronal injury and infarct size. Recent studies investigating neurotoxic mechanisms of COX-2 demonstrate both toxic and paradoxically protective effects of downstream prostaglandin receptor signaling pathways. We tested whether misoprostol, a PGE(2) receptor agonist that is utilized clinically as an anti-ulcer agent and signals through the protective PGE(2) EP2, EP3, and EP4 receptors, would reduce brain injury in the murine middle cerebral artery occlusion-reperfusion (MCAO-RP) model. Administration of misoprostol, at the time of MCAO or 2h after MCAO, resulted in significant rescue of infarct volume at 24 and 72h. Immunocytochemistry demonstrated dynamic regulation of the EP2 and EP4 receptors during reperfusion in neurons and endothelial cells of cerebral cortex and striatum, with limited expression of EP3 receptor. EP3-/- mice had no significant changes in infarct volume compared to control littermates. Moreover, administration of misoprostol to EP3+/+ and EP3-/- mice showed similar levels of infarct rescue, indicating that misoprostol protection was not mediated through the EP3 receptor. Taken together, these findings suggest a novel function for misoprostol as a protective agent in cerebral ischemia acting via the PGE(2) EP2 and/or EP4 receptors.


Combining identity by descent and association in genetic case-control studies.
In human case-control association studies, one of the chi-square tests typically carried out is based on a 2 x 3 table of genotypes (homogeneity of three genotype frequencies in case and control individuals). We formulate the two degrees of freedom associated with a given genotype distribution in terms of two biologically relevant parameters, (1) the probability F that an individual's two alleles are identical by descent (IBD) and (2) the frequency p of one of the alleles. Imposing the restriction, F > or = 0, makes some of the genotype frequencies invalid thereby reducing noise. We propose a new statistical association test, the FP test, by focusing on allele frequency differences between case and control individuals while allowing for suitable IBD probabilities. Power calculations show that (1) the practice of generally carrying out two association tests (allele and genotype test) has an increased type I error and (2) our test is more powerful than conventional genotype and allele tests under recessive trait inheritance, and at least as powerful as these conventional tests under dominant inheritance. For dominant and recessive modes of inheritance, any apparent power gain by an allele test when carried out in conjunction with a genotype test tends to be purchased entirely by an increased rate of false positive results due to omission of a multiple testing correction. As an alternative to these two standard association tests, our FP test represents a convenient and more powerful alternative.


HbA1c Identifies Subjects With Prediabetes and Subclinical Left Ventricular Diastolic Dysfunction.
Prediabetes is associated with subclinical cardiac changes associated with heart failure development. We investigated diastolic function and its association with markers of glycation and inflammation related to cardiovascular disease in patients with prediabetes. We focused on individuals with prediabetes identified only by glycated hemoglobin A1c [HbA1c; 5.7% to 6.4% and normal fasting glucose (NFG) and normal glucose tolerance (NGT) after an oral glucose tolerance test (OGTT)]. Cross-sectional study. Departments of Clinical and Experimental Medicine and Cardiology, University of Catania, Catania, Italy. HbA1c, OGTT, Doppler echocardiography, soluble receptor for advanced glycation end products (sRAGEs), and endogenous secretory RAGE (esRAGE) were evaluated. We recruited 167 subjects with NFG/NGT who were stratified according to HbA1c level: controls (HbA1c <5.7%) and HbA1c prediabetes (HbA1c 5.7% to 6.4%). Patients with HbA1c prediabetes (n = 106) showed a lower peak mitral inflow in early diastole (E wave) to late diastolic atrial filling velocity (A wave) ratio (E/A ratio) than controls (n = 61) (1.10 ± 0.24 vs 1.18 ± 0.23; P < 0.05). They showed a higher left atrium volume (LAV) (28.4 ± 5 vs 22.1 ± 3; P < 0.05) and sphericity index (SI) (0.6 ± 0.06 vs 0.5 ± 0.05; P < 0.05). After multiple regression analyses, HbA1c, sRAGE, and esRAGE were the major determinants of E/A ratio, LAV, and SI. Subjects with HbA1c prediabetes exhibited subclinical cardiac alterations associated with sRAGE, esRAGE, and HbA1c. These subjects would not have been classified as having prediabetes on the basis of fasting glycemia or post-OGTT values.


Molecularly targeted therapies in metastatic pancreatic cancer: a systematic review.
Pancreatic cancer is the fourth leading cause of cancer-related death. Most patients present with an advanced stage of disease that has a dismal outcome, with a median survival of approximately 6 months. Evidently, there is a clear need for the development of new agents with novel mechanisms of action in this disease. A number of biological agents modulating different signal transduction pathways are currently in clinical development, inhibiting angiogenesis and targeting epidermal growth factor receptor, cell cycle, matrix metalloproteinases, cyclooxygenase-2, mammalian target of rapamycin, or proteasome. This is the first systematic review of the literature to synthesize all available data coming from trials and evaluate the efficacy and safety of molecular targeted drugs in unresectable and metastatic pancreatic cancer. However, it should be stressed that although multiple agents have been tested, only 9 phase 3 trials have been conducted and one agent (erlotinib) has been approved by the Food and Drug Administration for use in clinical practice. As knowledge accumulates on the molecular mechanisms underlying carcinogenesis in the pancreas, the anticipated development and assessment of molecularly targeted agents may offer a promising perspective for a disease which, to date, remains incurable.


Molecular mechanisms of nitrovasodilator bioactivation.
All nitrovasodilators act intracellularly by a common molecular mechanism. This is characterized by the release of nitric oxide (NO). They are, thus, prodrugs or carriers of the active principle NO, responsible for endothelial controlled vasodilation. The rate of NO-formation strongly correlates with the activation of the soluble guanylate cyclase in vitro, resulting in a stimulation of cGMP synthesis. Nitrovasodilators thus are therapeutic substitutes for endogenous EDRF/NO. The pathways of bioactivation, nevertheless, differ substantially, depending on the individual chemistry of the nitrovasodilator. Besides NO, numerous other reaction products such as nitrite and nitrate anions are formed. The guanylate cyclase is only activated if NO is liberated. In the case of organic nitrates such as GTN, NO is only formed if certain thiol compounds are present as an essential cofactor. The rate of NO-formation correlates with the number of nitrate ester groups and proceeds with a simultaneous nitrite formation (with a ratio of 1:14 in the presence of cysteine). Nitrosamines such as molsidomine do not need thiol compounds for bioactivation. They directly liberate NO from the ring-open A-forms. This process basically depends on the presence of oxygen as electron acceptor from the sydnonimine molecule. Therefore, besides NO also superoxide radicals are formed, which may react with the generated NO under formation of nitrate ions. Organic nitrites (such as amyl nitrite) require the preceding interaction with a mercapto group to form a S-nitrosothiol intermediate, from which finally NO radicals are liberated. Nitrosothiols (like S-nitroso-acetyl-penicillamine) and sodium nitroprusside spontaneously release NO. The molecules themselves do not possess a direct enzyme activating potency. In the presence of thiol compounds organic nitrites (e.g., amyl nitrite) and nitrosothiols may act as intermediary products of NO generation.


The tissue kallikrein family of serine proteases: functional roles in human disease and potential as clinical biomarkers.
Prostate specific antigen (PSA) or human kallikrein 3 (hK3) has long been an effective biomarker for prostate cancer. Now, other members of the tissue kallikrein (KLK) gene family are fast becoming of clinical interest due to their potential as prognostic biomarkers. particularly for hormone dependent cancers. The tissue kallikreins are serine proteases that are encoded by highly conserved multi-gene family clusters in rodents and humans. The rat and mouse loci contain 10 and 25 functional genes, respectively, while the human locus at 19q 13.4 contains 15 genes. The structural organization and size of these genes are similar across species; all genes have 5 coding exons that encode a prepro-enzyme. Although the physiological activators of these zymogens have not been described, in vitro biochemical studies show that some kallikreins can auto-activate and others can activate each other, suggesting that the kallikreins may participate in an enzymatic cascade similar to that of the coagulation cascade. These genes are expressed, to varying degrees, in a wide range of tissues suggesting a functional involvement in a diverse range of physiological and pathophysiological processes. These include roles in normal skin desquamation and psoriatic lesions, tooth development, neural plasticity, and Alzheimer's disease (AD). Of particular interest is the expression of many kallikreins in prostate, ovarian, and breast cancers where they are emerging as useful prognostic indicators of disease progression.


Propagation of Gaussian Schell-model beams through a jet engine exhaust.
Theoretical predictions of light beam interactions with jet engine exhaust are of importance for optimization of various optical systems, including LIDARs, imagers and communication links operating in the vicinity of aircrafts and marine vessels. Here we extend the analysis previously carried out for coherent laser beams propagating in jet engine exhaust, to the broad class of Gaussian Schell-Model (GSM) beams, being capable of treating any degree of coherence in addition to size and radius of curvature. The analytical formulas for the spectral density (SD) and the spectral degree of coherence (DOC) of the GSM beam are obtained and analyzed on passage through a typical jet engine exhaust region. It is shown that for sources with high coherence, the transverse profiles of the SD and the DOC of the GSM beams gradually transition from initially circular to elliptical shape upon propagation at very short ranges. However, such transition is suppressed for sources with lower coherence and disappears in the incoherent source limit, implying that the GSM source with low source coherence is an excellent tool for mitigation of the jet engine exhaust-induced anisotropy of turbulence. The physical interpretation and the illustration are included.


Impact of protected mealtimes on ward mealtime environment, patient experience and nutrient intake in hospitalised patients.
Malnutrition is a common problem in hospitalised inpatients, resulting in a range of negative clinical, patient-centred and economic sequelae. Protected mealtimes (PM) aim to enhance the quality of the mealtime experience and maximise nutrient intake in hospitalised patients. The present study aimed to measure mealtime environment, patient experience and nutrient intake before and after the implementation of PM. PM were implemented in a large teaching hospital through a range of different approaches. Direct observations were used to assess ward-level mealtime environment (e.g. dining room use, removal of distractions) (40 versus 34 wards) and individual patient experience (e.g. assistance with eating, visitors present) (253 versus 237 patients), and nutrient intake was assessed with a weighed food intake at lunch (39 versus 60 patients) at baseline and after the implementation of PM, respectively. Mealtime experience showed improvements in three objectives: more patients were monitored using food/fluid charts (32% versus 43%, P = 0.02), more were offered the opportunity to wash hands (30% versus 40%, P = 0.03) and more were served meals at uncluttered tables (54% versus 64%, P = 0.04). There was no difference in the number of patients experiencing mealtime interruptions (32% versus 25%, P = 0.14). There was no difference in energy intake (1088 versus 837 kJ, P = 0.25) and a decrease in protein intake (14.0 versus 7.5 g, P = 0.04) after PM. Only minor improvements in mealtime experience were made after the implementation of PM and so it is not unexpected that macronutrient intake did not improve. The implementation of PM needs to be evaluated to ensure improvements in mealtime experience are made such that measurable improvements in nutritional and clinical outcomes ensue.


A large pericentric inversion of human chromosome 8.
A large pericentric inversion, inv(8) (p11q24), was ascertained in a male investigated because his wife had had repeated miscarriages. The inversion segregated in 3 generations of the family, and no chromosomally unbalanced offspring were detected. The miscarriage and the inversion could not be causally related.