The Antidepressant Duloxetine Inhibits Platelet Function and Protects against Thrombosis.
While cardiovascular disease (CVD) is the leading cause of death, major depressive disorder (MDD) is the primary cause of disability, affecting more than 300 million people worldwide. Interestingly, there is evidence that CVD is more prevalent in people with MDD. It is well established that neurotransmitters, namely serotonin and norepinephrine, are involved in the biochemical mechanisms of MDD, and consequently, drugs targeting serotonin-norepinephrine reuptake, such as duloxetine, are commonly prescribed for MDD. In this connection, serotonin and norepinephrine are also known to play critical roles in primary hemostasis. Based on these considerations, we investigated if duloxetine can be repurposed as an antiplatelet medication. Our results-using human and/or mouse platelets show that duloxetine dose-dependently inhibited agonist-induced platelet aggregation, compared to the vehicle control. Furthermore, it also blocked agonist-induced dense and α-granule secretion, integrin αIIbβ3 activation, phosphatidylserine expression, and clot retraction. Moreover duloxetine-treated mice had a significantly prolonged occlusion time. Finally, duloxetine was also found to impair hemostasis. Collectively, our data indicate that the antidepressant duloxetine, which is a serotonin-norepinephrine antagonist, exerts antiplatelet and thromboprotective effects and inhibits hemostasis. Consequently, duloxetine, or a rationally designed derivative, presents potential benefits in the context of CVD, including that associated with MDD.


Learning from plant movements triggered by bulliform cells: the biomimetic cellular actuator.
Within the framework of a biomimetic top-down approach, our study started with the technical question of the development of a hinge-free and compliant actuator inspired by plant movements. One meaningful biological concept generator was the opening and closing movements of the leaf halves of grasses. Functional morphological investigations were carried out on the selected model plant Sesleria nitida. The results formed the basis for further clarifying the functional movement principle with a particular focus on the role of turgor changes in bulliform cells on kinetic amplification. All findings gained from the investigations of the biological model were incorporated into a finite-element analysis, as a prerequisite for the development of a pneumatic cellular actuator. The first prototype consisted of a row of single cells positioned on a plate. The cells were designed in such a way that the entire structure bent when the pneumatic pressure applied to each individual cell was increased. The pneumatic cellular actuator thus has the potential for applications on an architectural scale. It has subsequently been integrated into the midrib of the facade shading system Flectofold in which the bending of its midrib controls the hoisting of its wings.


[Estimation by static SPECT of the radioactivity remaining after the first dose of two-fractionated 123I-IMP administration: the rate of increase in cerebral blood flow by ARG and the rate of increase in SPECT count].
We devised a method for estimating the radioactivity remaining after the first dose of two-fractionated tracer administration in the acetazolamide activation study in a facility where static SPECT is used for 123I-IMP cerebral blood flow scintigraphy. Three static SPECT scans were obtained over 9-min periods beginning 10 min after the first 123I-IMP administration, to estimate the remaining activity until after 60 min. In 72 patients at rest, 9-min SPECT was performed 6 times, and 11 different patterns of distribution and the fitting coefficient were obtained. The correlation between the actual measurement and the estimation at the mid scan time of 59.5 min could be expressed as y = 1.0064x - 1.9656, and r = 0.9972 (p < 0.01). The correlation between the first and second measurements of cerebral blood flow in 5 patients given two-fractionated administration at rest could be expressed as y = 0.9919x + 0.2978, r = 0.9976 (p < 0.01), indicating the usefulness of this method for estimating the radioactivity remaining after the first dose. In 57 patients with unilateral cerebrovascular disease, the cerebral blood flow on the unaffected side increased by 55.4 +/- 13.1%, whereas the blood flow in 19 regions exhibiting severe stenosis on the affected side was increased only 1.4 +/- 10.5% (p < 0.01). In addition, the correlation between the rate of increase in cerebral blood flow and the rate of increase in SPECT count could be expressed as y = 0.8415x + 0.291, and r = 0.9979 (p < 0.01). Thus, with this method, the cerebral hemodynamic reserve test using the rate of increase in cerebral blood flow by the ARG method or the rate of increase in SPECT count can be completed within a day while maintaining the image quality of static SPECT.


Protein Synthesis during Natural and Precocious Soybean Seed (Glycine max [L.] Merr.) Maturation.
Protein synthesis was studied during precocious and natural soybean seed (Glycine max [L.] Merr.) maturation. Developing seeds harvested 35 days after flowering were precociously matured through controlled dehydration. Total soluble proteins and proteins labeled with [(35)S]methionine were extracted from control, developing seeds and from precociously and naturally matured seeds and were analyzed by one-dimensional PAGE and fluorography. The results demonstrated that several polypeptides which were designated "mature polypeptides," were synthesized de novo during precocious and natural seed maturation. Two of these polypeptides, 31 and 128 kilodalton in mass, also stained intensely with Coomassie blue, suggesting their abundant accumulation during seed maturation. Results from in vitro translation experiments showed that the mRNAs corresponding to these "maturation polypeptides" accumulated during precocious maturation and in naturally matured seeds, but not in seeds freshly harvested 35 days after flowering (control). The role of the "maturation polypeptides" is currently unknown; however, their presence and that of their corresponding mRNAs was coincident with the ability of matured seeds to establish seedling growth. This study has demonstrated that precocious seed maturation treatments may be extremely useful for investigations of metabolic events and molecular control mechanisms affecting soybean seed maturation.


Transcriptome changes in DM1 patients' tissues are governed by the RNA interference pathway.
Myotonic dystrophy type 1 (DM1) is a multisystemic disease caused by pathogenic expansions of CTG repeats. The expanded repeats are transcribed to long RNA and induce cellular toxicity. Recent studies suggest that the CUG repeats are processed by the RNA interference (RNAi) pathway to generate small interfering repeated RNA (siRNA). However, the effects of the CTG repeat-derived siRNAs remain unclear. We hypothesize that the RNAi machinery in DM1 patients generates distinct gene expression patterns that determine the disease phenotype in the individual patient. The abundance of genes with complementary repeats that are targeted by siRNAs in each tissue determines the way that the tissue is affected in DM1. We integrated and analyzed published transcriptome data from muscle, heart, and brain biopsies of DM1 patients, and revealed shared, characteristic changes that correlated with disease phenotype. These signatures are overrepresented by genes and transcription factors bearing endogenous CTG/CAG repeats and are governed by aberrant activity of the RNAi machinery, miRNAs, and a specific gain-of-function of the CTG repeats. Computational analysis of the DM1 transcriptome enhances our understanding of the complex pathophysiology of the disease and may reveal a path for cure.


Developmental expression of genetically defined peptidases in maize.
The activities of genetically defined amino- and endopeptidases of maize were compared in pericarp, endosperm, and embryonic tissue of the maize kernel from 5 days postpollination until harvest. Activities were highest in the immature stages and declined as drying progressed. The expression of some of the peptidase genes contributed by the pollen parent was examined during early endosperm development in an F(1) cross. The paternally contributed peptidase variants could first be detected 7 days after pollination.During germination and early seedling growth the peptidase activities continued to decline in the endosperm but maintained high levels in the scutellum. The maize peptidases probably have no direct role in hydrolysis of endosperm storage proteins during germination and growth.The peptidase isozymes were associated with the soluble cytoplasmic fraction of the maize scutellum during early growth. Only one tissue specificity was found for the peptidases. Occurrence of a particular aminopeptidase isozyme, AMP2, is limited to endosperm tissue during kernel maturation.


Activity prediction of aminoquinoline drugs based on deep learning.
The results of the traditional prediction method for the activity of aminoquinoline drugs are inaccurate, so the prediction method for the activity of aminoquinoline drugs based on the deep learning is designed. The molecular holographic distance vector method was used to describe the molecular structure of 40 aminoquinoline compounds, and the principal component regression method was used for modeling and quantitative analysis. Two methods were used to predict the activity of aminoquinoline drugs. The correlation coefficients of the results obtained from the two sets of activity data and the cross test were 0.9438 and 0.9737, and 0.8305 and 0.9098, respectively. Our data suggested that method for the activity prediction of aminoquinoline drugs based on deep learning studied in this paper can better predict the activity of aminoquinoline drugs and provide a strong basis for the activity prediction of aminoquinoline drugs.


On the relationships between rural labor productivity, family investment in education, and population quality.
The authors consider "the relationships between labor productivity and population quality in order to demonstrate the importance of economic development, and of improving population quality in rural areas. This study is made on the basis of surveys conducted in 1979 and 1986 on the age-specific VP and VC [volume of production and volume of consumption] in certain areas of Jiangsu Province [China]." Population quality is defined primarily in terms of the educational status of the labor force.


In vitro DNA binding activity of Fos/Jun and BZLF1 but not C/EBP is affected by redox changes.
The leucine zipper family of proteins have a DNA binding domain composed of a leucine zipper dimerisation interface and a basic DNA binding structure. We show here that redox changes affect the in vitro DNA binding ability of a select subset of leucine zipper proteins. The bacterially expressed DNA binding domains of Fos/Jun and BZLF1 are unable to bind DNA under non-reducing conditions whereas binding of the C/EBP DNA binding domain is unaffected. Sensitivity to redox state is due to the presence of a conserved cysteine residue in the basic DNA binding motif of Fos, Jun and BZLF1 but not C/EBP. Under non-reducing conditions an intermolecular disulphide bridge is formed between the cysteine residues of each basic motif within a dimer, which prevents DNA binding. We show that oxidation of these C residues can be achieved enzymatically, using glutathione peroxidase, and that DNA binding protects them from oxidation. These data raise the possibility that intracellular changes in the redox state may differentially regulate the activity of leucine zipper family members. In addition the loss of DNA binding activity under non-reducing conditions has implications for the purification methods used to isolate proteins of the leucine zipper family for structural analysis.


Multimodal radiography: a new imaging technique and system for oral diagnosis.
The multimodal radiography system is based on the combined use of narrow-beam radiography and spiral tomography in one set of equipment. The essentials of a system are a multifunction unit, preprogrammed imaging procedures for various dentomaxillofacial objects and a co-ordinate system for object location. Four years of clinical experience have shown that multimodal radiography is effective for depicting dentomaxillofacial structures for diagnostic purposes. The detailed narrow beam imaging mode of multimodal radiography has also been compared with intraoral radiography as regards their abilities to reveal periodontal and periapical lesions. Preliminary results indicate that detailed narrow beam radiography is more accurate than intraoral radiography in the diagnosis of periodontal and periapical pathology.


A case-crossover study of risk factors for occupational eye injuries.
To study transient risk factors for occupational eye injuries. A case-crossover study was conducted among patients treated for occupational eye injuries in the emergency department at an eye hospital in Alabama. A questionnaire was administered to collect information regarding risk factors at the time of and prior to eye treatment. Incidence rate ratios were used to measure the relationship between each risk factor and injury occurrence. Protective eyewear reduced the risk of occupational eye injury, while increased risk was observed for the following: being distracted, use of tools, tool malfunction, performing an unfamiliar task, being rushed, working overtime, and feeling fatigued. Although use of protective eyewear can significantly reduce the risk of an eye injury, other factors are important contributors. Identification of potentially modifiable transient risk factors can be used to prevent occupational eye injuries.


Phosphorylation status determines the opposing functions of Smad2/Smad3 as STAT3 cofactors in TH17 differentiation.
Transforming growth factor-β (TGF-β) and interleukin-6 (IL-6) are the pivotal cytokines to induce IL-17-producing CD4(+) T helper cells (TH17); yet their signalling network remains largely unknown. Here we show that the highly homologous TGF-β receptor-regulated Smads (R-Smads): Smad2 and Smad3 oppositely modify STAT3-induced transcription of IL-17A and retinoic acid receptor-related orphan nuclear receptor, RORγt encoded by Rorc, by acting as a co-activator and co-repressor of STAT3, respectively. Smad2 linker phosphorylated by extracellular signal-regulated kinase (ERK) at the serine 255 residue interacts with STAT3 and p300 to transactivate, whereas carboxy-terminal unphosphorylated Smad3 interacts with STAT3 and protein inhibitor of activated STAT3 (PIAS3) to repress the Rorc and Il17a genes. Our work uncovers carboxy-terminal phosphorylation-independent noncanonical R-Smad-STAT3 signalling network in TH17 differentiation.


Characterization of the human cytochrome P4502D6 promoter. A potential role for antagonistic interactions between members of the nuclear receptor family.
The functional mapping of the human cytochrome P4502D6 (CYP2D6) promoter in HepG2 cells revealed the presence of both positive and negative regulatory elements. One of these regulatory elements overlapped a sequence that is highly conserved in most members of the CYP2 family. This element, which consists of a degenerate AGGTCA direct repeat spaced by 1 base pair (DR1) and is known to be a target for members of the steroid receptor superfamily, was found to bind in vitro translated hepatocyte nuclear factor 4 (HNF4) in gel retardation analysis. Using HepG2 nuclear extracts, three protein-DNA complexes were formed on the DR1 element, one of which was confirmed to be dependent on the binding of HNF4. The other DR1 complexes were shown to be due to the interaction of the orphan receptor chicken ovalbumin upstream promoter transcription factor I (COUP-TFI). Experiments in COS-7 cells showed that HNF4 could activate the CYP2D6 promoter 30-fold. Surprisingly, mutation of the DR1 element produced a relatively minor 23% decrease in activity in HepG2 cells. Additionally, COUP-TFI was shown to inhibit HNF4 stimulation of the CYP2D6 promoter in COS-7 cells, suggesting that COUP-TFI could attenuate the effect of HNF4 in HepG2 cells. However, when HNF4 levels were increased in HepG2 cells by co-transfection, it resulted in the enhancement of CYP2D6 promoter activity, indicating that HNF4 could overcome the repressive effect of COUP-TFI. Therefore, the contribution of the DR1 element in controlling the transcription of the CYP2D6 gene depends on the balance between positively and negatively acting transcription factors.


Reproductive toxicity evaluation of a new camptothecin anticancer agent, CKD-602, in pregnant/lactating female rats and their offspring.
CKD-602 is a camptothecin anticancer agent that was recently developed by the Chong Kun Dang Pharmaceutical Co. (Seoul, Korea). This study examined the potential adverse effects of CKD-602 on pregnancy, delivery, and lactation in female Sprague-Dawley rats as well as on the pre- and postnatal development of their offspring. One hundred pregnant females were divided into four groups: three treatment groups and a control group. CKD-602 was administered once daily by intravenous bolus injection to female rats at doses of 0, 5.7, 17, or 51 microg/kg/day from gestational day 6, through to parturition and throughout the period of lactation up to weaning [lactational day (LD) 21]. All the dams were sacrificed on LD 22 after weaning. The clinical signs, mortality, body weight change, food consumption, physical development, and behavioral function were evaluated in their progeny. When the exposed offspring reached maturity (postnatal day 70), their reproductive performance was assessed. In the high-dose group, suppressed body weight and a decrease in the amount of food consumption were observed in the dams during both the gestation and lactation periods. An increase in the incidence of thymic atrophy, decreased liver and ovary weight, and an increase in the weight of the spleen were also observed in the dams at the scheduled necropsy. In addition, an increase in the number of stillborn and postnatal mortality, a decrease in the live litter size, and a delay in physical development were observed in the F1 offspring. Teratological examinations showed an increase in the incidence of congenital anomalies in both the F1 offspring and F2 fetuses. In the medium dose group, only slight maternal toxicity including suppressed body weight and decreased food consumption was observed. There were no treatment-related effects on the maternal function and pre- and postnatal development in the low dose group. The no-observed-adverse-effect level (NOAEL) of CKD-602 for the dams are considered to be 5.7 microg/kg/day, however, the NOAEL for their offspring are estimated to be 17 microg/kg/day.


TLC-HEAT: Telephony-based self-care for overweight children.
Efforts to use information technology (IT) to link pediatric primary care patients in the home with their clinicians have been rudimentary to date. We have developed a model information system that uses a ubiquitous user-interface, the telephone, and the electronic health record (EHR) to support health behavior change and weight loss for overweight urban children and their parents. In this paper, we present the technical architecture and underlying psychological theories used to build the system.


A(3) adenosine receptor ligands: history and perspectives.
Adenosine regulates many physiological functions through specific cell membrane receptors. On the basis of pharmacological studies and molecular cloning, four different adenosine receptors have been identified and classified as A(1), A(2A), A(2B), and A(3). These adenosine receptors are members of the G-protein-coupled receptor family. While adenosine A(1) and A(2A) receptor subtypes have been pharmacologically characterized through the use of selective ligands, the A(3) adenosine receptor subtype is presently under study in order to better understand its physio-pathological functions. Activation of adenosine A(3) receptors has been shown to stimulate phospholipase C and D and to inhibit adenylate cyclase. Activation of A(3) adenosine receptors also causes the release of inflammatory mediators such as histamine from mast cells. These mediators are responsible for processes such as inflammation and hypotension. It has also been suggested that the A(3) receptor plays an important role in brain ischemia, immunosuppression, and bronchospasm in several animal models. Based on these results, highly selective A(3) adenosine receptor agonists and/or antagonists have been indicated as potential drugs for the treatment of asthma and inflammation, while highly selective agonists have been shown to possess cardioprotective effects. The updated material related to this field of research has been rationalized and arranged in order to offer an overview of the topic.


A structurally deviant member of the Euplotes raikovi pheromone family: Er-23.
Pheromones of Euplotes raikovi form a homologous family of proteins with 37- to 40-amino acid residues, including six cysteines that form three strictly conserved disulfide bridges. The determination of the primary structure of the pheromone Er-23, which was isolated from cells derived from natural populations of E. raikovi that secrete the other known pheromones, has now revealed a novel structure type. The polypeptide chain of this pheromone contains 51 residues, 10 of which are cysteines presumably involved in the formation of five disulfide bridges, and lacks a carboxyl-terminal tail following the last cysteine of the sequence. The elongation of the Er-23 molecule is presumed to result from multiple events of gene duplication starting from an ancestral motif Xxx(2-4)-Cys-Xxx(5-7)-Cys.


Pituitary apoplexy can mimic acute meningoencephalitis or subarachnoid haemorrhage.
Pituitary apoplexy is an uncommon but life-threatening condition that is often overlooked and underdiagnosed. We report a 45-year-old man who presented to our emergency department with a sudden onset headache, acute confusion, signs of meningeal irritation and ophthalmoplegia. An initial diagnosis of acute meningoencephalitis was made, which was amended to pituitary apoplexy following thorough investigation within the emergency department.A 45-year-old man was brought to our emergency department by ambulance with a history of sudden onset of frontal headache and acute confusion. His wife provided the history. There was no significant past medical history of diabetes, hypertension, recent travel abroad, exposure to sick contacts, involvement in outdoor pursuits such as hiking/cave diving, or trauma. He worked in a bank and had been well until 24 h prior to the onset of sudden headache, which was gradually worsening in nature and associated with increasing confusion. The patient's wife reported that he had neither experienced any fevers, night sweats, or coryzal symptoms nor received any recent vaccinations. He was not on any regular medications. He was a non-smoker and occasionally consumed alcohol. There was no significant family history. On examination in the ED, his temperature was 37.6°C, his pulse was 110/min, and he was normotensive and normoglycaemic. A macular blanching rash was noted over the patient's trunk. The patient was disoriented to time and place. Neurological examination revealed reduced GCS (11/15-E3, M6, V2), marked neck stiffness, a positive Kernig's sign and a right sixth nerve palsy.A provisional diagnosis of acute meningoencephalitis was made and the patient was started on a course of intravenous antibiotics with benzyl penicillin 1.2 g, cefotaxime 2 g and acyclovir 750 mg. Baseline blood investigations revealed hyponatraemia (122 mmol/l), a white-cell count of 11 × 109/l and a C-reactive protein > 250. Due to the sudden onset of the symptoms and lack of prodrome, an urgent CT head scan was performed to rule out a cerebrovascular event. The scan demonstrated an enlarged pituitary gland (3 cm in diameter) with impingement of the optic chiasm. The centre of the enlarged pituitary gland was noted to be hypodense in comparison to its periphery, which was consistent with a diagnosis of pituitary apoplexy. A subsequent MRI confirmed the diagnosis (Figure 1) of an enlarged sella containing abnormal soft tissue with increased signal intensity suggestive of haemorrhage (Figure 1A).Post-MRI a lumbar puncture was performed revealing glucose 3.4 mmol/l, protein 1.0 g/l, red cells of 53/mm3 and white cells of 174/mm3 with predominant neutrophilia. No organisms were seen, and CSF cultures and HSV DNA tests were found to be negative. Endocrinological investigations demonstrated low concentrations of thyroid hormones [TSH: 0.14 mIu/l (0.35-5.5 mlU/l), FT3: 1.1 nmol/l (1.2-3.0 nmol/l), FT4: 9.6 pmol/l (8-22 pmol/l)], gonadal hormones (LH: < 1 u/l) and prolactin: 16 u/l (<450 u/l). Serum FSH was 2.9 u/l (0.8-11.5 u/L) and cortisol 575 nmol/l (450-700 nmol/l). The patient was treated for hypopituitarism based on clinical and radiological findings with intravenous fluids, hydrocortisone (100 mg) and thyroxine (50 μg) as loading doses in the ED.Within 24 h of commencement of therapy the patient's GCS rose to 15, and within 48 h there was marked improvement in the right sixth cranial nerve palsy. Formal visual field assessment demonstrated temporal visual field loss in the left eye. The patient was discharged to his usual residence a week later and follow-up was organised with both the endocrinologists and ophthalmologists. Follow-up MRI demonstrated that there was no significant change in either size or signal characteristics of the pituitary fossa mass (Figure 1B).


Fertility and reproduction following inversion of uterus.
Puerperal inversion of the uterus is itself a rare occurrence. Records of fertility and reproduction following an episode of uterine inversion are even rarer. The reproductive outcome following correction of uterine inversion in 6 cases seen over a span of 35 years is being reported here. One case of acute inversion was managed by manual replacement and the remaining 5 of chronic inversion were corrected by Haultain's operation. The only patient managed by manual replacement conceived thrice. She aborted once, but delivered 2 healthy live babies subsequently by caesarean section done for uterine inertia each time. Out of the 5 patients treated by Haultain's technique, 3 conceived later. One did not come for follow-up after the 5th month. Each of the other 2 had full-term normal delivery of living baby under supervision. None of the cases had any complications. It is concluded that even after operative correction of inversion, uncomplicated delivery may be anticipated. Conservative surgical management is ideal even in apparently neglected and chronic cases, as most of these women were primipara or multipara with only one or 2 living children and had a desire of future child bearing.


Endometrial bacterial flora detected in patients with uterine endometrial cancer.
Certain bacteria produce some carcinogens such as N-nitro compounds, n-butyric acid and n-valeric acid. From this point of view, the examination of intrauterine bacterial flora in patients with uterine endometrial cancer may provide important information. Twenty patients with the diagnosis of uterine endometrial cancer and 20 patients without complications other than myoma uteri were enrolled in the study. Enterobacteriaceae, Streptococcus agalactiae and anaerobic bacteria were mainly detected. The products of these bacteria might be considered to contribute to the initiation of endometrial carcinogenesis. Mixed abnormal flora between aerobic and anaerobic bacteria were detected in all patients with uterine endometrial cancer. It is suggested that uterine endometrial cancer provides favorable conditions for bacterial growth. Mixed abnormal bacterial flora also might influence the onset and growth of uterine endometrial cancer.


Evidence for the decay Bs0-->Ds(*)Ds(*) and a measurement of DeltaGammasCP/Gammas.
We search for the semi-inclusive process Bs0-->Ds(*)Ds(*) using 2.8 fb(-1) of pp collisions at sqrt[s]=1.96 TeV recorded by the D0 detector operating at the Fermilab Tevatron Collider. We observe 26.6+/-8.4 signal events with a significance above background of 3.2 standard deviations yielding a branching ratio of B(Bs0-->Ds(*)Ds(*))=0.035+/-0.010(stat.)+/-0.011(syst.). Under certain theoretical assumptions, these double-charm final states saturate CP-even eigenstates in the Bs0 decays resulting in a width difference of DeltaGammasCP/Gammas=0.072+/-0.021(stat.)+/-0.022(syst.).


Use of the carbon dioxide laser for perianal and rectal surgery.
The skin of the perianal region is very thin and sensitive. The carbon dioxide laser is a very effective tool for removing tumors and treating conditions of this area. It offers a "no touch" method of treatment for conditions of the perianal region. There is less bleeding, less pain, less swelling with the use of the carbon dioxide laser. The carbon dioxide laser is an effective tool for excising perianal tumors, rectal tumors, performing anal sacculectomies and treating perianal fistulas.


Subdural recordings from an awake human brain for measuring current intensity during transcranial direct current stimulation.
Transcranial direct current stimulation (tDCS) is an emerging method, used for non-invasively stimulating the brain in normal healthy subjects and in patients with neurological disorders. However, the pattern of the spatial distribution of the current intensity induced by tDCS is poorly understood. In this study, we directly measured the spatial characteristics of the current intensity induced by tDCS using an intracranial strip electrode array implanted over the motor cortex in patients with Parkinson's disease undergoing deep brain stimulation lead placement surgery. We used a bilateral stimulation configuration for the tDCS electrode placement and measured the amount of electric current passing through the contacts along the implanted strip electrode contacts. Our results showed significant changes of the current flow induced by the tDCS in some of the contacts during stimulation with respect to baseline activities. These results may provide vital information regarding the biophysical effects of tDCS stimulation and might be potentially useful for developing more effective stimulation strategies.


[Croatian guidelines for perioperative enteral nutrition of surgical patients].
Nutritional status of patients significantly affects the outcome of surgical treatment, whether it's about being obese or malnutrition with loss of muscle mass. Inadequate nutritional support in the perioperative period compromises surgical procedures even in patients who are adequately nourished. In this paper, particular attention was paid to malnourished patients, and their incidence in population hospitalized in surgical wards can be high up to 30%. Special emphasis was paid to the appropriateness of preoperative fasting and to the acceptance of new knowledge in this area of treatment. The aim of this working group was to make guidelines for perioperative nutritional support with different modalities of enteral nutrition. The development of these guidelines was attended by representatives of Croatian Medical Association: Croatian Society for Digestive Surgery, Croatian Society for Clinical Nutrition, Croatian Society of Surgery, Croatian Society for Endoscopic Surgery, Croatian Trauma Society and the Croatian Society of Anesthesiology and Intensive Care. The guidelines are designed as a set of questions that arise daily in clinical practice when preparing patients for surgery and after the surgical treatment, which relate to the assessment of nutritional status, perioperative nutritional support, duration of preoperative fasting period and the selection of food intake route. Assessment of nutritional status and the use of different modes of enteral nutrition should enter into standard protocols of diagnosis and treatment in the Croatian hospitals.


Sensitivity and specificity of computed tomography hypodense sign when differentiating pulmonary inflammatory and malignant mass-like lesions.
Hypodense sign (HyS) reportedly is associated with pulmonary fungal infection, while it also common in many non-fungal lesions. This study aims to determine the significance of a HyS presented on contrast-enhanced computed tomography (CECT) when distinguishing pulmonary inflammatory from malignant mass-like lesions. From January 2013 to January 2021, we retrospectively evaluated the clinical and computed tomography (CT) data of patients with pathologically confirmed pulmonary inflammatory lesions (ILs) and malignant lesions (MLs). We analyzed and compared the CT features of the HyS in MLs and ILs, and then evaluated whether the HyS helped to differentiate MLs and ILs. There were significant differences in age and tumor markers between patients with ILs and MLs (both P<0.05). Compared with that in MLs, the occurrence of the HyS in ILs was higher (62.81% vs. 28.81%; P<0.0001). In ILs, more HyS were single, round or oval, well-defined, and had lower enhancement (ΔCT). Logistic regression analysis revealed that an ill-defined boundary, peripheral fibrosis, presence of a well-defined HyS, and a ΔCT value of the HyS <9.5 Hounsfield units (HU) were independent indicators for predicting ILs. After including the HyS CT features, the area under the curve (AUC) of the model predicting ILs increased from 0.953 to 0.986 with a sensitivity of 96.03% and a specificity of 94.03% (P=0.0027). The HyS is more common in ILs than in MLs. A single, regular, and well-defined HyS with a ΔCT value of <9.5 HU on CECT is highly suggestive of ILs. Combining the HyS with other morphological features could improve the diagnosis accuracy of pulmonary mass-like lesions.


In vivo skeletal muscle mitochondrial function in Leber's hereditary optic neuropathy assessed by 31P magnetic resonance spectroscopy.
We used 31P magnetic resonance spectroscopy (31P-MRS) to assess in vivo skeletal muscle mitochondrial function in 10 Leber's hereditary optic neuropathy patients/carriers with a mitochondrial DNA (mtDNA) mutation at one of three nucleotide positions, 11,778, 14,484, and 3,460. We studied one affected patient for each mutation and two unaffected carriers with the 11,778 or 3,460 mutation and three carriers with 14,484. All subjects were homoplasmic except the two 3,460 carriers, who showed 80% and 15% of mutated mtDNA. 31P-MRS at rest disclosed some abnormalities in all subjects. In particular, the phosphorylation potential was below the normal range in all cases. During recovery from exercise, the maximum rate of mitochondrial ATP production (Vmax) was reduced to 27% of normal in the 11,778 mutation and to 53% in the 14,484 mutation patient/carrier groups. Mitochondrial Vmax was within the normal range in all subjects with the 3,460 mutation but correlated inversely with the percentage of mutated mtDNA. This in vivo study shows that the 11,778 mutation causes a mitochondrial impairment more severe than the 14,484 and that the 3,460 mutation results in only a mild depression of muscle mitochondrial function.


Preconcentration of heavy metals in urine and quantification by inductively coupled plasma atomic emission spectrometry.
This paper describes a method for the determination of heavy metals (Co, Ni, Cu, Cd, Pb) in urine by inductively coupled plasma atomic emission spectrometry (ICP-AES). The method proposed requires purification of the samples with activated charcoal under acidic conditions before preconcentration by complexation with ammonium pyrrolidinedithiocarbamate (APDC). The formed complexes are extracted with methyl isobutyl ketone (MIBK) and the resulting residue is finally digested under acid oxidant conditions. Because of its low detection limit (below 10 micrograms/L), this procedure can be applied conveniently for toxicological diagnostic purposes.


Engineered 3D Immuno-Glial-Neurovascular Human Brain Model.
Patient-specific, human-based cellular models that integrate biomimetic BBB, immune, and myelinated neuron components are critically needed to enable translationally relevant and accelerated discovery of neurological disease mechanisms and interventions. By engineering a brain-mimicking 3D hydrogel and co-culturing all six major brain cell types derived from patient iPSCs, we have constructed, characterized, and utilized a multicellular integrated brain (miBrain) immuno-glial-neurovascular model with in vivo- like hallmarks. As proof of principle, here we utilized the miBrain to model Alzheimer's Disease pathologies associated with APOE4 genetic risk. APOE4 miBrains differentially exhibit amyloid aggregation, tau phosphorylation, and astrocytic GFAP. Unlike the co-emergent fate specification of glia and neurons in organoids, miBrains integrate independently differentiated cell types in a modular system with unique utility for elucidating cell-type specific contributions to pathogenesis. We here harness this feature to identify that risk factor APOE4 in astrocytes promotes tau pathogenesis and neuronal dysregulation through crosstalk with microglia. A novel patient-specific brain model with BBB, neuronal, immune, and glial components was developed, characterized, and harnessed to model Alzheimer's Disease-associated pathologies and APOE4 genetic risk.


Long-term results of Dana-Farber Cancer Institute ALL Consortium protocols for children with newly diagnosed acute lymphoblastic leukemia (1985-2000).
The Dana-Farber Cancer Institute (DFCI) acute lymphoblastic leukemia (ALL) Consortium has been conducting multi-institutional clinical trials in childhood ALL since 1981. The treatment backbone has included 20-30 consecutive weeks of asparaginase during intensification and frequent vincristine/corticosteroid pulses during the continuation phase. Between 1985 and 2000, 1457 children aged 0-18 years were treated on four consecutive protocols: 85-01 (1985-1987), 87-01 (1987-1991), 91-01 (1991-1955) and 95-01 (1996-2000). The 10-year event-free survival (EFS)+/-s.e. by protocol was 77.9+/-2.8% (85-01), 74.2+/-2.3% (87-01), 80.8+/-2.1% (91-01) and 80.5+/-1.8% (95-01). Approximately 82% of patients treated in the 1980s and 88% treated in the 1990s were long-term survivors. Both EFS and overall survival (OS) rates were significantly higher for patients treated in the 1990s compared with the 1980s (P=0.05 and 0.01, respectively). On the two protocols conducted in the 1990s, EFS was 79-85% for T-cell ALL patients and 75-78% for adolescents (age 10-18 years). Results of randomized studies revealed that dexrazoxane prevented acute cardiac injury without adversely affecting EFS or OS in high-risk (HR) patients, and frequently dosed intrathecal chemotherapy was an effective substitute for cranial radiation in standard-risk (SR) patients. Current studies continue to focus on improving efficacy while minimizing acute and late toxicities.


Comparison of DNA Polymerase of Rhizobium meliloti and Alfalfa Bacteroids.
DNA dependent-DNA polymerase activity was established and partially purified from extracts of cultured Rhizobium meliloti, F-28, and nodule bacteroids (R. meliloti, F-28) of alfalfa plants (Medicago sativa). Polymerase activity in the partially purified fractions showed characteristic dependence on Mg(2+), DNA, and a full complement of deoxyribonucleoside triphosphates. DNase activity, preference of "activated" double strand DNA, and inhibition by p-chloromercuribenzoate and MnCl(2) were responses common to both systems. The two systems however did exhibit some differences in pH, Mg(2+), and primer optima. Polymerase activity in crude extracts of the cultured bacteria was more stable and had 10- to 18-fold greater specific activity than the bacteroid extracts. Preliminary measurements of specific DNA polymerase activity in crude extracts of cultured Rhizobium japonicum were not significantly higher than that in the crude extracts of soybean nodule bacteroids. A possible correlation between DNA synthesis and the successful establishment of rhizobia-legume symbiosis is discussed.


[The changes of p-Akt/MuRF1/FoxO1 proteins expressions in the conditions of training and immobilization in rats' gastrocnemius muscle].
This study was aimed to investigate the changes of muscle protein synthesis and degradation under different movement conditions, so as to provide theoretical basis for muscle atrophy mechanism. Sprague Dawley (SD) rats were randomly divided into control, endurance training (treadmill training), hind limb overhanging and eccentric training (treadmill training, angle -16º) groups. The gastrocnemius muscles of rats were taken and weighed. The muscle was sectioned, and HE staining was employed to determine the cell's cross-sectional area. Protein expression of p-Akt was measured by immunohistochemistry; and the expressions of MuRF1 and FoxO1 were determined by Western blot. The results showed that, compared with control group, hind limb overhanging and eccentric training groups exhibited decreased muscle weight and cross-sectional area, but endurance training group did not show any changes. The expressions of p-Akt in endurance and eccentric training groups, not in hind limb overhanging group, were significantly higher than that in control group. Compared with that of control, MuRF1 protein remained unchanged in endurance training groups, but was increased in eccentric training and hind limb overhanging groups; FoxO1 protein was decreased in endurance training group, but was increased in eccentric training and hind limb overhanging groups. These results indicate that movement (endurance and eccentric training) can activate Akt expression, but does not increase muscle weight, whereas eccentric training and hind limb overhanging can increase the expressions of MuRF1 and FoxO1, and induce amyotrophy, suggesting MuRF1 and FoxO1 are major determinant factors in muscle atrophy.


Imaging the effects of prostaglandin analogues on cultured trabecular meshwork cells by coherent anti-stokes Raman scattering.
The aim of this study was to nondestructively monitor morphological changes to the lipid membranes of primary cultures of living human trabecular meshwork cells (hTMC) without the application of exogenous label. Live hTMC were imaged using two nonlinear optical techniques: coherent anti-Stokes Raman scattering (CARS) and two-photon autofluorescence (TPAF). The hTMC were treated with a commercial formulation of latanoprost (0.5 μg/mL) for 24 hours before imaging. Untreated cells and cells treated with vehicle containing the preservative benzalkonium chloride (BAK; 2 μg/mL) were imaged as controls. After CARS/TPAF imaging, hTMC were fixed, stained with the fluorescent lipid dye Nile Red, and imaged by conventional confocal microscopy to verify lipid membrane structures. Analysis of CARS/TPAF images of hTMC treated with latanoprost revealed multiple intracellular lipid membranes absent from untreated or BAK-treated hTMC. Treatment of hTMC with sodium fluoride or ouabain, agents shown to cause morphological changes to hTMC, also did not induce formation of intracellular lipid membranes. CARS microscopy detected changes in living hTMC morphology that were validated by subsequent histological stain. Prostaglandin-induced changes to hTMC involved rearrangement of lipid membranes within these cells. These in vitro results identify a novel biological response to a class of antiglaucoma drugs, and further experiments are needed to establish how this effect is involved in the hypotensive action of prostaglandin analogues in vivo.


Caring for cancer survivors: perspectives of oncologists, general practitioners and patients in Italy.
The present survey investigates the views of medical oncologists, general practitioners (GPs) and patients about the various surveillance strategies. An online survey was conducted in Italy on a population of 329 medical oncologists, 380 GPs and 350 patients. Most of GPs (n = 291; 76%) claim that follow-up should be provided by the collaboration between GPs and medical oncologists. Most medical oncologists report to have a poor relationship with GPs (n = 151; 46%) or no relationships at all (n = 14; 4%). Most patients believe there is no real collaboration between medical oncologists and GPs (n = 138; 54%). GPs, medical oncologists and patients share the idea that the collaboration between oncologists and GPs for surveillance of cancer survivors is poor and should be improved.


Where are all the men? Low energy availability in male cyclists: A review.
Most of the low energy availability (LEA) research has been conducted in female populations. The occurrence of LEA in male athletes is not well known, even with an understanding of the components involved in and contributing to LEA. Cycling is a major risk factor for LEA due to inherent sports characteristics: low impact, high energy demands, and a common perception that leanness is a performance advantage. The purpose of this review is to discuss the cycling-specific studies that have documented components of RED-S. The review demonstrates male cyclists (1) experience energy deficits daily, weekly and throughout a season; (2) exhibit lower bone mineral density at the spine compared to the hip, and low bone mineral density correlating with LEA and; (3) demonstrate downregulation of the endocrine system with elevated cortisol, reduced testosterone and insulin-like growth factor 1. The complexity of LEA is further explored by the socio-psychological contribution that may impact eating behaviours, and therefore increase the risk of developing LEA. Future research directions include applying multifaceted research methods to gain a greater understanding of this syndrome and the effect of LEA on male cyclists.


Medicine and environmental health.
Environmental threats to health and welfare are receiving much attention today and will become an even greater concern in the future. Since they often are the only group able to produce much of the basic data concerning environmental health effects, physicians must become more involved in environmental health problems, individually and organizationally. The Linton Report offers one proposal for attaining a liveable environment and should be reviewed by all health professionals.


A Role for NRF2-Signaling in the Treatment and Prevention of Solar Lentigines.
Photoaging is premature skin aging resulting from oxidative stress generated by exposure to solar radiation. A key clinical feature is solar lentigines, areas of hyperpigmentation on sun-exposed skin. Skin pigmentation is determined by cross-talk between keratinocytes and melanocytes, which is exquisitely sensitive to oxidative stress. Toll-like receptor (TLR) signaling and NF-E2-related factor 2 (NRF2) signaling, an endogenous antioxidant system, serve as a bridge between the oxidative stress response and immune regulation. Moreover, TLR-mediated induction of IL-6 production has been shown to prevent ultraviolet (UV)-induced hyperpigmentation. Shave biopsies of solar lentigines were obtained from 14 individuals. An additional 7 subjects applied broccoli sprout extract (BSE) containing sulforaphane daily or vehicle on photodamaged skin. Immunofluorescence staining was used to determine total and phosphorylated NRF2 in the lentiginous skin. Dermoscopy and Fontana & Masson staining were used to assess the effect of topical BSE on UV-induced pigmentation. Similar topical treatments were performed in a mouse model of UVB-induced hyperpigmentation utilizing WT, Nrf2-/-, or K14-Cre-ERT2IL-6Rαfl/fl C57BL/6 mice. NRF2 expression is altered in solar lentigines, and UV-induced skin pigmentation in humans could be ameliorated with topical BSE. Corresponding mouse models replicated the authors' clinical findings and identified a potential mechanistic link to IL-6Rα signaling in keratinocytes. The authors' findings suggest that dysregulation of NRF2 signaling is involved in the pathogenesis of UV-induced skin pigmentation and pharmacological activation of NRF2 may represent a potential therapeutic target in photoaging.


Patient Safety Culture Based on Medical Staff Attitudes in Khorasan Razavi Hospitals, Northeastern Iran.
Since establishing a safety culture in an organization is considered as the first step in patient safety improvement, there is always a need for updated field evaluation to better plan future decisions. We performed a cross-sectional, analytic-descriptive study in 25 hospitals related to Mashhad University of Medical Sciences (MUMS) during a 3-month period from April to June 2012. A questionnaire, designed by previous patient safety culture studies with confirmed validity and reliability, was used and distributed among a sample of 922 staff, chosen randomly from the mentioned hospitals. Data were analyzed by SPSS software version 16. "Organizational learning - continuous improvement" and "teamwork within unit" had the highest percentage of positive results as 79.85 ± 12.03% and 71.92 ± 17.08%, respectively; whereas "non-punitive response" to errors (21.57 ± 6.42) and "staffing" (26.36 ± 16.84) came out as the least important factors. There were no meaningful statistical relation between general features of the understudy hospitals including the number of beds, educational level or proficiency status with the general safety culture score. Most of the safety culture aspects were reported as low to moderate in terms of importance. If something needs to be modified interventionally in this respect, "the approach to confront errors" would be a wise choice. This could be achieved by establishing an atmosphere of open communication and continuous learning through elimination of the fear for reporting errors and installing a more acceptable approach in hospitals.


Indoleamine 2,3-Dioxygenase Activity-Induced Acceleration of Tumor Growth, and Protein Kinases-Related Novel Therapeutics Regimens.
Indoleamine 2,3-dioxygenase (IDO) is overexpressed in response to interferon-gamma (IFN-γ). IDO-mediated degradation of tryptophan (Trp) along the kynurenine (Kyn) pathway by immune cells is associated with the anti-microbial, and anti-tumor defense mechanisms. In contrast, IDO is constitutively expressed by various tumors and creates an immunosuppressive microenvironment around the tumor tissue both by depletion of the essential amino acid Trp and by formation of Kyn, which is immunosuppressive metabolite of Trp. IDO may activate its own expression in human cancer cells via an autocrine aryl hydrocarbon receptor (AhR)- interleukin 6 (IL-6)-signal transducer and activator of transcription 3 (STAT3) signaling loop. Although IDO is not a unique marker, in many clinical trials serum IDO activity is suggested to be an important parameter in the pathogenesis of cancer development and growth. Measuring IDO activity in serum seems to be an indicator of cancer growth rate, however, it is controversial whether this approach can be used as a reliable guide in cancer patients treated with IDO inhibitors. Thus, IDO immunostaining is strongly recommended for the identification of higher IDO producing tumors, and IDO inhibitors should be included in post-operative complementary therapy in IDO positive cancer cases only. Novel therapies that target the IDO pathway cover checkpoint protein kinases related combination regimens. Currently, multi-modal therapies combining IDO inhibitors and checkpoint kinase blockers in addition to T regulatory (Treg) cell-modifying treatments seem promising.


Mature teratoma of the placenta.
A rare case of a mature teratoma of the placenta is reported. The tumor lay between the amnion and the chorion and contained skin and its appendages, bone, cartilage, fat and ganglia, without organization. There was no evidence of a recognizable umbilical cord. We concluded that the tumor was a mature teratoma of the placenta. The distinction between a teratoma and a fetus acardius amorphus, and the possible origin of the tumor in this site, are discussed with a brief review of the literature.


The bend angle of water in ice Ih and liquid water: The significance of implementing the nonlinear monomer dipole moment surface in classical interaction potentials.
The implementation of the physically accurate nonlinear dipole moment surface of the water monomer in the context of the Thole-type, polarizable, flexible interaction potential results in the only classical potential, which, starting from the gas phase value for the bend angle (104.52 degrees), reproduces its experimentally observed increase in the ice Ih lattice and in liquid water. This is in contrast to all other classical potentials to date, which predict a decrease of the monomer bend angle in ice Ih and in liquid water with respect to the gas phase monomer value. Simulations under periodic boundary conditions of several supercells consisting of up to 288 molecules of water used to sample the proton disorder in the ice Ih lattice yield an average value of vartheta(HOH)(I(h))=108.4 degrees +/-0.2 degrees for the minimized structures (T=0 K) and 108.1 degrees +/-2.8 degrees at T=100 K. Analogous simulations for liquid water predict an average value of vartheta(HOH)(liquid)=106.3 degrees +/-4.9 degrees at T=300 K. The increase of the monomer bend angle of water in condensed environments is attributed to the use of geometry-dependent charges that are used to describe the nonlinear character of the monomer's dipole moment surface. Our results suggest a new paradigm in the development of classical interaction potential models of water that can be used to describe condensed aqueous environments.


The effect of molecular planarity on crystal non-centrosymmetry in benzylidene-aniline derivatives.
In the title compound, N-(2-methoxyphenyl)-4-nitrobenzylideneamine, C(14)H(12)N(2)O(3), the two phenyl rings make a dihedral angle of 48.0 (2) degrees and the nitro group is at an angle of 6.5 (1) degrees with respect to its attached phenyl ring. In the crystal structure, molecules are related as centrosymmetric pairs through pi-pi interactions and are further connected through strong C[bond]H...O hydrogen bonds [C...O 3.4259 (17) A and C[bond]H...O 167 degrees ], forming molecular stacks along [100]. These stacks associate further through longer C[bond]H...O interactions, forming two-dimensional networks. In the c direction, there are only weak van der Waals interactions. The relationship between the molecular planarity and its centrosymmetry is also briefly described.


SHORT-TERM ANNUAL VARIATIONS OF RADON CONCENTRATION IN WORKPLACES: SOME RESULTS IN A RESEARCH INSTITUTE.
Many international and national regulations on radon in workplaces, including the 2013/59/Euratom Council Directive, are based on the annual average of indoor radon concentration, assuming it is representative of the long-term average. However, a single annual radon concentration measurement does not reflect annual variations (i.e. year-to-year variations) of radon concentration in the same location. These variations, if not negligible, should be considered for an optimized implementation of regulations. Unfortunately, studies on annual variations in workplaces can be difficult and time-consuming and no data have been published on scientific journals on this issue. Therefore, we carried out a study to obtain a first evaluation of short-term annual variations in workplaces of a research institute in Rome (Italy). The radon concentration was measured in 120 rooms (mainly offices and laboratories) located in 23 buildings. In each room, two 1-year long measurements were performed, with an interval between the two measurements of up to 3 years. The results show variability between the two 1-year long measurements higher than the variability observed in a sample of dwellings in the same area. Further studies are required to confirm the results and to extend the study to other types of workplaces.


Near-Infrared Hyperspectral Imaging Pipelines for Pasture Seed Quality Evaluation: An Overview.
Near-infrared (800-2500 nm; NIR) spectroscopy coupled to hyperspectral imaging (NIR-HSI) has greatly enhanced its capability and thus widened its application and use across various industries. This non-destructive technique that is sensitive to both physical and chemical attributes of virtually any material can be used for both qualitative and quantitative analyses. This review describes the advancement of NIR to NIR-HSI in agricultural applications with a focus on seed quality features for agronomically important seeds. NIR-HSI seed phenotyping, describing sample sizes used for building high-accuracy calibration and prediction models for full or selected wavelengths of the NIR region, is explored. The molecular interpretation of absorbance bands in the NIR region is difficult; hence, this review offers important NIR absorbance band assignments that have been reported in literature. Opportunities for NIR-HSI seed phenotyping in forage grass seed are described and a step-by-step data-acquisition and analysis pipeline for the determination of seed quality in perennial ryegrass seeds is also presented.


The effect of disk type and cutting speed on the micro-tensile bond strength of ceramic specimens to resin cement.
The bond strength of dental materials has been evaluated by tensile testing of micro-specimens. The cutting process used to obtain specimens may influence the results. The objective of this study was to investigate the influence of different types of diamond disks and cutting speeds on the bond strength of ceramic specimens and on specimen integrity. Lithium disilicate-based ceramic cubes were bonded with resin cement to composite resin cubes, according to the manufacturers' instructions. The ceramic/cement/resin blocks thus obtained were divided into two groups to be cut with Buehler(®) or Extec(®) disks and then sectioned at cutting speeds of 200 rpm and 400 rpm. The results showed that the bond strength values were affected by the cutting speed and disk/speed interaction (p<0.05). SEM analysis revealed better specimen properties when the blocks were cut at 200 rpm. It was concluded that ceramic specimens must be cut at low speeds.


Hyperpolarization-activated cation currents: from molecules to physiological function.
Hyperpolarization-activated cation currents, termed If, Ih, or Iq, were initially discovered in heart and nerve cells over 20 years ago. These currents contribute to a wide range of physiological functions, including cardiac and neuronal pacemaker activity, the setting of resting potentials, input conductance and length constants, and dendritic integration. The hyperpolarization-activated, cation nonselective (HCN) gene family encodes the channels that underlie Ih. Here we review the relation between the biophysical properties of recombinant HCN channels and the pattern of HCN mRNA expression with the properties of native Ih in neurons and cardiac muscle. Moreover, we consider selected examples of the expanding physiological functions of Ih with a view toward understanding how the properties of HCN channels contribute to these diverse functional roles.


The piggyBac-Based Gene Delivery System Can Confer Successful Production of Cloned Porcine Blastocysts with Multigene Constructs.
The introduction of multigene constructs into single cells is important for improving the performance of domestic animals, as well as understanding basic biological processes. In particular, multigene constructs allow the engineering and integration of multiple genes related to xenotransplantation into the porcine genome. The piggyBac (PB) transposon system allows multiple genes to be stably integrated into target genomes through a single transfection event. However, to our knowledge, no attempt to introduce multiple genes into a porcine genome has been made using this system. In this study, we simultaneously introduced seven transposons into a single porcine embryonic fibroblast (PEF). PEFs were transfected with seven transposons containing genes for five drug resistance proteins and two (red and green) fluorescent proteins, together with a PB transposase expression vector, pTrans (experimental group). The above seven transposons (without pTrans) were transfected concomitantly (control group). Selection of these transfected cells in the presence of multiple selection drugs resulted in the survival of several clones derived from the experimental group, but not from the control. PCR analysis demonstrated that approximately 90% (12/13 tested) of the surviving clones possessed all of the introduced transposons. Splinkerette PCR demonstrated that the transposons were inserted through the TTAA target sites of PB. Somatic cell nuclear transfer (SCNT) using a PEF clone with multigene constructs demonstrated successful production of cloned blastocysts expressing both red and green fluorescence. These results indicate the feasibility of this PB-mediated method for simultaneous transfer of multigene constructs into the porcine cell genome, which is useful for production of cloned transgenic pigs expressing multiple transgenes.


Stability indicating LC method for the simultaneous determination of amlodipine and olmesartan in dosage form.
A simple, rapid, and precise method is developed for the quantitative simultaneous estimation of amlodipine (AM) and olmesartan (OL) in combined pharmaceutical dosage form. A chromatographic separation of the two drugs was achieved with an ACE 5 C(18) 25-cm analytical column using buffer-acetonitrile (60:40, v/v). The resolution between OL and AM was found to be more than 12. Theoretical plates for OL and AM were 6970 and 11,841, respectively. Tailing factor for OL and AM was 0.90 and 0.98, respectively. OL, AM, and combination drug product were exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions, and the stressed samples were analyzed by the proposed method. Peak homogeneity data of OL and AM is obtained by photodiode array detector in the stressed sample chromatograms, demonstrating the specificity of the method for their estimation in presence of degradation product. The described method shows excellent linearity over a range of 20-400 microg/mL for OL and 5-100 microg/mL for AM. The correlation coefficient for OL and AM are 0.9995 and 0.9998, respectively. The relative standard deviation for six measurements in two sets of each drug in tablets is always less than 2%. The proposed method was found to be suitable and accurate for quantitative determination and stability study of OL and AM in pharmaceutical preparations.


Pathological classification and follow-up of prostatic lesions initially diagnosed as "suspicious of malignancy".
Sections from a series of prostatic specimens (n = 44) in which the initial diagnosis implied a suspicion of malignancy were reviewed and the patients followed up (group 1). The aim was to categorise the morphological appearances according to current criteria and to determine the natural history of any pre-malignant lesions within this group. A "control" series of patients (n = 40) in whom the primary diagnosis was benign were also reviewed and their course followed (group 2). The heterogeneity of the abnormalities in group 1 was striking. Of the 44 cases, pre-malignant lesions were seen in 8, microinvasive adenocarcinoma in 11 and lesions not now considered pre-malignant in the remaining 25. None of the 8 patients with pre-malignant lesions developed clinical evidence of carcinoma over a follow-up period of 8 to 12 years. In contrast, 3 of the 11 diagnosed on review as having microscopic invasive adenocarcinoma developed clinically evident tumour within 5 years of surgery. Of the 25 patients whose prostatic lesions were considered benign, 2 developed carcinoma 12 and 13 years after surgery. Only 1 of the 40 patients in group 2 had a pre-malignant lesion and he did not develop tumour within 10 years. However, adenocarcinoma was diagnosed incidentally in 1 patient 3 years after surgery. It was concluded that there is a need to standardise interpretation and reporting of pre-malignant lesions and microscopic foci of adenocarcinoma in the prostate. The limited results on the course of pre-malignant lesions diagnosed incidentally in patients over 60 years of age do not indicate that follow-up would have prompted the early diagnosis of invasive disease.


Mothers with depressed mood: help-seeking from husbands and child-rearing behaviors.
Mothers with depressed mood tend not to seek help or support from others. Yet, there is no research providing a detailed examination of the processes that mothers with depressed mood undergo while seeking child-rearing support from their husbands. This study aimed to clarify the processes that mothers with depressed mood go through in seeking child-rearing support from their husbands and performing child-rearing duties. The participants were 10 mothers living in Japan who had given birth within the past three years and were suspected of having depression after screening using the Edinburgh Postnatal Depression Scale. Semi-structured interview data were analyzed based on the grounded theory. The responses revealed that the mothers felt as though they had insufficient time for themselves, which impelled them to seek support from their husbands, consequently leading them to conclude either that child-rearing and housework are difficult or that child-rearing can be managed some way or another. When the husbands fully cooperated in child-rearing or demonstrated their willingness to cooperate in child-rearing, despite difficulty, the wives accepted their child-rearing support. On the other hand, if the husbands did not recognize their wives' efforts, the wives did not accept their support, even if they had helped with child-rearing. In this way, the wives re-evaluated their relationship with the husbands based on their husbands' attitudes toward child-rearing.


Characteristics of patients and implantable defibrillators associated with failure to sense device alert systems.
In the era of increasing implantable cardioverter defibrillator (ICD) complexity, the ICD patient alert is deemed to be an important feature in the early detection of ICD system malfunction and is either an audible or a vibratory alert. We sought to evaluate the patient's ability to detect these ICD alerts in the device clinic setting as a surrogate endpoint of clinical utility. From 1 November 2006 to 31 March 2008, 563 patients with an ICD equipped with either an audible patient alert (APA, Medtronic and Guidant; n = 485) or a vibratory monitoring alert ([VMA, St Jude Medical; n = 78) had their alarm demonstrated in the quiet clinic setting. The ability to recognize the alert was analysed and then stratified by gender, age, manufacturer, type of alert, and pocket location. The average patient age was 63.3 (± 13.6) years and 82.8% of patients were male. Implantable cardioverter defibrillator manufacturers were Medtronic (n = 464), Boston Scientific (n = 21), and SJM (n = 78). The APA was heard in 86.0% of patients. This was less likely in patients who were older, male, and where the device was placed in the submuscular position. Every patient with a VMA sensed their alert. In the current ICD alert technology, the ability to sense the ICD alert in the device clinic appears to be higher for the VMA than for the APA. In particular, older patients and male patients are less likely to sense the APA.


Factors associated with the introduction of complementary feeding in the French ELFE cohort study.
The objectives of this study were to estimate the age of complementary feeding introduction (CFI) and investigate the related health, demographic, and socio-economic factors. Analyses were based on 10,931 infants from the French national birth cohort ELFE, born in 2011. Health, demographic, and socio-economic data concerning infants and parents were collected at birth (face-to-face interviews and medical records) and 2 months (telephone interviews). Data on milk feeding and CFI practices were collected at birth and 2 months then monthly from 3 to 10 months using online or paper questionnaires. The associations between both health and social factors and CFI age were tested by multivariable multinomial logistic regressions. The mean CFI age was 5.2 ± 1.2 months; 26% of the infants started complementary feeding before 4 months of age (CF < 4 months), 62% between 4 and 6 months of age, and 12% after 6 months of age (CF > 6 months). CF < 4 months was more likely when mothers smoked, were overweight/obese, younger (<29 years), and used their personal experience as an information source in child caregiving and when both parents were not born in France. CF < 4 months was less likely when the infant was a girl, second-born, when the mother breastfed longer, and had attended at least one birth preparation class. Mothers of second-born infants and who breastfed their child longer were more likely to introduce CF > 6 months. Couples in which fathers were born in France and mothers were not born in France were less likely to introduce CF > 6 months. CF < 4 months occurred in more than 25% of the cases. It is important to continue promoting clear CFI recommendations, especially in smoking, overweight, young, not born in France, and nonbreastfeeding mothers.


Ascorbic acid deficiency accelerates aging of hepatic stellate cells with up-regulation of PPARγ.
Senescent cells have been observed in certain aged or damaged tissues. However, the information about the effects of aging on liver cells is limited. In the present study, we have examined age-related histological changes in the livers of senescence marker protein knockout (SMP30-/-) mice, which are considered as a murine aging model due to the more sensitive response to apoptotic reagents and due to their shorter life span. In livers of old SMP30-/- mice, numerous hepatic stellate cells (HSCs) were hypertrophic and contained abundant microvesicular lipid droplets in cytoplasm. We have found that the expression of peroxisome proliferators-activated receptor γ (PPARγ), which is a protein related to lipid metabolism and HSC quiescence, was increased in hypertrophic HSCs by aging and vitamin C (VC) deficiency, whereas these phenomena were dramatically reduced by antioxidant treatment. Therefore, these prominent phenotypic changes can be considered as aging markers in the livers of animals which are subjected to antioxidant property evaluation.


Footprint evidence of early hominin locomotor diversity at Laetoli, Tanzania.
Bipedal trackways discovered in 1978 at Laetoli site G, Tanzania and dated to 3.66 million years ago are widely accepted as the oldest unequivocal evidence of obligate bipedalism in the human lineage1-3. Another trackway discovered two years earlier at nearby site A was partially excavated and attributed to a hominin, but curious affinities with bears (ursids) marginalized its importance to the paleoanthropological community, and the location of these footprints fell into obscurity3-5. In 2019, we located, excavated and cleaned the site A trackway, producing a digital archive using 3D photogrammetry and laser scanning. Here we compare the footprints at this site with those of American black bears, chimpanzees and humans, and we show that they resemble those of hominins more than ursids. In fact, the narrow step width corroborates the original interpretation of a small, cross-stepping bipedal hominin. However, the inferred foot proportions, gait parameters and 3D morphologies of footprints at site A are readily distinguished from those at site G, indicating that a minimum of two hominin taxa with different feet and gaits coexisted at Laetoli.


Relationship Between Dietary Omega-3 and Omega-6 Polyunsaturated Fatty Acids Level and Sarcopenia. A Meta-Analysis of Observational Studies.
Objective: This study investigates the relationship between dietary omega-3 and omega-6 polyunsaturated fatty acids (PUFAs) levels and sarcopenia. Methods: A comprehensive literature search in the databases of PubMed, Web of Science, and Embase (up to July 2021) were conducted to identify the observational studies on the relationship between dietary omega-3 and omega-6 PUFAs level and sarcopenia. The pooled odds ratio (OR) of sarcopenia for the highest vs. lowest dietary omega-3 and omega-6 PUFAs level and the standard mean difference (SMD) of dietary omega-3 and omega-6 PUFAs levels for sarcopenia vs. control subjects were calculated. Results: A total of six studies were identified in this meta-analysis. The overall multi-variable adjusted OR showed that dietary omega-3 PUFAs level was inversely associated with sarcopenia (OR = 0.41, 95% CI: 0.26-0.65; P = 0.0001). Moreover, the overall combined SMD showed that the dietary omega-3 PUFAs level in sarcopenia was lower than that in control subjects (SMD = -0.19, 95% CI: -0.32 to -0.07; P = 0.002). With regard to dietary omega-6 PUFAs level, the overall multi-variable adjusted OR suggested no significant relationship between dietary omega-6 PUFAs level and sarcopenia (OR = 0.64, 95% CI: 0.33-1.24; P = 0.19). However, the overall combined SMD showed that the dietary omega-6 PUFAs level in sarcopenia was slightly lower than that in control subjects (SMD = -0.15, 95% CI: -0.27 to -0.02; P = 0.02). Conclusion: Our results suggested that the dietary omega-3 PUFAs level was inversely associated with sarcopenia. However, current evidence is still insufficient to demonstrate the definite relationship between dietary omega-6 PUFAs levels and sarcopenia. More well-designed prospective cohort studies with the dietary omega-3/omega-6 PUFAs ratio are still needed.


Quality-adjusted life years: how useful in medico economic studies.
Cost-effectiveness analysis has evolved as a practical response to the need to allocate limited resources for health care. It can be used to compare interventions whose effects on health are different if the measure of effectiveness captures all the important health dimensions of the effects of the interventions. Using the quality-adjusted life year (QALY) as the unit of effectiveness attempts to approach this ideal and is currently the approach recommended by many consensus groups. Conventional QALYs represent time spend in a series of "quality-weighted" health states, where the quality weights reflect the desirability of living in the state. Many challenges arise when preferences are incorporated into an economic analysis. The purpose of this paper is to highlight some of the issues surrounding the use of QALYs and to encourage researchers to present their methodology in a clear and transparent way.


The medial breast margin sign: A new sign of pectus excavatum in adult females and comparison with classic signs.
Pectus excavatum (PX) denotes a concavity of the sternum and is the most common chest wall deformity. Many characteristic signs have been described on frontal chest radiographs including increased density of the inferomedial right lung, blurring of the right heart border, steeply angled anterior ribs, horizontal posterior ribs, straight or convex left heart border and loss of silhouette of the descending thoracic aorta. These signs are often subtle and rarely all present in any individual. We describe a further sign in women with PX, the medial breast margin sign. This sign consists of more sharply defined and more vertically oriented medial breast borders on frontal chest radiographs than in those of women with normal chest wall morphology. This study aims to document the sensitivity, specificity and inter-observer reliability of the new sign, and compare with that of the previously described signs in patients with varying degrees of PX. The frontal chest radiographs of 185 women with PX and 50 women with normal chest wall morphology were assessed by two reviewers for the presence of the new sign and previously described signs. All the signs had their sensitivity, specificity and inter-observer reliability calculated. The new sign had a sensitivity of 0.48, a specificity of 0.96 and inter-observer reliability of κ = 0.79, ranking third, second and first among all other signs, respectively. The medial breast margin sign is useful in diagnosing PX and is comparable to the previously described classic signs.


Childhood-Onset Multifocal Motor Neuropathy with IgM Antibodies to Gangliosides GM1: A Case Report with Poor Outcome.
Multifocal motor neuropathy (MMN) is a slowly progressive motor neuropathy characterized by asymmetric muscle weakness without sensory involvement. Typically, MMN respond completely to treatment with intravenous immunoglobulin (IVIg). MMN is even rarer in the pediatric population, where only five patients have been reported up to now. We discuss the 3-year follow-up of a 13-year-old girl with MMN who was positive for IgM antibodies to gangliosides GM1. She was diagnosed with MMN in accordance with the European Federation of Neurological Societies/Peripheral Nerve Society (EFNS/PNS) criteria. Serological studies revealed that she tested positive for IgM antibodies to GM1. She underwent intravenous methylprednisolone followed by an oral prednisone taper, intravenous immunoglobulin (IVIg), plasma exchange followed by IVIG and prednisone and Rituximab. No improvement was referred. At the present, she shows flaccid tetraplegia, facial diplegia, and bulbar cranial nerve palsy. Although childhood onset MMN is rare, most patients reported in literature respond to IVIg treatment. In a few cases, however, IVIg can be ineffective. In our patient, IVIg as well as treatment with prednisolone, plasma exchange and rituximab have failed.


The association of vascular endothelial growth factor related SNPs and circulating iron levels might depend on body mass index.
Vascular Endothelial Growth Factor (VEGF) is an essential regulator of vascular biology. In addition to the well-established role in angiogenesis, circulating VEGF levels were found elevated in severely anemic patients, pointing out that anemia might affect the progression of angiogenesis in malignant and benign diseases through the alteration of VEGF levels. Ten single nucleotide polymorphisms (SNPs) in VEGFA and other loci were shown to explain more than 50% of its circulating levels. This study investigated the association of those ten VEGF-related SNPs with serum iron levels in a general Lebanese population free of chronic diseases (N = 460). We found that the rs10738760 and the body mass index (BMI) were associated with decreased Iron levels (p = 0.002, and p < 0.001, respectively). When taken together, both variables, rs10738760 and BMI, interacted to reduce iron levels (p < 0.001). According to obesity status, the stratification revealed that the effect of rs10738760 was more pronounced in obese than non-obese individuals (p = 0.025). Conclusion: The intergenic SNP rs10738760 is associated with circulating iron levels, and this association depends on BMI status. Although of interest, these results need replication in larger populations from different ancestries.


Metabolomics: a tool for the diagnosis of GH deficiency and for monitoring GH replacement?
The diagnostic value of insulin-like growth factor 1 (IGF1) for GH deficiency (GHD) in adults is not optimal. Molecular profiling could be used for biomarker discovery. The aim of this pilot study was to compare the serum metabolome between GHD patients and healthy controls, and identification of potential markers for diagnosis and/or for individual GH dosing. A total of ten patients with GHD, median age of 55 years and BMI of 27 kg/m(2), were compared with ten healthy age- and gender-matched controls. The serum metabolic profiles were generated using gas chromatography-coupled mass spectroscopy on fasting samples taken in the morning from the controls and at baseline and during 6 months of GH replacement in the patients with GHD. The difference in low-molecular weight compounds (LMC) distinguished the healthy controls from GHD patients. Among 285 measured metabolites, 13 were identified as being most important in differentiating GHD patients from controls. Of these, 11 could not be structurally annotated but many were classified as lipids. The difference in the LMC pattern persisted despite normalisation of IGF1 following GH replacement. GH replacement increased the levels of specific fatty acid compounds and decreased the levels of certain amino acids. No metabolite changed in response to GH treatment, to the same extent as IGF1. The measurement of 285 metabolites resulted in a unique pattern in GHD, but changes in the metabolite patterns during GH treatment were limited. The utility of metabolomics to find new markers in GHD and GH replacement remains to be further elucidated.


Human common carotid wall shear stress as a function of age and gender: a 12-year follow-up study.
Hemodynamic forces play a role in the development of atherosclerosis. Their variations with age have been assessed in cross-sectional, but not longitudinal, studies. The aim of the present study was to investigate in both sexes the age-dependent change in wall shear stress and arterial stiffness in subjects studied twice 12 years apart. Forty-eight subjects (15 women and 33 men) were studied twice 12 years apart. Subjects underwent blood viscosity measurement and echo-Doppler of carotid arteries, from which the intima-media thickness (IMT) was measured and the wall shear stress and Peterson's elastic modulus were calculated. Blood viscosity increased in both sexes, more markedly in women (+13.2%) than men (+7.2%). Common carotid diameter increased in both sexes, but in men (+7.4%) more than in women (+5.5%). Peak and mean velocity decreased at follow-up by 10.7% and 18.9% in women and by 14.2% and 18.1% in men. Peak and mean shear stress significantly decreased in men by 13.0% and 17.5%, respectively, while in women only the mean shear stress was reduced (-11.8%). The IMT of the common carotid artery increased by 29% in women and 20% in men. Arterial stiffness significantly increased (+74.5% in women and +28.0% in men). The percent change in mean shear stress was significantly and inversely associated with the percent change in IMT. The data of this study show that, in a middle-aged population observed for almost 12 years, the mean shear stress decreases significantly in both sexes, while peak shear stress decreases significantly only in men. The change in mean shear stress is inversely associated with changes in IMT. Arterial stiffness, on the other hand, increases with aging.


Bioplastics from feather quill.
Poultry feather quills have been extruded in a twin screw extruder with sodium sulfite treatment as a reducing agent. The effect of four different plasticizers (ethylene glycol, propylene glycol, glycerol, and diethyl tartrate) on the thermoplastic properties was then investigated. Conformational changes and plasticizer-protein interactions in the extruded resins were assessed by Fourier transform infrared spectroscopy (FTIR), while viscoelastic behavior of the quill keratin plasticized with different plasticizers was investigated by dynamic mechanical analysis (DMA). Differential scanning calorimetry (DSC) was used to determine the effect of different plasticizers on protein denaturation. Thermal degradation patterns of the extrudates were studied by thermogravimetric analysis (TGA). The effect of plasticizers on the mechanical properties of resins was also assessed by tensile strength measurements. Results indicated that ethylene glycol was able to interact more effectively with quill keratin at the molecular level, exhibiting only one sharp glass transition, better mechanical properties, and higher transparency compared to other plasticized resins. The two phases found in glycerol plasticized material were attributed to glycerol-rich and protein-rich zones. Propylene glycol and diethyl tartrate exhibited lower H-bonding interactions and showed wide transition regions in DMA profiles during heating, suggesting weak and heterogeneous interactions between quill keratin and these plasticizers.


Attention deficit hyperactivity disorder and suicide: a review of possible associations.
To review the evidence of a possible association between attention deficit hyperactivity disorder (ADHD) and suicide. We searched the electronic data bases: Medline, Psych LIT, between 1966 and March 2003 looking for articles on ADHD, attention deficit disorder, hyperactivity and suicide. An association of ADHD and completed suicide was found, especially for younger males. However, the evidence for any direct or independent link was modest with an overall suicide rate from long-term follow-up studies of ADHD of 0.63-0.78%. The estimated relative risk ratio, compared with US national suicide rates (males 5-24 years) is 2.91 (95% confidence interval 1.47-5.7, chi2 = 9.3, d.f. = 1, P = 0.002). ADHD appears to increase the risk of suicide in males via increasing severity of comorbid conditions, particularly conduct disorder (CD) and depression. Identification of those at risk, particularly males with comorbid ADHD, depression and CD, may represent a useful clinical means of reducing completed suicide.


Highly Discriminative Physiological Parameters for Thermal Pattern Classification.
Infrared Thermography (IRT) is a non-contact, non-intrusive, and non-ionizing radiation tool used for detecting breast lesions. This paper analyzes the surface temperature distribution (STD) on an optimal Region of Interest (RoI) for extraction of suitable internal heat source parameters. The physiological parameters are estimated through the inverse solution of the bio-heat equation and the STD of suspicious areas related to the hottest spots of the RoI. To reach these values, the STD is analyzed by means: the Depth-Intensity-Radius (D-I-R) measurement model and the fitting method of Lorentz curve. A highly discriminative pattern vector composed of the extracted physiological parameters is proposed to classify normal and abnormal breast thermograms. A well-defined RoI is delimited at a radial distance, determined by the Support Vector Machines (SVM). Nevertheless, this distance is less than or equal to 1.8 cm due to the maximum temperature location close to the boundary image. The methodology is applied to 87 breast thermograms that belong to the Database for Mastology Research with Infrared Image (DMR-IR). This methodology does not apply any image enhancements or normalization of input data. At an optimal position, the three-dimensional scattergrams show a correct separation between normal and abnormal thermograms. In other cases, the feature vectors are highly correlated. According to our experimental results, the proposed pattern vector extracted at optimal position a=1.6 cm reaches the highest sensitivity, specificity, and accuracy. Even more, the proposed technique utilizes a reduced number of physiological parameters to obtain a Correct Rate Classification (CRC) of 100%. The precision assessment confirms the performance superiority of the proposed method compared with other techniques for the breast thermogram classification of the DMR-IR.


[Metastatic endometrial sarcoma of the right ventricular outflow tract associated with disseminated intravascular coagulopathy].
A case of myocardial metastasis of endometrial sarcoma to the right ventricular outflow tract is described. A 56-year-old woman was admitted with ecchymoses of lower limbs in September in 1995. Laboratory findings included thrombocytopenia and an increase of fibrinogen degradation products due to disseminated intravascular coagulopathy (DIC). Transesophageal echocardiography (TEE) demonstrated a large mass both in the right ventricular outflow tract and the proximal pulmonary artery. The mass was excised including base of the stalk. Cryoablation was employed on the endocardium where the stalk was originated. With pathological examination, the mass was identified to be endometrial sarcoma. The patient achieved an uneventful recovery and DIC was improved dramatically in a week after operation. Thirty-three months after the surgery, she has been well and free from local recurrent tumor.


Therapeutic potential of xanthones from Swertia chirata in breast cancer cells.
Medicinal plants like Swertia chirata are rich sources of different xanthones. This study was aimed to assess the cytotoxic potential of four most abundant xanthones present in S. chirata both in vivo and in vitro in Ehrlich ascites carcinoma (EAC), a mouse transplantable breast carcinoma cell line and two human breast carcinoma cell lines (MCF-7 and MDA-MB-231). Four xanthones derived from S. chirata namely 1-hydroxy-3,7,8-trimethoxyxanthone (XA), 1,8-dihydroxy-3,5-dimethoxyxanthone (XB), 1-hydroxy-3,5,8-trimethoxyxanthone (XC) and 1,5,8-trihydroxy-3-methoxyxanthone (XD) were used for determination of sub-lethal dose on the cell lines EAC, MCF-7, MDA-MB-231 and verified toxicity of sub-lethal dose on normal murine fibroblast cells. Cytotoxicity was measured in vivo and survivability of mice was plotted accordingly. Therapeutic efficacy of XD was evaluated both in vivo and in vitro by determination of lipid peroxidation (LPO), reactive oxygen species (ROS) generation and by quantitating the enzyme status (GSH, catalase, superoxide dismutase) in treated and untreated samples. DNA damage was evaluated using comet and DNA fragmentation assays. Furthermore, apoptotic effect was analyzed by flow cytometry and validated by TUNEL assay and Western blotting. Among all the xanthones tested XD showed IC50at the lowest dose, and normal cells were unaffected at this dose. Survivability of mice increased significantly when treated with XD compared to other xanthones and cisplatin. Significantly increased ROS and LPO were found in cancer cells as a result of XD treatment which was unaltered in normal cell line. XD induced DNA damage and apoptosis in cancer cell lines. Our experimental data indicate that XD may potentially act as a chemotherapeutic agent by enhancing ROS in breast cancer cells thereby leading to apoptosis.


Importance of algal biomass to growth and development of Anopheles gambiae larvae.
We conducted experiments to investigate the importance of algal food resources for larval growth and adult emergence of Anopheles gambiae Giles s.s. in simulated larval habitats in Kenya, and in greenhouse and laboratory microcosms in the United States. In the first experiment, we used shading to reduce algal biomass, and because algal production and larval development might be a function of underlying soil nutrients, we crossed sun-shade treatments with soils of two distinct types collected near larval habitats. Shading reduced pupation rates and total adult biomass of An. gambiae by approximately 50%. Soil type had no significant effect on mosquito production, but it did significantly affect concentrations of phosphorus and chlorophyll a in the surface microlayer. In a subsequent experiment conducted in the greenhouse to reduce temperature differences found between the shaded and sunlit treatments, <1% of larvae in the shaded treatments reached the pupal stage. There was a marked reduction of chlorophyll a levels as a function of shading and larval density. In a third experiment, larvae receiving material harvested from sunlit surface microlayers performed as well as those receiving liver powder, whereas those receiving surface microlayer from shaded habitats suffered >90% mortality and failed to pupate. In a fourth experiment, glucose was added to shaded microcosms to stimulate bacterial activity in the absence of algae. Bacterial growth rates were 2 to 3 times higher, and larval development was enhanced in glucose-amended treatments. However, pupation rates and adult weights in glucose-amended shaded microcosms were still poor compared with those in nonamended sunlit microcosms. Overall, these results demonstrate the importance of algal biomass in the surface microlayers of larval habitats to development and adult production of An. gambiae.


Socio-demographic correlates of physical activity and physical fitness in German children and adolescents.
Identifying factors that influence children's and adolescents' participation in physical activity (PA), as well as their physical fitness (PF), is essential for the development of effective intervention strategies. The aim of this study was to investigate and compare influential socio-demographic factors that affect PA such as socio-economic status (SES), rural-urban differences, immigration and age, as well as the effects of age and PA on PF as differentiated by gender. Subjects and German children between 6 and 9 years and adolescents between 10 and 17 years of age (n = 2574) participated in the representative, nationwide, cross-sectional 'Motorik'-Module study between 2003 and 2006. Results revealed that immigrant children and children with a lower SES background were less physically active and that this inactivity subsequently resulted in lower levels of PF as compared with non-immigrant children and children with a higher SES background. PA was further positively associated with age for children. All of these three socio-demographic factors were comparably meaningful for PA. In adolescents, the only PA-relevant socio-demographic parameter was SES with lower PA again resulting in lower PF levels observed in adolescents from lower income families. PF in childhood as well as adolescence was mostly positively affected by age, followed by PA, except for in female adolescents for whom PA and age were nearly equally significant. An intervention aiming to improve PA levels, and consequently PF levels, must, with respect to age and gender, refer to SES, as well as immigration background, but not to rural-urban differences.


Eradication of Salmonella Typhimurium infection in a murine model of typhoid fever with the combination of probiotic Lactobacillus fermentum ME-3 and ofloxacin.
The aim of the study was to detect whether in experimental Salmonella enterica Typhimurium infection the probiotic Lactobacillus fermentum ME-3 in combination with fluoroquinolone therapy would eradicate S. Typhimurium, prevent the development of liver and spleen granulomas and improve the indices of oxidative stress in the ileum mucosa. The selected bacteriological, histological and biochemical methods were applied. Combined treatment with L. fermentum ME-3 and ofloxacin eradicated Salmonella Typhimurium from blood, ileum and liver, decreased the number of animals with liver and spleen granulomas and reduced the value of lipid peroxides in the ileum mucosa. Higher total counts of intestinal lactobacilli in all experimental groups were associated with the absence of liver granulomas. The antimicrobial and antioxidative probiotic L. fermentum ME-3 combined with ofloxacin enhances the eradication of experimental S. Typhimurium infection. These observations on probiotic and antimicrobial co-action may serve as basis to develop new strategies for treatment of invasive bacterial infections of the gut.


Diseases caused by enterovirus 71 infection.
The purpose of this review was to explore the epidemiology, pathogenesis, virology, and management of enterovirus 71 (EV71) infection. Published literature was surveyed by Medline using the keyword "EV71." The reported incidence of cases of hand-foot-mouth disease/herpangina varied from year to year; seasonal variations in incidence were observed, with a peak in incidence during the summer season. Most cases of hand-foot-mouth disease/herpangina hospitalized for complications occurred in children less than 5 years old. The brainstem was the most likely major target of EV71 infection. Different enteroviruses cocirculate in the community annually. The emergence of the EV71 epidemic in the Asia Pacific region has been associated with the circulation of 5 genetic lineages (genotypes B3, B4, C1, C2, C4) that appear to be undergoing rapid evolutionary changes. The relationship between the gene structure of the EV71 virus and the factors that ensure its survival, ease of transmission, and evasion of immunity is still unclear. EV71 central nervous system involvement causes serious clinical illness, death, and long-term neurologic and psychiatric disorders in young children. EV71 infection has emerged as an important public health problem. Vaccine development is recommended for the prevention of EV71 infection in the future.


Differential secretion of cathepsins B and L from normal and tumor human lung cells stimulated by 12(S)-hydroxy-eicosatetraenoic acid.
Cathepsins B and L play roles in intracellular and extracellular proteolysis in normal and malignant processes. A directed extracellular proteolysis by regulated secretion could facilitate the process of invasion. We have therefore investigated the effect of the physiological signal mediator 12(S)-hydroxy-eicosatetraenoic acid on the release of cathepsins B and L in normal and malignant human lung cells. Quantitative determinations of cathepsin activities were done by flow cytometry and spectrofluorometry using synthetic dipeptidyl substrates coupled to fluorogens. Most interestingly, a difference in the secretion of cathepsins B and L was found: only release of active cathepsin B was detected. The effect was specific for 12(S)-hydroxy-eicosatetraenoic acid, 12(R)-hydroxy-eicosatetraenoic acid, and 5(S)-hydroxy-eicosatetraenoic acid were ineffective. The response was immediate but a substantial amount of nonreleasable activity remained cell bound. Alveolar macrophages, Wi-38 fibroblasts, and tumor cells derived from large cell carcinomas and adenocarcinomas were sensitive to 12(S)-hydroxy-eicosatetraenoic acid, but cells from undifferentiated squamous cell carcinomas were not. Sensitivity did not parallel malignancy but more likely the degree of differentiation of cells. The investigated tumor cell lines showed no detectable endogenous 12-lipoxy-genase activity to synthesize 12(S)-hydroxy-eicosatetraenoic acid from arachidonate; therefore, we assume a paracrine mechanism for 12(S)-hydroxy-eicosatetraenoic acid action. Protein kinase C alpha, a key enzyme involved in 12(S)-hydroxy-eicosatetraenoic acid-elicited responses, was expressed in all sensitive tumor cells, but insignificantly in a sensitive normal cell line and an insensitive tumor cell line. From our experiments we propose two separate intracellular pools of active cathepsin B: an unreleasable, lysosomal fraction and a fraction available for regulated secretion. Different processing and sorting mechanisms may be responsible for the generation of these cathepsin B-fractions in these pools.