The Role of Adipokines in Understanding the Associations between Obesity and Depression.
Objective. Two major causes of disability, major depression and obesity, share overlapping psychosocial and pathophysiological etiologies. Studies are now focused on biological mechanisms linking the two illnesses, and there is interest in the role that adipokines may have in mediating the association between obesity and depression. We reviewed the literature to look at what is currently known about this association, focusing on the adipokines leptin, adiponectin, and resistin. Methods. A MEDLINE search, citing articles from 1966 onward, supplemented by a review of bibliographies, was conducted to identify relevant studies. Results. This paper identified plausible pathways underlying a link between adipokines and depression. Only a few studies have yet been conducted specifically examining these biomarkers in patients with depression, but the results are intriguing. Conclusion. This paper is one of the first to examine the association between adipokines and depression. It provides an overview of the physiological role of adipokines and summarizes the data suggesting that they may be dysregulated in major depression. This area of research may become increasingly important as new treatment strategies are developed.


[Proteasomal degradation of ribonucleic acids of different NO-synthase isoforms].
Using a developed method of determination of the RNase activity of the proteasome in vitro with the application of reverse transcription followed by subsequent polymerase chain reaction it was shown that 26S proteasome from the proteasomal fraction II effectively cleaves RNA, encoding actin, myosin and all isoforms of NO synthase. The intensity of RNA degradation by proteasome and specific RNases is similar. It was also shown that clasto-lactacystin beta-lactone, a specific proteasome inhibitor significantly depresses RNase activity of the proteasome. Thus, proteasome is capable to degrade certain eukariotic RNA in vitro and the proposed method can be used in order to discover specific substances such as inhibitors of RNase activity of the proteasome.


Guidelines for pharmacotherapy in Alzheimer's disease - A primer on FDA-approved drugs.
The growing prevalence of dementia makes it important for us to better understand its pathophysiology and treatment modalities, to improve the quality of life of patients and caregivers. Alzheimer's disease (AD), a neurodegenerative disease, is the most common form of amnestic dementia in the geriatric population. Pathophysiology of AD is widely attributed to aggregation of amyloid-beta (Aβ) plaques and hyperphosphorylation of tau proteins. Initial treatment modalities aimed to increase brain perfusion in a non-specific manner. Subsequent therapy focused on rectifying neurotransmitter imbalance in the brain. Newer drugs modify the progression of the disease by acting against aggregated Aβ plaques. However, not all drugs used in therapy of AD have been granted approval by the United States Food and Drug Administration (FDA). This review categorizes and summarizes the FDA-approved drugs in the treatment of AD in a manner that would make it a convenient reference for researchers and practicing physicians alike. Drugs that mitigate symptoms of dementia may be categorized into mitigators of Behavioral and Psychological Symptoms of Dementia (BPSD), and mitigators of cognitive decline. BPSD mitigators include brexpiprazole, an atypical antipsychotic with a once-daily dosage suited to treat agitation in dementia patients, and suvorexant, an orexin receptor antagonist used to treat sleep disturbances. Cognitive decline mitigators include cholinesterase inhibitors such as donepezil, rivastigmine, and galantamine and glutamate inhibitors such as memantine. Donepezil is the most commonly prescribed drug. It is cheap, well-tolerated, and may be prescribed orally once daily, or as a transdermal patch once weekly. It increases ACh levels, enhances oligodendrocyte differentiation and also protects against Aβ toxicity. However, regular cardiac monitoring is required due to reports of cardiac conduction side effects. Rivastigmine requires a twice-daily oral dosage or once-daily replacement of transdermal patch. It has fewer cardiac side effects than donepezil, but local application-site reactions have been noted. Galantamine, in addition to improving cognitive symptoms in a short span of time, also delays the development of BPSDs and has minimal drug-drug interactions by virtue of having multiple metabolic pathways. However, cardiac conduction disturbances must be closely monitored for. Memantine, a glutamate regulator, acts as an anti-Parkinsonian agent and an antidepressant, in addition to improving cognition and neuroprotection, and requires a once-daily dosage in the form of immediate-release or sustained-release oral tablets. Disease-modifying drugs such as aducanumab and lecanemab reduce the Aβ burden. Both act by binding with fibrillary conformations of Aβ plaques in the brain. These drugs have a risk of causing amyloid-related imaging abnormalities, especially in persons with ApoE4 gene. Aducanumab is administered once every 4 weeks and lecanemab once every 2 weeks. The decision on the choice of the drug must be made after considering the availability of drug, compliance of patient (once-daily vs. multiple doses daily), cost, specific comorbidities, and the risk-benefit ratio for the particular patient. Other non-pharmacological treatment modalities must also be adopted to have a holistic approach toward the treatment of AD.


Studies on the source and release of collagenase in thermally burned corneas of vitamin A-deficient and control rats.
In previous studies we found that a mild thermal burn of the vitamin A-deficient rat cornea caused collagenase release into the medium of corneas placed in culture media 72 hr after applying the burn. Collagenase was released only on day 1 of culture and was not released from identically burned corneas of control rats. We now demonstrate that in deficient corneas, this collagenase release on day 1 of culture increased gradually with increasing time between burn and sacrifice, reaching a maximum at 16 hr after burning a remaining high up to 72 hr. In control rats day-1 collagenase release also increased to a maximum at 16 hr after the burn but then declined to almost zero at 72 hr. Trypsin treatment of day-1 media from both control and deficient corneas, taken at 72 hr after the burn, showed an almost complete absence of latent (inhibited) collagenase. Histologic observations revealed a close correlation between the presence of infiltrating polymorphonuclear neutrophils (PMNs) and the ulcerative lesions seen in burned, deficient corneas. When PMN infiltration was blocked by application of a tissue adhesive, no ulceration occurred and collagenase activity in the day-1 media dropped to almost zero. If burned and unburned areas of deficient corneas were separated and cultured separately, the burned area (containing most of the PMNs) was found to have 10 times the collagenase activity of the unburned area. In the controls, PMNs and some collagenase activity was detectable only 16 hr after the burn. We concluded that in the burned, vitamin A-deficient cornea there is increased attraction of PMNs to the lesion, resulting in collagenase release by these and possibly other cells, and ultimately resulting in ulceration.


Isolation and characterization of alveolar epithelial type II cells derived from mouse embryonic stem cells.
The use of embryonic stem cells (ESCs) to regenerate distal lung epithelia damaged by injuries or diseases requires development of safe and efficient methodologies that direct ESC differentiation into transplantable distal lung epithelial progenitors. Time-consuming culture procedure and low differentiation efficiency are major problems that are associated with conventional differentiation approaches via embryoid body formation. The use of a growth factor cocktail or a lung-specific cell-conditioned medium to enrich definitive endoderm for efficient differentiation of mouse ESCs (mESC) into alveolar epithelial progenitor type II cells (ATIICs) has been reported, but not yet successful for generating a homogenous population of ATIICs for tissue regeneration purpose, and it remains unclear whether or not those mESC-derived ATIICs possess normal biological functions. Here, we report a novel method using a genetically modified mESC line harboring an ATIIC-specific neomycin(R) transgene in Rosa 26 locus. We showed that ATIICs can be efficiently differentiated from mESCs as early as day 7 by culturing them directly on Matrigel-coated plates in DMEM containing 15% knockout serum replacement. With this culture condition, the genetically modified mESCs can be selectively differentiated into a homogenous population (>99%) of ATIICs. Importantly, the mESC-derived ATIICs (mESC-ATIICs) exhibited typical lamellar bodies and expressed surfactant protein A, B, and C as normal control ATIICs. When cultured with an air-liquid-interface culture system in Small Airway Epithelial Cell Growth Medium, the mESC-ATIICs can be induced to secrete surfactant proteins after being treated with dibutyryl cAMP+dexamethasone. These mESC-ATIICs can synthesize and secrete surfactant lipid in response to secretagogue, demonstrating active surfactant metabolism in mESC-ATIICs as that seen in normal control ATIICs. In addition, we demonstrated that the selected mESC-ATIICs can be maintained on Matrigel-coated plates for at least 4 days with robust proliferative capacity. When cultured in DMEM medium containing 10% FBS, mESC-ATIICs spontaneously differentiated into alveolar epithelial type I cells. Collectively, these data demonstrate that the genetically modified mESCs can be selectively differentiated into a homogenous population of functional ATIICs, providing a reliable cell source to explore their therapeutic potential in lung tissue regeneration.


Opioid modulation of scratching and spinal c-fos expression evoked by intradermal serotonin.
We investigated a spinal site for opioid modulation of itch-related scratching behavior in rats. Intradermal 5-HT (2%, 10 microl) elicited intermittent bouts of hindlimb scratching directed toward the injection site (nape of neck) beginning within minutes and lasting >1 hr. 5-HT-evoked scratching was significantly reduced by systemic administration of the opiate antagonist naltrexone but was not affected by systemic morphine at a dosage (3 mg/kg) that induces analgesia. Intradermal 5-HT elicited a significant increase in c-fos-like immunoreactivity (FLI) in superficial laminas I-III at the lateral aspect of the cervical C3-C6 dorsal horn compared with controls receiving intradermal saline. Neither systemic morphine nor naltrexone significantly affected counts of 5-HT-evoked FLI. The lack of effect of morphine suggests that intradermal 5-HT activates dorsal horn neurons, signaling itch but not pain. Attenuation of 5-HT-evoked scratching but not spinal FLI by naltrexone suggests a supraspinal site for its antipruritic action. In contrast, morphine significantly attenuated FLI elicited by intradermal capsaicin, a chemical that induces pain but not scratching.


An efficient and chemoselective method to generate arynes.
Arynes hold immense potential as reactive intermediates in organic synthesis as they engage in a diverse range of mechanistically distinct chemical reactions. However, the poor functional group compatibility of generating arynes or their precursors has stymied their widespread use. Here, we show that generating arynes by deprotonation of an arene and elimination of an "onium" leaving group is mild, efficient and broad in scope. This is achieved by using aryl(TMP)iodonium salts (TMP = 2,4,6-trimethoxyphenyl) as the aryne precursor and potassium phosphate as the base, and a range of arynophiles are compatible. Additionally, we have performed the first quantitative analysis of functional group compatibility for several methods to generate arynes, including the method developed here and the current state of the art. Finally, we show that a range of "sensitive" functional groups such as Lewis and Brønsted acids and electrophiles are compatible under our conditions.


Using electronic health records data to identify patients with chronic pain in a primary care setting.
To develop and validate an accurate method to identify patients with chronic pain using electronic health records (EHR) data at a multisite community health center. We identified patients with chronic pain in our EHR system using readily available data elements pertaining to pain: diagnostic codes (International Classification of Disease, revision 9; ICD-9), patient-reported pain scores, and opioid prescription medications. Medical chart reviews were used to evaluate the accuracy of these data elements in all of their combinations. We developed an algorithm to identify chronic pain patients more accurately based on these evaluations. The algorithm's results were validated for accuracy by comparing them with the documentation of chronic pain by the patient's treating clinician in 381 random patient charts. The new algorithm, which combines pain scores, prescription medications, and ICD-9 codes, has a sensitivity and specificity of 84.8% and 97.7%, respectively. The algorithm was more accurate (95.0%) than pain scores (88.7%) or ICD-9 codes (93.2%) alone. The receiver operating characteristic was 0.981. A straightforward method for identifying chronic pain patients solely using structured electronic data does not exist because individual data elements, such as pain scores or ICD-9 codes, are not sufficiently accurate. We developed and validated an algorithm that uses a combination of elements to identify chronic pain patients accurately. We derived a useful method that combines readily available elements from an EHR to identify chronic pain with high accuracy. This method should prove useful to those interested in identifying chronic pain patients in large datasets for research, evaluation or quality improvement purposes.


Integrating the interpersonal theory of suicide and the dualistic model of passion among adults at risk for suicide.
The dualistic model of passion proposes two passion types, harmonious and obsessive, representing adaptive and maladaptive passion, respectively. Studies suggest interpersonal experiences explain harmonious passion benefits and obsessive passion negative consequences. However, research has not examined passion among individuals with clinically elevated suicide risk, nor the associations between passion types and suicide-related outcomes. The present study presents a conceptual model linking the dualistic model of passion and the interpersonal theory of suicide constructs [specifically, thwarted belongingness (TB) and perceived burdensomeness (PB)]. U.S. adults with clinically elevated suicide risk (N = 484) completed online, cross-sectional assessments of harmonious and obsessive passion, TB, PB, and future dispositions (i.e., positive focus, negative focus, and suicide orientation). A mediation model indicated the effects of harmonious and obsessive passion on positive and negative focus and suicide orientation were largely explained by TB and PB. The present findings suggest engaging in a passion activity may be meaningfully related to suicide-related interpersonal perceptions (i.e., TB and PB).


The Last Mile-Community Engagement and Conditional Incentives to Accelerate Polio Eradication in Pakistan: Study Protocol for a Quasi-Experimental Trial.
Poliomyelitis is a condition of great concern and is endemic in only two countries of the world: Pakistan and Afghanistan. Community mobilization plays a vital role in raising awareness and can help reduce polio vaccine refusals. The objective of this study will be to decrease polio vaccine refusals and zero-dose vaccines by motivating behavior change through the provision of conditional-collective-community-based incentives (C3Is) based on a reduction in polio vaccine refusals. The project will adopt a pretest/post-test quasi-experimental design with two intervention high-risk union councils (HRUCs) and two control union councils (UCs) of peri-urban (Karachi) and rural (Bannu) settings in Pakistan. A participatory community engagement and demand creation strategy with trust-building community mobilization with C3Is, to reduce vaccine refusals and improve polio immunization coverage in two HRUCs, will be used. These UCs will be divided into clusters based on the polio program framework and community groups will be formed in each cluster. These community groups will carry out awareness activities and will be given serial targets to reduce vaccine refusals and those who qualify will be provided C3Is. The project intends to create a replicable model that the government can integrate within health systems for long-term sustainability until the goal of eradication of poliovirus is achieved. The evaluation will be carried out by an independent data collection and analysis team at baseline and endline (after 12 months of intervention). The trial is registered with linicalTrials.gov with number NCT05721274.


Relationship of the 1846G > A Polymorphism of the CYP2D6 Gene to the Equilibrium Concentration Levels of Haloperidol in Patients with Acute Alcoholic Hallucinosis.
Haloperidol is currently used in addictology for the treatment of acute psychotic disorders, including acute alcoholic hallucinosis. The use of haloperidol is often accompanied by the occurrence of adverse drug reactions (ADRs). There is evidence that CYP2D6 isoenzyme is involved in the biotransformation of haloperidol. The study aimed to evaluate the relationship of 1846G > A polymorphism of the CYP2D6 gene to the equilibrium concentration levels of haloperidol in patients with acute alcoholic hallucinosis. The study was conducted on 100 male patients with acute alcoholic hallucinosis (mean age 41.4 ± 14.4 years). The efficacy profile was evaluated using the PANSS (Positive and Negative Syndrome Scale) scale. The safety of therapy was assessed using the UKU Side-Effect Rating Scale and the SAS (Simpson-Angus Scale for Extrapyramidal Symptoms) scale. Genotyping was performed using the real-time polymerase chain reaction (Real-time PCR). Equilibrium plasma concentration levels of haloperidol were investigated using the high-performance liquid chromatography with mass spectrometry (HPLC with MS/MS). No statistically significant results were obtained during the therapy efficacy assessment (dynamics of the PANSS score: GG genotype (-13.00 [-16.00; -16.00; -11.00]), GA genotype (-15.00 [-16.75; -13.00], p = 0.728). There was a statistically significant difference in safety assessment scores (dynamics of the UKU score: GG genotype (8.00 [7.00; 10.00]), GA genotype (15.00 [9.25; 18.00], p < 0.001); dynamics of the SAS score: GG genotype (11.00 [9.00; 14.00]), GA genotype (14.50 [12.00; 18.00], p < 0.001). The pharmacokinetic study results showed a statistically significant difference: GG (3.13 [2.32; 3.95]), GA (3.89 [2.92; 5.26], p = 0.010). Thus, a study conducted on a group of 100 patients with acute alcoholic hallucinosis demonstrated an association between the 1846G > A polymorphism of the CYP2D6 gene (rs3892097) and the safety profile of haloperidol therapy. We also revealed the presence of statistically significant difference in the equilibrium concentration levels of haloperidol in patients with the GG and AG genotypes. It can be concluded that patients with the GA genotype have a higher risk of ADRs compared to patients carrying the GG genotype. It is shown that 1846G > A polymorphism of the CYP2D6 gene (rs3892097) has a statistically significant effect on the equilibrium concentration levels of haloperidol.


Red cell freezing and its impact on the supply chain.
Red blood cells (RBC) can be frozen in glycerol solutions and stored for many years. Thawed RBC must have the glycerol removed, but the recovered cells have normal survival in humans. Freezing has been used to store RBC of rare phenotypes for more than 40 years. In the 1960s and 1970s, when medical technology and blood use were expanding rapidly and liquid whole blood and RBC storage were limited to 3 weeks, many attempts were made to expand the use of frozen RBC for meeting the needs for a stable blood supply and to have RBC reserves for emergencies. These attempts have largely been abandoned because of the cost of freezing, storing and processing, better management of the larger and longer lived RBC inventory, concerns about the safety of stored RBC that have not received the most up-to-date testing and the losses associated with the short shelf life of thawed RBC. New automated frozen RBC processing systems will potentially allow extending the outdate of thawed RBC to 2 weeks, but will not materially effect the costs or losses associated with the use of frozen RBC. RBC freezing will have little effect on the logistics of blood supply.


Sigh Ventilation in Patients With Trauma: The SiVent Randomized Clinical Trial.
Among patients receiving mechanical ventilation, tidal volumes with each breath are often constant or similar. This may lead to ventilator-induced lung injury by altering or depleting surfactant. The role of sigh breaths in reducing ventilator-induced lung injury among trauma patients at risk of poor outcomes is unknown. To determine whether adding sigh breaths improves clinical outcomes. A pragmatic, randomized trial of sigh breaths plus usual care conducted from 2016 to 2022 with 28-day follow-up in 15 academic trauma centers in the US. Inclusion criteria were age older than 18 years, mechanical ventilation because of trauma for less than 24 hours, 1 or more of 5 risk factors for developing acute respiratory distress syndrome, expected duration of ventilation longer than 24 hours, and predicted survival longer than 48 hours. Sigh volumes producing plateau pressures of 35 cm H2O (or 40 cm H2O for inpatients with body mass indexes >35) delivered once every 6 minutes. Usual care was defined as the patient's physician(s) treating the patient as they wished. The primary outcome was ventilator-free days. Prespecified secondary outcomes included all-cause 28-day mortality. Of 5753 patients screened, 524 were enrolled (mean [SD] age, 43.9 [19.2] years; 394 [75.2%] were male). The median ventilator-free days was 18.4 (IQR, 7.0-25.2) in patients randomized to sighs and 16.1 (IQR, 1.1-24.4) in those receiving usual care alone (P = .08). The unadjusted mean difference in ventilator-free days between groups was 1.9 days (95% CI, 0.1 to 3.6) and the prespecified adjusted mean difference was 1.4 days (95% CI, -0.2 to 3.0). For the prespecified secondary outcome, patients randomized to sighs had 28-day mortality of 11.6% (30/259) vs 17.6% (46/261) in those receiving usual care (P = .05). No differences were observed in nonfatal adverse events comparing patients with sighs (80/259 [30.9%]) vs those without (80/261 [30.7%]). In a pragmatic, randomized trial among trauma patients receiving mechanical ventilation with risk factors for developing acute respiratory distress syndrome, the addition of sigh breaths did not significantly increase ventilator-free days. Prespecified secondary outcome data suggest that sighs are well-tolerated and may improve clinical outcomes. ClinicalTrials.gov Identifier: NCT02582957.


Serotyping for MLC gene products: I. Presumptive evidence that ABCIL may detect MLC factors.
Serum from a grand multiparous Inuit (Eskimo) woman, who is HL-A identical but stimulatory in MLC with cells of her husband, contains strong ABCIL reactivity. This serum is operationally monospecific against a cell membrane determinant on lymphocytes, but not platelets, of some unrelated persons. Cells of those who are ABCIL positive with this serum always give a lower MLC stimulation index compared with their MLC responsiveness to cells that are ABCIL negative. This serum also contains good MLC inhibitory activity but only in those one way MLC reactions in which it reacts, by ABCIL, with the stimulating cells. It is postulated that the ABCIL activity is directed against a common MLC factor on lymphocytes.


Reproductive decision making in women with multiple sclerosis.
Multiple sclerosis (MS), the most common acquired neurological disorder of young adults, often strikes young women in their childbearing years. Despite the overlap of MS onset with the childbearing years of women, little is known about how women with MS make decisions about pregnancy and childbearing. In an effort to understand the process of decision making in these women, an exploratory descriptive qualitative study was undertaken; 15 pregnant women with MS were interviewed about their decisions to become pregnant and the factors that entered into their decision-making processes. Content analysis was used to identify and describe the fears and concerns of women with MS around the process of making decisions and about how the presence of MS had influenced that process. The unpredictability of MS and the effect that pregnancy might have on MS was a strong theme that emerged from the analysis. Although participants in this study had proceeded to become pregnant despite their fears related to the effect of pregnancy on the course of their MS, they continued to perceive their decision as risky. The diagnosis of MS affected their previous plans for number of children as well as spacing of pregnancies.


Salivary mucins inhibit antibacterial activity of the cathelicidin-derived LL-37 peptide but not the cationic steroid CSA-13.
Cationic antimicrobial peptides (CAPs) are the effector molecules of innate immunity, similar in potency to classic antibiotics that function in the first-line of defence against infectious agents. The purpose of this study was to investigate the effects of negatively charged mucins on the antibacterial activity of the positively charged cathelicidin LL-37 peptide, its synthetic analogue WLBU2 and the antimicrobial cationic steroid CSA-13. Mucin, DNA, F-actin and hCAP-18/LL-37 in saliva samples were evaluated by microscopy or immunoblotting. Bacterial killing assays and determination of MICs were used to determine bactericidal activity. Binding of rhodamine-B-labelled LL-37 peptide to mucin was fluorimetrically assessed. Microscopic evaluation of saliva after addition of rhodamine-B-labelled LL-37 showed localization similar to that observed after the addition of a specific mucin-binding lectin. Immunoblotting confirmed the presence of hCAP-18/LL-37 in saliva samples and LL-37 peptide bound to isolated submaxillary gland mucin-coated plates. Mucin/LL-37 binding was partially prevented by treatment of mucin with neuraminidase, indicating involvement of sialic acid moieties. Decreased LL-37 and WLBU2 antibacterial activity was observed in the presence of mucin or dialysed human saliva, whereas CSA-13 antibacterial activity was significantly resistant to inhibition by mucins. This study shows that the antibacterial LL-37 peptide and its synthetic analogue WLBU2 are inhibited by salivary mucin and that the cationic steroid CSA-13 retains most of its function in the presence of an equal amount of mucin or saliva.


Abdominal Attenuation Values on Virtual and True Unenhanced Images Obtained With Third-Generation Dual-Source Dual-Energy CT.
The purpose of this study is to investigate the magnitude of differences between attenuation values measured on virtual unenhanced images and true unenhanced images obtained using third-generation dual-source dual-energy CT (DECT). A total of 83 patients requiring thoracoabdominal CT for cancer workup were included in this prospective study. CT examinations included true unenhanced acquisitions (tube potential, 120 kVp) and arterial and portal phase dual-energy CT (DECT) acquisitions (tube potential, 100 kVp and Sn 150 kVp [where Sn denotes the interposition of a tin filter in the high-energy beam]; tube current-exposure time product, 190 and 95 mAs). Virtual unenhanced images were created using two commercially available DECT postprocessing algorithms, one of which was designed to create liver images (hereafter referred to as VNC1 images) and the other of which was designed to create images of organs containing minor amounts of fat (hereafter referred to as VNC2 images). Attenuation values on the liver, spleen, paraspinal muscles, retroperitoneal fat, renal cortex and medulla, and gallbladder and bladder lumens were measured. The attenuation values of all tissues were significantly different between virtual unenhanced and true unenhanced images (p = < 0.001-0.042), except for the liver and spleen in the portal phase and muscles in both phases. When statistically significant, correlations between these differences and body mass index (weight in kilograms divided by the square of height in meters) depended on the tissue imaged and algorithm used. The percentage of cases in which these differences were 10 HU or greater was 1% for the liver and approximately 5% for the spleen and muscles, regardless of the algorithm and phase, but on VNC1 images it reached approximately 30% for the kidney, 70% for the gallbladder and bladder, and depending on the phase, 40-70% for fat. On VNC2 images, the percentage of cases in which these differences were 20 HU or greater was approximately 90% for fat. Abdominal virtual unenhanced images obtained with third-generation dual-source DECT still should not replace true unenhanced images because of substantial differences in attenuation measurements for fluid, fat, and renal tissues.


One bead per residue can describe all-atom protein structures.
Atomistic resolution is the standard for high-resolution biomolecular structures, but experimental structural data are often at lower resolution. Coarse-grained models are also used extensively in computational studies to reach biologically relevant spatial and temporal scales. This study explores the use of advanced machine learning networks for reconstructing atomistic models from reduced representations. The main finding is that a single bead per amino acid residue allows construction of accurate and stereochemically realistic all-atom structures with minimal loss of information. This suggests that lower resolution representations of proteins may be sufficient for many applications when combined with a machine learning framework that encodes knowledge from known structures. Practical applications include the rapid addition of atomistic detail to low-resolution structures from experiment or computational coarse-grained models. The application of rapid, deterministic all-atom reconstruction within multi-scale frameworks is further demonstrated with a rapid protocol for the generation of accurate models from cryo-EM densities close to experimental structures.


Non-TAL Effectors From Xanthomonas oryzae pv. oryzae Suppress Peptidoglycan-Triggered MAPK Activation in Rice.
Xanthomonas oryzae pv. oryzae, the causal pathogen of bacterial blight of rice, depends on its type III secretion system and associated effector proteins to grow and colonize the vascular tissues of rice plants. The type III effectors include a family of closely related transcription activator-like (TAL) effectors and the rest of diverse effectors, so-called non-TAL effectors. Our understanding of non-TAL effectors for pathogenesis in rice blight is still limited. Here we report a feasible method to rapidly detect the activation of mitogen-activated protein kinase pathway in rice mesophyll protoplasts by the X. oryzae pv. oryzae derived peptidoglycan and screen for virulent effectors that can suppress the pathogen-associated molecular pattern triggered immunity (PTI) response. Amongst 17 non-TAL effectors transiently expressed in rice cells, we found that three effectors (XopZ, XopN, and XopV) were able to suppress the peptidoglycan-triggered MAPK activation. The triple mutant of the X. oryzae pv. oryzae strain PXO99A lacking XopZ, XopN, and XopV showed additively reduced virulence. Adding back either of genes restored the virulence of the triple mutant. Our results demonstrate the collective and redundant ability of defense suppression by non-TAL effectors in causing bacterial blight of rice.


Acyl-coenzyme A dehydrogenases are localized on GLUT4-containing vesicles via association with insulin-regulated aminopeptidase in a manner dependent on its dileucine motif.
Insulin-regulated aminopeptidase (IRAP, also termed vp165) is known to be localized on the GLUT4-containing vesicles and to be recruited to the plasma membrane after stimulation with insulin. The cytoplasmic region of IRAP contains two dileucine motifs and acidic regions, one of which (amino acid residues 55-82) is reportedly involved in retention of GLUT4-containing vesicles. The region of IRAP fused with glutathione-S-transferase [GST-IRAP(55-82)] was incubated with lysates from 3T3-L1 adipocytes, leading to identification of long-chain, medium-chain, and short-chain acyl-coenzyme A dehydrogenases (ACDs) as the proteins associated with IRAP. The association was nearly abolished by mutation of the dileucine motif of IRAP. Immunoblotting of fractions prepared from sucrose gradient ultracentrifugation and vesicles immunopurified with anti-GLUT4 antibody revealed these ACDs to be localized on GLUT4-containing vesicles. Furthermore, 3-mercaptopropionic acid and hexanoyl-CoA, inhibitors of long-chain and medium-chain ACDs, respectively, induced dissociation of long-chain acyl-coenzyme A dehydrogenase and/or medium-chain acyl-coenzyme A dehydrogenase from IRAP in vitro as well as recruitment of GLUT4 to the plasma membrane and stimulation of glucose transport activity in permeabilized 3T3-L1 adipocytes. These findings suggest that ACDs are localized on GLUT4-containing vesicles via association with IRAP in a manner dependent on its dileucine motif and play a role in retention of GLUT4-containing vesicles to an intracellular compartment.


[Laparoscopic Proximal Gastrectomy as a Surgical Treatment for Upper Third Early Gastric Cancer].
Recently, the incidence of upper third gastric cancer has increased, and with it the number of endoscopic submucosal dissection (ESD) procedures performed has been increasing. However, if ESD is not indicated or non-curable, surgical treatment may be necessary. In the case of lower third gastric cancer, it is possible to preserve the upper part of the stomach; however, in the case of upper third gastric cancer, total gastrectomy is still the standard treatment option, regardless of the stage. This is due to the complications associated with upper third gastric cancer, such as gastroesophageal reflux after proximal gastrectomy rather than oncologic problems. Recently, the introduction of the double tract reconstruction method after proximal gastrectomy has become one of the surgical treatment methods for upper third early gastric cancer. However, since there has not been a prospective comparative study evaluating its efficacy, the ongoing multicenter prospective randomized controlled trial (KLASS-05) comparing laparoscopic proximal gastrectomy with double tract reconstruction and laparoscopic total gastrectomy is expected to be important for determining the future of treatment of upper third early gastric cancer.


Dendrimeric peptides can confer protection against foot-and-mouth disease virus in cattle.
Foot-and-mouth disease virus (FMDV) causes a highly contagious disease in cloven-hoofed animals. A synthetic vaccine candidate consisting of dendrimeric peptides harbouring two copies of a B-epitope [VP1(136-154)] linked to a T-cell epitope [3A(21-35)] of FMDV confers protection to type O FMDV challenge in pigs. Herein we show in cattle that novel dendrimeric peptides bearing a T-cell epitope [VP1(21-40] and two or four copies of a B-cell epitope [VP1(135-160)] from type O1 Campos FMDV (termed B2T and B4T, respectively) elicited FMDV specific immune responses to similar levels to a commercial vaccine. Animals were challenged with FMDV and 100% of vaccinated cattle with B2T or B4T were protected to podal generalization. Moreover, bovines immunized with B4T were completely protected (with no clinical signs) against FMDV challenge after three vaccine doses, which was associated with titers of viral neutralizing antibodies in serum higher than those of B2T group (p< 0.05) and levels of opsonic antibodies similar to those of animals immunized with one dose of FMDV commercial vaccine. Bovines vaccinated with both dendrimeric peptides presented high levels of IgG1 anti FMDV in sera and in mucosa. When IgA in nasal secretions was measured, 20% or 40% of the animals in B2T or B4T groups respectively, showed anti-FMDV IgA titers. In addition, B2T and B4T peptides evoked similar consistent T cell responses, being recognized in vitro by lymphocytes from most of the immunized cattle in the proliferation assay, and from all animals in the IFN-γ production assay. Taken together, these results support the potential of dendrimers B2T or B4T in cattle as a highly valuable, cost-effective FMDV candidate vaccine with DIVA potential.


Pathogenic features of CD4+CD28- T cells in immune disorders.
Aging of the immune system contributes to the increased morbidity and mortality of the elderly population and may occur prematurely in patients with immune disorders. One of the main characteristics of immunosenescence is the expansion of CD4(+)CD28(-) T cells in the blood. These cells are effector memory T cells with cytotoxic capacity, and have been recently described to have pathogenic potential in a variety of immune disorders. Interestingly, CD4(+)CD28(-) T cells have now been found to infiltrate target tissues of patients with multiple sclerosis, rheumatoid arthritis, myopathies, acute coronary syndromes, and other immune-related diseases. In this review, we discuss potential factors and mechanisms that may induce the expansion of these cells, as well as their putative pathogenic mechanisms in immune disorders.


Integrated analyses of long noncoding RNAs and mRNAs in the progression of breast cancer.
The objective was to explore the expression and potential functions of long noncoding RNA (lncRNA) and mRNAs in human breast cancer (BC). Differentially expressed lncRNAs and mRNAs were identified and annotated in BC tissues by using the Agilent human lncRNA assay (Agilent Technologies, Santa Clara, CA, USA) and RNA sequencing. After identification of lncRNAs and mRNAs through quantitative reverse transcription polymerase chain reaction, we conducted a series of functional experiments to confirm the effects of knockdown of one lncRNA, TCONS_00029809, on the progression of BC. We discovered 238 lncRNAs and 200 mRNAs that were differentially expressed in BC tissues and para-carcinoma tissue. We showed that differentially expressed mRNAs were related to biological adhesion and biological regulation and mainly enriched in cytokine-cytokine receptor interaction, metabolic pathways, and PI3K-Akt signaling pathway. We created a protein-protein interaction network to analyze the proteins enriched in these pathways. We demonstrated that silencing of TCONS_00029809 remarkably inhibited proliferation, invasion, and migration of BC cells, and accelerated their apoptosis. We identified a large number of differentially expressed lncRNAs and mRNAs, which provide data useful in understanding BC carcinogenesis. The lncRNA TCONS_00029809 may be involved in the development of BC.


Trauma appraisals, emotion regulation difficulties, and self-compassion predict posttraumatic stress symptoms following childhood abuse.
Experiencing traumatic events and abuse is unfortunately common in general, non-clinical samples. Recent research indicates that the ways in which individuals interpret traumatic experiences, as well as the ways that they manage challenging emotions in general, may statistically predict post-traumatic stress disorder (PTSD) symptoms to a greater extent than does trauma itself. Negative trauma appraisals, generalized emotion regulation (ER) difficulties, and low levels of self-compassion have each been shown to influence the connection between trauma exposure and subsequent PTSD symptoms. However, little is known regarding how these processes interact, or their relative contributions to mental health after trauma. The current study analyzed data from 466 university students who completed self-report measures of childhood abuse, PTSD symptoms, trauma appraisals, ER difficulties, and self-compassion. Childhood abuse exposure and PTSD symptoms were positively associated with negative trauma appraisals and ER difficulties, and negatively associated with self-compassion. Self-compassion was inversely associated with negative trauma appraisals and ER difficulties. Multiple mediation analyses demonstrated that negative trauma appraisals, ER difficulties, and levels of self-compassion fully explained the link between abuse exposure and PTSD symptoms via several specific pathways. These findings suggest that researchers, clinicians, and abuse survivors can benefit from addressing these interconnected domains during treatment and recovery processes.


Functional Characterization of Primordial Protein Repair Enzyme M38 Metallo-Peptidase From Fervidobacterium islandicum AW-1.
The NA23_RS08100 gene of Fervidobacterium islandicum AW-1 encodes a keratin-degrading β-aspartyl peptidase (FiBAP) that is highly expressed under starvation conditions. Herein, we expressed the gene in Escherichia coli, purified the recombinant enzyme to homogeneity, and investigated its function. The 318 kDa recombinant FiBAP enzyme exhibited maximal activity at 80°C and pH 7.0 in the presence of Zn2+. Size-exclusion chromatography revealed that the native enzyme is an octamer comprising a tetramer of dimers; this was further supported by determination of its crystal structure at 2.6 Å resolution. Consistently, the structure of FiBAP revealed three additional salt bridges in each dimer, involving 12 ionic interactions that might contribute to its high thermostability. In addition, the co-crystal structure containing the substrate analog N-carbobenzoxy-β-Asp-Leu at 2.7 Å resolution revealed binuclear Zn2+-mediated substrate binding, suggesting that FiBAP is a hyperthermophilic type-I IadA, in accordance with sequence-based phylogenetic analysis. Indeed, complementation of a Leu auxotrophic E. coli mutant strain (ΔiadA and ΔleuB) with FiBAP enabled the mutant strain to grow on isoAsp-Leu peptides. Remarkably, LC-MS/MS analysis of soluble keratin hydrolysates revealed that FiBAP not only cleaves the C-terminus of isoAsp residues but also has a relatively broad substrate specificity toward α-peptide bonds. Moreover, heat shock-induced protein aggregates retarded bacterial growth, but expression of BAP alleviated the growth defect by degrading damaged proteins. Taken together, these results suggest that the viability of hyperthermophiles under stressful conditions may rely on the activity of BAP within cellular protein repair systems.


Computational modelling of phonological dyslexia: how does the DRC model fare?
This paper investigates the patterns of reading impairment in phonological dyslexia using computational modelling with the dual-route cascaded model of reading (DRC, Coltheart, Rastle, Perry, Langdon, & Ziegler, 2001). Systematic lesioning of nonlexical and phonological processes in DRC demonstrates that different lesions and severity of those lesions can reproduce features of phonological dyslexia including impaired reading of nonwords, relatively spared reading of words, an advantage for reading pseudohomophones. Using the same stimuli for model and for patients, lesions to DRC were also used to simulate the reading accuracy shown by three individuals with acquired phonological dyslexia. No single lesion could replicate the reading performance of all three individuals. In order to simulate reading accuracy for one individual a phonological impairment was necessary (addition of noise to the phoneme units), and for the remaining two individuals an impairment to nonlexical reading procedures (increasing the time interval between each new letter being processed) was necessary. We argue that no single locus of impairment (neither phonological nor nonlexical) can account for the reading impairments of all individuals with phonological dyslexia. Instead, different individuals have different impairments (and combinations of impairments) that together provide the spectrum of patterns found in phonological dyslexia.


Venous thrombosis in children.
Venous thromboembolic (VTE) events are being increasingly diagnosed in systemic and cerebral vessels in children. Systemic VTE are increasing in children as a result of therapeutic advances and improved clinical acumen in primary illnesses that previously caused mortality. The epidemiology of systemic VTE has been studied in international registries. In children older than 3 months, teenagers are the largest group developing VTE. The most common etiologic factor is the presence of central venous lines. Clinical studies have determined the most sensitive diagnostic method for diagnosing upper system VTE are ultrasound for jugular venous thrombosis and venography for intrathoracic vessels. However, the most sensitive diagnostic methods for lower system VTE and pulmonary embolism (PE) have not been established. Treatment studies for VTE consist of inadequately powered randomized controlled trials or prospective cohort studies. The long-term outcome of systemic VTE, post-thrombotic syndrome, has been reported in children. Cerebral sinovenous thrombosis (CSVT) is becoming increasingly diagnosed in children due to the recognition of the associated subtle clinical symptoms and improved cerebrovascular imaging. The etiology of CSVT includes thrombophilia, head and neck infections, and systemic illness. Estimates of the incidence and outcome of childhood CSVT have recently become available through the Canadian Pediatric Ischaemic Stroke Registry. Clinical studies have not yet been carried out in children to determine the best method of diagnosis or treatment. There have only been case-series studies carried out in the treatment of CSVT. Properly designed clinical trials are urgently required in children with systemic VTE/PE and CSVT to define the best methods of diagnosis, treatment and long-term management.


CA9 expression as a prognostic factor in renal clear cell carcinoma.
We investigated whether CA9 protein could be used as a prognostic tumor marker as well as a diagnostic biomarker in renal clear cell carcinoma. Nephrectomy specimens from 92 patients were used in this study. 80 of these were renal cell carcinomas, 10 adenomas and 2 oncocytomas. Of the renal cell carcinomas, 67 were clear cell carcinomas. Immunohistochemical analysis using CA9 monoclonal antibody (M75) was performed on paraffin-embedded specimens. CA9 staining was correlated with tumor stage, grade, lymph node involvement, distant metastasis and cumulative survival time. CA9 was present in 91.2% of clear cell carcinomas. Low staining was a poor prognostic factor, and conversely high staining a good prognostic one. CA9 expression was found to be the best prognostic factor when compared with T stage and grade. Even in low-grade and stage tumors, the presence of low expression correlated with lowered survival times. On the basis of our study, CA9 is a significant molecular marker in renal clear cell carcinomas. Decreased CA9 expression is independently associated with poor survival. CA9 can be used to predict clinical outcome and identify high-risk patients in need for adjuvant immunotherapy and CA9 targeted therapies.


Net recharge vs. depth to groundwater relationship in the Platte River Valley of Nebraska, United States.
One-km resolution MODIS-based mean annual evapotranspiration (ET) estimates in combination with PRISM precipitation rates were correlated with depth to groundwater (d) values in the wide alluvial valley of the Platte River in Nebraska for obtaining a net recharge (Rn) vs. d relationship. MODIS cells with irrigation were excluded, yielding a mixture of predominantly range, pasture, grass, and riparian forest covers on sandy soils with a shallow groundwater table. The transition depth (dt ) between negative and positive values of the net groundwater recharge was found to be at about 2 (±1) m. Within 1 (±1) m of the surface and at a depth larger than about 7 to 8 (±1) m, the mean annual net recharge became independent of d at a level of about -4 (±12)% and 13 (±10)%, respectively, of the mean annual precipitation rate. The obtained Rn(d) relationship is based on a calibration-free ET estimation method and may help in obtaining the net recharge in shallow groundwater areas of negligible surface runoff where sufficient groundwater-depth data exist.


Preparticipation sports examination of the child and adolescent athlete: changing views of an old ritual.
The role of the pediatrician in the management of the child athlete has expanded in accord with the burgeoning number of children involved in sports competition. Growing information regarding physiologic responses to exercise in the pediatric age group has added to the importance and utility of this involvement. The scope of the preparticipation evaluation has enlarged in response to these changes; newer issues such as injury prevention, body composition assessment, and athlete education become important in not only assuring safe participation but also optimizing athletic performance. At the same time, the low yield of important findings during these examinations has prompted a reassessment of when, where, and by whom they might be most efficiently conducted.


Low reversed cyclic loading tests for integrated precast structure of lightweight wall with single-row reinforcement under a lightweight steel frame.
Given the development of precast structures for low-rise residential buildings, this study explores a new structure-namely, an integrated precast structure of lightweight recycled concrete wall with single-row reinforcement-under a lightweight steel frame filled with recycled concrete (integrated precast structure for short). The lightweight steel frame and lightweight wall cooperate to bear the forces. The applied concealed bracing, either a rebar bracing or a steel plate bracing, increases the shear resistance of the wall. The lightweight steel frame is designed to bear the vertical loading, whereas the seismic load in the horizontal direction is jointly borne by the frame and wall. This study presents the results of low reversed cyclic loading tests on nine specimens of integrated precast structures. An analysis is then carried out to investigate the mechanical properties of the specimens; based on these results, a formula for the force-bearing performance of the inclined section is developed. The results show satisfactory performance as an integrated piece; the proposed structure has two seismic lines of defence, with the lightweight wall restraint by the side frame being the first line and the steel frame being the second line. Because the failure of the wall can be categorized as shear failure, the restraint of the lightweight steel frame significantly reduces the potential damage of the wall. As the beams and columns of the steel frame tend to bend against failure, the wall filling helps resist sliding. Therefore, the reinforced joints of the connecting beams and columns show no visible signs of damage, indicating that the connection between the beams and columns is reliable. The narrow spacing of rebars and the setting of concealed bracing contribute to the increase in ductility and energy efficiency of the integrated structure and the evident reduction in the failure process. Furthermore, the recycled concrete increases the seismic resistance of the structure.


Identification of selective norbornane-type aspartate analogue inhibitors of the glutamate transporter 1 (GLT-1) from the chemical universe generated database (GDB).
A variety of conformationally constrained aspartate and glutamate analogues inhibit the glutamate transporter 1 (GLT-1, also known as EAAT2). To expand the search for such analogues, a virtual library of aliphatic aspartate and glutamate analogues was generated starting from the chemical universe database GDB-11, which contains 26.4 million possible molecules up to 11 atoms of C, N, O, F, resulting in 101026 aspartate analogues and 151285 glutamate analogues. Virtual screening was realized by high-throughput docking to the glutamate binding site of the glutamate transporter homologue from Pyrococcus horikoshii (PDB code: 1XFH ) using Autodock. Norbornane-type aspartate analogues were selected from the top-scoring virtual hits and synthesized. Testing and optimization led to the identification of (1R*,2R*,3S*,4R*,6R*)-2-amino-6-phenethyl-bicyclo[2.2.1]heptane-2,3-dicarboxylic acid as a new inhibitor of GLT-1 with IC(50) = 1.4 μM against GLT-1 and no inhibition of the related transporter EAAC1. The systematic diversification of known ligands by enumeration with help of GDB followed by virtual screening, synthesis, and testing as exemplified here provides a general strategy for drug discovery.


Nucleotide sequence of the human placental alkaline phosphatase gene. Evolution of the 5' flanking region by deletion/substitution.
Three closely related alkaline phosphatase (ALP) genes reside on the long arm of chromosome 2 in man. One of these genes (the placental ALP-1) encodes the classic heat-stable placental alkaline phosphatase. Another gene (the placental ALP-2) is closely related to the placental ALP-1 and may encode the so-called placental ALP-like enzyme of the testis and thymus. The third member of this gene family (the intestinal ALP gene) encodes the intestinal alkaline phosphatase. The expression of the placental ALP-1 and intestinal ALP genes is highly tissue-specific in spite of nearly 90% sequence similarity within their exons. To help determine the basis for this tissue specificity, the nucleotide sequence of the placental ALP-1 gene and some of its 5' flanking region has been determined and analyzed by comparison with placental ALP-2 and intestinal ALP gene sequences. The placental ALP-1 gene transcription unit has 4087 bases between the major cap site and the most distal of several reported 3' ends. The protein coding region is divided by 10 short introns varying in size from 74 to 241 nucleotides. Three of these introns bisect regions of the gene that encode residues conserved between the active site of the Escherichia coli enzyme and the human placental ALP. This result suggests that the human alkaline phosphatase genes have evolved in an intron-independent fashion. A comparison of the placental ALP-1 5' flanking sequence (up to -540) with the analogous sequence of the intestinal ALP gene revealed several deletion/substitutions which could be important in determining the tissue-specific expression of these genes.


The expression of NADPH oxidases and production of reactive oxygen species by human lung adenocarcinoma epithelial cell line A549.
Controlled production of reactive oxygen species (ROS) by NADPH oxidases in non-phagocytic cells has recently been suggested to participate in the regulation of cellular functions. Due to the role of ROS in control of cellular functions, precise and accurate detection of ROS is of essential importance. However, various methodological approaches currently used for ROS determination vary in sensitivity, specificity, as well as in requirements for specialized equipment. In this study, human lung epithelial cell line A549 was screened for expression of NADPH oxidases NOX1, NOX2, NOX4, NOX5, DUOX1 and DUOX2 by quantitative RT-PCR. Fluorometric, colorimetric, and chemiluminometric methods were applied to determine ROS production. A549 cells were found to significantly express NOX1, NOX2, DUOX1 and DUOX2. ROS production by A549 cells was detected with fluorometric probes 2',7'-dichlorofluorescein- diacetate, dihydroethidium, and amplex red or colorimetric probe nitrobluetetrazolium. The production of ROS detected by these probes was partially reduced by NADPH oxidase inhibitor diphenyleneiodonium. The inhibitory effect of diphenyleneiodonium was the most significant regarding amplex red detection of phorbol myristate acetateactivated ROS production. In contrast to other probes, neither cytochrome c colorimetric determination nor luminol- and L-012-amplified chemiluminescence, regardless of the addition of horseradish peroxidase, exerted sufficient sensitivity to detect ROS production by A549. The results revealed differences among methods used for ROS formation measurement by human lung epithelial cell line A549 and highlighted the sensitivity of fluorometric determination for this purpose.


[Influence of drugs on the effect of antitumor preparations in experiments and in clinic medicine].
Experimental and clinical data concerning the interaction of cytostatics with diuretics, anticoagulants, neurotropic drugs, prostaglandin inhibitors, etc. are reviewed. It is considered necessary to take into account the possible clinical effect when applying different drugs for treatment of the tumour patients. Side by side with intensification of the antitumour activity of the cytostatic or with a decrease of its toxic effect on haemopoiesis and on the function of parenchymatous organs some undesirable effects such as an increase of the toxicity or a decrease of the antitumour effect are possible. Interaction of cytostatics with different groups of drugs demands further study taking into account the "medicinal background" of the modern population.


A series of 18F-labelled pyridinylphenyl amides as subtype-selective radioligands for the dopamine D3 receptor.
Synthesis, biological activity, and structure-selectivity relationship (SSR) studies of a novel series of potential dopamine D3 receptor radioligands as imaging agents for positron emission tomography (PET) are reported. Considering a structurally diverse library of D3 ligands, SSR studies were performed for a new series of fluorinated pyridinylphenyl amides using CoMFA and CoMSIA methods. The in vitro D3 affinities of the predicted series of biphenyl amide ligands 9 a-d revealed single-digit to sub-nanomolar potencies (K(i)=0.52-1.6 nM), displaying excellent D3 selectivity over the D2 subtype of 110- to 210-fold for the test compounds 9 a-c. Radiofluorination by nucleophilic substitution of Br or NO(2) by (18)F led to radiochemical yields of 66-92 % for [(18)F]9 a-d. However, the specific activities of [(18)F]9 b and [(18)F]9 d were insufficient, rendering their use for in vivo studies impossible. Biodistribution studies of [(18)F]9 a and [(18)F]9 c using rat brain autoradiography revealed accumulation in the ventricles, thus indicating insufficient biokinetic properties of [(18)F]9 a and [(18)F]9 c for D3 receptor imaging in vivo.


Sustained swimming increases the mineral content and osteocyte density of salmon vertebral bone.
This study addresses the effects of increased mechanical load on the vertebral bone of post-smolt Atlantic salmon by forcing them to swim at controlled speeds. The fish swam continuously in four circular tanks for 9 weeks, two groups at 0.47 body lengths (bl) × s(-1) (non-exercised group) and two groups at 2 bl × s(-1) (exercised group), which is just below the limit for maximum sustained swimming speed in this species. Qualitative data concerning the vertebral structure were obtained from histology and electron microscopy, and quantitative data were based on histomorphometry, high-resolution X-ray micro-computed tomography images and analysis of bone mineral content, while the mechanical properties were tested by compression. Our key findings are that the bone matrix secreted during sustained swimming had significantly higher mineral content and mechanical strength, while no effect was detected on bone in vivo architecture. mRNA levels for two mineralization-related genes bgp and alp were significantly upregulated in the exercised fish, indicating promotion of mineralization. The osteocyte density of the lamellar bone of the amphicoel was also significantly higher in the exercised than non-exercised fish, while the osteocyte density in the cancellous bone was similar in the two groups. The vertebral osteocytes did not form a functional syncytium, which shows that salmon vertebral bone responds to mechanical loading in the absence of an extensive connecting syncytial network of osteocytic cell processes as found in mammals, indicating the existence of a different mechanosensing mechanism. The adaptive response to increased load is thus probably mediated by osteoblasts or bone lining cells, a system in which signal detection and response may be co-located. This study offers new insight into the teleost bone biology, and may have implications for maintaining acceptable welfare for farmed salmon.


New perspectives in antibiotic prophylaxis for intra-abdominal surgery.
Effective management of intra-abdominal infections requires a combination of preoperative preparation, antibiotic prophylaxis and appropriate surgical technique. Antibacterial prophylaxis should provide coverage of all likely pathogens, including aerobic and anaerobic organisms. Whereas antibacterial combination therapy is appropriate in certain situations, single-agent prophylaxis is appropriate for the majority of patients and ampicillin/sulbactam, with its broad-spectrum anti-aerobic/anti-anaerobic activity, is an attractive prophylactic option. Surgery involving the gastrointestinal tract provides a special challenge by virtue of its high, predominantly anaerobic, bacterial load. However, the requirement for prophylaxis varies depending upon the precise site of intervention. Biliary tract surgery requires prophylaxis in high-risk patients only, whereas hepatobiliary or pancreatic surgery requires prophylaxis in all patients. Gastroduodenal operations require prophylaxis in the presence of risk factors, such as abnormal gastric acidity or bleeding. Colorectal procedures present a high risk of anaerobic infection and sepsis, and require adequate prophylaxis combined with a thorough preoperative preparation designed to reduce considerably the bacterial load of the bowel. Where peritonitis does follow intra-abdominal surgery, patients should receive antibacterial therapy commensurate with the risk of serious infection. A small proportion of patients will be at risk of severe infection and will require triple-agent therapy. However, most patients are likely to develop mild-to-moderate infections only and can be treated with a single, broad-spectrum antibiotic agent, such as ampicillin/sulbactam, a beta-lactam/beta-lactamase inhibitor.


The role of cytology in the diagnosis and follow-up of patients with cervical adenocarcinoma.
A review was undertaken of the 121 patients with cervical adenocarcinoma, ranging in age from 10 to 91 years, who had cytology specimens examined in this laboratory. All but two had cervicovaginal smears. Fifty-eight patients had cervical smears taken prior to treatment; 91% of these contained malignant cells. The commonest histologic subtype was mucinous adenocarcinoma (61 patients; 52%), followed by endometrioid carcinoma (26 cases; 22%), adenosquamous carcinoma (16 cases; 14%) and clear-cell carcinoma (12 cases; 10%). The cytologic characteristics were evaluated and correlated with the histopathology. During the follow-up period after definitive treatment, 119 patients had gynecologic smears taken; 24% were positive, representing 64% of those cases with central recurrences. In two patients, the abnormal smear was the first indicator of recurrence. Nine patients had 11 nongynecologic specimens revealing metastatic disease. Notable in this study were the 18 asymptomatic women, of whom 15 also had either no visible cervical lesion or minimal changes associated with benign conditions; cytology was abnormal in all but two. Cytology was effective in detecting disease. Its usefulness continued during the follow-up period in identifying or confirming recurrent or metastatic disease.


Distribution and evolution of the lectin family in soybean (Glycine max).
Lectins are a diverse group of proteins that bind specific carbohydrates and are found throughout all kingdoms. In plants, lectins are involved in a range of important processes such as plant defense and stress signaling. Although the genome sequence of Glycine max (soybean) has been published, little is known about the abundance and expansion patterns of lectin genes in soybean. Using BLAST and hidden Markov models, a total of 359 putative lectin genes have been identified. Furthermore, these sequences could be classified in nine of the twelve plant lectin families identified today. Analysis of the domain organization demonstrated that most of the identified lectin genes encode chimerolectins, consisting of one or multiple lectin domains combined with other known protein domains. Both tandem and segmental duplication events have contributed to the expansion of the lectin gene family. These data provide a detailed understanding of the domain architecture and molecular evolution of the lectin gene family in soybean.


Morphological study of lumbricals - a cadaveric study.
The apparently simple human function of closing the hand to grasp an object or opening the palm to release it are in reality tasks of considerable mechanical complexity requiring the contraction of many individual muscles Lumbricals are important part of intrinsic musculature of hand. Though several deviations from normal have been described in standard textbooks of anatomy, a review of basic anatomy of lumbricals will be helpful. Proper knowledge about anatomy and variaitions prevalent in intrinsic musculature of hand is mandatory for various surgical procedure like cleft hand surgery, pollicization & other digital transposition procedures Present study was conducted to study morphology & variations prevalent in this region. The results were compiled and tried to find out commonest variations Phylogenetical & ontogenetical basis and clinical applications related to variations encountered were tried to know by correlating it with available literature. For this study 50 hands of formalin embalmed cadavers were dissected The limbs were labelled R OR L for right or left, M or F for male or female. Cunnigham's manual for dissection was followed for dissection. Origin and nerve supply of lumbricals was normal in all hands. The variations were encountered in insertion of the 3rd lumbrical (32%) and 4th lumbrical (24%) were more confined to the right hand. Variations encountered commonly were split and misplaced insertion of third & fourth lumbricals. These variations are critically analyzed in the light of accessible literature. Lumbricals are more variable in the insertion rather than the origin & nerve supply. The variation commonly seen were split & misplaced insertion Split insertion was common in the third lumbrical whereas misplaced insertion is more common in the fourth lumbrical Variations are more common on right hands.


Synthetic FLAIR as a Substitute for FLAIR Sequence in Acute Ischemic Stroke.
Background In acute ischemic stroke (AIS), fluid-attenuated inversion recovery (FLAIR) is used for treatment decisions when onset time is unknown. Synthetic FLAIR could be generated with deep learning from information embedded in diffusion-weighted imaging (DWI) and could replace acquired FLAIR sequence (real FLAIR) and shorten MRI duration. Purpose To compare performance of synthetic and real FLAIR for DWI-FLAIR mismatch estimation and identification of patients presenting within 4.5 hours from symptom onset. Materials and Methods In this retrospective study, all pretreatment and early follow-up (<48 hours after symptom onset) MRI data sets including DWI (b = 0-1000 sec/mm2) and FLAIR sequences obtained in consecutive patients with AIS referred for reperfusion therapies between January 2002 and May 2019 were included. On the training set (80%), a generative adversarial network was trained to produce synthetic FLAIR with DWI as input. On the test set (20%), synthetic FLAIR was computed without real FLAIR knowledge. The DWI-FLAIR mismatch was evaluated on both FLAIR data sets by four independent readers. Interobserver reproducibility and DWI-FLAIR mismatch concordance between synthetic and real FLAIR were evaluated with κ statistics. Sensitivity and specificity for identification of AIS within 4.5 hours were compared in patients with known onset time by using McNemar test. Results The study included 1416 MRI scans (861 patients; median age, 71 years [interquartile range, 57-81 years]; 375 men), yielding 1134 and 282 scans for training and test sets, respectively. Regarding DWI-FLAIR mismatch, interobserver reproducibility was substantial for real and synthetic FLAIR (κ = 0.80 [95% CI: 0.74, 0.87] and 0.80 [95% CI: 0.74, 0.87], respectively). After consensus, concordance between real and synthetic FLAIR was almost perfect (κ = 0.88; 95% CI: 0.82, 0.93). Diagnostic value for identifying AIS within 4.5 hours did not differ between real and synthetic FLAIR (sensitivity: 107 of 131 [82%] vs 111 of 131 [85%], P = .2; specificity: 96 of 104 [92%] vs 96 of 104 [92%], respectively, P > .99). Conclusion Synthetic fluid-attenuated inversion recovery (FLAIR) had diagnostic performances similar to real FLAIR in depicting diffusion-weighted imaging-FLAIR mismatch and in helping to identify early acute ischemic stroke, and it may accelerate MRI protocols. © RSNA, 2022 Online supplemental material is available for this article. See also the editorial by Carroll and Hurley in this issue.


The assembly of individual chaplin peptides from Streptomyces coelicolor into functional amyloid fibrils.
The self-association of proteins into amyloid fibrils offers an alternative to the natively folded state of many polypeptides. Although commonly associated with disease, amyloid fibrils represent the natural functional state of some proteins, such as the chaplins from the soil-dwelling bacterium Streptomyces coelicolor, which coat the aerial mycelium and spores rendering them hydrophobic. We have undertaken a biophysical characterisation of the five short chaplin peptides ChpD-H to probe the mechanism by which these peptides self-assemble in solution to form fibrils. Each of the five chaplin peptides produced synthetically or isolated from the cell wall is individually surface-active and capable of forming fibrils under a range of solution conditions in vitro. These fibrils contain a highly similar cross-β core structure and a secondary structure that resembles fibrils formed in vivo on the spore and mycelium surface. They can also restore the growth of aerial hyphae to a chaplin mutant strain. We show that cysteine residues are not required for fibril formation in vitro and propose a role for the cysteine residues conserved in four of the five short chaplin peptides.


Personal sun exposure and serum 25-hydroxy vitamin D concentrations.
Solar ultraviolet-B radiation (UVB) is essential for epidermal vitamin D production. We aimed to quantitate the relationship between personal solar UV exposure and serum 25hydroxy vitamin D (25[OH]D) concentration. Blood was collected for 25(OH)D analysis in 207 South Australian adults aged 27-61 years. At the time of blood collection, each participant completed a questionnaire, which included a calendar for recall of sun exposure in the preceding 16 weeks. We examined the association between solar UV exposure and serum 25(OH)D graphically from smoothed scatter plots, and modeled it using multiple linear regression, with age, sex and body mass index as covariates. Estimated erythemal solar UV exposure in the 6 weeks before blood collection best predicted serum 25(OH)D concentrations. Serum 25(OH)D rose with increasing personal solar UV exposure to a maximum of about 89 nmol L(-1) at an estimated mean weekly solar erythemal UV exposure of about 1230 mJ cm(-2). The maximum was the same after accounting for clothing coverage and was reached at an estimated whole body equivalent exposure to ambient UV of ca 700 mJ cm(-2). These results suggest that an average maximum serum 25(OH)D of ca 89 nmol L(-1) is achieved from sun exposure in a healthy Australian adult population.


Binding of G-CSF, GM-CSF, tumor necrosis factor-alpha, and gamma-interferon to cell surface receptors on human myeloid leukemia cells triggers rapid tyrosine and serine phosphorylation of a 75-Kd protein.
Granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), gamma-interferon (gamma-IFN), or tumor necrosis factor-alpha (TNF-alpha) triggered the rapid, stable phosphorylation of a 75-Kd protein (p75) when incubated with permeabilized HL60 human myeloid leukemia cells in the presence of [gamma-32P] ATP. Among several chemical inducers of HL60 cell differentiation, dimethyl sulfoxide also triggered p75 labeling, but retinoic acid or 12-O-tetradecanoylphorbol-13-acetate did not elicit this response. Pretreatment of cells with G-CSF or GM-CSF for more than 30 seconds before permeabilization rendered the p75 labeling undetectable, suggesting that ligand-stimulated labeling was rapidly completed within this time in intact cells. Phosphorylation of p75 occurred on serine and tyrosine residues. This conclusion was confirmed by direct phosphoamino acid analysis. Immunoblot analysis of lysates of intact HL60 cells that had been incubated with G-CSF, GM-CSF, IFN, or TNF confirmed that tyrosine phosphorylation of a p75 also occurred in response to these cytokines in intact cells. Pretreatment of intact HL60 cells with one biologic agent or dimethyl sulfoxide abolished p75 labeling in response to incubation of permeabilized cells with a second agent, strongly suggesting that the same protein was phosphorylated in response to these treatments. p75 labeling was strictly dependent on expression of the appropriate ligand receptor. Data suggest that activation of a tyrosine kinase system is an early response to the binding of G-CSF, GM-CSF, TNF, or IFN to their respective cell surface receptors, or to the addition of dimethyl sulfoxide, and that the resulting phosphorylation event(s) may play a role in securing common elements in the biologic responses to these agents.


Leaf structure in relation to solute transport and phloem loading in Zea mays L.
Small and intermediate (longitudinal) vascular bundles of the Zea mays leaf are surrounded by chlorenchymatous bundle sheaths and consist of one or two vessels, variable numbers of vascular parenchyma cells, and two or more sieve tubes some of which are associated with companion cells. Sieve tubes not associated with companion cells have relatively thick walls and commonly are in direct contact with the vessels. The thick-walled sieve tubes have abundant cytoplasmic connections with contiguous vascular parenchyma cells; in contrast, connections between vascular parenchyma cells and thin-walled sieve tubes are rare. Connections are abundant, however, between the thin-walled sieve tubes and their companion cells; the latter have few connections with the vascular parenchyma cells. Plasmolytic studies on leaves of plants taken directly from lighted growth chambers gave osmotic potential values of about-18 bars for the companion cells and thin-walled sieve tubes (the companion cell-sieve tube complexes) and about-11 bars for the vascular parenchyma cells. Judging from the distribution of connections between various cell types of the vascular bundles and from the osmotic potential values of those cell types, it appears that sugar is actively accumulated from the apoplast by the companion cell-sieve tube complex, probably across the plasmalemma of the companion cell. The thick-walled sieve tubes, with their close spatial association with the vessels and possession of plasmalemma tubules, may play a role in retrieval of solutes entering the leaf apoplast in the transpiration stream. The transverse veins have chlorenchymatous bundle sheaths and commonly contain a single vessel and sieve tube. Parenchymatic elements may or may not be present. Like the thick-walled sieve tubes of the longitudinal bundles, the sieve tubes of the transverse veins have plasmalemma tubules, indicating that they too may play a role in retrieval of solutes entering the leaf apoplast in the transpiration stream.


Molecular morphological approach to the pathological study of development and advancement of human breast cancer.
Since the concept of gene profile-based intrinsic subtypes was proposed, various studies on pathological characteristics have been performed. Particularly, triplenegative (TN) breast cancer, which is negative for all hormone receptors [estrogen receptor (ER) and/or progesterone receptor (PgR) and human epidermal growth factor 2 (HER2)], has been attracting attention because effects of endocrine and targeting therapies cannot be anticipated and thus selecting a treatment method is difficult. TN cancer accounts for about 10%-15% of all invasive breast cancer cases in Japanese, which is significantly lower than the incidence reported in the United States. Cytokeratin (CK) 5/6 or epidermal growth factor receptor (EGFR) is positive in 80%, being classified as basal-like carcinoma, but it should be understood that TN breast cancer and basal-like carcinoma are not necessarily the same. Criteria for positivity judgment of ER, PgR, and HER2 were established to select treatment in cases positive for each marker, and greater importance is attached to strict accuracy control. Inversely, the level of negative findings to judge TN varies among the judgment criteria. In any case, the prognosis of TN breast cancer is poor. Pathologically, TN breast cancer shows certain morphological characteristics, such as high grade and a pushing margin, and abnormalities of BRCA1 and p53 are frequently noted. At present, as no effective therapeutic strategy has been established for TN breast cancer, further clarification of the molecular biological characteristics of such cancers is needed. In addition, the incidence of TN-type ductal carcinoma in situ (DCIS) is low, suggesting that TN does not remain preinvasive DCIS for a prolonged period and that it transforms to invasive cancer in an early stage. Because mammary gland basal cells have characters of progenitor or stem cells that differentiate into both luminal epithelium and myoepithelial cells, these cells may be utilized for the differential diagnosis of the benignity or malignancy of intraductal lesions in routine pathological practice. As proliferation markers, such as Ki-67, and multiple gene arrays for gene signature are also utilized to select adjuvant therapy, analysis may progress further in the future.


Multistep prediction of physiological tremor for surgical robotics applications.
Accurate canceling of physiological tremor is extremely important in robotics-assisted surgical instruments/procedures. The performance of robotics-based hand-held surgical devices degrades in real time due to the presence of phase delay in sensors (hardware) and filtering (software) processes. Effective tremor compensation requires zero-phase lag in filtering process so that the filtered tremor signal can be used to regenerate an opposing motion in real time. Delay as small as 20 ms degrades the performance of human-machine interference. To overcome this phase delay, we employ multistep prediction in this paper. Combined with the existing tremor estimation methods, the procedure improves the overall accuracy by 60% for tremor estimation compared to single-step prediction methods in the presence of phase delay. Experimental results with developed methods for 1-DOF tremor estimation highlight the improvement.


Early depressive symptoms in cancer patients receiving interleukin 2 and/or interferon alfa-2b therapy.
Depressive symptomatology is frequently associated with interleukin (IL)-2 and interferon alfa-2b (INFalpha-2b) therapy in cancer patients. The objective of the present study was to evaluate the depressive and anxiety symptoms induced by IL-2 and/or INFalpha-2b in cancer patients during the first days of cytokine immunotherapy. The study included 48 patients with renal cell carcinoma or melanoma. Patients were treated either with subcutaneous IL-2, alone (n = 20) or in combination with INFalpha-2b (n = 6); or with INFalpha-2b alone, administered subcutaneously at a low dose (n = 8) or intravenously at a high dose (n = 14). Depressive symptoms were evaluated using the Montgomery and Asberg Depression Rating Scale (MADRS), and anxiety symptoms were evaluated using the Covi scale. Evaluations were performed just before initiation of treatment (day 1) and on days 3 and 5 of treatment. Patients treated with IL-2 alone or in association with INFalpha-2b had significantly higher MADRS scores after 5 days of cytokine therapy, and patients who received both cytokines had increased scores on day 3. In contrast, patients treated with INFalpha-2b alone did not have varying MADRS scores during the course of treatment. Cytokine therapy had no effect on anxiety, except in patients treated with IL-2 in combination with INFalpha-2b. In these patients, the enhancement in anxiety scores that was observed on day 5 was mainly attributable to increased somatic complaints. IL-2 and INFalpha-2b have differential effects on mood, and IL-2 therapy induces depressive symptoms early in treatment.


NatIonal Danish endocarditis stUdieS - Design and objectives of the NIDUS registry.
The National Danish endocarditis stUdieS (NIDUS) registry aims to investigate the mechanisms contributing to the increasing incidence of IE and to discover risk factors associated to the course, treatment and clinical outcomes of the disease. The NIDUS registry was created to investigate a nationwide unselected group of patients hospitalized for infective endocarditis (IE). The National Danish healthcare registries have been queried for validated IE diagnosis codes (International Classification of Disease, 10th edition (ICD-10): DI33, DI38, and DI398). Subsequently, a team of 28 healthcare professionals, including experts in endocarditis, will systematically review and evaluate all identified patient records using the modified Duke Criteria and the 2015 European Society of Cardiology (ESC) modified diagnostic criteria. The registry will contain all cases with definite or possible IE found in primary data sources in Denmark between January 1, 2016, and December 31, 2021. We will gather individual patient data, such as clinical, microbiological, and echocardiographic characteristics, treatment regimens, and clinical outcomes. A digital data collection form will be used to the gathering of data. A sample of approximately 4,300 individual patients will be evaluated using primary data sources. The NIDUS registry will be the first comprehensive nationwide IE registry, contributing critical knowledge about the course, treatment, and clinical outcomes of the disease. Additionally, it will significantly aid in identifying areas in which future research is needed.


Modeling Patient Treatment With Medical Records: An Abstraction Hierarchy to Understand User Competencies and Needs.
Health care is a complex sociotechnical system. Patient treatment is evolving and needs to incorporate the use of technology and new patient-centered treatment paradigms. Cognitive work analysis (CWA) is an effective framework for understanding complex systems, and work domain analysis (WDA) is useful for understanding complex ecologies. Although previous applications of CWA have described patient treatment, due to their scope of work patients were previously characterized as biomedical machines, rather than patient actors involved in their own care. An abstraction hierarchy that characterizes patients as beings with complex social values and priorities is needed. This can help better understand treatment in a modern approach to care. The purpose of this study was to perform a WDA to represent the treatment of patients with medical records. The methods to develop this model included the analysis of written texts and collaboration with subject matter experts. Our WDA represents the ecology through its functional purposes, abstract functions, generalized functions, physical functions, and physical forms. Compared with other work domain models, this model is able to articulate the nuanced balance between medical treatment, patient education, and limited health care resources. Concepts in the analysis were similar to the modeling choices of other WDAs but combined them in as a comprehensive, systematic, and contextual overview. The model is helpful to understand user competencies and needs. Future models could be developed to model the patient's domain and enable the exploration of the shared decision-making (SDM) paradigm. Our work domain model links treatment goals, decision-making constraints, and task workflows. This model can be used by system developers who would like to use ecological interface design (EID) to improve systems. Our hierarchy is the first in a future set that could explore new treatment paradigms. Future hierarchies could model the patient as a controller and could be useful for mobile app development.


The effects of Mpl-ligand, interleukin-6 and interleukin-11 on megakaryocyte and platelet alpha-granule proteins.
Thrombopoietin (Mpl ligand), interleukin-6 (IL-6), and interleukin-11 (IL-11) differ in their effects on megakaryocyte maturation and development. In the present study, the impact of these thrombocytopoietic cytokines on biochemical and structural granule and membrane components was examined. Western blotting was performed on equivalent amounts of isolated megakaryocyte or platelet protein and the relative intensities of the enhanced chemiluminescent-visualized bands were quantitated by densitometry. Megakaryocyte growth and development factor (MGDF), a recombinant thrombopoietin-related molecule, significantly increased megakaryocyte fibronectin (72%), thrombospondin (55%), von Willebrand factor (28%) and p-selectin (CD62p) (37%) when compared to equivalent amounts of protein from saline-treated controls (p<0.01). Megakaryocyte fibrinogen was not increased. Ultrastructurally, there was a marked increase in ribosomes and rough endoplasmic reticulum even in mature-appearing megakaryocytes. Platelets from MGDF-treated mice showed small increases in fibronectin (8%), and CD62p (18%), but did not show increases in other measured alpha-granule proteins. Neither IL-6 nor IL-11 increased megakaryocyte or platelet alpha-granule proteins over levels observed in saline controls. IL-11 treated megakaryocytes, while also exhibiting an increase in ribosomes, were characterized by prominent cytoplasmic fragmentation. The study demonstrates the impact of Mpl ligand in increasing synthesized megakaryocyte alpha-granule proteins and of IL-11 in promoting megakaryocyte fragmentation.


HOXA10 regulates p53 expression and matrigel invasion in human breast cancer cells.
HOX genes regulate cell differentiation during embryonic development. Here we demonstrate HOXA10 expression in both benign and malignant adult human breast tissue and in MCF-7, but not BT20 breast cancer cells. We have previously shown that HOXA10 mediates uterine differentiation in response to estrogens. The mechanism of action of estradiol and other estrogen receptor modulators on breast cancer cell growth is still poorly understood. MCF-7 cells, which are ER (+) and express HOXA10, were used to assay the effect of estradiol and tamoxifen on HOXA10 expression. Semi-quantitative RT-PCR and northern analysis revealed that treatment with either estradiol or tamoxifen increased HOXA10 mRNA expression. BT20 cells, which are ER (-) and do not endogenously express HOXA10, were used to assay the effect of increased HOXA10 expression on p53 expression and on the invasive phenotype. Constitutively expressing HOXA10 in BT20 cells increased p53 protein expression. Increased HOXA10 also reduced invasiveness through matrigel. The mechanism by which estrogen and other estrogen receptor modulators influence both normal breast development as well as breast cancer may involve the regulation of developmental control genes such as HOXA10; HOXA10 in turn regulates expression of key downstream genes such as p53 and regulates tumor cell functional phenotype.


Antithrombotic therapy for secondary stroke prevention.
: Antithrombotic therapy is a key component of any strategy for the secondary prevention of ischemic stroke. A better understanding of the various therapeutic options will lead to improved stroke prevention, better medication adherence, and fewer complications. : Antiplatelet agents and anticoagulants are the two major classes of antithrombotic therapy used for stroke prevention. The etiology and mechanism of the stroke must be considered in order to make the best decision regarding which agent(s) to use for secondary stroke prevention. The recent Prevention Regimen for Effectively Avoiding Second Strokes (PRoFESS) study showed that clopidogrel and aspirin plus extended-release dipyridamole had similar event rates in terms of recurrent stroke, but clopidogrel was better tolerated, with fewer bleeding events. Several new anticoagulants are poised to replace warfarin for stroke prevention in the setting of atrial fibrillation. These include dabigatran (a new oral direct thrombin inhibitor) and possibly apixaban (a new oral factor Xa inhibitor). These new medications are much easier to use than warfarin and may be more effective and safer, with fewer drug and food interactions and no need for routine blood monitoring. Thus, these new medications may improve adherence as well as clinicians' inclination to treat with anticoagulation. : Because each antiplatelet agent or anticoagulant has certain advantages and disadvantages, clinicians must choose an agent that the patient can afford and tolerate in terms of side effects and adherence. The hope and expectation is that the proper use of these medications in accordance with current guidelines will reduce the risk of a recurrent stroke.


IL-17A is produced by Th17, gammadelta T cells and other CD4- lymphocytes during infection with Salmonella enterica serovar Enteritidis and has a mild effect in bacterial clearance.
T(h)17 cells represent a new pro-inflammatory T(h) cell lineage distinct from T(h)1 and T(h)2 cells. T(h)17 cells have been shown to be involved in extracellular bacterial infection but their role in intracellular infection remains unclear. We found antigen-specific IL-17A production during a systemic infection of mice with the facultative intracellular bacterium Salmonella enterica serovar Enteritidis (S. Enteritidis) and examined the function and cellular source of IL-17A during the adaptive immune response to S. Enteritidis. Infected IL-17A-/- mice survived completely after inoculation with the highest infection dose found to be sub-lethal for wild-type (WT) C57BL/6 mice. However, at 20 and 80 days post-infection (d.p.i.), we repeatedly found mildly elevated bacterial burden in spleen and liver of IL-17A-/- mice as compared with WT mice. Overall, IL-17A-/- mice showed reduced clearance of S. Enteritidis. S. Enteritidis-specific IL-17A production was induced in splenocytes and lymph node cells of infected WT mice at both time points, 20 and 80 d.p.i. Classical CD4+ T(h)17 cells developed upon infection with Salmonella. CD4- gammadelta TCR+ and CD4- gammadelta TCR- cells were found to be additional IL-17A-producing cell populations. In infected IL-17A-/- mice, a normal T(h)1 cytokine profile was observed consistent with the overall subtle phenotype. Nevertheless, in the absence of IL-17A, recruitment of neutrophils and delayed-type hypersensitivity (DTH) reactivity was significantly compromised. Our data indicate that IL-17A responses are induced by Salmonella and mildly contribute to protective immunity during S. Enteritidis infection. Thus, IL-17A complements the IL-12/IFN-gamma axis which is essential for protective immunity against salmonellosis in mice and men.


Shift of DNA polymerase alpha nuclear distribution and activity in apoptotic human leukaemia cells.
The changes in the distribution of DNA polymerase alpha in nuclei from HL-60 cells treated with Methotrexate (MTX) for up to 15 hr. were checked by means of both confocal analysis and electron microscopy analysis. The results provided evidence that, relative to controls, in the MTX treated cells the enzyme undergoes a topographical rearrangement throughout the nucleus, showing a pattern of distribution which calls to mind the nuclear matrix structure. The "in vitro" analysis of DNA polymerases alpha, beta, and gamma activities revealed that, in nuclei from control cells, DNA polymerase alpha was the principal DNA polymerase driving this "in vitro" system, while after 15 hr. of MTX treatment its activity was largely decreased and replaced by DNA polymerase beta, which is believed to be associated with DNA repair. Taken together, these results suggest that among the intracellular processes elicited by MTX-induced apoptosis in HL60 cells, the redistribution of DNA polymerase alpha and the stimulation of DNA polymerase beta activity might represent an extreme attempt of the cell to preserve the replicative machinery during fragmentation and chromatin margination.


A role for thrombin in liver fibrosis.
Several lines of evidence incriminate the serine proteinase thrombin in liver fibrogenesis either through its procoagulant function or its signaling via cell-surface receptors. The aim of this study was therefore to evaluate the effect of thrombin inhibition on experimental liver fibrosis. Fibrosis was induced in rats by administration of CCl4 for either three or seven weeks. Oral administration of the thrombin antagonist SSR182289 started one week after the start of CCl4 intoxication. Fibrosis and the area occupied by alpha smooth muscle actin (ASMA) positive cells were quantified with histomorphometry. Expression of fibrosis related genes was measured by real time RT-PCR. After three weeks of CCl4, treatment with SSR182289 did not significantly decrease the area of fibrosis but significantly decreased the area of ASMA positive cells by 22% (p = 0.03) and the expression of TIMP-1 mRNA by 52% (p = 0.02). There was no effect on gene expression of collagen I, MMP-2, or TIMP-2. After seven weeks of CCl4, treatment with SSR182289 resulted in a significant decrease in fibrosis (-30%, p = 0.04) and ASMA positive areas (-35%, p = 0.05). SSR182289 alone had no effect on the measured parameters. Additionally, it did not alleviate the acute toxicity of CCl4 as shown by measuring levels of serum aminotransferases and the area of necrosis. These data provide evidence that thrombin antagonism can reduce liver fibrogenesis. The early effect of SSR182289 on ASMA and TIMP-1 expression suggests that it is beneficial in reducing fibrogenic cell activation.


Evaluating the impact of sustainable comprehensive primary health care on rural health.
To develop a conceptual framework for monitoring the relationship between health services and health outcomes in rural Australia. Development of an evaluation framework for a rural comprehensive primary health service in Victoria. Evidence regarding essential components for successful primary health care, and objective health service and health status measures were combined to develop a conceptual health service evaluation framework. Application of the framework is illustrated using a case study of a rural primary health service in Victoria. Inadequate health services limit access to health care, delay use at times of need and result in poor health outcomes. Currently, there is a lack of evidence from rigorous health service evaluations to indicate which rural health services work well, where and why that could inform rural health policies and funding. Although the nature of health service models will vary across communities in order to meet their differing geographic circumstances, there is considerable scope for the translation and generalisation of evidence gained from health service models that are shown to be sustainable, responsive and able to deliver local quality health care. This framework can guide future health service evaluation research and thereby provide a better understanding of a health service's impact on the health of the community and its residents.


Impact of time-interleaving on directly-detected 100-Gb/s POLMUX RZ-DQPSK robustness to PDL-induced crosstalk.
The impact of PDL-induced crosstalk on 100-Gb/s POLMUX RZ-DQPSK performance is investigated both experimentally and through numerical simulations in direct detection systems. We found that RZ time-interleaving, contrary to what happens with PMD, features a strong robustness to PDL. The detrimental effect of optical narrow filtering on PDL-induced penalty is also analyzed; RZ time-interleaving still proves the best solution to deal with the PDL issue.


Immunization of cystic fibrosis patients with a Pseudomonas aeruginosa O-polysaccharide-toxin A conjugate vaccine.
Healthy, non-colonized cystic fibrosis (CF) patients (N = 26) were immunized with an octavalent Pseudomonas aeruginosa O-polysaccharide-toxin A conjugate vaccine. Vaccination was well tolerated and induced anti-lipopolysaccharide (LPS) antibodies of a high affinity capable of promoting the opsonophagocytic killing of P. aeruginosa by human peripheral lymphocytes. In contrast, anti-LPS antibodies acquired after natural infection possessed a very low affinity and were non-opsonic. To determine if immunization could prevent or delay infections due to P. aeruginosa, the infection rate among immunized patients was compared retrospectively to age and gender-matched controls. After 6 years of clinical follow-up, 15/20 (75%) of control and 8/23 (35%) of immunized subjects were classified as infected (p = 0.022). The persistence of high-affinity antibodies among immunized patients correlated with a significantly lower rate of infection after 4-6 years of observation. Infection of immunized patients was correlated with a dramatic decline in total antibody titer between year 2 and 3 of follow-up. Smooth, typeable strains of P. aeruginosa predominated among immunized patients. In contrast, rough, nontypeable strains were most frequently isolated from nonimmunized patients. Mucoid P. aeruginosa strains were isolated from 6 nonimmunized patients versus only I immunized subject.