Individual and dyadic network analyses of depressive symptoms in Chinese postpartum couples: A cross-sectional study.
The network approach to psychopathology is symptom oriented and may open new possibilities for intervention development and health care practices in postpartum depression. To investigate the individual and dyadic symptom network of postpartum depression in Chinese mothers and fathers in the very early postpartum period. A cross-sectional study was conducted. A total of 457 couples in the 2∼3 days postpartum period was recruited consecutively from a hospital in Guangzhou, China from September 2020 to April 2021. The Edinburgh Postnatal Depression Scale and socio-demographic and obstetric data sheet were used to collect data. We estimated the individual symptom networks of postpartum mothers and fathers separately and a dyadic symptom network that consisted of symptoms of both spouses. Network characteristics including global strength and node centralities were analyzed and systematically compared. Strength centralities in the individual networks showed acceptable stability [Correlation stability coefficient (CS) for mothers = 0.60; CS for fathers = 0.52]. The central depressive symptoms in mothers were Crying (Zstrength = 1.32), Overwhelmed (Zstrength = 1.01) and Sad mood (Zstrength = 0.93). The central depressive symptom in fathers was Sad Mood (Zstrength = 1.35). The symptom "Crying" had a distinctive link to thoughts of self-harm in fathers. The symptom network of mothers (global strength = 4.15) was more interconnected than that of fathers (global strength = 3.74). There was a statistically significant but unstable within-couple connection of thoughts of self-harm (CS = 0.21). Postpartum mothers are more vulnerable to activation spreads of depressive symptoms than postpartum fathers. Symptoms including "Sad mood", "Overwhelmed" and "Crying" warrant the attention of health care providers. Investigations with larger sample sizes and gender-sensitive instruments are needed to further unfold the individual and dyadic symptom dynamics of postpartum depression.


Preparation of tamarind fruit seed activated carbon by microwave heating for the adsorptive treatment of landfill leachate: A laboratory column evaluation.
The preparation of tamarind fruit seed granular activated carbon (TSAC) by microwave induced chemical activation for the adsorptive treatment of semi-aerobic landfill leachate has been attempted. The chemical and physical properties of TSAC were examined. A series of column tests were performed to determine the breakthrough characteristics, by varying the operational parameters, hydraulic loading rate (5-20 mL/min) and adsorbent bed height (15-21 cm). Ammonical nitrogen and chemical oxygen demand (COD), which provide a prerequisite insight into the prediction of leachate quality was quantified. Results illustrated an encouraging performance for the adsorptive removal of ammonical nitrogen and COD, with the highest bed capacity of 84.69 and 55.09 mg/g respectively, at the hydraulic loading rate of 5 mL/min and adsorbent bed height of 21 cm. The dynamic adsorption behavior was satisfactory described by the Thomas and Yoon-Nelson models. The findings demonstrated the applicability of TSAC for the adsorptive treatment of landfill leachate.


[Acute syndromes of the brain stem in newborn infants with birth injuries--case report].
The acute syndroms of the brainstem of cerebral injuried newborns by the birth trauma (casuistry). The neurologic clinical symptomatology of the acute syndroms of the brainstem are described in brief. Three cases of damaged newborn with this syndroms are particular published. Especially performed are: the possibility of repair, the changing of the symptoms, the danger of manual transformation of abnormous cephalic positions and the need of topological diagnosis in follow-up of special care.


Perinatal outcomes with intrahepatic cholestasis of pregnancy in twin pregnancies.
To describe perinatal outcomes of twin pregnancies complicated by intrahepatic cholestasis of pregnancy (ICP). We conducted a retrospective cohort study of women delivered at a large tertiary obstetric center in Shanghai, China from January 2006 to May 2014. Delivery data were abstracted from medical records of all twin gestations delivered at the hospital. A total of 129/1922(6.7%) twin and 1190/92 273 singleton (1.3%) pregnancies were complicated by ICP. An increased risk of stillbirth among twin pregnancies was observed (3.9% and 0.8% in the ICP and non-ICP groups, respectively; aOR 5.75, 95% CI 2.00-16.6). Stillbirths with ICP and twins occurred between 33 and 35 weeks gestation compared to 36-38 weeks gestation among singletons. ICP in twins was also associated with an increased risk of preterm birth (<37 weeks) with an aOR of 4.17 (95% CI 2.47-7.04) and an aOR of 1.89 (95% CI 1.26-2.85) for delivery <35 weeks. Twin pregnancies complicated by ICP also had increased meconium staining of amniotic fluid and lower birth weight. Twin pregnancies with ICP have significantly increased risks of adverse perinatal outcomes including stillbirth and preterm birth. Stillbirth occurs at an earlier gestational age in twin gestation compared to singletons, suggesting that earlier scheduled delivery should be considered in these women.


Unhealthy Glasgow: a case for ecological public health?
Ecological public health has been proposed as an approach appropriate for addressing the multiple transitions that currently affect human health and sustainability. The paper draws on the author's experience in public health in Glasgow to analyse the health challenges faced by this postindustrial Scottish city. Like other such cities, it not only faces multiple health challenges but also demonstrates a currently unexplained excess mortality that has been dubbed the 'Glasgow Effect'. To explore this troubled mixture, the paper outlines four historical waves of public health challenge and response in Glasgow over the last century, and proposes that a fifth is emerging. The challenge now is how to negotiate environmental sustainability with social, political and economic sustainability to enhance health for all. The paper suggests that gains made by past approaches still need to be protected and can be included within ecological public health, but they lack the wider vision, coherence and capacity required if cities are to address the scale and range of contemporary conditions. A number of lessons are offered for the ecological public health perspective.


Isolation and partial characterization of a mitochondrial deoxyribonucleic acid-protein complex from sea urchin embryos.
Lysis of mitochondria from sea urchin embryos with Triton X-100 led to a complete conversion of DNA-containing mitochondrial residues into protein-DNA complex with a density higher than 1.22 g/cm3 in sucrose solutions. This complex banded isopycnically in metrizamide gradients at a density of 1.26 g/cm3. Exposure to mixtures of Triton X-100 with Tween 80 resulted in progressively less delipitated and disorganized mitochondria over Tween/Triton weight ratios from 1 to 2, with the retention of the starting buoyant density in sucrose of approximately 1.16 g/cm3 at Tween/Triton ratios above 2.5. The DNA-internal protein complex sedimented with the bulk of the surviving mitochondrial structure under all conditions studied. No free DNA could be detected under any conditions of membrane removal.


Determination of silica-purified plasma calcitonin for the detection and management of medullary thyroid carcinoma: comparison of two provocative tests.
Radioimmunoassays for human calcitonin in whole plasma are limited in sensitivity and specificity; basal values of calcitonin are often undetectable in normal plasma, and nonspecific increases are occasionally found in seemingly healthy persons. We avoided these problems by applying a silica-cartridge extraction-concentration technique for calcitonin assay, and the effectiveness of two calcitonin stimulation tests in healthy volunteers and patients with medullary thyroid carcinoma was compared. The radioimmunoassay was improved by using a new antiserum and a sequential incubation procedure that reduced the previously used sample-volume requirement and incubation period substantially. This method was used to measure mean basal plasma levels of calcitonin (+/-SD) in 45 normal men (8.2 +/- 5 pg/ml) and 47 normal women (4.8 +/- 4 pg/ml) (P less than 0.001). Calcium infusions (2 mg/kg over 5 minutes) in 18 normal men and 37 normal women yielded a significantly greater secretory response than did pentagastrin (0.5 micrograms/kg as a bolus). Among 12 patients with medullary thyroid carcinoma, pentagastrin seemed to be a better secretagogue than calcium (P less than 0.001). We recommend routine measurement of plasma silica-extractable calcitonin and pentagastrin injection as the provocative test of choice for the detection and management of medullary thyroid carcinoma. The short calcium-infusion test is a good alternative.


The posterior subtemporal keyhole approach combined with the transchoroidal approach to the ambient cistern: microsurgical anatomy and image-guided quantitative analysis.
The aim of this study is to describe microsurgical anatomy and to quantitatively analyze exposure using the posterior subtemporal keyhole (PSK) approach combined with the transchoroidal keyhole (TCK) approach to the ambient cistern. We determined the proper location of craniotomy of such combined keyhole approach on 16 sides of cadaver heads. The PSK approach and the TCK approach were performed in the same minicraniotomy to observe microanatomic features and to quantitatively measure exposure limits of the ambient cistern and related structures using image-guided system. Some superficial and bone landmarks could be used to find the proper location of such combined minicraniotomy. In the PSK approach, the exposure distances of the trigeminal nerve and the anterior portion of the P2 segment (P2a) were 10.02 ± 0.76 mm and 16.32 ± 2.02 mm, respectively. The superior, inferior, anterior, and posterior exposure limit of brainstem from the intersection point of the lateral mesencephalic sulcus and pontomesencephalic sulcus was 7.5 ± 0.19 mm, 11.04 ± 0.27 mm, 15.72 ± 0.52 mm, and 10.16 ± 0.38 mm, respectively. In the TCK approach, the vertical distances between the taenia fimbriae and the lateral geniculate body without and with mild caudal retraction of the hippocampus were 5.28 ± 0.46 mm and 11.18 ± 0.57 mm, respectively. The linear exposure distances of the posterior portion of the P2 segment (P2p) or P3 segment were 12.14 ± 1.88 mm. Except of one case, the P2p could be exposed using the TCK approach. The midpoint of the medial edge of the parahippocampal gyrus on the coronal magnetic resonance images provides landmark to choose the appropriate approach. The PSK combined with the TCK approach can simultaneously expose the lower and upper ambient cistern in a proper minicraniotomy.


The association of endogenous Go alpha with the purified omega-conotoxin GVIA receptor.
Modulation of the neuronal omega-conotoxin GVIA-sensitive N-type voltage-dependent calcium channel (VDCC) by neurotransmitters and guanine nucleotides suggests a dynamic interaction between activated G-protein alpha subunits and the N-type VDCC. Our previous report on the purification of the N-type VDCC (McEnery, M. W., Snowman, A. M., Sharp, A. H., Adams, M. E., and Snyder, S. H. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 11095-11099), has led us to investigate a possible association of CTXR with an endogenous G alpha subunit. The addition of the G-protein activator AIF4- modulated the 125I-CTX binding characteristics of the solubilized CTXR. Further immunological analyses employing G alpha subunit-specific antibodies to monitor the cofractionation of G alpha with 125I-CTX binding activity throughout the purification procedure indicate the selective recovery of Go alpha in the purified CTXR preparation, as neither Gs alpha, Gi alpha, nor G beta gamma could be detected. Furthermore, Go alpha associated with CTXR acted as a substrate for pertussis toxin-dependent ADP-ribosylation only upon the addition of exogenous G beta gamma subunits. These results strongly indicate a high affinity complex between an activated Go alpha and CTXR maintained throughout biochemical purification of the 125I-CTX receptor.


Could nephrotoxicity due to colistin be ameliorated with the use of N-acetylcysteine?
The protective effect of N-acetylcysteine (NAC) on nephrotoxicity due to contrast nephropathy and reperfusion-induced ischemia has been reported in experimental models. However, its efficacy on colistin-induced nephrotoxicity has not been elucidated yet. The primary aim of this study was to evaluate the nephrotoxic effect of colistin and to investigate the possible protective effect of NAC on colistin-induced nephrotoxicity. The secondary aim was to research the systemic effects of nephrotoxicity-induced oxidative stress on the lung. Eighteen female Sprague-Dawley rats were randomly assigned and were given (a) 1 ml/kg sterile saline, (b) 300,000 IU/kg/day colistin, and (c) 300,000 IU/kg/day colistin and 150 mg/kg NAC for six consecutive days. Plasma blood urea nitrogen (BUN), creatinine, urinary creatinine, urinary protein, plasma TNF-alpha levels, renal tissue superoxide dismutase (SOD) and malondialdehyde (MDA) activity and immunocytochemical findings were evaluated. Colistin exerted nephrotoxicity and achieved a significant increase in plasma BUN and creatinine levels and renal tissue SOD levels. NAC exhibited no significant effect on biochemical parameters but reduced renal tissue SOD level and reversed immunocytochemical staining of inducible nitric oxide synthase (i-NOS) and neurotrophin-3. Increased oxidative stress in the lung tissue of the rats treated with colistin has also been documented. Additionally, NAC significantly reduced the immunostaining of endothelial NOS (e-NOS) and i-NOS in the lung tissue. Colistin-induced renal toxicity may be attributable to oxidative damage. The combined treatment of colistin plus NAC seems to have a beneficial role in restoration of the oxidant injury which may be related to its antioxidant effect.


DNA binding proteins in chromatin structure and function.
The binding of chromosomal proteins to cloned mouse and human globin genes can be determined by transferring proteins from one dimensional electrophoretic separations on to nitrocellulose paper and exposing them to nick translated cloned globin DNAs. The usefulness of this technique is assessed in the case of RNA polymerases isolated from mouse Ascites tumor cells where a specific initiation site for transcription of mouse beta globin DNA can be demonstrated.


Suppression of an eplieptiform type of electrocortical activity in the rat by stimulation in the vicinity of locus coeruleus.
Stimulation of the locus coeruleus, or in the vicinity of this nucleus or of its ascending tracts, could markedly suppress the appearance of epileptiform-like ECoG bursts. The latter were induced in rats by a subconvulsive dose of pentylenetetrazol. Electrode sites were identified histologically. A unilateral stimulus suppressed bursts bilaterally. An individual burst already in progress could be aborted, stopping within less than 0.5-1 sec after onset of a stimulus train. The antiepileptiform actions occurred with no evidence of any desynchronizing effect of the stimulus on the resting ECoG; they appear to be different in sites of origin and nature from those reported for stimulation of the reticular activating system. It is proposed that stimulation of the ascending noradrenergic system in the brain stem may limit the development and spread of hyperexcitatory, epileptiform states.


Hypovitaminosis D: a novel risk factor for coronary heart disease in type 2 diabetes?
Vitamin D (25(OH)D) levels have been associated with cardiovascular disease. Thus, the aim of our study was to investigate the association of 25(OH)D levels with coronary heart disease (CHD) in 698 consecutive type 2 diabetic outpatients. 698 consecutive type 2 diabetic outpatients (25.2 % men, age 66 ± 9 years) and 100 (90 % men, age 65 ± 13 years) age-matched non-diabetic volunteers were enrolled. 25(OH)D assay and the main cardiovascular risk factors were explored. 25(OH)D concentration was 22 ± 10 ng/ml in control subjects and 18.23 ± 10 ng/ml in diabetic patients (p < 0.01). The prevalence of hypovitaminosis D was higher in diabetic patients than in control subjects (90 vs. 83 %, p < 0.01). Diabetic subjects with hypovitaminosis D had higher prevalence of high values of A1C (p < 0.01), BMI (p < 0.01), LDL cholesterol (p < 0.01), triglycerides (p < 0.01), and glycemia (p < 0.01) than their vitamin D-sufficient counterparts. 25(OH)D and HDL cholesterol were lower (p < 0.01), while BMI (p < 0.01), age (p < 0.01), systolic (p < 0.01) and diastolic blood pressure (p < 0.01), diabetes duration (p < 0.01), A1C (p < 0.01), glycemia (p < 0.01), fibrinogen (p < 0.01), triglycerides (p < 0.01), and total (p < 0.01) and LDL cholesterol (p < 0.01) were higher in diabetic subjects with CHD than diabetic subjects without CHD. At the logistic regression analysis, the association of vitamin D with CHD was lost, while sex (p = 0.026), diabetes duration (p = 0.023), and age (p = 0.024) were the most powerful predictors of CHD. The current study demonstrates that 25(OH)D does not have a direct effect on CHD but may have an indirect effect mediated by cardiovascular risk factors such as diabetes duration, age, and sex.


Peroxisomal disorders.
The peroxisomal disorders represent a group of genetic diseases in man in which there is an impairment in one or more peroxisomal functions. The peroxisomal disorders are subdivided into three subgroups comprising: (1) the peroxisome biogenesis disorders (PBDs); (2) the single peroxisomal (enzyme-) protein deficiencies; and (3) the single peroxisomal substrate transport deficiencies. The PBD group comprises four different disorders that include Zellweger syndrome (ZS), neonatal adrenoleukodystrophy (NALD), infantile Refsum disease (IRD), and rhizomelic chondrodysplasia punctata (RCDP). ZS, NALD, and IRD are clearly distinct from RCDP and are usually referred to as the Zellweger spectrum with ZS being the most severe, and IRD the less severe disorder, with sometimes onset in adulthood. The single peroxisomal enzyme deficiency group comprises seven different disorders, of which D-bifunctional protein and phytanoyl-CoA hydroxylase (adult Refsum disease) deficiencies are the most frequent. The single peroxisomal substrate transport deficiency group consists of only one disease, X-linked adrenoleukodystrophy. It is the purpose of this chapter to describe the current state of knowledge about the clinical, biochemical, cellular, and molecular aspects of peroxisomal diseases, and to provide guidelines for their post- and prenatal diagnosis. Therapeutic interventions are mostly limited to X-linked adrenoleukodystrophy.


Structural basis for the specificity and catalysis of human Atg4B responsible for mammalian autophagy.
Reversible modification of Atg8 with phosphatidylethanolamine is crucial for autophagy, the bulk degradation system conserved in eukaryotic cells. Atg4 is a novel cysteine protease that processes and deconjugates Atg8. Herein, we report the crystal structure of human Atg4B (HsAtg4B) at 1.9-A resolution. Despite no obvious sequence homology with known proteases, the structure of HsAtg4B shows a classical papain-like fold. In addition to the papain fold region, HsAtg4B has a small alpha/beta-fold domain. This domain is thought to be the binding site for Atg8 homologs. The active site cleft of HsAtg4B is masked by a loop (residues 259-262), implying a conformational change upon substrate binding. The structure and in vitro mutational analyses provide the basis for the specificity and catalysis of HsAtg4B. This will enable the design of Atg4-specific inhibitors that block autophagy.


Human leukocyte antigen evolutionary divergence influences outcomes of paediatric patients and young adults affected by malignant disorders given allogeneic haematopoietic stem cell transplantation from unrelated donors.
High genetic heterogeneity in the human leukocyte antigen (HLA) increases the likelihood of efficient immune response to pathogens and tumours. As measure of HLA diversity, HLA evolutionary divergence (HED) has been shown to predict the response of tumours to immunotherapy and haematopoietic stem cell transplantation (HSCT) in adults. We retrospectively investigated the association of HED with outcomes of 153 paediatric/young adults patients, treated for malignant disorders with HSCT from 9-10/10 HLA-matched unrelated donors. HED was calculated as pairwise genetic distance between alleles in patient HLA-A, -B, -C, -DRB1, -DQB1 and -DPB1, using the locus median to stratify patients with 'high' or 'low' HED. Patients with high HED-B and -DRB1 showed significantly improved disease-free survival (DFS), especially when combined (70.8% vs 53.7% p = 0.008). High HED-B + -DRB1 was also associated with improved overall survival (OS) (82.1 vs 66.4% p = 0.014), and concomitant reduction of non-relapse-mortality (5.1% vs 21.1% p = 0.006). The impact on OS and DFS of combined HED-B + -DRB1 was confirmed in multivariate analysis [hazard ratio (HR) 0.39, p = 0.009; and HR 0.45, p = 0.007 respectively]. Only high HED scores for HLA-DPB1 were associated, in univariate analysis, with reduced incidence of relapse (15.9% vs 31.1%, p = 0.03). These results support HED as prognostic marker in allogeneic HSCT and, if confirmed in larger cohorts, would allow its use to inform clinical risk and potentially influence clinical practice.


Coastal blue carbon in China as a nature-based solution toward carbon neutrality.
To achieve the Paris Agreement, China pledged to become "Carbon Neutral" by the 2060s. In addition to massive decarbonization, this would require significant changes in ecosystems toward negative CO2 emissions. The ability of coastal blue carbon ecosystems (BCEs), including mangrove, salt marsh, and seagrass meadows, to sequester large amounts of CO2 makes their conservation and restoration an important "nature-based solution (NbS)" for climate adaptation and mitigation. In this review, we examine how BCEs in China can contribute to climate mitigation. On the national scale, the BCEs in China store up to 118 Tg C across a total area of 1,440,377 ha, including over 75% as unvegetated tidal flats. The annual sedimental C burial of these BCEs reaches up to 2.06 Tg C year-1, of which most occurs in salt marshes and tidal flats. The lateral C flux of mangroves and salt marshes contributes to 1.17 Tg C year-1 along the Chinese coastline. Conservation and restoration of BCEs benefit climate change mitigation and provide other ecological services with a value of $32,000 ha-1 year-1. The potential practices and technologies that can be implemented in China to improve BCE C sequestration, including their constraints and feasibility, are also outlined. Future directions are suggested to improve blue carbon estimates on aerial extent, carbon stocks, sequestration, and mitigation potential. Restoring and preserving BCEs would be a cost-effective step to achieve Carbon Neutral by 2060 in China despite various barriers that should be removed.


Double-negative metamaterial square enclosed Q.S.S.R For microwave sensing application in S-band with high sensitivity and Q-factor.
Metamaterials have gained much attention due to their exciting characteristics and potential uses in constructing valuable technologies. This paper presents a double negative square resonator shape metamaterial sensor to detect the material and its thickness. An innovative double-negative metamaterial sensor for microwave sensing applications is described in this paper. It has a highly sensitive Q-factor and has good absorption characteristics approximately equal to one. For the metamaterial sensor, the recommended measurement is 20 by 20 mm. Computer simulation technology (C.S.T.) microwave studios are used to design the metamaterial structure and figure out its reflection coefficient. Various parametric analyses have been performed to optimize the design and size of the structure. The experimental and theoretical results are shown for a metamaterial sensor that is attached to five different materials such as, Polyimide, Rogers RO3010, Rogers RO4350, Rogers RT5880, and FR-4. A sensor's performance is evaluated using three different thicknesses of FR-4. There is a remarkable similarity between the measured and simulated outcomes. The sensitivity values for 2.88 GHz and 3.5 GHz are 0.66% and 0.19%, respectively, the absorption values for both frequencies are 99.9% and 98.9%, respectively, and the q-factor values are 1413.29 and 1140.16, respectively. In addition, the figure of merit (FOM) is analyzed, and its value is 934.18. Furthermore, the proposed structure has been tested against absorption sensor applications for the purpose of verifying the sensor's performance. With a high sense of sensitivity, absorption, and Q-factor, the recommended sensor can distinguish between thicknesses and materials in various applications.


Aspergillus detection in airways of ICU COVID-19 patients: To treat or not to treat?
It is now well known that patients with severe COVID-19 are at risk for developing invasive pulmonary aspergillosis (IPA). Nevertheless, the symptomatology of IPA is often atypical in mechanically ventilated patients and the radiological aspects of SARS CoV-2 pneumonia and IPA are difficult to differentiate. In this context, the significance of the presence of Aspergillus in respiratory tract samples (detected by culture, galactomannan antigen, or specific PCR) is not yet fully understood. Here we report two cases of intubated and mechanically ventilated ICU patients with SARS-CoV-2 pneumonia, in whom Aspergillus was detected in respiratory samples, who had a favorable outcome in the absence of antifungal treatment. These two cases highlight the difficulty of using the new definitions of COVID-19 associated pulmonary aspergillosis for routine management of patients.


Primacy of lowered baseline glomerular filtration rate as a risk for incident chronic kidney disease: A longitudinal study in Japanese subjects.
Risk profile for incident chronic kidney disease (CKD) in Japanese subjects has not been established. Our aim was to identify risk factors for CKD in Japanese. Consecutive 171 536 health examinees (median age 49 years and estimated glomerular filtration rate (eGFR) 78.2 mL/min per 1.73 m2 ) without CKD were re-examined after a median period of 6.2 years. Results of Cox proportional hazards models in randomly assigned two thirds (Derivation cohort) were verified in the rest (Validation cohort). CKD was defined as eGFR <60 mL/min per 1.73 m2 or positive dipstick proteinuria. In the Derivation cohort, CKD developed in 1002 (5.8%) subjects. Seven variables such as lower eGFR, male gender, higher uric acid concentration, lower red cell count and higher age and systolic blood pressure were identified as significant risks for CKD, with lowered eGFR being an overwhelmingly strong risk: adjusted hazard ratio for those with the baseline eGFR <70 mL/min per 1.73 m2 was as high as 90.1. Performance of prediction of CKD by the probability on the basis of the seven risk factors combined was only marginally preferable to eGFR alone. The area under the receiver operating characteristic curve (95% CI) for the prediction was 0.846 (0.826-0.864) and 0.822 (0.802-0.840) (P < 0.01), the kappa statistic was 0.263 and 0.250 (n.s.), and the mean absolute difference between "predicted probability" and "observed" CKD was 1.4% and 1.9% (P = 0.14) by the combined model and eGFR alone, respectively. Seven risk factors for incident CKD were identified in Japanese health examinees. However, lowered baseline eGFR outweighed other risks to the degree that eGFR alone was suffice for CKD prediction.


Endoscopic repair of anterior or middle skull base cerebrospinal fluid leaks after tumour resection.
This study analysed the factors influencing the risk of recurrence following endoscopic repair of CSF leaks of the anterior and middle skull base after endonasal skull base tumour resection. A retrospective review was conducted on 17 patients operated between May 2007 and December 2010 by endonasal endoscopic resection of anterior or middle skull base tumour who presented an intraoperative CSF leak. Epidemiological data (body mass index [BMI], age, gender) and type of leak (site, size) were studied. CSF leaks involved the roof of the ethmoid sinus (one patient), cribriform plate (three), posterolateral wall of the sphenoid sinus (six) or the sella turcica (seven). The CSF leak recurrence rate after the first endoscopic procedure was 29.4% (5/17). Failures were not influenced by gender, age, BMI or size of the leak. All recurrences involved the sella turcica or the lateral wall of the sphenoid sinus. The success rate after a second endoscopic procedure was 88.2%. Endonasal endoscopic repair of anterior and middle skull base meningeal injuries after tumour resection is a minimally invasive and effective technique. The main challenge of this method consists of ensuring effective control of the postoperative defect after tumour resection in the sphenoidal region, as this region was the only potential risk factor for recurrence identified in this study.


Expression and localization of Tmie in adult rat cochlea.
Loss-of function mutations in transmembrane inner ear expressed (Tmie/TMIE) gene have been shown to cause deafness in mice and humans (DFNB6). However, the functional roles of TMIE in the cochlea remain unclear. A primary step toward the understanding of the role of TMIE in hearing and its dysfunction is the documentation of its cellular and sub-cellular location within the cochlea, the auditory organ. In this study, we located and determined the cellular expression of Tmie within the rat cochlea using a polyclonal anti-Tmie antibody. The anti-Tmie antibody identified a specific band of 17 kDa in a variety of rat tissues by using Western blot analyses. The expression products of Tmie were also detected in the spiral limbus, spiral ligament, organ of Corti, and stria vascularis by immunohistochemistry analysis and RT-PCR. Our results point out the presence and localization of Tmie products in the cochlea of rat. Knowledge of spatial distribution of Tmie will provide important insight into the mechanisms that lead to deafness due to mutations in the TMIE gene.


Pinning down proline-directed phosphorylation signaling.
The reversible phosphorylation of proteins on serine or threonine residues preceding proline (Ser/Thr-Pro) is a major cellular signaling mechanism. Although it is proposed that phosphorylation regulates the function of proteins by inducing a conformational change, there are few clues about the actual conformational changes and their importance. Recent identification of the novel prolyl isomerase Pin1 that specifically isomerizes only the phosphorylated Ser/Thr-Pro bonds in certain proteins led us to propose a new signaling mechanism, whereby prolyl isomerization catalytically induces conformational changes in proteins following phosphorylation to regulate protein function. Emerging data indicate that such conformational changes have profound effects on catalytic activity, dephosphorylation, protein-protein interactions, subcellular location and/or turnover. Furthermore, this post-phosphorylation mechanism might play an important role in cell growth control and diseases such as cancer and Alzheimer's.


Total knee replacement after failed tibial osteotomy: results of a matched-pair study.
Sixty-three total knee replacements were performed after a failed tibial osteotomy. The goal of this study was to compare the perioperative problems and the outcome of this group of patients (study group) to a group of patients with primary arthroplasties matched for age, gender, length of follow-up, weight, and preoperative Charnley class. Operative problems were more frequently encountered in the study group, with 7 tibial tubercle elevations and 15 lateral retinaculum releases needed, whereas lateral retinaculum release was necessary for only 1 knee in the control group. Outcome was assessed using both the International Knee Society (IKS) scoring system and Hospital for Special Surgery (HSS) knee score. The follow-up period averaged 4.6 years. The IKS score of the control group was significantly higher, averaging 80.9 +/- 13.8, whereas it was 74.4 +/- 14.8 for the study group (P = .0001). Among the parameters included in the knee score, only pain was significantly different with the control group (P = .03). The IKS function score and the HSS score were not statistically different. Conversion of a failed tibial osteotomy is a technically demanding procedure. Careful preoperative planning is needed. Results, especially on pain, appeared to be inferior to those for primary arthroplasties.


Cardio-Obstetrics: A Focused Review.
Cardiovascular disease is the leading cause of maternal mortality in the USA. All cardiovascular care providers should have a foundational knowledge on the management of pregnant individuals with heart disease. This focused review touches on several key cardio-obstetric themes. Many individuals with cardiovascular disease can safely undergo pregnancy, but should have counseling preconception to optimize cardiac status. There are several cardiovascular conditions that are high risk for maternal mortality and morbidity. These individuals should be adequately counseled preconception and offered reliable birth control. The approach to a high-risk pregnant patient with cardiac disease is best managed by a multidisciplinary team to address potential maternal and fetal complications. Identification of at risk individuals can be estimated preconception with several risk scores. The development of risk scores to stratify and identify those at elevated risk during pregnancy is an area of continued research and development.


Radioautographic localization of radioactivity in rat brain after intraventricular or intracarotid injection of 3H-L-prolyl-L-leucyl glycinamide.
Tritiated L-prolyl-L-leucyl-glycinamide was injected into the carotid artery or lateral ventricle of rats and the radioactivity localized by radioautography. After intracarotid administration, the radioactivity labeled the meninges, ependymal cells bordering the ventricles, and cells of the choroid plexus, as well as the median eminence and subfornical organ and thus may have reached the cerebrospinal fluid. After intraventricular injection, globus pallidus, indusium griseum, hippocampus, corpus callosum, and meninges. The results after intracarotid injection suggest that L-prolyl-L-leucyl-glycinamide may cross the blood brain barrier and the results after intraventricular injection suggest sites of localization which might be correlated with the previously reported effects of the tripeptide on the brain.


Multiparametric MR Imaging of Breast Cancer.
Breast MR imaging has increased in popularity over the past 2 decades due to evidence of its high sensitivity for cancer detection. Current clinical MR imaging approaches rely on the use of a dynamic contrast-enhanced acquisition that facilitates morphologic and semiquantitative kinetic assessments of breast lesions. The use of more functional and quantitative parameters holds promise to broaden the utility of MR imaging and improve its specificity. Because of wide variations in approaches for measuring these parameters and the considerable technical challenges, robust multicenter data supporting their routine use are not yet available, limiting current applications of many of these tools to research purposes.


Extensive angiosarcoma on chronically sun-damaged skin.
Cutaneous angiosarcoma of the face and scalp is a rare malignant vascular tumor that most often occurs in the white elderly males. No clear etiologic factors have been associated with the development of this aggressive neoplasm. Due to delay in diagnosis, inability to adequately define clinical and surgical margins, and its high local recurrence and metastatic rates, angiosarcomas generally have a poor prognosis. Histologic evaluation of angiosarcoma can be somewhat difficult due to the varying patterns of differentiation. We present a case of angiosarcoma arising on chronically sun-damaged skin and review the histopathology and prognostic factors important in this malignancy.


Preparation and characterization of insulin nanoparticles using chitosan and its quaternized derivatives.
Insulin (INS), like other peptides, has low therapeutic activity when administered orally due to degradation by proteolytic enzymes. Polymeric nanoparticles have been introduced as a useful carrier for peptide oral delivery, because they can protect these compounds from degradation. The objective of the present study is to develop an INS nanoparticulate system by using chitosan (CS), triethylchitosan (TEC), and dimethyl-ethylchitosan (DMEC, a new quaternized derivative of CS). INS-polymer nanoparticles were prepared by the polyelectrolyte complexation method. The physicochemical properties of the nanoparticles including particle size distribution, zeta potential, and polydispersity index were determined by using dynamic light scattering technique. Transmission electron microscopy was also used to observe the morphology of the nanoparticles. The amount of INS loaded into the nanoparticles was determined by measuring the association efficiency and also the content of INS in the nanoparticles. In vitro release studies showed a relatively small burst effect at the beginning and then a sustained release characteristic for 5 hours.


Family bereavement care interventions during the COVID-19 pandemic: a scoping review protocol.
The COVID-19 pandemic has caused significant disruptions to daily social routines and to the lived experience of bereaved families. This article outlines the protocol for a scoping review of published studies to evaluate psychosocial and psychotherapeutic interventions intended to help family carers adjust to grief, loss and bereavement due to COVID-19. This review addresses one broad research question: 'What do we know about bereavement support interventions for family carers of COVID-19 victims?' The seminal framework by Arksey and O'Malley will guide the review process, which will cover both the qualitative and quantitative scientific literature on grief support during COVID-19. We will search for relevant studies in several databases, namely PubMed, Web of Science, CINAHL Complete, PsycINFO, Scopus and Directory of Open Access Journals. Moreover, we will search the reference lists of included studies and grey literature sources. The database search will be limited to studies from February 2020 (first death by COVID-19) to 1 January 2022. Only literature written in English, Portuguese and Spanish shall be included. Two independent reviewers will screen the literature, select articles and extract data, in an iterative process. Any disagreements will be solved through consensus-based discussion. Results will be reported with descriptive statistics, accompanied by a thematic analysis. The study will use information acquired from previously published papers and hence does not require ethical approval. This protocol is registered with the Open Science Framework (https://osf.io/bw7fn/). Scoping results will be disseminated via posters and oral presentations to both academic and clinical audiences, as well as through peer-reviewed journals.


An unusual buried polar cluster in a family of fungal lipases.
The stability of globular proteins arises largely from the burial of non-polar amino acids in their interior. These residues are efficiently packed to eliminate energetically unfavorable cavities. Contrary to these observations, high resolution X-ray crystallographic analyses of four homologous lipases from filamentous fungi reveal an alpha/beta fold which contains a buried conserved constellation of charged and polar side chains with associated cavities containing ordered water molecules. It is possible that this structural arrangement plays an important role in interfacial catalysis.


Systemic responses to inhaled ozone in mice: cachexia and down-regulation of liver xenobiotic metabolizing genes.
Rats or mice acutely exposed to high concentrations of ozone show an immediate and significant weight loss, even when allowed free access to food and water. The mechanisms underlying this systemic response to ozone have not been previously elucidated. We have applied the technique of global gene expression analysis to the livers of C57BL mice acutely exposed to ozone. Mice lost up to 14% of their original body weight, with a 42% decrease in total food consumption. We previously had found significant up-regulation of genes encoding proliferative enzymes, proteins related to acute phase reactions and cytoskeletal functions, and other biomarkers of a cachexia-like inflammatory state in lungs of mice exposed to ozone. These results are consistent with a general up-regulation of different gene families responsive to NF-kappaB in the lungs of the exposed mice. In the present study, we observed significant down-regulation of different families of mRNAs in the livers of the exposed mice, including genes related to lipid and fatty acid metabolism, and to carbohydrate metabolism in this tissue, consistent with a systemic cachexic response. Several interferon-dependent genes were down-regulated in the liver, suggesting a possible role for interferon as a signaling molecule between lung and liver. In addition, transcription of several mRNAs encoding enzymes of xenobiotic metabolism in the livers of mice exposed to ozone was decreased, suggesting cytokine-mediated suppression of cytochrome P450 expression. This finding may explain a previously controversial report from other investigators more than 20 years ago of prolongation of pentobarbital sleeping time in mice exposed to ozone.


Mapping cis-regulatory elements in human neurons links psychiatric disease heritability and activity-regulated transcriptional programs.
Genome-wide association studies (GWASs) have identified hundreds of loci associated with psychiatric diseases, yet there is a lack of understanding of disease pathophysiology. Common risk variants can shed light on the underlying molecular mechanisms; however, identifying causal variants remains challenging. We map cis-regulatory elements in human neurons derived from pluripotent stem cells. This system allows us to determine enhancers that activate the transcription of neuronal activity-regulated gene programs, which are thought to be critical for synaptic plasticity and are not possible to identify from postmortem tissues. Using the activity-by-contact model, we create variant-to-gene maps to interpret the function of GWAS variants. Our work nominates a subset of variants to elucidate the molecular mechanisms involving GWAS-significant loci. It also highlights that in vitro human cellular models are a powerful platform for identifying and mechanistic studies of human trait-associated genetic variants in cell states that are inaccessible from other types of human samples.


ECG as a risk stratification tool in patients with wearable cardioverter-defibrillator.
The wearable cardioverter defibrillator (WCD) is increasingly used in patients at elevated risk for ventricular arrhythmias but not fulfilling the indications for an implantable cardioverter defibrillator (ICD). Currently, there is an insufficient risk prediction of fatal arrhythmias in patients at risk. In this study, we assessed the prognostic role of baseline electrocardiogram (ECG) in WCD patients. WCD patients from diverse clinical institutions in Germany (n = 227) were retrospectively enrolled and investigated for the incidences of death or ventricular arrhythmias during WCD wearing. In addition, the widely accepted ECG predictors of adverse outcome were analyzed in patients with arrhythmic events. Life-threatening arrhythmias occurred in 22 (9.7 %) patients, mostly in subjects with ischemic heart disease (15 of 22). There was no difference in baseline left ventricular ejection fraction (LVEF) in subjects with and without arrhythmic events (31.3 ± 7.9 % vs. 32.6 ± 8.3 %; p = 0,24). Patients with arrhythmia exhibited significantly longer QRS duration (109.5 ± 23.1 ms vs. 100.6 ± 22.3 ms, p = 0,04), Tpeak-Tend (Tp-e) (103.1 ± 15.6 ms vs. 93.2 ± 19.2 ms, p = 0,01) and QTc (475.0 ± 60.0 ms vs. 429.6 ± 59.4 ms, p < 0,001) intervals. In contrast, no significant differences were found for incidences of fragmented QRS (27.3 % vs. 24 %, p = 0.79) and inverted/biphasic T-waves (16.6 % vs. 22.7 %, p = 0,55). In multivariate regression analysis both Tp-e (HR 1.03; 95 % CI 1.001-1.057; p = 0.02) and QTc (HR 1.02; 95 % CI 1.006-1.026; p < 0.001) were identified as independent predictors of ventricular arrhythmias. After WCD use, the prophylactic ICD was indicated in 76 patients (33 %) with uneventful clinical course but persistent LVEF ≤35 %. The ECG analysis in these subjects did not reveal any relevant changes in arrhythmogenesis markers. ECG repolarization markers Tp-e and QTc are associated with malignant arrhythmias in WCD patients and may be used - in addition to other established risk markers - to identify appropriate patients for ICD implantation.


Differential effects of interleukin-4 on superoxide anion production by human alveolar macrophages stimulated with lipopolysaccharide and interferon-gamma.
The effect of interleukin-4 (IL-4) on the activation state of human alveolar macrophages (AMs) and blood monocytes induced by lipopolysaccharide (LPS) or recombinant interferon-gamma (IFN-gamma) was investigated on the basis of their ability to produce superoxide anion (O2-). AMs were obtained from healthy donors by bronchoalveolar lavage, and O2- productions of these cells were assayed by a cytochrome c reduction method after incubation with stimulants for 24 h. AMs produced more O2- than autologous blood monocytes when stimulated with LPS. IL-4 alone had little effect on O2- production by unstimulated AMs but down-regulated O2- production by LPS-stimulated AMs in a dose-dependent manner. IL-4 also suppressed O2- production by AMs induced by the synergistic actions of muramyl dipeptide (norMDP) and IFN-gamma. Maximum suppression by IL-4 of O2- production by AMs was observed when IL-4 was added within 1 h after initiation of LPS stimulation. AMs also showed high O2- production when stimulated with IFN-gamma alone. In contrast to its suppression of O2- production by LPS-stimulated AMs, IL-4 enhanced O2- production by AMs stimulated with IFN-gamma. These data suggest that IL-4 is an important regulator of O2- production by macrophages through different pathways depending on the stimulus.


Anterior capsule re-attachment in terrible triad elbow injury with coronoid tip fracture.
During the surgical treatment of terrible triad elbow injury (TTEI), the usefulness of re-attaching the anterior joint capsule when the coronoid tip is fractured remains unclear. The primary objective of this study was to assess potential benefits during surgery for TTEI of re-attaching the joint capsule when the coronoid tip is fractured. Re-attaching the anterior joint capsule in TTEI with a fractured coronoid tip improves clinical and radiological outcomes and decreases the complication and revision rates. This single-centre retrospective study included patients who underwent surgery at the acute phase of TTEI with a fractured coronoid tip. In all patients, a physical examination and elbow radiographs were performed at least 1year after surgery. A statistical analysis was done to compare the groups with vs. without re-attachment of the anterior capsule and coronoid tip. The study included 30 patients, 16 females and 14 males, with a mean age of 51years (range: 21-84years). Among them, 11 did and 19 did not undergo re-attachment. The two groups were comparable regarding demographic features and follow-up duration. No significant differences were found at last follow-up for flexion-extension motion arc (p=0.75), pronation-supination motion arc (p=0.3051), or the Mayo Elbow Performance Score (p=0.19). Radiographic evidence of humero-radial osteoarthritis was significantly more common in the absence of re-attachment (p=0.04), whereas no differences were evidenced regarding humero-ulnar osteoarthritis (p=0.73), the occurrence of subluxation or dislocation (p=0.43), or loosening of the radial head implant (p=0.47). The complication and revision rates were similar in the two groups. In our experience, re-attaching the anterior capsule during the surgical treatment of TTEI with a coronoid tip fracture did not improve the clinical or radiographic outcomes after a mean follow-up of 54months. IV, retrospective study.


Facile microfluidic channels for acoustophoresis on a budget.
Acoustophoresis is a powerful yet gentle technique for manipulating cells and particles that has quickly earned a place in the lab-on-a-chip toolkit. However, traditional construction techniques for acoustophoretic resonators have typically required prohibitively expensive and laborious processing methods. Here, we propose a highly cost-effective and cleanroom-free construction technique for transversal acoustophoretic resonators. Channels with two different widths of 750 and 300 μm were constructed using a simple glass and polyimide sandwiching technique. Half and full wavelength resonators were then established using 1 and 5 MHz ultrasound respectively and polystyrene beads were successfully manipulated in both types of resonators. This construction technique was then utilized to demonstrate a bifurcation and trifurcation microchannel with 600 μm widths and 2.5 MHz ultrasound. Our approach addresses some of the key drawbacks of acoustophoretic devices by drastically simplifying the fabrication and prototyping of transversal resonators and will assist in expanding this technology from laboratory benches and into the broader market.


Effects of thermoneutrality on food intake, body weight, and body composition in a Prader-Willi syndrome mouse model.
Prader-Willi syndrome (PWS) is a multisystem genetic disorder. Unfortunately, none of several mouse models carrying PWS mutations emulates the entirety of the human PWS phenotype, including hyperphagia plus obesity. To determine whether housing at thermoneutrality (TN, 30 °C) permits the development of hyperphagia and obesity in the Snord116del PWS mouse model, the effects of housing three different ages of Snord116del and wild-type (WT) littermates at TN versus room temperature (RT, 22-24 °C) for 8 weeks were compared. Snord116del mice born and maintained at TN exhibited lower body weight curves, lower percentage fat mass, and lower food intake than WT mice at RT. In 4- to 6-month-old high-fat diet-fed female mice, TN raised the Snord116del body weight curve closer to that of RT-housed WT mice although the TN-housed Snord116del mice did not gain more adiposity or exhibit greater food intake. In 6- to 8-month-old high-fat diet-fed male mice, body weight, adiposity, and food intake of TN-housed Snord116del mice remained far below levels in RT-housed WT mice. TN elicited hypotonia in Snord116del adults and exacerbated mortality of Snord116del newborns. In none of three tested TN protocols were greater food intake, body weight, or adiposity induced in Snord116del mice compared with RT-housed WT mice.


In vivo application of tissue-engineered blood vessels of bacterial cellulose as small arterial substitutes: proof of concept?
Tissue-engineered blood vessels (TEBVs) represent an innovative approach for overcoming reconstructive problems associated with vascular diseases by providing small-caliber vascular grafts. This study aimed to evaluate a novel biomaterial of bacterially synthesized cellulose (BC) as a potential scaffold for small-diameter TEBV. Small-diameter blood vessels with a supramolecular fiber network structure consisting of tubular hydrogels from biodesigned cellulose were created using Gluconacetobacter strains and Matrix reservoir technology. BC tubes (length: 100 mm, inner diameter: 4.0-5.0 mm) were applied to replace the carotid arteries of 10 sheep for a period of 3 mo to gain further insights into (a) functional (in vivo) performance, (b) ability of providing a scaffold for the neoformation of a vascular wall and (c) their proinflammatory potential, and the (d) technical feasibility of the procedure. Preoperative analysis revealed a bursting strength of the grafts of approximately 800 mm Hg and suture retention strength of 4-5 N. Postexplantation analysis showed a patency rate of 50% (n = 5) and physiological performance of the patent grafts at 4, 8, and 12 wk postoperatively, compared with native arteries. Histologic analysis revealed a neoformation of a vascular wall-like structure along the BC scaffold consisting of immigrated vascular smooth muscle cells and a homogeneous endothelialization of the inner graft surface without signs of prothrombogenic or inflammatory potential. Scanning electron microscopy revealed a confluent luminal endothelial cell layer and the immigration of vascular smooth muscle cells into the BC matrix. BC grafts provide a scaffold for the neoformation of a three-layered vascular wall exhibit attractive properties for their use in future TEBV programs for cardiovascular surgery.


The protective effect of tadalafil on IMA (ischemia modified albumin) levels in experimental renal ischemia-reperfusion injury.
To investigate the effect of the tadalafil in experimental renal I/R injury and to evaluate these changes with IMA (nonspesific early biomarker of ischemia), NO and MDA levels. Twenty four female Wistar rats were randomly divided into 3 groups (n=8): Group I, sham; Group II, 60 min I/R; Group III, 60 min I/R plus tadalafil. Tadalafil was administered via an orogastric tube (10 mg/kg) 24 h prior to the procedure. After ischemia of the left kidney and 1 h of reperfusion, blood samples were obtained, and the kidney was removed. Statistically significant histopathologic changes were exist between groups, with the most severe injury was determined in group II in comparison to the others (X(2)=21,803, P=0.000). Also mean serum IMA levels were higher in group II, but not statistically significant (19.83±7.81 U/ml, 22.26±7.14 U/ml and 19.82±7.77 U/ml, P=0.613). In addition, NO values were lower in I/R groups (P=0.049). There were no differences among the groups in terms of MDA. IMA may be used as a nonselective biomarker for IR injury before the occurrence of necrosis. Decreased IMA levels may indicate the nephroprotective effect of tadalafil in renal IR injury.


Efficacy of etanercept in the treatment of a patient with Behçet's disease.
Behçet's disease is a systemic inflammatory disorder without a specific treatment that is chosen on the basis of the type and severity of manifestation in the organ involved. More recently, biological agents like etanercept have emerged as possible therapeutic alternatives in patients resistant to conventional therapy. We describe the successful treatment for 1 year of resistant Behçet's disease with etanercept. After the administration of this drug, a resolution of the clinical, laboratory and instrumental picture was achieved with a suspension of immunosuppressive treatments and a reduction of steroid dependency (5 mg/day). No side effects were observed.


A UPLC-MS/MS method for the determination of oxidative stress biomarkers in amniotic fluid.
Oxidative stress in the fetal period is associated with preterm birth as well as short and long-term adverse clinical outcomes. Here, an Ultra-Performance Liquid Chromatography-tandem Mass Spectrometry (UPLC-MS/MS) method for the simultaneous quantification of biomarkers of oxidative stress-derived damage to proteins and DNA in amniotic fluid (AF) samples is presented. Appropriate accuracy and precision levels, as well as sensitivity with limits of detection in the low nanomolar (<2 nM) range were achieved. The analytical method was applied to a set of AF samples and reference ranges of the biomarker panel are presented. Median concentrations of biomarkers of protein oxidation (ortho-, 3-chloro-, and 3-nitrotyrosine) and their precursors (para-tyrosine and phenylalanine) ranged between 0.6 and 3 nM and 23 and 30 μM, respectively, while levels of a biomarker of DNA-oxidation (8-hydroxydeoxyguanosine, 8OHdG) and its precursor (2'-deoxyguanosine) were found to be 0.18 and 3 nM, respectively. Detection frequencies of all metabolites were 100% with exception of 3-chlorotyrosine (3Cl-Tyr) and 8OHdG, that were only detected in 8% of samples. The developed method may be applied in research studies focusing on oxidative stress-related complications during pregnancy.


Isolation and characterization of gallic acid and methyl gallate from the seed coats of Givotia rottleriformis Griff. and their anti-proliferative effect on human epidermoid carcinoma A431 cells.
Gallic acid (GA) and its derivative methyl gallate (MG) are well studied plant phenolics. They have exhibited anticancer effects in several cancer cell lines. However, the presence of GA/MG in the seed coats of Givotia rottleriformis and their inhibitory effect on human epidermoid carcinoma (A431) skin cancer cells were not reported. In this study we have isolated and chemically characterized the bioactive compounds GA and MG from the bioassay guided methanolic (MeOH) seed coat extracts of G. rottleriformis. The fractions obtained from open silica column chromatography were subjected to in vitro enzymatic assays. Among seven fractions we found that only fractions 5 and 6 showed significant inhibition activity toward COX-1 with an IC50 value of 28 μg/mL and 9.3 μg/mL and COX-2 with an IC50 value of 35 μg/mL and 7.0 μg/mL respectively. However, we could not find 5-LOX enzyme inhibition activity. MG (10 mg/g DW) and GA (6 mg/g DW) were the major compounds of seed coats. Cell viability was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, which showed that GA/MG significantly reduced the growth of A431 cells with an IC50 value of 25 μg/mL and 53 μg/mL and 11 μg/mL and 43 μg/mL at 24 h and 48 h, respectively. However the cytotoxic effect of GA/MG on HaCaT normal skin keratinocyte cell line was found to be less. Western blot analysis has shown that GA/MG treatment down regulated Bcl-2 and up regulated cleaved caspase-3 with respect to increasing doses. Our results conclude that GA and MG have potential anticancer effects and can be used as therapeutic agents for skin cancers.


The facultative human oral pathogen Prevotella histicola in equine cheek tooth apical/ periapical infection: a case report.
Prevotella histicola is a facultative oral pathogen that under certain conditions causes pathologies such as caries and periodontitis in humans. Prevotella spp. also colonize the oral cavity of horses and can cause disease, but P. histicola has not yet been identified. A 12-year-old Tinker mare was referred to the clinic for persistent, malodorous purulent nasal discharge and quidding. Conservative antibiotic (penicillin), antiphlogistic (meloxicam), and mucolytic (dembrexine-hydrochloride) treatment prior to referral was unsuccessful and symptoms worsened. Oral examination, radiography, sino-/ rhinoscopy, and standing computed tomography revealed severe apical/ periapical infection of the upper cheek tooth 209 with accompanying unilateral sinonasal inflammation and conchal necrosis. The tooth exhibited extensive subocclusal mesial infundibular cemental hypoplasia and caries, and an occlusal fissure fracture. After mechanical debridement and thermoplastic resin filling of the spacious subocclusal carious infundibular lesion, the tooth was extracted intraorally. The sinusitis and conchal necrosis were treated transendoscopically. Selective bacteriological swab cultures of affected tooth roots and subsequent matrix-assisted laser desorption ionization-time of flight mass spectrometry showed an infection with the obligate anaerobic, Gram-negative bacterium P. histicola. Surgical intervention and adapted antibiotic therapy led to normal healing without complications. This study provides the first documented case of dental infection in a horse caused by P. histicola at once indicating necessity of more sufficient microbiological diagnostics and targeted antibiotic treatment in equine dental practice. This finding is also conducive to understand species-specific Prevotella diversity and cross-species distribution.


3D Rosa centifolia-like CeO2 encapsulated with N-doped carbon as an enhanced electrocatalyst for Zn-air batteries.
Reasonable design and synthesis of high-efficiency rare earth oxides-based materials as alternatives to noble-metal catalysts are of great significance for oxygen electrocatalysis. Herein, we report three-dimension (3D) Rosa centifolia-like CeO2 encapsulated with N-doped carbon (NC) composites (CeO2@NC) for enhancing oxygen reduction reaction (ORR) and oxygen evolution reaction (OER) activities. This synthetic method allows CeO2 to tune the oxygen vacancy concentration and electronic structure of a series of CeO2@NC catalysts due to its large oxygen-storage-capacity (OSC) property. Moreover, benefiting from the exposed active sites in NC as well as the synergy between CeO2 and NC, among as-prepared samples, the resultant CeO2@NC-900 delivers a half-wave potential (E1/2) of 0.854 V, which is more positive compared with counterpart of NC-900 (0.806 V) and even comparable to that of commercial Pt/C catalyst (0.855 V). This indicates that the ORR electrocatalytic activity of CeO2@NC-900 is significantly improved. Meanwhile, CeO2@NC-900 exhibits satisfactory performance toward OER. For practical application, the CeO2@NC-900 involved rechargeable Zn-air battery possesses excellent energy efficiency, superior stability, and large energy density (666.1 Wh kgZn-1 at 5 mA cm-2). This approach provides a valid way to develop advanced rare earth oxides-based materials for energy applications.


Colloid cysts of the third ventricle. A review of 36 cases.
Thirty-six colloid cysts were treated from 1949 to 1983. There were 26 male and 10 female patients, ranging in age from 12 to 65 years old, 60% between 31 and 40 years. Headache or disturbed mental function was the most frequent complaint, papilloedema the most frequent sign. The patients were classified into 3 groups by symptoms and signs; Group I (17 patients): Headache, papilloedema and no neurological signs. Group II (6 patients): Fluctuating or progressive dementia. Group III: Twelve cases with "classical" features, episodic headache and drop attacks. One patient could not be classified in any of these groups. Seventeen of 36 patients were diagnosed by ventriculography, 19 patients were diagnosed by CT scan. CT scan was the most reliable diagnostic study, but was unavailable in the earlier part of the series. All patients have been operated by transventricular exposure of the right foramen of Munro with incision of the middle frontal gyrus in an antero-posterior linear manner. Twenty-five of 36 patients showed an excellent operative result, and nine had a good result, one of the 9 patients dying of an unrelated intracerebral haemorrhage 4 years after operation. Two patients had a poor result, characterized by memory loss and confusion. One of these died of an unknown cause 5 years after operation.


Clinical features do not identify risk of progression from isolated postcapillary pulmonary hypertension to combined pre- and postcapillary pulmonary hypertension.
Pulmonary hypertension is a common sequelae of left heart failure and may present as isolated postcapillary pulmonary hypertension (Ipc-PH) or combined pre- and postcapillary pulmonary hypertension (Cpc-PH). Clinical features associated with progression from Ipc-PH to Cpc-PH have not yet been described. We extracted clinical data from patients who underwent right heart catheterizations (RHC) on two separate occasions. Ipc-PH was defined as mean pulmonary pressure >20 mmHg, pulmonary capillary wedge pressure >15 mmHg, and pulmonary vascular resistance (PVR) < 3 WU. Progression to Cpc-PH required an increase in PVR to ≥3 WU. We performed a retrospective cohort study with repeated assessments comparing subjects that progressed to Cpc-PH to subjects that remained with Ipc-PH. Of 153 patients with Ipc-PH at baseline who underwent a repeat RHC after a median of 0.7 years (IQR 0.2, 2.1), 33% (50/153) had developed Cpc-PH. In univariate analysis comparing the two groups at baseline, body mass index (BMI) and right atrial pressure were lower, while the prevalence of moderate or worse mitral regurgitation (MR) was higher among those who progressed. In age- and sex-adjusted multivariable analysis, only BMI (OR 0.94, 95% CI 0.90-0.99, p = 0.017, C = 0.655) and moderate or worse MR (OR 3.00, 95% CI 1.37-6.60, p = 0.006, C = 0.654) predicted progression, but with poor discriminatory power. This study suggests that clinical features alone cannot distinguish patients at risk for development of Cpc-PH and support the need for molecular and genetic studies to identify biomarkers of progression.


Hypoglycaemia is associated with the absence of a decrease in diurnal macular thickness in patients with diabetic macular oedema.
Spontaneous diurnal variations measured by optical coherence tomography (OCT) have been reported in diabetic macular oedema (DME) together with a daytime decrease in central macular thickness (CMT). For this reason, this study aimed to investigate the influence of acute glucose and blood pressure changes on daytime variations in CMT in patients with DME. In this prospective observational study of type 1 (n=4) and type 2 (n=18) diabetic patients with DME, OCT scans, capillary blood glucose, and systolic and diastolic blood pressure measurements were performed at 9 a.m., 12 a.m., 3 p.m., 6 p.m. and again at 9 a.m. the day after. At the same time, the study protocol included simultaneous ambulatory blood pressure and glucose monitoring over a 24-h period. Hypoglycaemic episodes, defined as glucose values<60mg/dL, were also recorded. CMT decreased consistently between 9 a.m. and 6 p.m. in 10 patients (from 374±82μm to 337±72μm; P=0.01) and increased or remained steady in 12 others (from 383±136μm to 390±149μm; P=0.58), with a significant difference in CMT absolute change between the two groups (P<0.001). In the study population as a whole, the lower the mean diurnal blood glucose, the smaller the decrease in CMT during the day (P=0.027). Also, eight (67%) of the 12 patients with a flat CMT profile experienced a diurnal hypoglycaemic event whereas none of those with a CMT decrease had hypoglycaemia (P=0.002). Hypoglycaemic events may explain the lack of diurnal CMT decrease in diabetic patients with DME. However, further studies need to be conducted to evaluate whether having no diurnal CMT decrease is associated with a poorer visual prognosis and whether it can be modified by better glucose control.


Cerebrospinal Fluid Leak Repair Simulation Model: Face, Content, and Construct Validation.
Cerebrospinal fluid leak (CSFL) repair simulation models are scarce; however, these models are valuable tools for skull base reconstruction surgery training. This study aims to assess the face, content, and construct validity of a CSFL repair simulation model. Eight novices (residents- PGY3) and eight experts have performed skull base reconstruction in multiple sites in twelve human cadaveric heads in simulated surgical environment. The experts completed a post-study 21-item questionnaire to assess the face and content validity. The performances of the participants were recorded and scored by two independent investigators who were blinded to the participant's level. Global Rating Scale of Operative Performance (GRSOP) and a Specific Skull Base Reconstruction Checklist (SBRC) were used to score the performances. The responses from the expert group for the 21-item questionnaire were high for all items (4.13-4.88 out of 5). The internal consistency reliability of the questionnaire and the intraclass correlation, which was derived by Cronbach's Alpha, were 0.913 and 0.941 respectively. Differences in construct validity between the two groups were statistically significant for both the GRSOP and SBRS (P-value < 0.001). We demonstrated the face, content, and construct validity of the CSFL repair simulation model, which facilitates the acquisition of technical skills necessary for skull base reconstruction surgery. The model includes realistic features that make it useful in educational courses.


Searching for genes influencing a complex disease: the case of coeliac disease.
Recently, a few genes have been reported to be causative in inflammatory diseases. Still, we are waiting for the vast majority to be discovered. New tools for genotyping and statistical analysis have been developed and emphasis has been put on study design. Coeliac disease (CD) is a disorder, where prolamins in dietary wheat gluten and related proteins from rye or barley are not tolerated. It is one of the most common chronic diseases in humans exceeding a population prevalence of 1%. In this article, we will summarise what is currently known about the genetics influencing CD with the emphasis on the non-HLA genetic component. We will discuss some difficulties when searching for susceptibility genes in disorders with complex inheritance patterns.


Morphological characterization and expression analysis of genes for antioxidants enzymes in gamma rays induced M3 mutants of sunflower (Helianthus annuus L.) cultivar TNAUSUF-7.
Objectives of the present investigation were to investigate the performance of mutant lines (mutant generation 3 (M3) generation) of sunflower obtained through gamma irradiations for yield and yield attributing traits including oil content (%), oil quality in terms of antioxidant activity and to investigate the expression of genes encoding antioxidant enzymes in control and gamma irradiated M3 lines. Mutant lines (M3 generation) of sunflower variety TNAUSUF-7 obtained from 100 and 130 Gray (Gy) doses of gamma irradiation were evaluated for several traits such as days to 50% flowering, days to maturity, head diameter, number of filled seeds and unfilled seeds per head, fertility %, oil content %, antioxidant activity of oil and expression of genes encoding antioxidant enzymes. All the data were analyzed using different statistical tools. Our results showed that the gamma irradiation dose of 100 and 130 Gy induced significant variations in yield and yield attributing traits especially for days to 50% flowering, days to maturity, fertility %, antioxidant property of sunflower seed oil and expression of genes encoding antioxidant enzymes. From our results, it can be concluded that the gamma irradiations were effective in creating variations in terms of several traits in sunflower. Many desirable traits like reduced days to 50% flowering and maturity, increased fertility %, increased antioxidant activity of oil were observed in M3 lines. These M3 lines have the potential to be utilized as an inbred line in sunflower hybrid development or it could be used as a source of desirable traits in sunflower breeding programs.


MAP3K1 rs889312 genotypes influence survival outcomes of Chinese gastric cancer patients who received adjuvant chemotherapy based on platinum and fluorouracil regimes.
For patients with gastric cancer (GC), adjuvant chemotherapy is a standard therapy. However, the responses to the treatment are quite different. Mitogen-activated protein kinase (MAPK) pathway is a core pathway that modulates the efficacy of anticancer drugs. The purpose of our study was to investigate the clinical significance of one pivotal functional gene polymorphism in the MAPK pathway - MAP3K1 rs889312 - in patients with stage II GC to stage III GC. The genotypes of MAP3K1 rs889312 were analyzed in 591 GC patients enrolled in this study who had received radical gastrectomy. Among them, 204 patients accepted adjuvant chemotherapy based on platinum and fluorouracil (PF) regimens after an operation. Cox regression analysis, log-rank test and Kaplan-Meier method were used to explore the link between MAP3K1 rs889312 variant and overall survival (OS) of GC. Compared with the AA genotype (mean OS of 68.12 months), MAP3K1 rs889312 AC/CC significantly reduced the mean OS of 56.83 months in patients who received adjuvant chemotherapy only. In addition, AC/CC genotype had a negative impact on OS of patients who received oxaliplatin-based therapy (HR, 8.253; 95% CI: 1.119-60.853, log-rank p=0.013). Stratification analysis showed that MAP3K1 rs889312 AC/CC significantly reduced OS of patients with tumors smaller than or equal to 5 cm in size (HR, 3.706; 95% CI: 1.329-10.335, p=0.012), poorly differentiated tumors (HR, 3.002; 95% CI: 1.076-8.377, p=0.036) and intestinal tumors (HR, 4.780; 95% CI: 1.138-20.073, p=0.033). Our findings suggested that MAP3K1 rs889312 single-nucleotide polymorphism may be considered as a biomarker for adjuvant chemotherapy reaction and can predict prognosis of GC patients who received PF-based therapy.


Hemoretroperitoneum associated with liver bare area injuries: CT evaluation.
In hepatic injury restricted to the postero-superior region of segment VII (bare area), hemoperitoneum may be absent and this condition may be associated with hemoretroperitoneum. The aim of this paper is to present the association between bare area injuries and hemoretroperitoneum evaluated by CT. The CT examinations of 32 patients with blunt liver trauma were reviewed and the number and location of lesions were evaluated. Right lobe involvement was identified, focusing on the bare area lesions. The presence of hemoperitoneum and hemoretroperitoneum were determined. In the 32 patients 44 parenchymal lesions were detected. Segment VII was involved in 16 cases: 5 patients presented an intraparenchymal lesion, 11 patients a lesion emerging to the liver surface. In 8 cases the lesion was localized in the bare area. In the 16 patients presenting a segment-VII lesion, hemoperitoneum was detected in 3 cases, hemoretroperitoneum in 4 cases, and both conditions in 4 cases. A traumatic hepatic lesion may be associated with hemoretroperitoneum rather than hemoperitoneum. This justifies the absence of clinical signals of peritoneal irritation; the negativity of both US scan and peritoneal lavage may cause an inappropriate therapeutic management. Computed tomography yields both the detection of the parenchymal damage and the correct localization of the intraperitoneal and retroperitoneal hemorrhage.


Biomarkers of severe dengue disease - a review.
Dengue virus infection presents a wide spectrum of manifestations including asymptomatic condition, dengue fever (DF), or severe forms, such as dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) in affected individuals. The early prediction of severe dengue in patients without any warning signs who may later develop severe DHF is very important to choose appropriate intensive supportive therapy since available vaccines for immunization are yet to be approved. Severe dengue responses include T and B cell activation and apoptosis, cytokine storm, hematologic disorders and complement activation. Cytokines, complement and other unidentified factors may transiently act on the endothelium and alter normal fluid barrier function of the endothelial cells and cause plasma leakage. In this review, the host factors such as activated immune and endothelial cells and their products which can be utilized as biomarkers for severe dengue disease are discussed.


Consistent functional abnormalities in patients with postpartum depression.
Postpartum depression (PPD) is a common public health concern. A wide range of functional abnormalities in various brain regions have been reported in fMRI studies on PPD, however, a consistent functional changing pattern is still lacking. Herein, we obtained functional Magnetic Resonance Imaging (fMRI) data from 52 patients with PPD and 24 healthy postpartum women (HPW). Functional indexes (low-frequency fluctuation, degree centrality, and regional homogeneity) were calculated and compared among these groups to explore the functional changing patterns of PPD. Then, correlation analyses were performed to investigate the relationship between changed functional indexes and clinical measurements in the PPD. Finally, support vector machine (SVM) was performed to test whether these abnormal features can be used to distinguish PPD from HPW. As a result, we identified significantly and consistently functional changing pattern characterizing by increased functional activity in the left inferior occipital gyrus and decreased functional activity right anterior cingulate cortex in the PPD as compared to HPW. These functional values in the right anterior cingulate cortex were significantly correlated with depression symptoms in the PPD, and can be used as features to distinguish PPD from HPW. In conclusion, our results suggested that the right anterior cingulate cortex could be served as a functional neuro-imaging biomarker for PPD, which might be used as a potential target for neuro-modulation.


Improved biocompatibility of dendrimer-based gene delivery by histidine-modified nuclear localization signals.
Polyamidoamine (PAMAM) dendrimers have been explored as an alternative to polyethylenimine (PEI) as a gene delivery carrier because of their relatively low cytotoxicity and excellent biocompatibility. The transfection efficiency of PAMAM dendrimers can be improved by the addition of nuclear localization signal (NLS), a positively charged peptide sequence recognized by cargo proteins in the cytoplasm for nuclear transport. However, increased positive charges from NLS can cause damage to the cytoplasmic and mitochondrial membranes and lead to reactive oxygen species (ROS)-induced cytotoxicity. This negative effect of NLS can be negated without a significant reduction in transfection efficiency by adding histidine, an essential amino acid known as a natural antioxidant, to NLS. However, little is known about the exact mechanism by which histidine reduces cytotoxicity of NLS-modified dendrimers. In this study, we selected cystamine core PAMAM dendrimer generation 2 (cPG2) and conjugated it with NLS derived from Merkel cell polyomavirus large T antigen and histidine (n = 0-3) to improve transfection efficiency and reduce cytoxicity. NLS-modified cPG2 derivatives showed similar or higher transfection efficiency than PEI 25 kDa in NIH3T3 and human mesenchymal stem cells (hMSC). The cytotoxicity of NLS-modified cPG2 derivatives was substantially lower than PEI 25 kDa and was further reduced as the number of histidine in NLS increased. To understand the mechanism of cytoprotective effect of histidine-conjugated NLS, we examined ROS scavenging, hydroxyl radical generation and mitochondrial membrane potential as a function of the number of histidine in NLS. As the number of hisidine increased, cPG2 scavenged ROS more effectively as evidenced by the hydroxyl radical antioxidant capacity (HORAC) assay. This was consistent with the reduced intracellular hydroxyl radical concentration measured by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) assay in NIH3T3. Finally, fluorescence imaging with JC-1 confirmed that the mitochondrial membranes of NIH 3T3 were well-protected during the transfection when NLS contained histidine. These experimental results confirm the hypothesis that histidine residues scavenge ROS that is generated during the transfection process, preventing the excessive damage to mitochondrial membranes, leading to reduced cytotoxicity.


Application and validation of the barrow neurological institute screen for higher cerebral functions in a control population and in patient groups commonly seen in neurorehabilitation.
To determine whether the Barrow Neurological Institute Screen for Higher Cerebral Functions (BNIS) can differentiate brain-dysfunctional patients from controls. A case-control study. A total of 92 controls and 120 patients from a neuro-rehabilitation clinic with a diagnosis of: right and left hemisphere stroke, traumatic brain injury, Parkinson's disease or anoxic brain damage. The BNIS has a maximum total score of 50 points, < 47 indicates cognitive dysfunction. Group comparisons and exploration of variables influencing the BNIS total score were made. A significant difference was found between the control group and the total patient group for the BNIS total score and for the subscales (p < 0.0005). Sensitivity was 88% and specificity 78%. Presence of disease and educational level had the greatest influence on the results of the BNIS. Patients with Parkinson's disease were shown to be the least cognitively affected and those with anoxic brain damage the most affected. The BNIS has potential value as a screening instrument for cognitive functions and is sufficiently sensitive to differentiate brain-dysfunctional patients from a control population. It appears to be applicable in a neurological rehabilitation setting, and can be used early in the process, giving a baseline cognitive functional level.


Higher Dispositional Optimism Predicts Lower Pain Reduction During Conditioned Pain Modulation.
Optimism is associated with lower pain sensitivity, positive adjustment to chronic pain, and greater reduction of pain thresholds in a conditioned pain modulation (CPM) paradigm. We hypothesized that participants with higher levels of optimism would experience greater inhibition of suprathreshold pain during CPM. Seventy-seven healthy adults completed a test of optimism, the Life Orientation Test-Revised, as well as measures of depression, pain catastrophizing, and neuroticism. Participants also underwent psychophysical tests of heat pain tolerance, heat pain threshold, and CPM. CPM magnitude was calculated as the change in heat pain ratings when applied alone and simultaneously with painful pressure. Greater optimism was significantly correlated with reduced CPM magnitude (P = .013). Regression analysis was performed using optimism as a predictor of CPM magnitude while controlling for pain catastrophizing, neuroticism, depression, and age. The overall model was significant (P = .003). Significant positive coefficients were found for depression (P = .014) and optimism (P < .001) scores. These results suggest that greater optimism predicts less inhibition of suprathreshold pain, the opposite of our hypothesis. This unexpected finding may be due to factors such as perceived stress and coping differences, and suggests that modulation of threshold-level and suprathreshold pain involves different underlying mechanisms. PERSPECTIVE: This article reports that greater optimism predicts less inhibition of suprathreshold pain, in contrast with previous work showing that optimism correlates positively with pain threshold reductions. These findings suggest that the association between optimism and the function of endogenous pain modulatory systems is complex and differs for threshold-level and suprathreshold pain.


Calibration Marker Position in Digital Templating of Total Hip Arthroplasty.
We report a mathematical method to assess the vertical and horizontal positions of spherical radiopaque objects of known size in conventional radiographs. The reliability and validity of the method were tested in an experimental setting and applied to 100 anteroposterior pelvic radiographs with external calibration markers and unilateral total hip arthroplasty (THA). We found excellent reliabilities; intraclass correlation coefficients for interobserver and intraobserver reliabilities were 0.999-1.000 (P = .000). The mean normal height of THA was 198 mm (range: 142-243 mm, standard deviation: 18 mm) above the detector. Vertical and horizontal external marker positions differed significantly from the true hip center (THA; P < .001 and P = .017). This method could enhance patient safety by enabling automated detection of malpositioned calibration markers by templating software.


An Endogenously Tagged Fluorescent Fusion Protein Library in Mouse Embryonic Stem Cells.
Embryonic stem cells (ESCs), with their dual capacity to self-renew and differentiate, are commonly used to study differentiation, epigenetic regulation, lineage choices, and more. Using non-directed retroviral integration of a YFP/Cherry exon into mouse ESCs, we generated a library of over 200 endogenously tagged fluorescent fusion proteins and present several proof-of-concept applications of this library. We show the utility of this library to track proteins in living cells; screen for pluripotency-related factors; identify heterogeneously expressing proteins; measure the dynamics of endogenously labeled proteins; track proteins recruited to sites of DNA damage; pull down tagged fluorescent fusion proteins using anti-Cherry antibodies; and test for interaction partners. Thus, this library can be used in a variety of different directions, either exploiting the fluorescent tag for imaging-based techniques or utilizing the fluorescent fusion protein for biochemical pull-down assays, including immunoprecipitation, co-immunoprecipitation, chromatin immunoprecipitation, and more.


Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing--a continuous attempt at species and serovar differentiation.
Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize Leptospira spp. isolated from various hosts by genotyping and broth microdilution susceptibility testing in an attempt to differentiate Leptospira species, serogroups and serovars. Forty-seven isolates were studied. They were previously serotyped, and species confirmation was performed by 16S rRNA sequencing. Single-enzyme amplified fragment length polymorphism (SE-AFLP) and pulsed-field gel electrophoresis (PFGE) analysis enabled the distinction of L. interrogans from L. santarosai, L. meyeri and L. borgpetersenii in two main clusters. Among L. interrogans, it was possible to differentiate into two new clusters the serogroup Icterohaemorrhagiae from the serogroups Canicola and Pomona. L. santarosai isolates presented higher genetic variation than the other species in both techniques. Interestingly, the minimum inhibitory concentration (MIC) cluster analysis also provided Leptospira serogroup differentiation. Further studies are necessary regarding serovar Bananal isolates, as they presented the highest MIC values for most of the antimicrobials tested. All studied techniques successfully distinguished Leptospira species and serogroups. Despite being library-dependent methods, these approaches are less labor intensive and more economically viable, particularly SE-AFLP, and can be implemented in most reference laboratories worldwide to enable faster Leptospira typing.


Gene mapping with pooled samples on three genotyping platforms.
We have evaluated three genotyping techniques to detect allelic association between single nucleotide polymorphisms (SNPs) and disease loci by using pooled DNA samples. The use of DNA pools instead of individual samples saves considerably time, DNA quantity, and reagent costs. The methods that were compared were PCR-restriction fragment length polymorphism (PCR-RFLP), Single nucleotide primer extension, and chip-based mass spectrometry. As a model disease we studied an autosomal recessive disorder, congenital chloride diarrhea (CLD), caused by mutations in the human gene SLC26A3 on chromosome 7q31. Patient, carrier, and control pools were genotyped with eight SNP markers located within 10 cM around the target gene. Measured allele frequencies were consistent between the three platforms and we detected disease association with a cluster of several SNPs, the most significant being the marker located closest to the target gene (p=1.56 x 10(-16)). The largest source of error in pooled genotyping was shown to be the volumetric error in preparing the pools. The PCR-RFLP method was further evaluated by applying the pooling strategy to an ongoing multicenter-study. Measured allele frequencies of six polymorphisms agreed well with those measured individually and confirmed that multiple platforms are suitable for pooled genotyping.


Genotype-phenotype correlation in hepatocellular adenoma: new classification and relationship with HCC.
Hepatocellular adenomas are benign tumors that can be difficult to diagnose. To refine their classification, we performed a comprehensive analysis of their genetic, pathological, and clinical features. A multicentric series of 96 liver tumors with a firm or possible diagnosis of hepatocellular adenoma was reviewed by liver pathologists. In all cases, the genes coding for hepatocyte nuclear factor 1alpha (HNF1alpha) and beta-catenin were sequenced. No tumors were mutated in both HNF1alpha and beta-catenin enabling tumors to be classified into 3 groups, according to genotype. Tumors with HNF1alpha mutations formed the most important group of adenomas (44 cases). They were phenotypically characterized by marked steatosis (P < 10(-4)), lack of cytological abnormalities (P < 10(-6)), and no inflammatory infiltrates (P < 10(-4)). In contrast, the group of tumors defined by beta-catenin activation included 13 lesions with frequent cytological abnormalities and pseudo-glandular formation (P < 10(-5)). The third group of tumors without mutation was divided into two subgroups based on the presence of inflammatory infiltrates. The subgroup of tumors consisting of 17 inflammatory lesions, resembled telangiectatic focal nodular hyperplasias, with frequent cytological abnormalities (P = 10(-3)), ductular reaction (P < 10(-2)), and dystrophic vessels (P = .02). In this classification, hepatocellular carcinoma associated with adenoma or borderline lesions between carcinoma and adenoma is found in 46% of the beta-catenin-mutated tumors whereas they are never observed in inflammatory lesions and are rarely found in HNF1alpha mutated tumors (P = .004). In conclusion, the molecular and pathological classification of hepatocellular adenomas permits the identification of strong genotype-phenotype correlations and suggests that adenomas with beta-catenin activation have a higher risk of malignant transformation.


The DNA demethylating 5-azaC induces endoreduplication in cultured Chinese hamster cells.
We have investigated the possible influence of 5-azacytidine (5-azaC) substitution for cytidine into DNA on topoisomerase II (topo II) function in chromosome segregation. The endpoint chosen has been the induction of endoreduplicated cells at mitosis showing diplochromosomes. Experiments were performed in the presence and absence of the cytidine analogue to assess the degree of 5-azaC-induced DNA hypomethylation, using differential cutting by restriction endonucleases Hpa II and Msp I. Using the pulsed-field gel electrophoresis (PFGE) technique, we have also observed a protective effect provided by 5-azaC treatment against DNA breakage induced by the topo II poison m-AMSA. Concentrations of 5-azaC shown as able to induce extensive DNA hypomethylation and capable to protect DNA from double-strand breaks induced by m-AMSA were used for our cytogenetic experiments to analyze chromosome segregation. Our results seem to indicate that the presence of 5-azaC in DNA induces a dose-dependent increase in the yield of endoreduplicated cells that parallels the levels of hypomethylation observed.


Molecular Recognition in an Aqueous Medium Using Water-Soluble Prismarene Hosts.
The synthesis of water-soluble prism[n]arenes (n = 5 and 6) bearing anionic carboxylato groups on the rims is described. The binding properties of this novel class of water-soluble hosts are studied by nuclear magnetic resonance and calorimetry. The complexation of singly and doubly charged ammonium guests with the more rigid pentamer is enthalpically driven by secondary interactions, while the thermodynamic fingerprint for the larger hexamer reveals driving forces that strongly depend on the guest charge and/or size.


A new donor system for the patients with thalassemia: "Blood mother and blood father".
Donor recruitment programs differ in countries depending on local conditions and causes. Regularly voluntary blood donation rate should be 5% of the population but it is extremely low in Turkey. In 1998, "Thalassemia flowers don't fade" campaigning was started to get regular voluntary blood for patients with thalassemia. We would like to present results of our campaigning. The Thalassemia center was established in Antalya on the 1(st) June 1994 by Ministry of Health, Turkey, because the incidence of thalassemia is very high in the Antalya region. A total of 388 patients with thalassemia were followed up regularly in the center. The annually blood requirement was approximately 5000 units per year. In 1998, a new program of blood donation for patients with thalassemia called "BLOOD MOTHER and BLOOD FATHER" was started with the support of Governor of Antalya and health management system in Antalya. Between year 1998 and 2006, a total 3000 voluntary blood donors between age 18 and 65 years, of which 2160 males (72%) and 840 were females (28%), had participated in this program. "Blood Mother and Blood Father" campaign was successful donor recruitment program for thalassemic care. After 2006, this program is now adopted and run by Turkish Red Crescent and Thalassemia Federation of Turkey for all thalassemics in Turkey.


The postgenomic age: characterization of proteomes.
Global analysis of biological systems is becoming increasingly feasible as technologies that facilitate genome-wide analyses of gene expression are developed. Proteomics is the global analysis of expressed proteins (including posttranslational modifications) and seeks to establish the relationship between genome sequence, expressed proteins, protein-protein interactions, and cell and tissue phenotype. While the relative abundance of transcripts can be quantified using gene expression microarrays, the identification and quantitation of expressed proteins is more challenging. Nevertheless, the potential payoff for global protein analyses is immense because identification of distinctive protein signatures associated with cell function may provide novel therapeutic targets, molecular markers of disease, and increased understanding of determinants of cell phenotype. The challenges and promises of applications of established and emerging proteome strategies to detect and quantify differentially expressed proteins in culture cells are discussed.


Current indications for axillounifemoral and axillobifemoral bypass grafts.
Revascularization of the lower extremities may require an axillofemoral bypass when an aortobifemoral bypass is contraindicated. Thirty-one patients underwent axillounifemoral and 59 had an axillobifemoral bypass, with a mortality rate of 9%. The indication for operation was limb salvage in 67%, intra-abdominal sepsis in 21%, and disabling claudication in 12%. Cumulative survival, patency, and limb salvage rates were determined by life-table analysis. The cumulative patency and limb salvage rates (with standard errors) at 3 years were 68% +/- 8% and 78% +/- 9%, respectively. When stratified for type of operation, axillobifemoral bypass had a superior patency rate compared with axillounifemoral bypass (log rank = 3.882, p less than 0.05). There was no significant difference when patients were stratified for diabetes (log rank = 2.213, p = no significance [NS]), operative indication (disabling claudication vs. limb salvage) (log rank = 0.0005, p = NS), or outflow (no profundaplasty vs. profundaplasty) (log rank = 2.011, p = NS). We conclude that axillofemoral bypass is a reasonable alternative for revascularization in high-risk patients or in those patients in whom a transabdominal approach is contraindicated. We recommend aggressive use of the profunda femoris artery when the superficial femoral artery is occluded to achieve optimal results.


Pharmacological characterization and antidiabetic activity of a long-acting glucagon-like peptide-1 analogue conjugated to an antithrombin III-binding pentasaccharide.
To examine the biological characteristics of a novel glucagon-like peptide-1 (GLP-1) conjugate, in which an antithrombin III (ATIII)-binding pentasaccharide is conjugated to d-Ala(8) GLP-1 using a tetraethylene glycol linker. We assessed GLP-1 receptor binding, cAMP generation and insulin secretory activity of the GLP-1 conjugate in vitro. Circulating half-life, glucose homeostatic and subchronic therapeutic effectiveness were then examined in vivo. The half-life of the GLP-1 conjugate in mice was ∼11 h. In vitro insulin secretion from clonal β cells and islets was increased (p < 0.001) by the conjugate. The conjugate had half maximum effective concentration values of 1.3 × 10(-7) and 9.9 × 10(-8) M for displacement of (125) I-GLP-1 in competitive GLP-1 receptor binding and cAMP generation, respectively. Glucose tolerance in normal mice, immediately and 4 h after conjugate injection, resulted in significant (p < 0.001) improvements in blood glucose. These effects persisted for >48 h after administration. Daily treatment (21 days) of high-fat-fed and ob/ob mice with 25 nmol/kg conjugate resulted in significant improvement in glucose tolerance (p < 0.001) and reductions in glycated haemoglobin (HbA1c; p < 0.01) equivalent to or better than with exenatide or liraglutide. Treatment of C57BL/KsJ db/db mice for 15 days with 100 nmol/kg conjugate significantly (p < 0.001) reduced glucose and raised plasma insulin. Oral glucose tolerance was significantly (p < 0.001) improved and both 24-h glucose profile (p < 0.001) and HbA1c levels (p < 0.001) were reduced. Islet size (p < 0.001) and pancreatic insulin content were increased without change of islet cell proliferation or apoptosis. These data show that d-Ala(8) GLP-1(Lys(37) ) pentasaccharide exerts significant antidiabetic actions and has a projected pharmacokinetic/pharmacodynamic profile that merits further evaluation in humans for a possible once-weekly dosing regimen.