Incidence of anencephalus and spina bifida in Greece.
A cross-sectional national survey of 11,048 births in the whole of Greece in April 1984 identified 7 cases of anencephalus and 9 of spina bifida. There was thus an incidence of 1.45 neural tube defects per 1,000 total births. This is the first geographically defined population study from southern Europe. In comparison with hospital-based data from the 2 countries on its borders, the Greek rates are rather similar to those of Yugoslavia but much lower than reports from Turkey.


Phenotypic and genetic virulence and antibiotic resistance markers in Escherichia coli strains isolated from hospital surfaces.
The aim of this study was the phenotypic and genotypic analysis of the antibiotic resistance and virulence markers in enterobacterial strains isolated from the hospital environment. In this purpose, 100 enterobacterial strains isolated from hospital surfaces were investigated for their susceptibility patterns, for the ability to colonize the cellular (HeLa) and inert substrate and for the production of soluble, enzymatic factors. The bacterial strains were also investigated for the presence of resistance and virulence genes (aggR, aggA, EaggEC, EAST1, hlyA). The enterobacterial strains isolated from the hospital surfaces exhibited high levels resistance rates to beta-lactams, including 3'd generation cephalosporins, teteracyclines, sulphametoxazole and nalidixic acid. The PCR analysis demonstrated the presence of TEM1, tetA, tetB, tetC, dfrA12, sulI and sulII in a low percentage of the resistant strains. The majority of the tested strains exhibited ability to colonize the inert and cellular substrate and also the ability to produce a series of soluble enzymes implicated in enteric and extra-intestinal pathogenesis (pore forming enzymes, proteases, mucinases, iron chelating agents). The presence of beta-haemolysis on sheep blood agar was well correlated with the presence of hlyA gene, while the aggregative adherence pattern with the presence of aggA, aggR and EAST/1 genes, in different combinations. Our results are demonstrating that the E. coli strains isolated from the hospital environment harbor phenotypic and genetic virulence markers, thus contributing to the development of resistance and virulence genes reservoirs with potential implication for the human health in the hospital environment.


Healing of the anterior attachment of the rabbit meniscus to bone.
In a rabbit model the healing process of the anterior attachment of the medical meniscus was observed during the first 12 weeks after sharp transection and refixation in a tibial bone channel. Evaluations of the healing tissue were histologic analysis, application of immunohistochemical methods to show collagen types and nerve regeneration, and mechanical load to failure tests. Secondary changes to knee joint cartilage, as signs of eventual dysfunction of the refixed meniscus, were evaluated by analysis of proteoglycan fragment concentration in joint fluid and histologic analysis of knee joint articular cartilage and synovium. The healing tissue between the refixed attachment and bone matured from highly cellular, nonspecific granulation tissue at 1 week, to bone, fibrocartilaginous, and fibrous tissues, which at some sites developed an insertion specific tissue arrangement within a 12-week period. However, the irregular interface between the fibrocartilaginous tissue and the underlying bone, which is typical for a normal insertion, was not reestablished. Labeling for collagen Types I and II in the newly formed insertion did not return to normal. In addition a few collagen fibers connected the refixed attachment tissue to bone. New bone formation turned the initially cancellous bone tunnel walls into more solid cortical bone. However, new bone formation did not fill the distal part of the channel. The refixed meniscal attachment underwent necrosis and was revitalized by cell ingrowth from the periphery. Nerve fibers were found in the newly formed insertion by 12 weeks. The failure load at tensile testing never reached more than 20% that of a normal attachment. Degeneration of articular cartilage and increased proteoglycan fragment in the joint fluid were common after this procedure. These data suggest that, despite the focal appearance of insertion specific tissues and healing of collagen fibers to bone, the tissue architecture of a normal meniscal insertion and a normal meniscal joint protective function were not reestablished.


Anatomy and motor function of extra-laryngeal branching patterns of the recurrent laryngeal nerve; an electrophysiological study of 1001 nerves at risk.
Safe thyroid surgery depends on a deep knowledge of human neck anatomy, including the recurrent laryngeal nerve (RLN). Anatomic variations such as extra-laryngeal terminal branching (ETB) are common. We studied the ETB pattern of 1001 RLNs at risk in 596 patients. We identified and exposed the location of division points on the cervical part of bifid RLN. The function of nerve branches was assessed through intraoperative nerve monitoring (IONM). Bifid RLNs was identified in 39.6% of patients. The nerve-based prevalence of ETB was 28.5%. The prevalence of ETB for the right and left RLN was 21.8% and 35.5%, respectively (p < 0.001). The location of the division point was found in the middle, distal, and proximal segments in 48.8%, 33.3%, and 18% of bifid RLNs, respectively. Electrophysiological monitoring revealed motor functions in all anterior and in 7% of posterior branches. The rate of injury was 0.4%, and 1.1% in single trunk and bifid nerves, respectively (p = 0.360), and 3.9% in nerves with proximal branching (p = 0.084). The ETB prevalence is high and showing division points in different cervical segments of the RLN. All anterior branches and some posterior branches contain motor fibers. Knowledge and awareness of these anatomic and functional variations are mandatory for every thyroid surgeon to avoid misidentification and misinterpretation of human RLN anatomy.


Acidic Centers on the Surface of a Crystalline α-Sn(IV) Phosphate Characterized by the Solid-State 1H, 2H, 31P, and 119Sn MAS NMR Techniques.
A layered crystalline phosphate α-Sn(HPO4)2·H2O (1), prepared and characterized in the present study by the multinuclear solid-state nuclear magnetic resonance (NMR), powder X-ray diffraction, and thermogravimetric analysis techniques, was treated with D2O and HOD imitating the reaction conditions in a water medium. The 2H solid-echo magic angle spinning NMR spectra of the products have revealed on their surface low mobile water molecules and hydronium ions, forming a structure close to the Zundel cation, [D2O···D-OD2]+. All the deuterons in the hydronium ions are tangled by hydrogen bonds with the water and the surface phosphate groups and stabilized by ionic interactions.


Triorthocresyl phosphate poisoning--a review of human cases.
Since the end of the nineteenth century, numerous cases of triorthocresyl phosphate (TOCP) poisoning due to accidental contamination of drink, food or drugs have been reported. Following the ingestion of preparations contaminated by TOCP, gastrointestinal symptoms may occur and after an interval of ten to twenty days, a well-known delayed neurotoxicity gradually develops. In general, the initial symptoms are pain and paresthesia in the lower extremities. In most cases, muscle weakness progresses rapidly developing into a striking paralysis of the lower extremities with or without an involvement of the upper extremities. Severe cases show pyramidal signs. The histopathological findings show axonal degeneration in the peripheral nerves and degenerative changes in the anterior horn cells. Degenerative change also occurs in the lateral and dorsal tracts of the spinal cord. The cardinal therapy is physical rehabilitation.


Enchondroma of the cervical spine in young woman: A rare case report.
Enchondroma is a type of benign cartilaginous bone tumor. Enchondroma of the spine is very rare. There are only a few cases of enchondromas located in the lamina of the cervical spine have been reported. Therefore, we report a case of enchondroma in the cervical spine. A 24-year-old female patient presented with a history of neck pain, restriction of neck movement, pain and numbness along the right scapula, and weakness accompanied by wasting of the right hand. Presumptive diagnoses included bony tumors such as an aneurysmal bone cyst or a giant cell tumor. Radiologic examinations revealed a round tumor in the right lamina of C4 with extracortical extension and foramen of C4-5. C4 right hemilaminectomy and facetectomy were performed with near complete removal of the tumor. On histological examination, the tumor was confirmed to be an enchondroma. At the 6-month follow-up, a computed tomography scan showed no recurrence with good alignment.


[Hybrid osteosynthesis in orthognathic surgery: 28 cases of Le Fort I osteotomy].
The use of resorbable materials in orthognathic surgery is presently the object of numerous clinical and experimental studies. Their main drawbacks are their lack of rigidity and induced chronic inflammatory reactions. The purpose of this study was to propose and to test an original system of hybrid osteosynthesis associating a single titanium plate with three resorbable plates in Le Fort I osteotomies in the absence of maxillary expansion, or two titanium plates with two resorbable plates in case of maxillary expansion. Our retrospective study concerned 28 patients operated between 2002 and 2005. A clinical follow-up of over one year detected the specific complications of the hybrid system and a cephalometric study analyzed the secondary relapse 5 months after surgery. These results were compared to the literature. One case of mobility and a case of instability of the jaw were observed, along with 5 cases of local chronic inflammatory reaction. The radiological results found a secondary sub-clinical relapse. The removal of the titanium material under local anesthesia was performed without difficulties in ambulatory care, which avoided a new hospitalization for the patient and a new general anesthesia. The hybrid system allowed obtaining a postoperative stability of the jaw comparable to that obtained by the exclusive use of titanium plates with an acceptable morbidity. This method of osteosynthesis combined the advantages of the resorbable material (removal of titanium plates under local anesthesia) and of titanium material (stability of the procedure). However this method requires a good experience of working with resorbable material, a good follow-up, and cooperative patients. These encouraging results urge us to extend the indications to mandibular sagittal osteotomy and to bi-maxillary osteotomies.


Evaluation of HPV DNA positivity in colorectal cancer patients in Kerman, Southeast Iran
Background: The HPV virus is known to be oncogenic and associations with many cancers has been proven. Although many studies have been conducted on the possible relationship with colorectal cancer (CRC), a definitive role of the virus has yet to be identified. Method: In this cross-sectional study, the frequency of HPV positivity in CRC samples in Kerman was assessed in 84 cases with a mean age of 47.7 ± 12.5 years over two years. Qualitative real time PCR was performed using general primers for the L1 region of HPV DNA. Results: Out of 84 CRC samples, 19 (22.6%), proved positive for HPV DNA. Genotyping of positive samples showed all of these to be of high risk HPV type. Prevalence of HPV infection appears to depend geographic region, life style, diet and other factors. Conclusion: In our location frequency of CRC is low, and this limited the sample size for evaluation of HPV DNA. The most prevalent types were HPV types 51 and 56. While HPV infection may play an important role in colorectal carcinogenesis, this needs to be assessed in future studies.


[The role of oxidative stress in experimental diabetic neuropathy].
To study the role of oxidative stress in the initiation and development of diabetic neuropathy. The diabetic rats were induced with streptozotocin (STZ). The malondialdehyde (MDA) level, total superoxide dismutase (SOD) and Na(+) -K(+) -ATPase activity were measured in the sciatic nerves at various stages of diabetes. The correlation of the MDA level and Na(+) -K(+) -ATPase activity was analyzed in diabetic rats. The pathological changes of sciatic nerve at diabetic various stages were examined by light microscopy. The MDA level increased significantly in diabetic sciatic nerves as compared to controls at all time intervals. Total SOD activity increased significantly in diabetic sciatic nerves as compared to controls at one month of diabetes and progressively decreased at three/six months of diabetes. Na(+) -K(+) -ATPase activity progressively decreased at three/six months of diabetes. The correlation analysis indicated that the Na(+) -K(+) -ATPase activity was negative correlation with the MDA level in the diabetic rats. Histopathological study of the diabetic sciatic nerves showed that the pathological changes were observed at 3 months of diabetes, the changes were more serious as the diabetic duration was longer. Oxidative stress is found to occur during the early stages of STZ-induced diabetes (no neuropathy) and this state is maintained after initiation of neuropathy. The decreased Na(+) -K(+) -ATPase activity is associated with oxidative stress in the diabetic rats. Therefore, oxidative stress plays an important role in the initiation and development of diabetic neuropathy.


Shape-Programmable Liquid-Crystalline Polyurethane-Based Multimode Actuators Triggered by Light-Driven Molecular Motors.
In recent years, light-driven soft actuators have been rapidly developed as enablers in the fabrication of artificial robots and biomimetic devices. However, it remains challenging to amplify molecular isomerization to multiple modes of macroscopic actuation with large amplitude and complex motions. Here, a strategy is reported to build a light-responsive liquid-crystalline polyurethane elastomer by phototriggered overcrowded alkene-based molecular motors. A trifunctional molecular motor modified with an ethylene glycol spacer on the rotor and stator functions as a crosslinker and unidirectional stirrer that amplifies molecular motion into macroscopic movement. The shape-programmable polymeric film presents superior mechanical properties and characteristic shape-memory effect. Furthermore, diverse modes of motions including bending, unwinding, and contracting with tunable actuation speed over a wide range are achieved. Such research is hoped to pave a new way for the design of advanced light-responsive soft actuators and robots.


[A Case of Cervical Lymph Node Metastasis from an Unknown Primary Cancer Controlled with Immunotherapy, Chemotherapy, and Surgery].
We present a case of cervical lymph node metastasis from an unknown primary cancer that was controlled with immunotherapy, chemotherapy, and surgery. The patient, a 61-year-old man, was referred to our department for treatment of a lesion in the left cervical lateral area. At the initial visit, the mass was covered by reddened skin and was elastic, hard, and immobile on palpation. The presence of a malignant disease such as malignant lymphoma or lymphadenitis because of infection by tubercle bacillus or Epstein-Barr virus was suspected on the basis of the clinical and magnetic resonance imaging findings. Biopsy and resection of the cervical mass was performed under general anesthesia. Because the pathological diagnosis during surgery indicated squamous cell carcinoma, the surgical approach was changed to neck dissection. Head, neck, and thoracic computed tomography and other examinations were performed to locate the primary cancer, but its origin remained unknown. Postoperative therapy consisted of chemotherapy and immunotherapy. The patient has been followed up for 4 years and 10 months without any evidence of recurrence.


[Statistical analysis of the activity of loxapine and thioridazine in a double-blind study].
49 patients have been treated for 12 weeks with loxapine and thioridazine in a double-blind study. The evaluation was carried out with a covariance analysis with B.P.R.S. and NOSIE as bases and a variance analysis concerning the general clinical improvement. With 36 schizophrenics, the two treatments have shown significant and similar general improvement as early as the second week, and this in particular with all the factors and most of the items of the B.P.R.S. with 13 psychotic not schizophrenic patients, the covariance analysis showed that the results (improvement of all the factors, with most of the items and in the total score of the B.P.R.S.), while comparable, were slightly in favour of loxapine for the item "disorientation", for "disturbance of thought", and "excitement-disorientation", for the final score of B.P.R.S., for the general and clinical improvement and for the alteration of the initial degree of severity. The analysis of the NOSIE showed a significant improvement which is similar for both treatments.


Climate Warming-Driven Changes in the Molecular Composition of Soil Dissolved Organic Matter Across Depth: A Case Study on the Tibetan Plateau.
Dissolved organic matter (DOM) is critical for soil carbon sequestration in terrestrial ecosystems. DOM molecular composition varies with soil depth. However, the spatial heterogeneity of depth-dependent DOM in response to climate warming remains unclear, especially in alpine ecosystems. In this study, the DOM of alpine meadow soil samples was characterized comprehensively by using spectroscopy and mass spectrometry, and open-top chambers (OTCs) were employed to simulate warming. It was found that climate warming had the greatest impact on the upper layer (0-30 cm), followed by the lower layer (60-80 cm), while the middle layer (30-60 cm) was the most stable among the three soil layers. The reasons for the obvious changes in DOM in the upper and lower layers of soil were further explained based on biotic and abiotic factors. Specifically, soil nutrients (NH4+-N, NO3--N, TC, and TP) affected the molecular composition of DOM in layer L1 (0-15 cm), while pH affected layer L5 (60-80 cm). Gemmatimonadetes, Proteobacteria, and Actinobacteria played important roles in the composition of DOM in the L5 layer (60-80 cm), while the dominant fungal groups affecting the DOM composition increased in the L1 layer (0-15 cm) under warming. In summary, this research has contributed to a deeper understanding of depth-dependent changes in DOM molecular composition in alpine ecosystems.


Gene Regulation in and out of Equilibrium.
Determining whether and how a gene is transcribed are two of the central processes of life. The conceptual basis for understanding such gene regulation arose from pioneering biophysical studies in eubacteria. However, eukaryotic genomes exhibit vastly greater complexity, which raises questions not addressed by this bacterial paradigm. First, how is information integrated from many widely separated binding sites to determine how a gene is transcribed? Second, does the presence of multiple energy-expending mechanisms, which are absent from eubacterial genomes, indicate that eukaryotes are capable of improved forms of genetic information processing? An updated biophysical foundation is needed to answer such questions. We describe the linear framework, a graph-based approach to Markov processes, and show that it can accommodate many previous studies in the field. Under the assumption of thermodynamic equilibrium, we introduce a language of higher-order cooperativities and show how it can rigorously quantify gene regulatory properties suggested by experiment. We point out that fundamental limits to information processing arise at thermodynamic equilibrium and can only be bypassed through energy expenditure. Finally, we outline some of the mathematical challenges that must be overcome to construct an improved biophysical understanding of gene regulation.


A tailored compassion-focused therapy program for sexual minority young adults with depressive symotomatology: study protocol for a randomized controlled trial.
Lesbian, gay, and bisexual (LGB) men and women represent one of the highest-risk populations for depressive symptomatology and disorders, with young LGB adults being at greatest risk. To date, there have been no randomized controlled trials (RCT) to specifically target depressive symptoms in young LGB adults. This is despite research highlighting unique predictors of depressive symptomatology in this population. Here we outline a protocol for an RCT that will test the preliminary efficacy of a tailored compassion-focused therapy (CFT) intervention for young LGB adults compared with a self-directed cognitive behavioral therapy (CBT) program with no specific tailoring for LGB individuals. The CFT intervention consists of 8 units with self-directed reading and activities tailored to LGB young adults, and 8 x weekly 1-hour consultations with a therapist. The CBT intervention consists of 8 units with self-guided reading and activities, with 1 x 1-hour session with a therapist at the mid-point of therapy. Fifty LGB individuals with scores of 13 or above on the Beck Depression Inventory-II will be randomized to either the CFT or CBT condition. The primary outcome measure is depressive symptomatology. Secondary outcome measures are symptoms of anxiety, suicidal ideation, internalized homophobia, self-compassion, and shame and guilt proneness. Assessments will occur at pre-intervention, post-intervention, and at 3-month post-intervention. This study is an RCT to test the preliminary efficacy of an LGB-tailored compassion-focused intervention for young LGB adults with depressive symptomatology. If this intervention is efficacious, this could begin to address the substantial mental health disparities amongst sexual minorities. ACTRN12616001018404 . Prospective registration, registered 02/08/2016.


Preclinical Pharmacological Development of Chlorcyclizine Derivatives for the Treatment of Hepatitis C Virus Infection.
Hepatitis C virus (HCV) is a small, single-stranded, positive-sense RNA virus that infects more than an estimated 70 million people worldwide. Untreated, persistent HCV infection often results in chronic hepatitis, cirrhosis, or liver failure, with progression to hepatocellular carcinoma. Current anti-HCV regimens comprising direct acting antivirals (DAAs) can provide curative treatment; however, due to high costs there remains a need for effective, shorter-duration, and affordable treatments. Recently, we disclosed anti-HCV activity of the cheap antihistamine chlorcyclizine, targeting viral entry. Following our hit-to-lead optimization campaign, we report evaluation of preclinical in vitro absorption, distribution, metabolism, and excretion properties, and in vivo pharmacokinetic profiles of lead compounds. This led to selection of a new lead compound and evaluation of efficacy in chimeric mice engrafted with primary human hepatocytes infected with HCV. Further development and incorporation of this compound into DAA regimens has the potential to improve treatment efficacy, affordability, and accessibility.


Oxadiazon Derivatives Elicit Potent Intracellular Growth Inhibition against Toxoplasma gondii by Disrupting Heme Biosynthesis.
Infections of Toxoplasma gondii can cause severe and sometimes fatal diseases in immunocompromised individuals. The de novo heme biosynthesis pathway is required for intracellular growth and pathogenesis, making it an appealing therapeutic target. We synthesized a small library of derivatives of the herbicide oxadiazon, a known inhibitor of the penultimate reaction within the heme biosynthesis pathway in plants, catalyzed by protoporphyrinogen oxidase (PPO). Seven of the 18 analogs exhibit potent intracellular growth inhibition of wild-type T. gondii (IC50 = 1 to 2.4 μM). An assay of the compounds against Toxoplasma PPO knockout and complementation strains confirmed the mode of action to be due to the potent inhibition of PPO. The most potent compounds have no detectable cytotoxicity against human foreskin fibroblast cells up to 100 μM. This study suggests that oxadiazon derivatives may represent a new molecular scaffold for the effective treatment of T. gondii infections.


Infant botulism.
Although the worldwide incidence of infant botulism is rare, the majority of cases are diagnosed in the United States. An infant can acquire botulism by ingesting Clostridium botulinum spores, which are found in soil or honey products. The spores germinate into bacteria that colonize the bowel and synthesize toxin. As the toxin is absorbed, it irreversibly binds to acetylcholine receptors on motor nerve terminals at neuromuscular junctions. The infant with botulism becomes progressively weak, hypotonic and hyporeflexic, showing bulbar and spinal nerve abnormalities. Presenting symptoms include constipation, lethargy, a weak cry, poor feeding and dehydration. A high index of suspicion is important for the diagnosis and prompt treatment of infant botulism, because this disease can quickly progress to respiratory failure. Diagnosis is confirmed by isolating the organism or toxin in the stool and finding a classic electromyogram pattern. Treatment consists of nutritional and respiratory support until new motor endplates are regenerated, which results in spontaneous recovery. Neurologic sequelae are seldom seen. Some children require outpatient tube feeding and may have persistent hypotonia.


LncRNA XR_351665 Contributes to Chronic Pain-Induced Depression by Upregulating DNMT1 via Sponging miR-152-3p.
Chronic pain is frequently comorbid with depression. However, the mechanisms underlying chronic pain-induced depression remain unclear. Here, we found that DNA methyltransferase 1 (DNMT1) was upregulated in the central amygdala (CeA) of spared nerve injury (SNI)-induced chronic pain-depression rats, and knockdown of DNMT1 could improve the depression-like behaviors in SNI rats. Additionally, a panel of differentially expressed lncRNAs, including 38 upregulated and 12 downregulated lncRNAs, were identified by microarray analysis. Bioinformatics analysis suggested that the upregulated lncRNA XR_351665 was the upstream molecule to regulate DNMT1 expression. The knockdown of XR_351665 significantly alleviated the depression-like behaviors in SNI rats, whereas overexpression of XR_351665 induced the depression-like behaviors in naïve rats. Further mechanism-related researches uncovered that XR_351665 functioned as a competing endogenous RNA (ceRNA) to upregulate DNMT1 by competitively sponging miR-152-3p, and subsequently promoted the development of chronic pain-induced depression. Our findings suggest that lncRNA XR_351665 is involved in the development of chronic pain-induced depression by upregulating DNMT1 via sponging miR-152-3p. These data provide novel insight into understanding the pathogenesis of chronic pain-induced depression and identify a potential therapeutic target. PERSPECTIVE: LncRNA XR_351665 in CeA functions as a ceRNA to block the inhibitory effect of miR-152-3p on DNMT1 and contributes to the development of chronic pain-induced depression. These data suggest that manipulation of XR_351665/miR-152-3p/DNMT1 axis may be a potential method to attenuate chronic pain-induced depression.


Two sensory channels mediate perception of fingertip force.
In two experiments we examined the ability of humans to exert forces accurately with the fingertips, and to perceive those forces. In experiment 1 participants used visual feedback to apply a range of fingertip forces with the distal pad of the thumb. Participants made magnitude discriminations regarding these forces, and their just noticeable differences were calculated at a series of standards by means of a two-interval, forced-choice tracking paradigm. As the standard increased, participants demonstrated a relative improvement in force discrimination; and the presence of a possible inflection point, at approximately 400 g, suggested that two sensory channels may contribute to performance. If this is the case, the operative channel at low forces is almost certainly the slowly adapting type I (SA-I) channel, while another mechanoreceptor class, the SA-II nail unit, is a plausible mediator of the more accurate performance seen at high force levels. To test this two-channel hypothesis in experiment 2, we hydrated participants' thumbnails in order to reduce nail rigidity and thus prevent stimulation of underlying SA-II mechanoreceptors. This technique was found to reduce sensory accuracy in a force-matching task at high forces (1000 g) while leaving low force matching (100 g) unimpaired. Taken together, these results suggest that two sensory channels mediate the perception of fingertip forces in humans: one channel predominating at low forces (below approximately 400 g) and another responsible for perceiving high forces which is likely mediated by the SA-II nail unit.


Modulation of RNA primer formation by Mn(II)-substituted T7 DNA primase.
Lagging strand DNA synthesis by DNA polymerase requires RNA primers produced by DNA primase. The N-terminal primase domain of the gene 4 protein of phage T7 comprises a zinc-binding domain that recognizes a specific DNA sequence and an RNA polymerase domain that catalyzes RNA polymerization. Based on its crystal structure, the RNA polymerase domain contains two Mg(II) ions. Mn(II) substitution leads to elevated RNA primer synthesis by T7 DNA primase. NMR analysis revealed that upon binding Mn(II), T7 DNA primase undergoes conformational changes near the metal cofactor binding site that are not observed when the enzyme binds Mg(II). A machine-learning algorithm called linear discriminant analysis (LDA) was trained by using the large collection of Mn(II) and Mg(II) binding sites available in the protein data bank (PDB). Application of the model to DNA primase revealed a preference in the enzyme's second metal binding site for Mn(II) over Mg(II), suggesting that T7 DNA primase activity modulation when bound to Mn(II) is based on structural changes in the enzyme.


The 1.7 A crystal structure of Dronpa: a photoswitchable green fluorescent protein.
The green fluorescent protein (GFP), its variants, and the closely related GFP-like proteins possess a wide variety of spectral properties that are of widespread interest as biological tools. One desirable spectral property, termed photoswitching, involves the light-induced alteration of the optical properties of certain GFP members. Although the structural basis of both reversible and irreversible photoswitching events have begun to be unraveled, the mechanisms resulting in reversible photoswitching are less clear. A novel GFP-like protein, Dronpa, was identified to have remarkable light-induced photoswitching properties, maintaining an almost perfect reversible photochromic behavior with a high fluorescence to dark state ratio. We have crystallized and subsequently determined to 1.7 A resolution the crystal structure of the fluorescent state of Dronpa. The chromophore was observed to be in its anionic form, adopting a cis co-planar conformation. Comparative structural analysis of non-photoactivatable and photoactivatable GFPs, together with site-directed mutagenesis of a position (Cys62) within the Dronpa chromophore, has provided a basis for understanding Dronpa photoactivation. Specifically, we propose a model of reversible photoactivation whereby irradiation with light leads to subtle conformational changes within and around the environment of the chromophore that promotes proton transfer along an intricate polar network.


Body impedance studies in end-stage heart failure.
This study adds another category of patients to those amenable to body impedance analysis (BIA). BIA measurements were obtained for the first time in 23 male patients with end-stage heart failure who were waiting for heart transplantation, and the data were compared with those obtained in 69 healthy controls matched for age, sex, height and weight. The data indicate that in end-stage heart failure there is an increased reactance (p<0.01) and an altered intracellular water/extracellular water ratio (p<0.03) due to the increased intracellular water (p<0.01) and decreased extracellular water (p<0.01).


Synergistic association between underweight and type 2 diabetes on the development of laryngeal cancer: a national population-based retrospective cohort study.
Although cigarette smoking is the most significant risk factor for laryngeal cancer, other risk factors might also be associated with the development of laryngeal cancer. We investigated whether underweight and type 2 diabetes are associated with laryngeal cancer in a Korean population. A total of 9,957,059 participants (≥20 years) without prior history of cancer who underwent a National Health Insurance Service health checkup in 2009 were followed up until December 31, 2018. Newly diagnosed laryngeal cancer was identified using claim data, and underweight was defined as body mass index (BMI) < 18.5 kg/m2. A Cox proportional-hazards models with multivariable adjustment were used to estimate hazard ratios (HRs) and corresponding 95% confidence intervals (95% CIs). During the median follow-up period of 8.3 years, 3504 cases of laryngeal cancer occurred. Underweight was associated with increased risk of laryngeal cancer after adjusting for potential confounders (HR: 1.43, 95% CI: 1.22-1.69) compared to those who were not underweight. Underweight and type 2 diabetes were synergistically associated with higher risk of laryngeal cancer (HR: 2.33, 95% CI: 1.54-3.51), compared to those without either condition. This relationship was stronger in those with an age < 65 years (HR: 3.33, 95% CI: 1.88-5.87) and alcohol consumption (HR: 2.72, 95% CI: 1.64-4.53). These results suggest that underweight may be a significant risk factor for laryngeal cancer and that underweight and type 2 diabetes might synergistically increase the risk of laryngeal cancer.


Influence of the vitrification solution on the angiogenic factors in vitrificated mouse ovarian tissue.
To investigate the effect of the dimethyl sulfoxide (DMSO) and EFS-40 during vitrification on the expression of angiogenic factors in vitrified mouse ovarian tissue. The ovarian tissues were obtained from 5 or 6 weeks aged ICR mouse. Ovarian tissues were divided into four groups: ovarian tissue without cryopreservation (control, group I), ovarian tissue vitrified with 15% DMSO (group II), ovarian tissue vitrified with EFS-40 (group III), and ovarian tissue slowly frozen with 10% DMSO (group IV). Thawing was carried out at room temperature. Levels of messenger RNA (mRNA) and protein for vascular endothelial growth factor-A (VEGF-A) and angiopoietin-2 (Angpt-2) were checked in ovarian tissues of four groups recovered on day 7 after cryopreservation. Reverse transcription-polymerase chain reaction and Western blot analysis were used to identify the levels of angiogenic factors in mouse ovarian tissues. Levels of mRNA and protein for VEGF-A and Angpt-2 were significantly decreased in cryopreserved group (group II, III and IV) than control group (group I) (P< 0.05). The significant differences of levels of mRNA and protein for VEGF-A and Angpt-2 between cryopreservation methods were observed (P< 0.05). Group III showed highest expression of mRNA and protein for VEFG-A and Angpt-2 than other cryopreservation groups (P< 0.05). These findings suggest that EFS-40 is more efficient vitrification solution for preservation of angiogenic factors than 15% DMSO during vitrification of mouse ovarian tissue. Future studies should investigate to improve the vitrification solution for ovarian tissue vitrification.


Differential capacities of CD4+, CD8+, and CD4-CD8- T cell subsets to express IL-18 receptor and produce IFN-gamma in response to IL-18.
IL-12 and IL-18 have the capacity to stimulate IFN-gamma production by T cells. Using a T cell clone, we reported that IL-18 responsiveness is generated only after exposure to IL-12. Here, we investigated the induction of IL-18 responsiveness in resting CD8+, CD4+, and CD4-CD8- T cells. Resting T cells respond to neither IL-12 nor IL-18. After stimulation with anti-CD3 plus anti-CD28 mAbs, CD8+, CD4+, and CD4-CD8- T cells expressed IL-12R, but not IL-18R, and produced IFN-gamma in response to IL-12. Cultures of T cells with anti-CD3/anti-CD28 in the presence of rIL-12 induced IL-18R expression and IL-18-stimulated IFN-gamma production, which reached higher levels than that induced by IL-12 stimulation. However, there was a substantial difference in the expression of IL-18R and IL-18-stimulated IFN-gamma production among T cell subsets. CD4+ cells expressed marginal levels of IL-18R and produced small amounts of IFN-gamma, whereas CD8+ cells expressed higher levels of IL-18R and produced more IFN-gamma than CD4+ cells. Moreover, CD4-CD8- cells expressed levels of IL-18R comparable to those for CD8+ cells but produced IFN-gamma one order higher than did CD8+ cells. These results indicate that the induction of IL-18R and IL-18 responsiveness by IL-12 represents a mechanism underlying enhanced IFN-gamma production by resting T cells, but the operation of this mechanism differs depending on the T cell subset stimulated.


Familial benign chronic neutropenia associated with periodontal disease. A case report.
A rare case report of periodontal disease associated with familial benign chronic neutropenia is presented. The medical, dental and family histories as well as clinical and histologic observations are described and discussed.


RobiNA: a user-friendly, integrated software solution for RNA-Seq-based transcriptomics.
Recent rapid advances in next generation RNA sequencing (RNA-Seq)-based provide researchers with unprecedentedly large data sets and open new perspectives in transcriptomics. Furthermore, RNA-Seq-based transcript profiling can be applied to non-model and newly discovered organisms because it does not require a predefined measuring platform (like e.g. microarrays). However, these novel technologies pose new challenges: the raw data need to be rigorously quality checked and filtered prior to analysis, and proper statistical methods have to be applied to extract biologically relevant information. Given the sheer volume of data, this is no trivial task and requires a combination of considerable technical resources along with bioinformatics expertise. To aid the individual researcher, we have developed RobiNA as an integrated solution that consolidates all steps of RNA-Seq-based differential gene-expression analysis in one user-friendly cross-platform application featuring a rich graphical user interface. RobiNA accepts raw FastQ files, SAM/BAM alignment files and counts tables as input. It supports quality checking, flexible filtering and statistical analysis of differential gene expression based on state-of-the art biostatistical methods developed in the R/Bioconductor projects. In-line help and a step-by-step manual guide users through the analysis. Installer packages for Mac OS X, Windows and Linux are available under the LGPL licence from http://mapman.gabipd.org/web/guest/robin.


Augmentation of hospital critical care capacity after bioterrorist attacks or epidemics: recommendations of the Working Group on Emergency Mass Critical Care.
The Working Group on Emergency Mass Critical Care was convened by the Center for Biosecurity of the University of Pittsburgh Medical Center and the Society of Critical Care Medicine to provide recommendations to hospital and clinical leaders regarding the delivery of critical care services in the wake of a bioterrorist attack resulting in hundreds or thousands of critically ill patients. In these conditions, traditional hospital and clinical care standards in general, and critical care standards in particular, likely could no longer be maintained, and clinical guidelines for U.S. hospitals facing these situations have not been developed. The Working Group offers recommendations for this situation.


Failed TAVI in TAVI Implantation: TAVI Dislocation Followed by Ensuing Surgical Graft Resection.
We are presenting a case report of failed valve-in-valve treatment of severe aortic stenosis. A control ultrasonography after TAVI implantation revealed a severe aortic regurgitation of the graft which was subsequently unresolved with postimplantation dilatation. Second TAVI was implanted with cranial dislocation to the aortic root. Patient underwent a CT examination to clarify the TAVI in TAVI position. Patient underwent a surgical resection of TAVI with implantation of biological aortic valve prosthesis. In situations where TAVI treatment fails or is complicated beyond the possibility of endovascular repair, surgical intervention despite its higher risks is the preferred choice.


Structure of 2-deoxy-scyllo-inosose synthase, a key enzyme in the biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics, in complex with a mechanism-based inhibitor and NAD+.
A key enzyme in the biosynthesis of clinically important aminoglycoside antibiotics is 2-deoxy-scyllo-inosose synthase (DOIS), which catalyzes carbocycle formation from D-glucose-6-phosphate to 2-deoxy-scyllo-inosose through a multistep reaction. This reaction mechanism is similar to the catalysis by dehydroquinate synthase (DHQS) of the cyclization of 3-deoxy-D-arabino-heputulosonate-7-phosphate to dehydroquinate in the shikimate pathway, but significant dissimilarity between these enzymes is also known, particularly in the stereochemistry of the phosphate elimination reaction and the cyclization. Here, the crystal structures of DOIS from Bacillus circulans and its complex with the substrate analog inhibitor carbaglucose-6-phosphate, NAD+, and Co2+ have been determined to provide structural insights into the reaction mechanism. The complex structure shows that an active site exists between the N-terminal and C-terminal domains and that the inhibitor coordinates a cobalt ion in this site. Two subunits exist as a dimer in the asymmetric unit. The two active sites of the dimer were observed to be different. One contains a dephosphorylated compound derived from the inhibitor and the other includes the inhibitor without change. The present study suggested that phosphate elimination proceeds through syn-elimination assisted by Glu 243 and the aldol condensation proceeds via a boat conformation. Also discussed are significant similarities and dissimilarities between DOIS and DHQS, particularly in terms of the structure at the active site and the reaction mechanism.


[The problem of comorbidity of epilepsy and psychogenic paroxysms].
A review of publications over the last two decades is presented. Psychogenic paroxysms develop in approximately 12% of patients with epilepsy. The analysis of social and demographic data, history details, semiological features and results of electrophysiological and neuroimaging studies does not unequivocally support the comorbidity of epilepsy and psychogenic paroxysms. The pathogenetic mechanisms of the development of comorbidity are various and depend on the presence of pharmacoresistance, psychological traumas in the past, intellectual disability etc. Video-EEG-monitoring is the gold standard in the diagnosis of comorbidity of epilepsy and psychogenic paroxysms. Treatment of such cases includes anticonvulsants and cognitive-behavioral therapy.


Prognostic Factors Associated With Decreased Survival in Patients With Acinic Cell Carcinoma of the Parotid Gland.
Acinic cell carcinoma (ACC) of the parotid gland is relatively rare. This study aimed to estimate 5-year survival and to identify prognostic factors associated with survival of ACC of the parotid gland. Thirty-seven patients with ACC of the parotid gland were included in this retrospective cohort study. They were treated in Beijing Stomatological Hospital from January 2000 to December 2011. Predictor variables, including age, gender, tumor stage, nodal stage, perineural invasion, and margin status, were analyzed. The Cox regression model was used to determine prognostic factors for overall survival (OS) and disease-free survival (DFS). Of the 37 patients, 20 were men (54.1%). Mean age was 42 years (range, 13 to 73 yr). Six patients (16.2%) had T3 or T4 tumors and 5 (13.5%) had positive neck nodes. The 5-year OS and DFS were 86.5 and 78.3%, respectively. Positive margin status was associated with worse OS. Patients older than 60 years with a fixed mass, high-grade tumor and nodal stage, perineural invasion, and angiolymphatic invasion had adverse OS and DFS (P < .05). The present study identified several important prognostic factors associated with OS and DFS in patients with ACC. These prognostic variables include symptoms at presentation, clinical tumor stage and pathologic nodal status, and histopathology-related factors. All these factors were important in patient therapy and could prolong survival rate.


[Third operation for breast hypertrophy].
The authors report an original technique for correction of synmastia with loss of the intermammary sulcus during a third revision for mammary hypertrophy. The technique consists of an intermammary incision with revision of the previous scars and dissection of the entire thoracic cutaneo-glandular tissue. The creation of the new intermammary sulcus is achieved by defatting a presternal "gutter" and fixation of the skin by sutures trans-fixing the sternum and tied over a pad.


Viral-mediated RdCVF and RdCVFL expression protects cone and rod photoreceptors in retinal degeneration.
Alternative splicing of nucleoredoxin-like 1 (Nxnl1) results in 2 isoforms of the rod-derived cone viability factor. The truncated form (RdCVF) is a thioredoxin-like protein secreted by rods that promotes cone survival, while the full-length isoform (RdCVFL), which contains a thioredoxin fold, is involved in oxidative signaling and protection against hyperoxia. Here, we evaluated the effects of these different isoforms in 2 murine models of rod-cone dystrophy. We used adeno-associated virus (AAV) to express these isoforms in mice and found that both systemic and intravitreal injection of engineered AAV vectors resulted in RdCVF and RdCVFL expression in the eye. Systemic delivery of AAV92YF vectors in neonates resulted in earlier onset of RdCVF and RdCVFL expression compared with that observed with intraocular injection using the same vectors at P14. We also evaluated the efficacy of intravitreal injection using a recently developed photoreceptor-transducing AAV variant (7m8) at P14. Systemic administration of AAV92YF-RdCVF improved cone function and delayed cone loss, while AAV92YF-RdCVFL increased rhodopsin mRNA and reduced oxidative stress by-products. Intravitreal 7m8-RdCVF slowed the rate of cone cell death and increased the amplitude of the photopic electroretinogram. Together, these results indicate different functions for Nxnl1 isoforms in the retina and suggest that RdCVF gene therapy has potential for treating retinal degenerative disease.


The blood sparing effect and the safety of aprotinin compared to tranexamic acid in paediatric cardiac surgery.
Recently, the safety of aprotinin administration during open-heart surgery has been debated. The aim of the study was to compare the blood sparing effect and the side effects of aprotinin and tranexamic acid in paediatric cardiac surgery patients. Perioperative data of 199 consecutive patients weighing less than 20kg undergoing open-heart cardiac surgery were prospectively collected between September 2005 and June 2006. During the first 5 months, 85 patients received aprotinin (group A); in the next 5 months, 114 patients were treated with tranexamic acid (group T). Except for antifibrinolytic therapy, the anaesthesiological and surgical protocols remained unchanged. Postoperative complications and in-hospital and 1-year mortality were considered as outcome parameters. The descriptive parameters and the intraoperative parameters were well comparable in the two groups. The blood loss was significantly lower in group A compared to group T at 6h [55 (35-82.5) vs 70 (45-100)ml, p=0.031], but not at 12 and 24h after operation. The incidence [9 (11%) vs 25 (22%), p=0.035] and the amount of red blood cell transfusion during the first 24h after surgery were also significantly lower in group A (0.1+/-0.4 vs 0.3+/-0.6 unit, p=0.036). There were significantly less rethoracotomies in group A [2 (2.4%) vs 11 (9.6%), p=0.039]. We found no difference in the incidence of the postoperative complications and in-hospital and 1-year mortality. There was a tendency for a higher incidence of seizures in group T [4 (3.5%) vs 0 (0%), p=0.14]. Aprotinin administration bears no additional risks compared to tranexamic acid and it has a stronger blood sparing effect in paediatric cardiac surgery. There were fewer rethoracotomies and less postoperative red blood cell transfusion in patients who received aprotinin.


Effects of prenatal substance exposure on neurocognitive correlates of inhibitory control success and failure.
Adolescents with prenatal substance (drug and alcohol) exposure exhibit inhibitory control (IC) deficits and aberrations in associated neural function. Nearly all research to date examines exposure to individual substances, and a minimal amount is known about the effects of heterogeneous exposure-which is more representative of population exposure levels. Using functional magnetic resonance imaging (fMRI), we investigated IC (Go/NoGo) in heterogeneously exposed (n = 7) vs. control (n = 7) at-risk adolescents (ages 13-17). The fMRI results indicated multiple IC processing differences consistent with a more immature developmental profile for exposed adolescents (Exposed > Nonexposed: NoGo > Go: right ventrolateral prefrontal cortex, right cuneus, and left inferior parietal lobe; NoGo > false alarm: occipital lobe; Go > false alarm: right anterior prefrontal cortex). Simple effects suggest exposed adolescents exhibited exaggerated correct trial but decreased incorrect trial activation. Results provide initial evidence that prenatal exposure across substances creates similar patterns of atypical brain activation to IC success and failure.


An amiloride-sensitive H+-gated Na+ channel in Caenorhabditis elegans body wall muscle cells.
About 30 genes are predicted to encode degenerin/epithelial sodium channels (DEG/ENaCs) in Caenorhabditis elegans but the gating mode of these channels has not been determined. Using the whole-cell configuration of the patch-clamp technique in acutely dissected C. elegans, we investigated the effects of H+ as a potential activating factor of DEG/ENaCs on electrical properties of body wall muscle cells. Under current-clamp conditions, decreasing external pH from 7.2 to 6.1 led to a reversible depolarization of muscle cells associated with a decrease in input resistance which was partially inhibited by amiloride. Under voltage-clamp conditions, extracellular acidification activated an inward desensitizing current at -60 mV. In the absence of external Ca2+, H+ -gated channels were found to be slightly more permeable to Na+ than to K+ and were blocked by amiloride with a K0.5 of 31 microm at -60 mV. An inward current could be also activated by protons in a GABA receptor null mutant in the presence of D-tubocurare and in an unc-105 null mutant. These results demonstrate that ion channels sharing common properties with mammalian acid-sensing ion channels (ASICs) are functional in C. elegans muscle which should prove useful for understanding proton sensing in animals.


Possible health effects of liquefied petroleum gas on workers at filling and distribution stations of Gaza governorates.
Liquefied petroleum gas (LPG) is widely used in the Gaza Strip for domestic purposes, in agriculture and industry and, illegally, in cars. This study aimed to identify possible health effects on workers exposed to LPG in Gaza governorates. Data were collected by a questionnaire interview, and haematological and biochemical analyses of venous blood samples were made from 30 workers at filling and distribution stations and 30 apparently healthy controls. Statistically significant differences were found in all self-reported health-related complaints among LPG workers versus controls. LPG workers had significantly higher values of red blood cell counts, haemoglobin, haematocrit mean corpuscular haemoglobin and platelet counts. They also had significantly higher values of kidney function tests (urea, creatinine and uric acid) and liver function enzyme activities (aspartate aminotransferase and alanine aminotransferase). LPG workers at Gaza Strip petroleum stations are at higher risk for health-related symptoms and clinical abnormalities.


Effect of retinoids on hyperproliferation-associated keratins K6 and K16 in cultured human keratinocytes: a quantitative analysis.
The keratin patterns of human epidermal keratinocytes cultured on a 3T3-feeder layer in the presence of 10(-8) M non-aromatic (all-trans retinoic acid and 13-cis retinoic acid) and polyaromatic (arotinoid, arotinoid-sulfone, and free arotinoic-acid) retinoids were analyzed by high resolution one- and two-dimensional gel electrophoresis and immunoblotting. Laser densitometric evaluation of one-dimensional gels allowed to quantitate the changes within the keratin patterns and revealed an increase in the expression of keratins K13, K15, and K19 as induced by both non-aromatic and polyaromatic retinoids, except for the parent compound arotinoid. This would then indicate that such keratinocytes are pursuing a more embryonic type of differentiation. In evaluating the data for the hyperproliferation-associated keratins K6 and K16 we noticed an unexpected result: except for all-trans retinoic acid, these two keratins showed opposite responses. As compared to control cultures, the amount of K6 did generally increase, while K16 was reduced, with arotinoid acid being the most effective retinoid. The apparently uncoupled expression of K6 and K16 appeared also to be concentration dependent when 13-cis retinoic acid at concentrations of 10(-9), 10(-8), and 10(-7) M was analyzed. Considering the overall antiproliferative potency of retinoids, we therefore conclude that K16 alone, rather than the pair K6/K16, should be regarded as a proliferation-related keratin and as such may be used as a sensitive marker to evaluate keratinocyte proliferation.


Effect of verapamil and trifluoperazine upon glucose-induced calcium distribution within B cells.
Insulin secretion and B-cell calcium distribution, assessed with the pyroantimonate precipitation technique, were studied in rat pancreases perfused with glucose (3.3 or 16.6 mM) alone or together with verapamil or trifluoperazine (TFP). Total calcium pyroantimonate precipitates (CPP), and those bound to every B cell structure, at every sampling period, were larger at 16.6 mM glucose concentration. The largest percentage of CPP was located, at early stages of the glucose stimulatory period, mainly within the clear halo of the B granules, while later on they shifted to the plasma membrane. Verapamil and TFP diminished the second phase of glucose-induced insulin release and greatly affected the above mentioned pattern of B cell CPP distribution. The main changes consist in a diminution in the total number of CPP all throughout the perfusion-time as well as an alteration in the percentage distribution of the CPP within the different B cell organelles, i.e., an early diminution in the CPP present in the B granules and of those attached to the plasma membrane and to the mitochondria at the end of the perfusion, were the most striking changes observed. The results suggest that during the glucose stimulus, different B cell structures take over the control of available calcium within the cell, following a chronological sequence. Such sequence might be determined by the different Ca2(+)-set point of those structures. Intracellular provision of calcium might be sufficient to maintain the early phase of insulin secretion when the cation entrance is blocked. Conversely, this substitution might not be enough to sustain the second phase of insulin release. The different amounts of CPP in every B cell organelle, besides their buffering capacity to control free Ca2+ availability, might also be coupled to a regulatory role of the cation upon their respective metabolic functions. In some cases, this latter effect may be the main role for the cation distribution. Our results support the concept that the level of free calcium, acting as the coupler for the stimulus:secretion process, might be regulated by different calcium pools located in the B cell.


The Body as a Tool for Anger Awareness--Differential Effects of Angry Facial and Bodily Expressions on Suppression from Awareness.
Emotional signals are perceived whether or not we are aware of it. The evidence so far mostly came from studies with facial expressions. Here, we investigated whether the pattern of non-conscious face expression perception is found for whole body expressions. Continuous flash suppression (CFS) was used to measure the time for neutral, fearful, and angry facial or bodily expressions to break from suppression. We observed different suppression time patterns for emotions depending on whether the stimuli were faces or bodies. The suppression time for anger was shortest for bodily expressions, but longest for the facial expressions. This pattern indicates different processing and detection mechanisms for faces and bodies outside awareness, and suggests that awareness mechanisms associated with dorsal structures might play a role in becoming conscious of angry bodily expressions.


Click synthesis of podand triazole-linked gold nanoparticles as highly selective and sensitive colorimetric probes for lead(II) ions.
Podand triazole-linked gold nanoparticles were designed via an in situ'click' reaction. The resulting gold nanoparticles were characterized by UV-vis spectroscopy, FT-IR spectroscopy, transmission electron microscopy (TEM). Podand triazole-linked gold nanoparticles were utilized as colorimetric probes for Pb(2+)via Pb(2+)-induced gold nanoparticle aggregation which incorporates two podand molecules from different nanoparticles forming an intermolecule type of association. Moreover, the application of the resulting gold nanoparticles to evaluate the exceeding standard of lead ions in drinking water and leaded paint was investigated.


The influence of methylpropionic acid and pyridazinone-3 derivatives on some immunologic and hemopoietic functions.
There was studied the influence on the cell-mediated and humoral response in vivo manifested by selected methylpropionic acid and pyridazinone-3 derivatives which had been found to possess strong immunotropic effects in the in vitro screening previously. It was shown that the compounds were generally poorly tolerated by animals, and they exerted only weak suppressive effects on antibody production, the contact hypersensitivity and survival of skin grafts. This immunosuppressive activity was accompanied by a slight decrease in the number of spleen colony forming cells (CFU-s). Only limited correlation between the biological activity of the preparations and their chemical structure was found.


Undercarboxylated matrix GLA protein levels are decreased in dialysis patients and related to parameters of calcium-phosphate metabolism and aortic augmentation index.
Vascular calcifications are related to cardiovascular mortality and morbidity in dialysis patients. Limited data exist on the role of calcification inhibitors, such as matrix-carboxyglutamic acid protein (MGP) in dialysis patients. In 120 dialysis patients and 41 age-matched healthy controls, circulating undercarboxylated (uc) MGP levels were measured with a novel ELISA-based competitive assay. The association between ucMGP levels and determinants of bone mineral metabolism, including the calcification inhibitor fetuin-A, was studied. Moreover, the relation between ucMGP levels and arterial stiffness was investigated. The ucMGP level was significantly lower in dialysis patients compared to controls (173 +/- 70 vs. 424 +/- 126 nmol/l; p < 0.0001). After adjustment for age, sex and duration of dialysis an independent negative association between time-averaged phosphate levels [regression coefficient beta with 95% confidence interval = -64 (-107 to -21)] and a positive association between serum ucMGP and fetuin-A [131 (55-208)] was observed. Duration of dialysis was inversely correlated with ucMGP (r = -0.24, p = 0.007). ucMGP levels were not related to high-sensitivity C-reactive protein or time-averaged calcium levels. After adjustment for age, sex, cardiovascular disease, diabetes, height and mean arterial pressure, ucMGP level was negatively associated with the aortic augmentation index [-0.036 (-0.061 to -0.010)] but not with pulse wave velocity or pulse pressure. Significantly lower serum ucMGP levels were observed in dialysis patients compared to healthy controls. ucMGP levels were inversely associated with phosphate and positively associated with serum fetuin-A levels. Furthermore, ucMGP levels were inversely associated with the aortic augmentation index. These data suggest that low ucMGP levels may be a marker of active calcification.


Effects of gender on stress ulcer formation in rats.
In the experimental stress literature, the results of investigations have not shown a specific sex-dependent vulnerability to stress ulceration. The aim of this study was to evaluate the importance of sex differences on stress ulcer development. Related to gender, the contributing factors for stress ulcer production such as luminal acidity, sialic acid as an marker of gastric mucosal protection, oxygen (O2)-derived free radicals and endogenous antioxidant defence mechanisms were also investigated. Fifty Wistar Albino rats weighing about 230 g and aged 7 or 8 months were divided equally into five groups: Group I normal male rats, group II castrated male rats, group III normal female rats in estrus phase, group IV normal female rats in diestrus phase and group V castrated female rats. Cold restraint model was used for 6 hours to produce stress ulcer. No statistically significant difference was found out between groups in view of gross and histopathologic damage. There was no significant difference between groups according to gastric luminal acidity, gastric mucosal sialic acid, gastric malonaldehyde (MDA) and catalase values. Gastric superoxide dismutase (SOD) activity was significantly lower in Group I in comparison to those of Group III and IV. Sex differences do not interfere stress ulcer formation. SOD activity in rat gastric tissue has varied significantly by hormonal milieu.


[Inhibition of peroxidase oxidation of aromatic amines by substituted phenols].
Peroxidase-catalyzed oxidation of o-phenylene diamine (OPD) was competitively inhibited by trimethylhydroquinone (TMHQ), 4-tert-butylpyrocatechol (In5), and 4,6-di-tert-butyl-3-sulfanyl-1,2-dihydroxybenzene (In6). In6 was the most efficient inhibitor (Ki = 11 microM at 20 degrees C in 0.015 M phosphate-citrate buffer, pH 6.0). The effects of In5 and In6 were not preceded by periods of induction of OPD oxidation products (contrary to TMHQ). Peroxidase-catalyzed oxidation of tetramethylbenzidine (TMB) was non-competitively inhibited by In6 and 3-(2-hydroxyethylthio)-4,6-di-tert-butylpyrocatechol (In4), whereas o-aminophenol (OAP) acted as a mixed-type inhibitor. The effects of all three inhibitors were preceded by an induction period, during which TMB oxidation products were formed. Again, In6 was the most efficient inhibitor (Ki = 16 microM at 20 degrees C in 0.015 M phosphate-citrate buffer supplemented with 5% ethanol, pH 6.0). Judging by the characteristics of the inhibitors, taken in aggregate, it is advisable to use the pairs OPD-In5 and OPD-In6 in systems for testing the total antioxidant activity of biological fluids of humans.


Focal seizures and EEG: prognostic considerations.
The goal of this study was to determine whether the EEG could predict if patients with focal seizures would eventually be uncontrolled (U), more than two seizures per month, or be controlled (C), fewer than two seizures per year. Using these latter criteria, U and C patients were randomly selected from our files, 150 in each of these two groups; 804 EEGs were found in the U and 674 in the C group. Excluded were patients with generalized epilepsy and also the benign epilepsies of childhood. Age was an important factor since patients 5-18 yrs old were more often under good control (C group), and those 21-48 yrs of age were more often under poor control (U group). With rare exceptions, focal spikes and focal slow waves in every area were much more often seen in U patients than C patients. No spikes or rare spikes appeared especially in the controlled patients, and many or very many spikes in the uncontrolled patients, mainly on the first or second EEG. No slow waves or a mild degree of slowing was seen especially in the C patients, while greater degrees of slow wave abnormalities were noted much more often in the U patients. For conditions to identify the C group, the best predictors were no spikes or rare spikes, especially on the first or second EEG, properly identifying two-thirds of the C patients and misidentifying only one-fourth of the U group. To identify the U patients on any EEG, many or very many spikes at any location or frontal spikes correctly designated a U patient in 84%, but incorrectly predicted a C patient as uncontrolled in 29%. For the first or second EEG, these values were 61% and 21%. Thus, the EEG, especially by its first or second record, can predict well the probable future of the uncontrolled patient, so that extraordinary means may be instituted early to avoid a deteriorating condition. When complete EEGs are done with sleep records and the results are quantified, reasonable prediction of eventual outcome can usually occur that is much more timely than waiting over 9 yrs, using only clinical data.


Progressive orbital granular cell tumour associated with medial rectus.
Granular cell tumour is a rare soft tissue tumour that can occur in any part of the body, but seldom in ocular adnexa. It usually behaves in a benign fashion. We report a case of a 54-year-old man with a well-demarcated, solitary, slow-growing orbital tumour which lead to significant ocular symptoms. The case was a diagnostic and therapeutic challenge due to its location and difficulty in obtaining tissue for a histological diagnosis. Surgical biopsy attempts were made but they all failed to uncover the true identity of the lesion. A definitive diagnosis was revealed with complete surgical excision of the tumour, which was challenging due to its size and close association with rectus muscle. This case has highlighted that orbital granular cell tumour may result in significant ocular symptoms. Adequate exposure to the anatomical site is the key to obtaining diagnosis and complete excision of a lesion.


[Quantitative mineralogical analyzes of kidney stones and diagnosing metabolic disorders in female patients with calcium oxalate urolithiasis].
To conduct a complex examination of female patients with calcium oxalate urolithiasis to detect metabolic disorders, leading to stone formation. The study was carried out using complex physical and chemical methods, including quantitative X-ray phase analysis of urinary stones, pH measurement, volumetry, urine and blood spectrophotometry. Quantitative mineralogical composition of stones, daily urine pH profile, daily urinary excretion of ions of calcium, magnesium, oxalate, phosphate, citrate and uric acid were determined in 20 female patients with calcium oxalate stones. We have shown that most of the stones comprised calcium oxalate monohydrate or mixtures of calcium oxalate dihydrate and hydroxyapatite. Among the identified abnormalities, the most frequent were hypocitraturia and hypercalciuria - 90 and 45%, respectively. Our findings revealed that the daily secretion of citrate and oxalate in patients older than 50 years was significantly lower than in younger patients. In conclusion, daily urinary citrate excretion should be measured in female patients with calcium oxalate stones. This is necessary both to determine the causes of stone formation, and to monitor the effectiveness of citrate therapy.


Development and Assessment of Postcranial Sex Estimation Methods for a Guatemalan Population.
This study tests whether postcranial sex estimation methods generated from Hispanic, and mainly Mexican samples, can be successfully applied to other increasingly common migrant populations from Central America. We use a sample of postcranial data from a modern (1980s) Guatemalan Maya sample (n = 219). Results indicate a decrease in classification accuracies for previously established univariate methods when applied to the Guatemalan study sample, specifically for males whose accuracies ranged from 30 to 84%. This bias toward inaccuracies for Guatemalan males is associated with the smaller skeletal sizes for the Guatemalan sample as compared to the samples used in the tested sex estimation methods. In contrast, the tested multivariate discriminant function classification yielded less sex bias and improved classification accuracies ranging from 82 to 89%. Our results highlight which of the tested univariate and multivariate methods reach acceptable levels for accuracy for sex estimation of cases where the region of origin may include Guatemala.


Genetic variations in sterol regulatory element binding protein cleavage-activating protein (SCAP) are associated with blood pressure in overweight/obese Chinese children.
Previous studies demonstrated a role of variations in sterol regulatory element binding protein (SREBP) cleavage-activating protein (SCAP) in obesity and blood lipids. But the associations between SCAP polymorphisms and blood pressure (BP) are not clear. This study aimed to investigate the relationship between genetic variations in SCAP and BP phenotypes in a Chinese pediatric population. A case-control study on 702 high blood pressure (HBP) children and 1319 controls was conducted to explore the correlation between single nucleotide polymorphism markers (rs12487736 and rs12490383) of SCAP and BP phenotypes. The associations with continuous and categorical variables were examined by linear regression and logistic regression models under a dominant genetic model for the minor rs12487736 A allele and rs12490383 T allele. The rs12487736 polymorphism was significantly associated with systolic BP (SBP) (β = 1.66, P = 0.003) and diastolic BP (DBP) (β = 1.35, P = 0.024) with age, age-squared, sex, study population and body mass index (BMI) adjusted under the dominant genetic model. The rs12490383 polymorphism was significantly associated with SBP (β = 1.71, P = 0.004) and SHBP (OR = 1.39, 95%CI: 1.04-1.86, P = 0.027). When analyzed by BMI categories, in the normal-weight children, no significant association between the SCAP polymorphisms and BP phenotypes was observed (all P > 0.05). However, in the overweight/obese children, rs12487736 was significantly associated with SBP (β = 1.6, P = 0.019) and SHBP (OR = 1.36, 95%CI: 1.02-1.82; P = 0.037), rs12490383 was associated with SBP (β = 2.04, P = 0.004) and SHBP (OR = 1.50, 95%CI: 1.10-2.05; P = 0.01). This study demonstrated that SCAP rs12487736 and rs12490383 were significantly associated with SBP and SHBP in overweight/obese Chinese children. It provided the evidence for association of SCAP with SBP.


Linking calcium-channel isoforms to potential therapies.
Voltage-gated calcium channels underlie a plethora of physiological functions in the cardiovascular and nervous systems. Calcium channels are considered important targets for the treatment of conditions such as absence epilepsy and hypertension, and significant efforts are increasing to discover novel calcium-channel modulators for the treatment of pain. An overview of advances in the development of calcium-channel therapeutics in the areas of pain, epilepsy and disorders associated with cardiovascular function are discussed.


Does the serum uric acid level have any relation to arterial stiffness or blood pressure in adults with congenital renal agenesis and/or hypoplasia?
The relationship between serum uric acid and arterial stiffness or blood pressure is not clear. The serum uric acid level and its association with cardiovascular risk is not well known in patients with reduced renal mass. We aimed to investigate the relation between serum uric acid levels and arterial stiffness and also blood pressure in patients with congenital renal agenesis and/or hypoplasia. In this single center, cross-sectional study, a total of 55 patients (39 (% 70.9) with unilateral small kidney and 16 (%29.1) with renal agenesis) were included. The median age was 35 (21-50) years. The study population was divided into tertiles of serum uric acid (according to 2.40-3.96, 3.97-5.10, and 5.11-9.80 mg/dl cut-off values of serum uric acid levels). Official and 24-h ambulatory non-invasive blood pressures of all patients were measured. The arterial stiffness was assessed by pulse wave velocity (PWV). PWV values were increased from first to third tertile (5.5 ± 0.6, 5.7 ± 0.8, 6.1 ± 0.7, respectively), but this gradual increase between tertiles did not reach significance. Linear regression analyses showed a positive correlation between serum uric acid levels and PWV (β = 0.40, p = 0.010), but no correlation was found between uric acid and daytime systolic blood pressure (β = 0.24, p = 0.345). In congenital renal agenesis/hypoplasia, the serum uric acid level was positively correlated with arterial stiffness, but there was no correlation with blood pressure.


Serum Concentrations of Ghrelin and Leptin according to Thyroid Hormone Condition, and Their Correlations with Insulin Resistance.
Thyroid hormones can influence energy metabolism and insulin sensitivity via their interaction with adipocytokines and gut hormones. The aims of this study were to evaluate differences in serum ghrelin and leptin concentrations according to thyroid hormone levels, and to investigate the correlation of insulin resistance. A total of 154 patients (57 hyperthyroid patients, 61 euthyroid patients, and 36 hypothyroid patients; mean age, 47.9 years) were enrolled. Serum leptin, ghrelin, and insulin levels were measured and insulin resistance was calculated using the formula of the homeostasis model assessment of insulin resistance (HOMA-IR). There were no differences in mean concentrations of ghrelin or leptin among the three groups. There were no significant differences in insulin levels between the groups (P=0.06), although hyperthyroid patients had borderline statistically significantly higher levels of insulin than did euthyroid subjects by post hoc test (26.4 μIU/mL vs. 16.1 μIU/mL, P=0.057). Regarding HOMA-IR index, the mean levels were highest in the hyperthyroid group among those of the three groups (hyperthyroid vs. euthyroid vs. hypothyroid, 6.7 vs. 3.8 vs. 4.4, P=0.068). Plasma levels of ghrelin were significantly negatively correlated with age, insulin, glucose, body mass index (BMI), and HOMA-IR. Plasma levels of leptin showed significant positive correlation with BMI and triglyceride. There were no significant correlations among thyroid hormone, thyrotropin, ghrelin, leptin, or insulin. The present study found that serum ghrelin, leptin, and insulin levels didn't differ according to thyroid function conditions. Further studies with larger numbers of patients are required to establish a direct relationship between plasma ghrelin, leptin, and thyroid hormone.


Geographic stomatitis: an oral manifestation of psoriasis?
Geographic stomatitis is an uncommon oral lesion that presents similar clinical, histopathological and genetic features as those of psoriasis. These findings suggest that this lesion may actually represent an oral manifestation of psoriasis. We report one case of geographic stomatitis and discuss a possible connection between this condition and psoriasis. A 37-year-old woman presented with red patches, surrounded by a white border on the labial mucosa and a positive family history of psoriasis. Histopathological examination, immunohistochemical analysis with antibodies against CD4, CD8, CD20, CD68, CD31, and Ki-67 and HLA-A*, -B*, -C*, -DRB1*, -DQA1* and -DQB1* genotyping were performed. Histopathological examination revealed parakeratosis, marked elongation of rete ridges with acanthosis and clubbing, exocytosis, Munro microabscesses, pustule of Kogoj, dilated tortuous vessels at the tip of dermal papillae, and predominant superficial and perivascular lymphocytic chronic inflammatory cell infiltrate. Immunohistochemistry analysis revealed a predominant T-cell subepithelial infiltrate. Based on the referred clinicopathological findings and in the absence of cutaneous lesions, the diagnosis of geographic stomatitiswas confirmed. This case and theoretical data indicate that geographic stomatitis may be an oral manifestation of psoriasis. Moreover, to improve our understanding, psoriatic patients should routinely undergo a detailed oral examination and patients with geographic stomatitis should routinely be submitted to a cutaneous routine examination.


Tissue-specific accumulation and regulation of zeaxanthin epoxidase in Arabidopsis reflect the multiple functions of the enzyme in plastids.
The enzyme zeaxanthin epoxidase (ZEP) catalyzes the conversion of zeaxanthin to violaxanthin, a key reaction for ABA biosynthesis and the xanthophyll cycle. Both processes are important for acclimation to environmental stress conditions, in particular drought (ABA biosynthesis) and light (xanthophyll cycle) stress. Hence, both ZEP functions may require differential regulation to optimize plant fitness. The key to understanding the function of ZEP in both stress responses might lie in its spatial and temporal distribution in plant tissues. Therefore, we analyzed the distribution of ZEP in plant tissues and plastids under drought and light stress by use of a ZEP-specific antibody. In addition, we determined the pigment composition of the plant tissues and chloroplast membrane subcompartments in response to these stresses. The ZEP protein was detected in all plant tissues (except flowers) concomitant with xanthophylls. The highest levels of ZEP were present in leaf chloroplasts and root plastids. Within chloroplasts, ZEP was localized predominantly in the thylakoid membrane and stroma, while only a small fraction was bound by the envelope membrane. Light stress affected neither the accumulation nor the relative distribution of ZEP in chloroplasts, while drought stress led to an increase of ZEP in roots and to a degradation of ZEP in leaves. However, drought stress-induced increases in ABA were similar in both tissues. These data support a tissue- and stress-specific accumulation of the ZEP protein in accordance with its different functions in ABA biosynthesis and the xanthophyll cycle.


Donor strains of the soft-rot bacterium Erwinia chrysanthemi and conjugational transfer of the pectolytic capacity.
Donor strains of Erwinia chrysanthemi ICPB EC16, a member of the soft-rot (pectolytic) section of the enterobacterial genus Erwinia, were obtained by chromosomal integration of an F'lac(+) plasmid originating from Escherichia coli. These stable donor strains, selected from an unstable F'lac(+) heterogenote by repeated platings of single Lac(+) colonies on lactose minimal agar, do not segregate (as does the parent F'lac(+) heterogenote) into Lac(-) or F(-) clones, in either the presence or absence of acridine orange. One representative donor strain (from the 12 that have been selected) has been examined in more detail; it can transfer ade(+), gal(+), gtu(+) (utilization of galacturonate), his(+), lac(+), leu(+), lys(+), mcu(+) (multiple carbohydrate utilization), pat(+) (production of polygalacturonic acid trans-eliminase), thr(+), and trp(+) in a polarized manner to appropriate recipient strains of E. chrysanthemi; the frequencies of ade(+), leu(+), and thr(+) transfer were higher than those of the other markers tested to date. This donor strain transfers lac(+) genes during a 6-h mating on membranes; most of the Lac(+) recombinants are donors of chromosomal markers. The kinetics of entry as well as the frequencies of transfer of chromosomal markers indicate that thr(+) and leu(+) enter the recipient as proximal markers and that lac(+) enters as a distal marker. Analysis of the recombinants demonstrates close linkage between thr and leu, ade and thr, his and pat, and his and trp loci. The results suggest that the integration of F'lac(+) into the chromosome of E. chrysanthemi has occurred at a region adjacent to the leu-thr loci, and that the chromosome is transferred in the following sequence: origin----leu--thr--ade--lys--mcu--pat--his--trp--gal--gtu--lac--F. Plant-tissue maceration occurs in Pat(+) recombinants and not in Pat(-) recombinants, even though both form another pectolytic enzyme, hydrolytic polygalacturonase. This genetic evidence supports the idea that the E. chrysanthemi polygalacturonic acid trans-eliminase plays an essential role in bringing about plant-tissue maceration.


Dose-response effects of dietary protein on muscle protein synthesis during recovery from endurance exercise in young men: a double-blind randomized trial.
Protein ingestion increases skeletal muscle protein synthesis rates during recovery from endurance exercise. We aimed to determine the effect of graded doses of dietary protein co-ingested with carbohydrate on whole-body protein metabolism, and skeletal muscle myofibrillar (MyoPS) and mitochondrial (MitoPS) protein synthesis rates during recovery from endurance exercise. In a randomized, double-blind, parallel-group design, 48 healthy, young, endurance-trained men (mean ± SEM age: 27 ± 1 y) received a primed continuous infusion of l-[ring-2H5]-phenylalanine, l-[ring-3,5-2H2]-tyrosine, and l-[1-13C]-leucine and ingested 45 g carbohydrate with either 0 (0 g PRO), 15 (15 g PRO), 30 (30 g PRO), or 45 (45 g PRO) g intrinsically l-[1-13C]-phenylalanine and l-[1-13C]-leucine labeled milk protein after endurance exercise. Blood and muscle biopsy samples were collected over 360 min of postexercise recovery to assess whole-body protein metabolism and both MyoPS and MitoPS rates. Protein intake resulted in ∼70%-74% of the ingested protein-derived phenylalanine appearing in the circulation. Whole-body net protein balance increased dose-dependently after ingestion of 0, 15, 30, or 45 g protein (mean ± SEM: -0.31± 0.16, 5.08 ± 0.21, 10.04 ± 0.30, and 13.49 ± 0.55 μmol phenylalanine · kg-1 · h-1, respectively; P < 0.001). 30 g PRO stimulated a ∼46% increase in MyoPS rates (%/h) compared with 0 g PRO and was sufficient to maximize MyoPS rates after endurance exercise. MitoPS rates were not increased after protein ingestion; however, incorporation of dietary protein-derived l-[1-13C]-phenylalanine into de novo mitochondrial protein increased dose-dependently after ingestion of 15, 30, and 45 g protein at 360 min postexercise (0.018 ± 0.002, 0.034 ± 0.002, and 0.046 ± 0.003 mole percentage excess, respectively; P < 0.001). Protein ingested after endurance exercise is efficiently digested and absorbed into the circulation. Whole-body net protein balance and dietary protein-derived amino acid incorporation into mitochondrial protein respond to increasing protein intake in a dose-dependent manner. Ingestion of 30 g protein is sufficient to maximize MyoPS rates during recovery from a single bout of endurance exercise.This trial was registered at trialregister.nl as NTR5111.


Empathy for others versus for one's child: Associations with mothers' brain activation during a social cognitive task and with their toddlers' functioning.
Caregivers who are higher in dispositional empathy tend to have children with better developmental outcomes; however, few studies have considered the role of child-directed (i.e., "parental") empathy, which may be relevant for the caregiver-child relationship. We hypothesized that mothers' parental empathy during their child's infancy will be a stronger predictor of their child's social-emotional functioning as a toddler than will mothers' dispositional empathy. We further explored whether parental and dispositional empathy have shared or distinct patterns of neural activation during a social-cognitive movie-watching task. In 118 mother-infant dyads, greater parental empathy assessed when infants were 6 months old was associated with more social-emotional competencies and fewer problems in the children 1 year later, even after adjusting for dispositional empathy. In contrast, dispositional empathy was not associated with child functioning when controlling for parental empathy. In a subset of 20 mothers, insula activation was positively associated with specific facets of both dispositional and parental empathy, whereas right temporoparietal junction activation was associated only with parental empathy. Thus, dispositional and parental empathy appear to be dissociable by both brain and behavioral metrics. Parental empathy may be a viable target for interventions, especially for toddlers at risk for developing social-emotional difficulties.


Predator-induced changes of male and female mating preferences: innate and learned components.
While many mating preferences have a genetic basis, the question remains as to whether and how learning/experience can modify individual mate choice decisions. We used wild-caught (predator-experienced) and F1 laboratory-reared (predator-naïve) invasive Western mosquitofish Gambusia affinis from China to test whether mating preferences (assessed in a first mate choice test) would change under immediate predation threat. The same individuals were tested in a second mate choice test during which 1 of 3 types of animated predators was presented: 1) a co-occurring predator, 2) a co-evolved but not currently co-occurring predator, and 3) a non-piscivorous species as control. We compared preference scores derived from both mate choice tests to separate innate from experiential effects of predation. We also asked whether predator-induced changes in mating preferences would differ between sexes or depend on the choosing individual's personality type and/or body size. Wild-caught fish altered their mate choice decisions most when exposed to the co-occurring predator whereas laboratory-reared individuals responded most to the co-evolved predator, suggesting that both innate mechanisms and learning effects are involved. This behavior likely reduces individuals' risk of falling victim to predation by temporarily moving away from high-quality (i.e., conspicuous) mating partners. Accordingly, effects were stronger in bolder than shyer, large- compared with small-bodied, and female compared with male focal individuals, likely because those phenotypes face an increased predation risk overall. Our study adds to the growing body of literature appreciating the complexity of the mate choice process, where an array of intrinsic and extrinsic factors interacts during decision-making.


Phonomyogram of the diaphragm during unilateral and bilateral phrenic nerve stimulation and changes with fatigue.
As previously demonstrated in humans, contracting skeletal muscles vibrate laterally and generate sound. The amplitude of this acoustic signal, called the phonomyogram (PMG), is related to muscle force production. As in limb muscles, the diaphragm might produce mechanical tissue vibrations and a PMG. We recorded diaphragm twitches during phrenic nerve stimulation in normal subjects at FRC. Diaphragmatic compound motor action potentials (CMAP) were recorded on each side of the chest by surface electrodes over the eighth intercostal space. Condenser microphones were fixed to the skin close to the electrodes, and esophageal and gastric balloons employed to measure transdiaphragmatic pressure (Pdi). For unilateral and bilateral supramaximal single shocks (0.1 ms), PMG onset latencies from either side (12.4 +/- 0.6 ms) were intermediate between those of CMAP (7.3 +/- 0.7 ms) and of Pdi (20.3 +/- 2.8 ms). By varying stimulation intensity, significant linear relationships were obtained between CMAP and PMG amplitudes on each side. A significant linear relationship was also found between PMG for each side and Pdi peak values. PMG and Pdi both decreased with fatigue of the diaphragm. We conclude that PMG in response to single phrenic nerve shocks is a noninvasive index of electromechanical coupling, recruitment, and transdiaphragmatic pressure generation by the diaphragm.


Development of a nucleic acid lateral flow strip for detection of hepatitis C virus (HCV) core antigen.
The object of this study was to develop a simple, rapid, specific, and highly sensitive method to detect HCV core antigen. A nucleic acid aptamer was designed with the high specificity and sensitivity in a nucleic acid lateral flow strip to compete with HCV core antigen and DNA probes. The lower detection limit of the test strip was calculated to be 10 pg/mL with the scanner and 100 pg/mL with naked eyes. Results showed that there were no cross-interactions with other proteins such as HCV NS3, E1/E2 antigens, HIV p24 antigens, or BSA proteins (HCV unrelated protein). When the viral load exceeded 10(4) copies/mL, the positive coincidence rates of ELISA and strip detection, when compared with the HCV RNA assay, were 98.44% and 97.28%, respectively. The results indicated that the ELISA detection and strip assay were in good agreement with the measured value. The results indicated that a nucleic acid lateral flow strip was a simple, rapid, specific, highly sensitive, and cost-effective field-based method for detecting HCV core antigen. The strip assay is an acceptable alternative to diagnose HCV core antigen and to investigate its epidemiology in clinical laboratories lacking specialized equipment and skills.


A cytotoxic monoclonal anti-leukemia antibody binds to histone H1.
Monoclonal antibody (MAb) AP64 is a mouse IgM MAb raised against human acute non-lymphocytic leukemia (ANLL) cells. It has been shown to bind to a wide variety of cell lines and is capable of initiating complement (C) dependent cytotoxicity. Other studies indicated that MAb AP64 can effect long term cure in a leukemia minimal residual disease model. By using various techniques we have determined the identity of a protein which is bound by this MAb. Immunofluorescent studies have shown that MAb AP64 stains the nuclei of fixed cells as well as metaphase chromosomes, indicating that this MAb binds to a component of chromatin. Biochemical characterization revealed that MAb AP64 western blots a 31 and 32 kilodalton doublet from NP-40 extracts from both rat and human leukemia cells. The mobility of this doublet is identical under reducing and non-reducing conditions. Further studies have shown that the bands detected by western blot analysis using MAb AP64 as a probe have a similar migration to those of bovine histone H1. Also, 1 nanogram of bovine histone H1 can be detected by MAb AP64 when spotted onto nitrocellulose. These data demonstrate that MAb AP64 binds to a conserved epitope present on molecules coded for by the histone H1 gene family.


A mathematical model for lambda dv plasmid replication: analysis of wild-type plasmid.
A mathematical model for lambda dv plasmid replication in a growing single cell of Escherichia coli has been formulated and solved numerically. Quantitative description of the molecular control mechanism for initiation of lambda dv replication presumes regulatory functions of repressor and initiator proteins and transcriptional activation of the origin region. Random selection of a single plasmid for activation and replication is assumed, as is regular plasmid segregation to daughter cells. The model is capable of simulating the periodic changes in each regulatory element and the plasmid copy number during the cell cycle. The calculated average copy number, repressor concentration, and timing of plasmid replication agree well with experimental data. The simulated lambda dv plasmid replication rate is controlled primarily by transcription frequency. Initiation of plasmid replication is not related to variations in the levels of repressor or initiator proteins during the cell cycle. Simulation studies of perturbations in plasmid and repressor segregation indicate that replication regulation of the lambda dv plasmid compensates to readjust copy number to normal values in a few generations. Implications of these studies relative to the molecular mechanisms of replication control are discussed.


Use of equine chorionic gonadotropin in lactating dairy cattle: a rapid review.
To use an evidence-based approach to evaluate the available research data on the efficacy of equine chorionic gonadotropin (eCG) to improve reproductive performance of dairy cows when added to oestrus synchronisation programmes for the treatment of cows not detected in oestrus. A rapid literature review was conducted to summarise the existing trial data, primarily the effect of eCG on conception rate (CR) to fixed time artificial insemination (FTAI) from oestrus synchronisation programmes in lactating dairy cows. Relevant papers were identified via databases and auxiliary search strategies, then information was collected using a standardised data collection form. Similar studies were grouped together to create a descriptive summary of CR to FTAI and secondary outcome measures, with the creation of forest plots to visualise results. A total of 15 studies were identified for review: nine examined the administration of 400-500 IU eCG as part of an oestrus synchronisation programme based on intra-vaginal progesterone-releasing devices, gonadotropin-releasing hormone and prostaglandin (P4-GPG programme). The variable methodologies in the trials evaluated posed difficulties in grouping studies for further analysis and meant that there was insufficient data for a full meta-analysis. A single study reported a statistically significant increase in CR to FTAI from the addition to eCG to a P4-GPG programme in cows with anovulatory anoestrous. Four studies evaluated the addition of eCG to P4-GPG programmes in cows without visible oestrous signs; risk differences for CR to FTAI were between -1.7% and 28.8%, with three out of four studies reporting a positive effect of eCG on CR to FTAI, but all 95% CI values crossed the line of no effect. Two studies reported on the effect of eCG on 42-day pregnancy rate following a P4-GPG programme; one reported a risk difference of -4.9% (95% CI = -10.6-0.8%), while the other reported a risk difference of 7.9% (95% CI = 0.8-15%). The results of published studies describing the effect of eCG in P4-GPG programmes on CR to FTAI are inconsistent and further well-designed, adequately powered studies with standardised outcome measures are required to investigate its effect.


Expression and clinical contribution of MRGD mRNA in non-small cell lung cancers.
MAS-related G protein-coupled receptor, member D (MRGD) has been reported to be involved in tumorigenesis in vivo. However, the clinical role of MRGD in non-small cell lung cancer (NSCLC) remains unclarified. The purpose of the current study was to detect the expression of MRGD mRNA in NSCLC formalin-fixed (FF), paraffin-embedded (PE) tissues and to investigate the clinicopathological significance of the MRGD level in NSCLC patients. The expression of MRGD mRNA was examined in 125 NSCLC tissue samples together with paired para-noncancerous FF/PE tissues by using real time quantitative PCR (qRT-PCR). Furthermore, the relationship between MRGD level and clinicopathological parameters of NSCLC was analyzed. The average level of MRGD in NSCLC tumor tissues (1.0682±0.6096) was remarkably higher than that in the adjacent non-cancerous lung tissue (0.3994±0.2838, p<0.001). The area under curve (AUC) of receiver operating characteristic curve (ROC) of MRGD mRNA was 0.853 (95% CI: 0.808-0.898, p(0.001). Moreover, the level of MRGD mRNA was found to be correlated to lymph node metastasis (r=0.219, p=0.014), tumor size (r=0.221, p=0.013) and clinical TNM stage (r=0.187, p=0.037). Finally, the survival of patients in high MGRD expression group was 7.94±9.85 months, remarkably shorter than that of the low expression group (20.84±1.19 months, p=0.049). MRGD may be a vital diagnostic and prognostic factor in NSCLC. MRGD possesses the potential to become a new target for the molecular therapy of NSCLC.


Superionic adjustment leading to weakly temperature-dependent ZT values in bulk thermoelectrics.
Thermoelectric (TE) materials are of worldwide interest for energy sustainability through direct waste-heat-to-electricity conversion. Practically, a TE power generator requires a large working temperature gradient; to achieve high efficiency, key TE materials with high ZT values are necessary and, furthermore, their ZT values should decline as little as possible over the imposed temperature range. Unfortunately, sharp ZT declines in all of the known materials are inevitable. Here we found the bulk superionic α-Ag(1-x)CuSe material exhibits unusual weakly temperature-dependent ZT values in the range of 480-693 K with the smallest ZT-T slope known to date. These result from the Seebeck coefficient balance of the countercontributions of holes and electrons and the weakly temperature-dependent thermal conductivity.


Daunorubicin and cytarabine for certain types of poor-prognosis acute myeloid leukemia: a systematic literature review.
Induction chemotherapy based on anthracyclines and cytarabine (Ara-C) combination remains the standard of care for acute myeloid leukemia (AML) patients who are considered candidate for intensive and curative approaches. However, the toxicity of this regimen is high, with disappointing clinical outcomes among the so-called poor-prognosis AML subsets, which generally refer to patients with adverse cytogenetic risk, secondary AML including therapy-related AML, poor-prognosis mutations, especially FLT3-ITD, and relapse/refractory AML. Areas covered: To the best of our knowledge, the role and efficacy of 7 + 3 schedules containing daunorubicin (DNR) and Ara-C for certain types of poor-prognosis AML has not been systematically assessed. A critical approach to the role of DNR and Ara-C induction could be relevant to establish which patients should be enrolled in clinical trials using novel therapies. Expert commentary: In this regard, a recent randomized clinical trial (RCT) showed improved results in older patients with sAML or high-risk cytogenetics who received CPX-351 compared with standard 7 + 3 combination. We perform a systematic literature review to analyze the clinical outcomes reported with DNR plus Ara-C regimens in adult patients with poor-prognosis AML, the use of liposomal formulations of DNR and Ara-C and the RCTs which compared standard 7 + 3 with the addition of a third drug.