Integration of hospital and community care for paediatric ophthalmology: A mixed-methods study.
To explore collaborative care models for paediatric eye care that integrate hospital and community-based care to address access blocks. Sequential referrals to a tertiary paediatric ophthalmology clinic between April and October 2019 and subsequent encounters up until July 2020 at a major metropolitan public children's hospital in Sydney, Australia, were reviewed to identify those cases suitable for community care. Semi-structured phone interviews were conducted with eye health service providers, including ophthalmologists, orthoptists and optometrists, as well as service users to explore their perspectives on potential changes to service delivery. Qualitative data were analysed deductively using the Levesque model for access to healthcare and Consolidated Framework for Implementation Research (CFIR) to inform implementation strategies for future models of care. One-third of the 439 audited referrals (30.5%; 134/439) were identified as suitable for community management. Interviews revealed five themes relating to potential models of care, which would support and promote access: integrated health systems, standardised quality of care, interprofessional trust, multidisciplinary governance and patient-centred care. Key recommendations for future implementation included: (i) identifying and preparing clinical champions, (ii) conducting educational meetings, (iii) conducting local needs assessments and (iv) informing local opinion leaders. This audit highlights access blocks and poor targeting of referrals to tertiary paediatric ophthalmology services in a metropolitan hospital. Integration with community practitioners was identified as an acceptable way to streamline services, and strategies that may support successful implementation in this setting were identified.


The interplay of predictive and postdictive components of experienced selfhood.
Objects that we affect by our body movements can be experienced as being controlled by (agency) and belonging to the own body (ownership). Such impressions of minimal selfhood arise when objects move as predicted prior to the action (predictive component). But they can also arise when otherwise unpredictable object movements turn out to be consistent with (e.g. spatially compatible to) preceding actions (postdictive component). Here we studied how the impact of postdictive components of inferred minimal selfhood in terms of action-object compatibility is shaped by different levels of predictability of these object movements. We found that compatibility between actions and object movements, and to a lesser extent predictability of object movements, affected reported agency while only compatibility affected reported ownership. Importantly, predictive and postdictive factors influenced these measures in an independent manner. We discuss these results against the background of models that assume multiple components of experienced minimal selfhood.


Corticospinal tract transection prevents operantly conditioned H-reflex increase in rats.
Operant conditioning of the H-reflex, the electrical analog of the spinal stretch reflex, in freely moving rats is a relatively simple model for studying long-term supraspinal control over spinal cord function. Motivated by food reward, rats can gradually increase (i.e., up-condition) or decrease (i.e., down-condition) the soleus H-reflex. Earlier work showed that corticospinal tract transection prevents acquisition and maintenance of H-reflex down-conditioning while transection of other major spinal cord tracts does not. This study explores the effects on acquisition of up-conditioning of the right soleus H-reflex of mid-thoracic transection of: the right lateral column (LC, five rats) (containing the rubrospinal, vestibulospinal, and reticulospinal tracts); the entire dorsal column (DC, six rats) [containing the main corticospinal tract (CST) and the dorsal ascending tract (DA)]; the CST alone (five rats); or the DA alone (seven rats). After initial (i.e., control) H-reflex amplitude was determined, the rat was exposed for 50 days to the up-conditioning mode in which reward was given when the H-reflex was above a criterion value. H-reflex amplitude at the end of up-conditioning was compared to initial H-reflex amplitude. An increase > or =20% was defined as successful up-conditioning. In intact rats, H-reflex amplitude at the end of up-conditioning averaged 164% (+/-10%, SE), and 81% were successful. In the present study, LC and DA rats were similar to intact rats in final H-reflex amplitude and percent successful. In contrast, results for DC and CST rats were significantly different from those of intact rats. In the six DC rats, final H-reflex amplitude averaged 105% (+/-3)% of control and none was successful; and in the five CST rats, final H-reflex amplitude averaged 94% (+/-3)% and none was successful. The results indicate that the main CST, located in the dorsal column, is essential for H-reflex up-conditioning as it is for down-conditioning, while the dorsal column ascending tract and the ipsilateral lateral column (containing the main rubrospinal, vestibulospinal, and reticulospinal tracts) do not appear to be essential.


A combined strategy improves the solubility of aggregation-prone single-chain variable fragment antibodies.
Recombinant single-chain variable fragment (scFv) antibodies have wide applications in the areas of biotechnology and medicine. However, there is currently no universal expression-purification system for generating different soluble scFvs. In this study, A15 and E34, two genes coding scFvs against human IL-17A, were fused with N-terminal signal peptide sequences pelB or STII, or with highly hydrophilic tags Trx, NusA, or MBP, respectively. These constructs were expressed in Escherichia coli. We found that the scFvs fused with either NusA or MBP showed a higher solubility than fused with signal peptides or Trx. The scFvs were aggregated when the NusA or MBP was removed by thrombin. Interestingly, we observed a reduction of precipitation when the fusion proteins were expressed in Origami B(DE3)pLysS cells but not in BL21(DE3)pLysS. Because cleaving the tags resulted in the aggregation of scFvs, several solubility-enhancing additives were added in the digestion buffer and only L-arginine (Arg) or Tween20 promoted the solubility. After an affinity chromatography, the scFvs were separated from the tags with the purity up to 90%. The final yield of scFvs from the scFv-MBP system was approximately 8.9 mg/L of culture medium and 1.5 mg/g of wet weight cells, which was 1.6-fold higher than the yield from the scFv-NusA system. The obtained scFvs exhibited normal binding affinities and activities after endotoxin removal. In conclusion, we describe a strategy combining the fusion tags, the Escherichia coli with oxidizing bacterial cytoplasm, and the solubility-enhancing additives for expressing and purifying the soluble and functional scFvs.


Combined exposure to pyrene and fluoranthene and their molecular effects on the Sydney rock oyster, Saccostrea glomerata.
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitously detected in the water column, associated with particulate matter or in the tissue of marine organisms such as molluscs. PAH exposure and their resultant bioaccumulation in molluscs can cause a range of serious physiological effects in the affected animals. To examine the molecular response of these xenobiotics in bivalves, Sydney rock oysters (Saccostrea glomerata) were exposed to pyrene and fluoranthene for seven days. Chemical analysis of the soft-tissue of PAH stressed S. glomerata confirmed that pyrene and fluoranthene could be bioaccumulated by these oysters. RNA-Seq analysis of PAH-exposed S. glomerata showed a total of 765 transcripts differentially expressed between control and PAH-stressed oysters. Closer examination of the transcripts revealed a range genes encoding enzymes involved in PAH detoxification (e.g. cytochrome P450), innate immune responses (e.g. pathogen recognition, phagocytosis) and protein synthesis. Overall, pyrene and fluoranthene exposure appears to have resulted in a suppression of pathogen recognition and some protein synthesis processes, whereas transcripts of genes encoding proteins involved in clearance of cell debris and some transcripts of genes involved in PAH detoxification were induced in response to the stressors. Pyrene and fluoranthene exposure thus invoked a complex molecular response in S. glomerata, with results suggesting that oysters focus on removing the stressors from their system and dealing with the downstream effects of PAH exposure, potentially at the exclusion of other, less immediate concerns (e.g. protection from infection).


Characterizing the relationship between lesion-activation distance using fMRI and verbal measures in brain tumor patients.
Functional resonance magnetic imaging (fMRI) allows for identification of eloquent cortex in pre-treatment planning. Previous studies have shown a correlation among lesion to activation distance (LAD) measures and morbidity and mortality. This study investigates the relationship between LAD, well-established language centers (Wernicke's and Broca's), and language performance measures. We included a sample population of brain tumor patients that received language fMRI (verbal fluency and sentence verification) for pre-treatment assessment (n = 51). LAD to the nearest language area was measured and divided into groups ≤ 10 mm and > 10 mm. Verbal fluency scores were compared between these groups. Additionally, patients were divided into similar groups based on LAD to either Broca's or Wernicke's areas, and the verbal fluency scores and sentence verification accuracy (n = 29) were subsequently compared between groups. Brain tumor patients with LAD ≤ 10 mm to either language area had significantly lower verbal fluency scores (p = 0.028). The difference in verbal fluency scores between groups with LAD ≤ 10 mm and > 10 mm to Wernicke's area trends toward significance (p = 0.067). The sentence verification accuracy was significantly lower in patients with LAD ≤ 10 mm to either language area (p = 0.039). These findings suggest that there exists a significant relationship between LAD to language centers and measures; greater language deficits are seen when LAD ≤ 10 mm.


Understanding transcriptional regulatory networks using computational models.
Transcriptional regulatory networks (TRNs) encode instructions for animal development and physiological responses. Recent advances in genomic technologies and computational modeling have revolutionized our ability to construct models of TRNs. Here, we survey current computational methods for inferring TRN models using genome-scale data. We discuss their advantages and limitations. We summarize representative TRNs constructed using genome-scale data in both normal and disease development. We discuss lessons learned about the structure/function relationship of TRNs, based on examining various large-scale TRN models. Finally, we outline some open questions regarding TRNs, including how to improve model accuracy by integrating complementary data types, how to infer condition-specific TRNs, and how to compare TRNs across conditions and species in order to understand their structure/function relationship.


On the formation of circulating patterns of excitation in anisotropic excitable media.
We present a model of excitable media with the feature that it has a vulnerable phase during which a premature current stimulus will result in the formation of a reentrant self-sustained wave of excitation. The model exploits anisotropic coupling of identical cells, and is therefore useful as a model for the myocardium. We give rigorous verification that there is a vulnerable phase, and demonstrate numerically that permanently rotating waves are formed. Finally, it is shown that the direction of fastest propagation in myocardium is not necessarily the direction of highest safety factor, contrary to commonly accepted opinion.


The N-terminal cytokine binding domain of LIFR is required for CNTF binding and signaling.
Ciliary neurotrophic factor (CNTF) forms a functional receptor complex containing the CNTF receptor, gp130, and the leukemia inhibitory factor receptor (LIFR). However, the nature and stoichiometry of the receptor-mediated interactions in this complex have not yet been fully resolved. We show here that signaling by CNTF, but not by LIF or oncostatin M (OSM), was abolished in cells overexpressing a LIFR mutant with the N-terminal cytokine binding domain deleted. Our results illustrate molecular differences between the CNTF active receptor complex and those of LIF and OSM and provide further support for the hexameric model of the CNTF receptor complex.


Ultrastructure of bacteriophage phi 6: arrangement of the double-stranded RNA and envelope.
Pseudomonas phaseolicola cells synchronously infected with phi 6 were fixed and embedded by several procedures. The diameter of phi 6 measured 75 nm and its nucleocapsid 60 nm. Virus nucleocapsids were icosahedral and surrounded by a double membrane. In planar sections the nucleic acid appeared as a hexagonal ring and in cross section as two angular structures.


Parallel Doppler assessment of yolk sac and intervillous circulation in normal pregnancy and missed abortion.
This study assessed yolk sac morphology and vascularity and intervillous blood flow in normal early pregnancy and missed abortion. Transvaginal colour and pulsed Doppler were used in a prospective analysis of 87 normal pregnancies and 48 missed abortions between 6 and 12 weeks gestation. The Kruskal-Wallis rank test was used to calculate the difference in yolk sac diameter and vascularity visualization rate between gestational weeks. Repeated measures analysis of variance was used for comparison of the intervillous circulation between groups. The growth of the yolk sac was considered statistically significant between gestational weeks 6 and 9, being most prominent between 9 and 10 weeks of gestation. Vascularity of the yolk sac, characterized by low velocity and absence of diastolic flow, was demonstrated in 67 per cent of normal pregnancies. Yolk sac blood flow was detected in 19 per cent of the patients with missed abortion. Doppler analysis of the intervillous circulation demonstrated decreased peak velocity of the continuous flow in patients with missed abortion for gestational weeks 11 and 12. It is concluded that progressive decrease of yolk sac vascularity coincides with visualization of more prominent colour-coded areas within the intervillous space. In patients with missed abortion, such changes do not occur.


[Development and test of a new plantar pressure control device for foot unloading].
To validate a new plantar pressure control device able to detect excessive plantar pressure on-line and to improve the subject's awareness of the gait pattern in order to correct and optimize the load distribution patterns. The aim of this study was to verify the feasibility of a deliberate and partial unload of the first metatarsal head. Eight healthy subjects were equipped with the plantar pressure sensors integrated in customized insoles. They were told to unload the first metatarsal head of the right foot by 5%. During the trials, the auditory and visual signals inform the subject of an excessive, insufficient or correct unload. Five subjects over eight succeeding in modifying significantly their gait pattern and in unloading by 5% the first metatarsal head. The unload is effective in 70.7% of the trials. However, the subjects were spontaneously inclined to reduce peak pressure under the first metatarsal head beyond the critical peak plantar pressure threshold (48.2% of the trials). The results showed the feasibility of a partial and deliberate foot unloading in using biofeedback device but emphasized the subject's difficulty to control a discriminated and accurate unload of the first metatarsal head. That points out the necessity of learning period. The baropodometric biofeedback rehabilitation focuses on the subject's capacities to modify its own locomotor pattern. It would be used as primary and secondary prevention means of diabetic foot ulceration.


Performance of the AIDRScreening system in detecting diabetic retinopathy in the fundus photographs of Chinese patients: a prospective, multicenter, clinical study.
Diabetic retinopathy (DR) is the leading cause of blindness in the working-age population worldwide, and there is a large unmet need for DR screening in China. This observational, prospective, multicenter, gold standard-controlled study sought to evaluate the effectiveness and safety of the AIDRScreening system (v. 1.0), which is an artificial intelligence (AI)-enabled system that detects DR in the Chinese population based on fundus photographs. Participants with diabetes mellitus (DM) were recruited. Fundus photographs (field 1 and field 2) of 1 eye in each participant were graded by the AIDRScreening system (v. 1.0) to detect referable DR (RDR). The results were compared to those of the masked manual grading (gold standard) system by the Zhongshan Image Reading Center. The primary outcomes were the sensitivity and specificity of the AIDRScreening system in detecting RDR. The other outcomes evaluated included the system's diagnostic accuracy, positive predictive value, negative predictive value, diagnostic accuracy gain rate, and average diagnostic time gain rate. Among the 1,001 enrolled participants with DM, 962 (96.1%) were included in the final analyses. The participants had a median age of 60.61 years (range: 20.18-85.78 years), and 48.2% were men. The manual grading system detected RDR in 399 (41.48%) participants. The AIDRScreening system had a sensitivity of 86.72% (95% CI: 83.39-90.05%) and a specificity of 96.09% (95% CI: 94.14-97.54%) in the detection of RDR, and a false-positive rate of 3.91%. The diagnostic accuracy gain rate of the AIDRScreening system was 16.57% higher than that of the investigator, while the average diagnostic time gain rate was -37.32% lower. The automated AIDRScreening system can detect RDR with high accuracy, but cannot detect maculopathy. The implementation of the AIDRScreening system may increase the efficiency of DR screening.


Effects of the structure of classmates' perceptions of peers' academic abilities on children's perceived cognitive competence, peer acceptance, and engagement.
This study examined the effects of classroom indegree for ability (the degree to which peer nominations as academically capable show high consensus and focus on a relatively few number of children in a classroom) on first grade children's peer acceptance, teacher-rated classroom engagement, and self-perceived cognitive competence. Participants were 291 children located in 84 classrooms. Participating in sociometric interviews were 937 classmates. Consistent with social comparison theory, classroom indegree moderated the associations between children's achievement and classroom engagement and peer liking. Children with lower ability, relative to their classmates, were less accepted by peers and less engaged in classrooms in which students' perceptions of classmates' abilities converged on a relatively few number of students than in classrooms in which peers' perceptions were more dispersed. High indegree was associated with lower self-perceived cognitive competence regardless of ability level.


Differential analysis of phosphorylated proteins in resting and thrombin-stimulated human platelets.
Blood platelets are important components of haemostasis. After their activation they cause healing of wounds by forming plugs and initiate repair processes. One important event in regulating this activation is the phosphorylation/dephosphorylation of multiple proteins on various tyrosine, serine and threonine residues. To understand the exact molecular mechanisms in platelet activation it is essential to identify proteins involved in the signalling pathways and to localise and characterise their phosphorylation sites. After treatment with (32)P and separation by 2D-PAGE using different pI ranges, phosphorylated platelet proteins were detected by autoradiography. Phosphotyrosine-containing proteins were assigned by immunoblotting with an anti-phosphotyrosine antibody. Another approach for the identification of phosphorylated proteins was immunoprecipitation of tyrosine-phosphorylated proteins using an anti-phosphotyrosine antibody. Protein spots/bands of interest were excised from the gel, digested with trypsin and analysed by MALDI-TOF-MS and nano-LC-ESI-MS/MS, respectively. Several phosphorylated proteins could be identified and the localisation of some in vivo phosphorylation sites was possible.


Unexpected metachronous hernia development in children following laparoscopic unilateral hernia repair with negative evaluation for contralateral patent processus vaginalis.
Metachronous hernia has been reported to develop unexpectedly in children after negative evaluation for contralateral patent processus vaginalis (CPPV) by transinguinal laparoscopy. Scarce data exist regarding such phenomena following laparoscopic unilateral hernia repair and negative findings for CPPV in transumbilical laparoscopy. A retrospective study was performed to investigate metachronous hernia development in a cohort of consecutive children who had undergone laparoscopic unilateral hernia repair with negative findings of CPPV by transumbilical laparoscopy 5 or more years ago. Study subjects included 293 children 1 month to 15 years old at the time of laparoscopic unilateral hernia repair (left, n=116; right, n=177). There were 246 boys and 47 girls. Nine children (3.1%), all boys, developed metachronous hernia at a median of 24 months (range, 6-42 months) from the time of negative laparoscopic evaluation for CPPV. There was no statistical difference between the genders and laterality of the initial hernia in the development of metachronous hernia. False-negative CPPV evaluation by transumbilical laparoscopy during laparoscopic unilateral hernia repair can occur and result in unexpected metachronous hernia development. Further prospective studies are warranted to develop effective maneuvers in addition to inspection alone to reduce false-negative laparoscopic assessment.


Complement system deregulation in SAPHO syndrome revealed by proteomic profiling.
SAPHO syndrome is an inflammatory disease invading the skin and bones, whose diagnosis has been difficult due to its low incidence and diversified manifestation. We investigated the serum proteomic profile of SAPHO patients to identify key proteins associated with SAPHO syndrome, trying to find clinical biomarkers or functional molecules for this rare disease. Blood samples from 8 SAPHO patients and 8 healthy controls were detected and analyzed using data independent acquisition (DIA) method to identify differentially expressed proteins (DEPs) specific to SAPHO. A total of 57 differentially expressed proteins were identified (p < 0.05, fold change >1.2), in which 27 proteins were upregulated and 30 downregulated. DEPs may participate in GO terms such as "lipid particle" and "Notch signaling pathway", as well as KEGG pathways including "complement and coagulation cascades" and "mTOR signaling pathway". The overexpression of inhibitors of the complement system (CFH and C4BP), were verified in a larger cohort (16 SAPHO patients, 8 AS patients and 24 healthy controls) with ELISA, and the combined diagnostic ability of CFH and C4BP was predicted by ROC curve with an AUC of 0.91, which may be molecular candidates for further study on diagnosis and pathology of this rare disease. SIGNIFICANCE: Our research provided the first insight into plasma proteomic profile for SAPHO patients,offering potential biomarkers for disease diagnosis. We found that inhibitors of complement system such as CFH and C4BP were up-regulated in SAPHO syndrome, which may play important roles in the pathogenesis of SAPHO syndrome.


Health Risk Assessment for Human Exposure to Trace Metals Via Bushmeat in Ghana.
Consumption and trade in bushmeat are entrenched in sub-Saharan Africa amid growing land degradation and environmental pollution that raise safety concerns, yet sustainability of bushmeat extraction and zoonoses considerations have been the focus of bushmeat research. Here, we evaluated the health risk of trace metals in the skeletal muscle, bone, liver, and skin of the black duiker (Cephalophus niger), brush-tailed porcupine (Atherurus africanus), bushbuck (Tragelaphus scriptus), cane rat (Thryonomys swinderianus), and Maxwell's duiker (Cephalophus maxwelli) being the five most hunted and consumed species of bushmeat in Ghana. Sample tissues of the five species were analyzed for Cu, Zn, Se, Cd, As, Pb, and Hg. Targeted hazard quotient (THQ) of Se for black duiker, bushback, and Maxwell's duiker as well as that of Pb for black duiker, Brush-tailed porcupine, and Maxwell's duiker were concerning (> 1) for individuals foraging daily on bushmeat. Thus, cane rat was the only species that recorded both estimated daily intake (EDI) and THQ of Se and Pb within safe limits for daily consumption. In respect of Cu, Zn, Cd, As, and Hg, EDI and THQ values indicated no perceived risk to consumers for all five species. Lifetime cancer risk (CR) of As was negligible for all species. At the rate of bushmeat consumption in sub-Saharan Africa (0.104 kg per day), the cane rat was the only species considered safe for daily consumption among the five species.


Scanning electron microscopy of biosynthetic wound dressings Biocol.
The surface of wound dressing Biocol was studied by scanning electron microscopy. This composite system consists of latex matrix with incorporated water-soluble polysaccharide. The peculiarities of the surface are important for manufacturing of the dressing and for modification of its surface upon contact with fluids, e.g. during de novo tissue reconstruction. The method for studying the fine structure of the polymeric film surface was developed. The relief of the wound dressing changes during interaction with the fluid and nanopores appear on the surface. Thus, scanning electron microscopy is an informative method for studying the surface of biosynthetic films.


Naltrexone aversion and treatment efficacy are greatest in humans and rats that actively consume high levels of alcohol.
The opioid antagonist naltrexone is the standard pharmacotherapy for alcoholism, although compliance is often low. The mechanism by which naltrexone reduces drinking is yet unclear. Here we show that in active alcoholics the magnitude of naltrexone treatment efficacy is correlated with the level of naltrexone-induced aversive side effects. This correlation is not observed when subjects are sober, but emerges following alcohol administration, when subjects are intoxicated. In contrast, there is no correlation following placebo administration. To clarify these results, naltrexone was administered to ethanol-consuming rats prior to quantification of naltrexone aversion. Ethanol consumption preceding naltrexone treatment was again correlated with subsequent naltrexone-induced aversion, and this aversion correlated with subsequent decrease in ethanol consumption. In contrast, when naltrexone was given to ethanol-free rats, aversion was not predictive of ethanol consumption. We conclude that naltrexone treatment efficacy is greater during active ethanol consumption and may be partly due to aversive side effects.


Auxiliary screening COVID-19 by computed tomography.
The 2019 novel coronavirus (COVID-19) pandemic remains rampant in many countries/regions. Improving the positive detection rate of COVID-19 infection is an important measure for the control and prevention of this pandemic. This meta-analysis aims to systematically summarize the current characteristics of the computed tomography (CT) auxiliary screening methods for COVID-19 infection in the real world. Web of Science, Cochrane Library, Embase, PubMed, CNKI, and Wanfang databases were searched for relevant articles published prior to 1 September 2022. Data on specificity, sensitivity, positive/negative likelihood ratio, area under curve (AUC), and diagnostic odds ratio (dOR) were calculated purposefully. One hundred and fifteen studies were included with 51,500 participants in the meta-analysis. Among these studies, the pooled estimates for AUC of CT in confirmed cases, and CT in suspected cases to predict COVID-19 diagnosis were 0.76 and 0.85, respectively. The CT in confirmed cases dOR was 5.51 (95% CI: 3.78-8.02). The CT in suspected cases dOR was 13.12 (95% CI: 11.07-15.55). Our findings support that CT detection may be the main auxiliary screening method for COVID-19 infection in the real world.


Predictive power of progesterone receptor status in early breast carcinoma.
Progesterone receptor status (PgR) and oestrogen receptor status (ER) were examined in 332 women with stage I-II breast carcinoma treated at a single centre either by simple mastectomy with (n = 283) or without (n = 6) axillary clearance, or by segmental mastectomy with (n = 18) or without (n = 25) axillary clearance and radiotherapy; median follow-up time was 40 (range 10-60) months. Neither the concentration nor the presence of PgR related to either disease-free interval or survival. However both the presence and the concentration of ER were found to be associated with a longer disease-free interval amongst patients with stage II tumours (P less than 0.01). From this study it appears that progesterone receptor status does not predict outcome in early breast carcinoma.


Revealing the anticancer potential of candidate drugs in vivo using Caenorhabditis elegans mutant strains.
Drug repurposing is used as a strategy for finding new drugs for cancer. The process is a faster and a more cost-effective way of providing new indications for drugs that can address emerging drug resistance and numerous side effects of chemotherapeutic drugs. In this study, the in vivo anticancer potential of itraconazole, disulfiram, etodolac, and ouabain were assessed using five different C. elegans mutant strains. Each strain contains mutations in genes involved in different signaling pathways such as Wnt (JK3476), Notch (JK1107 and BS3164), and Ras-ERK (SD939 and MT2124) that result to phenotypes of sterility, infertility, and multivulva formation. These same signaling pathways have been shown to be defective in several human cancer types. The four candidate drugs were tested on the C. elegans mutant strains to determine if they rescue the mutant phenotypes. Both ouabain and etodolac significantly reduced the sterile and infertile phenotypes of JK3476, JK1107, BS3164, and SD939 strains (p=0.0010). Finally, itraconazole and etodolac significantly reduced multivulva formation (p=0.0021). The degrees of significant phenotypic rescues of each mutant were significantly higher than vehicle only (1% DMSO). Therefore, this study demonstrated that the four candidate drugs have anticancer potential in vivo, and etodolac had the highest anticancer potential.


Clinicopathological characteristics of non-diabetic renal disease in patients with type 2 diabetes mellitus in a northeastern Chinese medical center: a retrospective analysis of 273 cases.
To investigate the clinical and histopathological features of non-diabetic renal disease (NDRD) superimposed on diabetic nephropathy (DN) in northeastern Chinese patients with type 2 diabetes mellitus (T2D), and compare the changes with those of pure DN and isolated NDRD. Single-center retrospective analysis based on medical records of 273 patients (172 men, mean age: 51.1 ± 12.4 years) with T2D who underwent renal biopsy between February 2000 and October 2015. All patients were diagnosed as cases of pure DN, isolated NDRD or NDRD superimposed on DN. Out of the 273 T2D patients, 68 (24.9 %) had DN, 175 (64.1 %) had NDRD, and 30 (11.0 %) had NDRD superimposed on DN. Idiopathic membranous nephropathy (IMN, 29.7 %) was the most common NDRD followed by IgA nephropathy (IgAN, 22.9 %), and hypertensive renal arteriolar sclerosis was the most common lesion in patients diagnosed as NDRD superimposed on DN. Patients with NDRD had a shorter duration of diabetes and lower frequencies of diabetic retinopathy (DR, 6.9 %) and renal failure (28.0 %), which is consistent with higher estimated glomerular filtration rates (eGFR) and lower systolic blood pressure (SBP). No significant between-group differences were observed with respect to proteinuria and hematuria. Renal biopsy is strongly recommended for T2D patients to distinguish DN, NDRD and NDRD superimposed on DN, especially in patients with no signs of DR. This approach may help in early diagnosis and treatment of NDRD and improve renal outcomes in northeastern Chinese T2D patients.


Prevalence of renal failure and use of antithrombotic prophylaxis among medical inpatients at increased risk of venous thromboembolic events.
Evidence-based guidelines recommend the use of antithrombotic prophylaxis in medical patients at risk of venous thromboembolism (VTE). Low molecular weight heparins (LMWHs) are usually preferred to unfractionated heparin. However, when prophylactic doses of LMWH are administered, patients with renal failure (RF) are exposed to the risk of excessive accumulation, and thus to an increased risk of bleeding. We aimed to assess the prevalence of RF among medical inpatients at increased risk of VTE and the use and dosage of antithrombotic prophylaxis in these patients. In a cross sectional study carried out at three different hospitals, information on all medical inpatients was collected. Patients were defined at increased risk of VTE according to the American College of Chest Physicians guidelines. Data on the proportion of patients with renal RF, on the use and dosage of antithrombotic prophylaxis, and on the presence of contraindications to antithrombotic therapy were ascertained. 439 hospital charts were examined; 158 patients (36.0%) were defined at increased risk of VTE and had no contraindications to antithrombotic treatment. Thromboprophylaxis was prescribed to 61.4% of these patients. Eighty (50.7%) of them also had moderate or severe RF. There was no difference in the rate of prescription nor in the doses of antithrombotic prophylaxis between patients with and without RF (p=0.81 and p=0.94, respectively). RF is frequently present in medical patients at risk of VTE. A considerable proportion of these patients may not receive the optimal type or dose of antithrombotic prophylaxis.


Dissociation of proteinase-inhibitor complexes by trichloroacetate.
It was demonstrated that the addition of high concentrations of the chaotrope, sodium trichloroacetate, to proteinase assays provided for a dissociation of proteinase-inhibitor complexes. The complexes evaluated contained a heat-stable, polypeptide inhibitor of cysteine proteinases isolated from the cellular slime mold, Dictyostelium discoideum. The proteinases that were present in separate complexes included either D. discoideum proteinases or the plant proteinase papain. The general assay procedures described may be useful in detection of endogenous proteinase-inhibitor complexes in many systems.


Publisher Correction: Large Chinese land carbon sink estimated from atmospheric carbon dioxide data.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Aspects of matrix effects in applications of liquid chromatography-mass spectrometry to forensic and clinical toxicology--a review.
In the last decade, liquid chromatography coupled to (tandem) mass spectrometry (LC-MS(-MS)) has become a versatile technique with many routine applications in clinical and forensic toxicology. However, it is well-known that ionization in LC-MS(-MS) is prone to so-called matrix effects, i.e., alteration in response due to the presence of co-eluting compounds that may increase (ion enhancement) or reduce (ion suppression) ionization of the analyte. Since the first reports on such matrix effects, numerous papers have been published on this matter and the subject has been reviewed several times. However, none of the existing reviews has specifically addressed aspects of matrix effects of particular interest and relevance to clinical and forensic toxicology, for example matrix effects in methods for multi-analyte or systematic toxicological analysis or matrix effects in (alternative) matrices almost exclusively analyzed in clinical and forensic toxicology, for example meconium, hair, oral fluid, or decomposed samples in postmortem toxicology. This review article will therefore focus on these issues, critically discussing experiments and results of matrix effects in LC-MS(-MS) applications in clinical and forensic toxicology. Moreover, it provides guidance on performance of studies on matrix effects in LC-MS(-MS) procedures in systematic toxicological analysis and postmortem toxicology.


Five-Year Results of a Randomized Controlled Trial Comparing Patients Rehabilitated with Immediately Loaded Maxillary Cross-Arch Fixed Dental Prosthesis Supported by Four or Six Implants Placed Using Guided Surgery.
To compare the 5-year clinical and radiological outcomes of patients rehabilitated with four or six implants placed using guided surgery and immediate function concept. Forty patients randomly received four (All-on-4) or six (All-on-6) immediately loaded implants, placed using guided surgery, to support a cross-arch fixed dental prosthesis. Outcome measures were survival rates of implants and prostheses, complications, peri-implant marginal bone loss, and periodontal parameters. No drop-out occurred. Seven implants failed at the 5-year follow-up examination: six in the All-on-6 group (5%) and one in the All-on-4 group (1.25%), with no statistically significant differences (p = .246). No prosthetic failure occurred. Both group experienced some technical and biologic complications with no statistically significant differences between groups (p = .501). All-on-4 treatment concept demonstrated a trend of more complications during the entire follow-up period. A trend of more implant failure was experienced for the All-on-6 treatment concept. Marginal bone loss (MBL) from baseline to the 5-year follow-up was not statistically different between All-on-4 (1.71 ± 0.42 mm) and All-on-6 (1.51 ± 0.36 mm) groups (p = .12). For periodontal parameters, there were no differences between groups (p > .05). Both approaches may represent a predictable treatment option for the rehabilitation of complete edentulous patients in the medium term. Longer randomized controlled studies are needed to confirm these results.


Treating Parents to Reduce NICU Transmission of Staphylococcus aureus (TREAT PARENTS) trial: protocol of a multisite randomised, double-blind, placebo-controlled trial.
More than 33,000 healthcare-associated infections occur in neonatal intensive care units (NICUs) each year in the USA. Parents, rather than healthcare workers, may be a reservoir from which neonates acquire Staphylococcus aureus (S. aureus) colonisation in the NICU. This study looks to measure the effect of treating parents with short course intranasal mupirocin and topical chlorhexidine antisepsis on acquisition of S. aureus colonisation and infection in neonates. The TREAT PARENTS trial (Treating Parents to Reduce Neonatal Transmission of S. aureus) is a multicentre randomised, masked, placebo-controlled trial. Shortly after a neonate is admitted to the NICU, parents will be tested for S. aureus colonisation. If either parent screens positive for S. aureus, then both parents as a pair will be enrolled and randomised to one of the two possible masked treatment arms. Arm 1 will include assignment to intranasal 2% mupirocin plus topical antisepsis with chlorhexidine gluconate impregnated cloths for 5 days. Arm 2 will include assignment to placebo ointment and placebo cloths for skin antisepsis for 5 days. The primary outcome will be neonatal acquisition of an S. aureus strain that is concordant to the parental baseline S. aureus strain as determined by periodic surveillance cultures or a culture collected during routine clinical care that grows S. aureus. Secondary outcomes will include neonatal acquisition of S. aureus, neonatal S. aureus infection, eradication of S. aureus colonisation in parents, natural history of S. aureus colonisation in parents receiving placebo, adverse reactions to treatment, feasibility of intervention, and attitudes and behaviour in consented parents. Four hundred neonate-parent pairs will be enrolled. The study was approved by Johns Hopkins University IRB in June 2014 (IRB number 00092982). Protocol V.7 was approved in November 2014. Findings will be published in peer-reviewed journals. NCT02223520.


Recombinant N-terminal fragments of chromogranin-A modulate cardiac function of the Langendorff-perfused rat heart.
In this study we tested the hypothesis that vasostatins could act as myocardial modulators in the mammalian heart. Using the Langendorff-perfused rat heart, the cardiac effects of the two recombinant human CGA N-terminal fragments STA-CGA1-78 and STA-CGA1-115, containing the vasostatin-1 (CGA 1-76) and vasostatin-2 (CGA 1-113) sequences, respectively, were evaluated at concentrations of 11 / 165 nM. Cardiac performance was evaluated by analyzing left ventricular pressure (LVP) and the rate pressure product (RPP: HR x LVP), used as indexes of contractile activity and cardiac work, respectively. Under basal conditions, STA-CGA1-78 at all concentrations tested elicited a dose-dependent negative inotropism (LVP variations ranging from -9.6% +/- 2 to -23% +/- 2.9) without affecting coronary pressure (CP). In contrast, STA-CGA1-115 increased CP at 110 and 165 nM without affecting inotropism. Both STA-CGA1-78 and STA-CGA1-115 counteracted the cardio-stimulatory effects of isoproterenol (ISO). The ISO-dependent positive chronotropism was unaffected by STA-CGA1-78, while being reduced by STA-CGA1-115. Both peptides abolished the ISO-induced positive inotropism without modifying either the beta-adrenergic-dependent coronary dilation or the ouabain-induced positive inotropism. The analysis of the percentage of variations of RPP in terms of EC50 values of ISO alone (-8.5 +/- 0.3; r2 = 0.88) and in presence of STA-CGA1-78 (11, or 33, or 65 nM: -7.7 +/- 0.15, r2 = 0.97; -7.7 +/- 0.15, r2 = 0.97; -7.8 +/- 0.78, r2 = 0.55, respectively) revealed a non-competitive type of antagonism of STA-CGA1-78. Taken together, these data suggest vasostatins as novel cardioregulatory peptides in mammals.


Solution structure and calcium-binding properties of EF-hands 3 and 4 of calsenilin.
Calsenilin is a member of the recoverin branch of the EF-hand superfamily that is reported to interact with presenilins, regulate prodynorphin gene expression, modulate voltage-gated Kv4 potassium channel function, and bind to neurotoxins. Calsenilin is a Ca+2-binding protein and plays an important role in calcium signaling. Despite its importance in numerous neurological functions, the structure of this protein has not been reported. In the absence of Ca+2, the protein has limited spectral resolution that increases upon the addition of Ca+2. Here, we describe the three-dimensional solution structure of EF-hands 3 and 4 of calsenilin in the Ca+2-bound form. The Ca+2-bound structure consists of five alpha-helices and one two-stranded antiparallel beta-sheet. The long loop that connects EF hands 3 and 4 is highly disordered in solution. In addition to its structural effects, Ca+2 binding also increases the protein's propensity to dimerize. These changes in structure and oligomerization state induced upon Ca+2 binding may play important roles in molecular recognition during calcium signaling.


Quality of life after unilateral acoustic neuroma surgery via middle cranial fossa approach.
Patients with acoustic neuroma experienced reduced quality of life (QOL) after surgery. Individual factors did not have a significant effect on QOL. In the future, QOL should be a basic factor in the outcome evaluation of different therapeutic regimens in the treatment of acoustic neuroma. To measure the QOL of patients who underwent unilateral acoustic neuroma surgery via the middle cranial fossa approach. The Short Form-36 (SF-36) Health Survey and a self-designed disease-specific questionnaire were used during follow-up examinations to assess health-related QOL. The pure-tone average was used to specify hearing ability. Facial nerve function was described using the House-Brackmann grading system. A total of 28 male and 14 female patients who underwent surgery between 1997 and 2001 were included in the study. Patients' QOL scores revealed significant reductions in QOL in comparison to normative German QOL data. Gender, age, tumor size or location and clinical symptoms such as hearing loss and restricted facial nerve function did not have an effect on QOL. The SF-36 scales physical functioning, role functioning-physical, bodily pain, general health, social functioning and role functioning-emotional demonstrated significant QOL reductions.


Live-Cell Imaging of Protease Activity: Assays to Screen Therapeutic Approaches.
Methodologies to image and quantify the activity of proteolytic enzymes have been developed in an effort to identify protease-related druggable pathways that are involved in malignant progression of cancer. Our laboratory has pioneered techniques for functional live-cell imaging of protease activity in pathomimetic avatars for breast cancer. We analyze proteolysis in the context of proliferation and formation of structures by tumor cells in 3-D cultures over time (4D). In order to recapitulate the cellular composition and architecture of tumors in the pathomimetic avatars, we include other tumor-associated cells (e.g., fibroblasts, myoepithelial cells, microvascular endothelial cells). We also model noncellular aspects of the tumor microenvironment such as acidic pericellular pH. Use of pathomimetic avatars in concert with various types of imaging probes has allowed us to image, quantify, and follow the dynamics of proteolysis in the tumor microenvironment and to test interventions that impact directly or indirectly on proteolytic pathways. To facilitate use of the pathomimetic avatars for screening of therapeutic modalities, we have designed and fabricated custom 3D culture chambers with multiple wells that are either individual or connected by a channel to allow cells to migrate between wells. Optical glass microscope slides underneath an acrylic plate allow the cultures to be imaged with an inverted microscope. Fluid ports in the acrylic plate are at a level above the 3D cultures to allow introduction of culture media and test agents such as drugs into the wells and the harvesting of media conditioned by the cultures for immunochemical and biochemical analyses. We are using the pathomimetic avatars to identify druggable pathways, screen drug and natural product libraries and accelerate entry of validated drugs or natural products into clinical trials.


Industrial agglomeration and carbon neutrality in China: lessons and evidence.
This study explored the impact of industrial agglomeration on carbon neutrality and its spatial spillover effects utilizing the expanded the output density theoretical model of Ciccone and Hall. The main findings are as follows: (1) In terms of long-term effects, industrial output has significantly reduced regional carbon emissions, while industrial labor and capital agglomeration have aggravated carbon emissions, and industrial technology agglomeration has no significant impact on carbon emissions. (2) From the perspective of lagging effects, industrial output agglomeration has significantly increased regional carbon emissions, while industrial technology agglomeration has effectively reduced carbon emissions. The lagging effects of industrial labor and capital agglomeration are not significant. (3) From the perspective of regional differences, there are significant regional differences in the impact of industrial output and capital agglomeration on regional carbon emissions, while this differential influence does not exist on industrial labor and technological agglomeration. Therefore, in carbon emissions governance, it is necessary to analyze the long-term impact of industrial agglomeration on carbon emissions but also to pay attention to the lag and regional differences of the impact.


Testosterone stimulation of mitochondrial aspartate aminotransferase levels and biosynthesis in rat ventral prostate.
The effects of testosterone on mitochondrial aspartate aminotransferase (mAAT) synthesis in rat ventral prostate was investigated. Procedures for the isolation, purification and characterization of AAT isozymes were developed and described. Purified mAAT preparations contained no demonstrable contaminating proteins. Prostatic mAAT was characterized as a cationic protein with an estimated mol. wt of 120,000. Cytoplasmic AAT (cAAT) isozyme was identified as an anionic protein with an estimated mol. wt of 132,000. A cytosolic cationic isozyme, similar to mAAT, was also identified as pre-mAAT. Testosterone administration to castrated rats resulted in significant increases in leucine incorporation into mAAT, in the level of mAAT, and in mAAT activity. These effects of testosterone were observed within 2 h of administration. Conversely, testosterone administration had none of these effects on cAAT or on non-AAT protein pool. Testosterone treatment did appear to increase leucine incorporation into pre-mAAT. Testosterone treatment in organ cultures and in prostate epithelial cell cultures resulted in the same stimulatory effects on mAAT as observed in the in vivo studies. The hormone was effective at the physiological concentration of 2 X 10(-9) M. These results indicated that testosterone has a rapid and specific effect on the biosynthesis of mAAT. This continues to support our proposal that testosterone regulates prostate citrate production via a stimulatory effect on mAAT which results in increased mitochondrial synthesis of citrate from aspartate.


Further experience with laparoscopic nephrectomy for Wilms' tumour after chemotherapy.
To report the results of laparoscopic nephrectomy for unilateral Wilms' tumour in children treated with chemotherapy before surgery. Eight children with unilateral nonmetastatic Wilms' tumour included in the International Society of Pediatric Oncology 2001 protocol were treated with vincristine/actinomycin D and then had laparoscopic nephrectomy and lymph-node sampling. A Veress needle puncture was made and a four-trocar transperitoneal approach was used in all cases. The tumour was extracted with no morcellation through a Pfannenstiel incision. All eight tumours were completely removed, with lymph node samples; intraoperative bleeding was minimal (50 mL). There were no complications after surgery and patients were discharged after 2-3 days. No recurrences of disease, port-site implantation or long-term complications were detected. Laparoscopic nephrectomy for unilateral Wilms' tumour is feasible in children after chemotherapy; it is safe and allows the complete surgical approach required for treating this tumour. Although the patients had a good long-term follow-up, more patients are needed to compare the results of laparoscopic techniques with open surgery.


The Shu complex prevents mutagenesis and cytotoxicity of single-strand specific alkylation lesions.
Three-methyl cytosine (3meC) are toxic DNA lesions, blocking base pairing. Bacteria and humans express members of the AlkB enzymes family, which directly remove 3meC. However, other organisms, including budding yeast, lack this class of enzymes. It remains an unanswered evolutionary question as to how yeast repairs 3meC, particularly in single-stranded DNA. The yeast Shu complex, a conserved homologous recombination factor, aids in preventing replication-associated mutagenesis from DNA base damaging agents such as methyl methanesulfonate (MMS). We found that MMS-treated Shu complex-deficient cells exhibit a genome-wide increase in A:T and G:C substitutions mutations. The G:C substitutions displayed transcriptional and replicational asymmetries consistent with mutations resulting from 3meC. Ectopic expression of a human AlkB homolog in Shu-deficient yeast rescues MMS-induced growth defects and increased mutagenesis. Thus, our work identifies a novel homologous recombination-based mechanism mediated by the Shu complex for coping with alkylation adducts.


Finite element analysis of the hip and spine based on quantitative computed tomography.
Quantitative computed tomography (QCT) provides three-dimensional information about bone geometry and the spatial distribution of bone mineral. Images obtained with QCT can be used to create finite element models, which offer the ability to analyze bone strength and the distribution of mechanical stress and physical deformation. This approach can be used to investigate different mechanical loading scenarios (stance and fall configurations at the hip, for example) and to estimate whole bone strength and the relative mechanical contributions of the cortical and trabecular bone compartments. Finite element analyses based on QCT images of the hip and spine have been used to provide important insights into the biomechanical effects of factors such as age, sex, bone loss, pharmaceuticals, and mechanical loading at sites of high clinical importance. Thus, this analysis approach has become an important tool in the study of the etiology and treatment of osteoporosis at the hip and spine.


The prevalence and associated factors of metabolic syndrome in Chinese aging population.
Metabolic syndrome (MetS) is hitting high notes in the aging society in China. However, the prevalence and associated factors in Chinese aging population lack clarity to some extent. In the present study, we projected to inquire into the prevalence of MetS and its associated factors by analyzing datasets downloaded from the China Health and Retirement Longitudinal Study (CHARLS). Data comprising age, gender, socioeconomic status, lifestyle and health behaviors as well as blood biomarkers were subjected to descriptive statistics followed by univariate logistic regression and multivariate logistic regression. The overall prevalence of MetS was 33.38% (95% CI 32.42-34.34%). With age augments, prevalence increased during 40-70 years, while declined in participants aged 70 years above. Females had 2.94 times of risks (95% CI 2.55-3.39, P < 0.001). Marital status and alcohol consumption contributed nothing to the suffering of MetS. Participants with GDP per capita > 10,000 RMB and a non-agricultural hukou sustained higher risks than other participants (P < 0.05). Participants under education of middle school suffered 1.16 times of risks than other level of education (95% CI 1.01-1.34, P < 0.05). Smokers, participants with high low-density lipoprotein (LDL) or hyperuricemia or high glycosylated hemoglobin HbA1c sustained increased risks (P < 0.05). In Chinese aging population, with the augment of age, the prevalence ascended in men, while descended in women and was interfered by socioeconomic status, lifestyle and health behaviors as well as blood biomarkers, but not marital status and alcohol consumption.


"It's just more personal": Using multiple methods of qualitative data collection to facilitate participation in research focusing on sensitive subjects.
Often researchers use only one interview method within a single study. However, it is increasingly common for a variety of interview methods to be employed within a project. Providing choice to participants may facilitate recruitment, particularly when research focuses on a sensitive subject. This paper aims to explore participants' reasons for their choice of interview method in qualitative research focusing on a sensitive subject. Qualitative data was collected from 12 participants regarding their choice of interview method (face-to-face, Skype, telephone or email) in a wider study investigating parents' experiences following their child's burn-injury. Interview data was transcribed verbatim and thematic analysis was used to analyse responses. Participants most frequently chose to take part in an interview by telephone (n = 5), followed by email (n = 4), then Skype (n = 2), and face-to-face (n = 1). Four themes emerged suggesting that participants' decisions to participate via a particular method were determined by personal convenience, their belief in their ability to be open with the researcher despite potential upset caused by the topic, their ability to get a "feel" for the researcher, and concern about giving adequate depth in responses. Flexibility regarding the ways in which participants can take part in qualitative research may improve participant access to research, recruitment, and response-rate. Although, the depth and quantity of data gathered using different methods can vary. It is important that researchers consider the procedures and appropriateness of using different methods of interviewing and what impact such methods might have upon themselves and their participants.


Modulation of immune responses with monoclonal antibodies. I. Effects on regional lymph node morphology and on anti-hapten responses to haptenized monoclonal antibodies.
Repeated injections of monoclonal antibody (mAb) culture supernatants into rat footpads increased the weights of the draining lymph nodes. Immunostained freeze sections showed that injection of MRC OX2, a mAb reacting with rat follicular dendritic cells and MRC OX7 (anti-Thy-1.1), led to gross hypertrophy primarily of the follicular areas, whereas MRC OX6 (anti-rat major histocompatibility complex class II molecules) resulted in selective stimulation of the paracortex. These findings indicate that mAb, when conjugated to certain antigens, would modulate the immune response to these antigens. Consequently, the mAb were conjugated with fluorescein isothiocyanate (FITC) and the humoral response against the hapten measured. The primary anti-FITC antibody response was tenfold stronger than after stimulation with FITC conjugated to a conventional carrier such as ovalbumin, and had some characteristics of a secondary response: a fast increase of IgG level to very high titers and a long duration without further amplification at later antigen challenges.


O(6) -methylguanine-DNA methyltransferase (MGMT) promoter methylation and low MGMT-encoded protein expression as prognostic markers in glioblastoma patients treated with biodegradable carmustine wafer implants after initial surgery followed by radiotherapy with concomitant and adjuvant temozolomide.
O(6) -methylguanine-DNA methyltransferase (MGMT) promoter methylation status was proposed as a prognostic biomarker for patients with glioblastoma. However, the prognostic impact of MGMT in patients with newly diagnosed glioblastoma who receive carmustine-releasing wafers (Gliadel) along with temozolomide (TMZ) is still unknown. MGMT promoter methylation status and protein expression were analyzed in formalin-fixed, paraffin-embedded tumor specimens obtained from 111 French patients with newly diagnosed glioblastoma. Patients received the Gliadel wafers followed by radiotherapy plus concomitant and adjuvant TMZ chemotherapy while they were enrolled in a French multicenter prospective study. For the whole cohort, the median overall survival (OS) was 17.5 months, and the progression-free survival was 10.3 months. Patients with tumors that harbored MGMT methylation had a significantly longer OS compared with patients who had wild-type MGMT (21.7 months vs 15.1 months; P = .025). Similarly, patients who had low MGMT protein expression (≤15%) had a significantly improved OS compared with patients who had high MGMT expression (27.0 months vs 15.1 months; P = .021). The extent of resection was the strongest clinical predictor of outcome. In multivariate Cox models that were adjusted for sex, performance status, and extent of surgery, both MGMT methylation and protein expression were identified as independent prognosticators, and the finding was validated internally using a bootstrap resampling technique. Discrepancies were identified between protein expression and MGMT methylation status, thus suggesting that the 2 assays probably assess different biologic features. MGMT promoter methylation status and low MGMT expression both were identified as positive prognosticators in patients with newly diagnosed glioblastoma who underwent surgical resection and received Gliadel wafer implants followed by adjuvant radiotherapy and concomitant oral TMZ chemotherapy (the Stupp protocol).


Evaluation of the bioactive potential of foods fortified with fish protein hydrolysates.
Protein hydrolysates bioactivity makes them suitable for functional food ingredients. However, their chemical reactivity with different molecules, mainly reducing sugars, might lead to a modification of their structure, and thus function, after their inclusion into food matrices. The effect of food matrix and the storage time on the antihypertensive and antioxidant capacity of sardine hydrolysates was evaluated. Furthermore, bioactivity remaining after simulated gastrointestinal digested samples was also analyzed to verify its potential to be administrated orally. Sugar containing matrix improved the ACE inhibitory and DPPH radical scavenging activity of hydrolysates after thermal treatment and storage up to 80%. Maillard reaction products were responsible for the changes observed in the bioactivity of enriched food. Digested samples showed different but still adequate bioactivity depending on the matrix containing the hydrolysate. These results support the potential of fish protein hydrolysates to be used as ingredient in food formulation.


Fate of the rectum in patients undergoing total colectomy for Crohn's disease.
This study aimed to analyse disease recurrence and fate of the rectum in patients who had a total colectomy for Crohn's disease. One hundred and forty-four patients who had a total colectomy for Crohn's colitis were reviewed retrospectively. Ileorectal anastomosis (IRA) was performed in 118 patients, while 26 never had an IRA after colectomy because of severe anorectal lesions. Factors associated with recurrence and rectal preservation failure were studied. The probability of clinical recurrence after IRA was 58 and 83 per cent at 5 and 10 years respectively. The probability of rectal preservation at 5 and 10 years was 70 and 63 per cent after colectomy, and 86 and 86 per cent after IRA, respectively. Patients with extraintestinal manifestation had a higher risk of recurrence and of rectal preservation failure. Previous ileal involvement was associated with a higher rate of ileal recurrence after IRA. After IRA, prophylactic treatment with 5-aminosalicylic acid was associated with a lower rate of recurrence and of failure to preserve the rectum. Overall, 63 per cent of patients had a functioning IRA 10 years after total colectomy. Absence of extraintestinal manifestation and prophylactic treatment with 5-aminosalicylates after IRA were the main factors associated with long-term rectal preservation.


Mucocele of the paranasal sinuses. A retrospective clinical and histological study.
Clinical and radiological findings in 112 patients with muco/pyocele in the paranasal sinuses are presented. The modified Lynch-Howarth operation gave a primary cure rate of 82% which is satisfactory compared with other reports. Histological examination of the mucocele membranes and anamnestic information support the theory that a mucocele develops secondary to obstruction of the sinus outlet.


Fever and aging: central nervous system prostaglandin E2 in response to endotoxin.
The pathophysiology of the blunted febrile response often seen in elderly individuals with infection is not well understood. In this study, we attempted to determine the impact of aging on prostaglandin E2 release from the brain in response to endotoxin (LPS) stimulation. Eight young (4-6 month) and eight old (24-28 month) BALB/c mice were studied. Right and left half brains from old and young mice were either stimulated with LPS or control solution. Each mouse provided a stimulated and a control value. Results were reported for each mouse as the difference (stimulated minus control) in picograms of PGE2 released per milligram tissue and as the percent of baseline (control). Significant stimulation was demonstrated in the young mice, mean difference being +3.7 pg/mg, SD = 2.2 (Student's paired t, p less than 0.01) or +44% of control. In the old mice the mean difference was +2.9 pg/ml, SD = 6.7, or +22%, which was not statistically significant. Moreover, in three of eight old mice, there was a lack of PGE2 stimulation. The authors conclude that in a select group of old mice ("nonresponders"), the failure to mount a febrile response to an infection may be related to diminished release of PGE2 from the brain.


Rationale and design of EXPLORE: a randomized, prospective, multicenter trial investigating the impact of recanalization of a chronic total occlusion on left ventricular function in patients after primary percutaneous coronary intervention for acute ST-elevation myocardial infarction.
In the setting of primary percutaneous coronary intervention, patients with a chronic total occlusion in a non-infarct related artery were recently identified as a high-risk subgroup. It is unclear whether ST-elevation myocardial infarction patients with a chronic total occlusion in a non-infarct related artery should undergo additional percutaneous coronary intervention of the chronic total occlusion on top of optimal medical therapy shortly after primary percutaneous coronary intervention. Possible beneficial effects include reduction in adverse left ventricular remodeling and preservation of global left ventricular function and improved clinical outcome during future coronary events. The Evaluating Xience V and left ventricular function in Percutaneous coronary intervention on occLusiOns afteR ST-Elevation myocardial infarction (EXPLORE) trial is a randomized, prospective, multicenter, two-arm trial with blinded evaluation of endpoints. Three hundred patients after primary percutaneous coronary intervention for ST-elevation myocardial infarction with a chronic total occlusion in a non-infarct related artery are randomized to either elective percutaneous coronary intervention of the chronic total occlusion within seven days or standard medical treatment. When assigned to the invasive arm, an everolimus-eluting coronary stent is used. Primary endpoints are left ventricular ejection fraction and left ventricular end-diastolic volume assessed by cardiac Magnetic Resonance Imaging at four months. Clinical follow-up will continue until five years. The ongoing EXPLORE trial is the first randomized clinical trial powered to investigate whether recanalization of a chronic total occlusion in a non-infarct related artery after primary percutaneous coronary intervention for ST-elevation myocardial infarction results in a better preserved residual left ventricular ejection fraction, reduced end-diastolic volume and enhanced clinical outcome. trialregister.nl NTR1108.


Fetal cardiac dysfunction in pregnancies affected by intrahepatic cholestasis of pregnancy: A cohort study.
To analyze the presence of fetal myocardial dysfunction in intrahepatic cholestasis of pregnancy (ICP) at diagnosis. This prospective cohort study included 49 pregnant participants with ICP at diagnosis and 49 nonaffected controls from a single public hospital. ICP was diagnosed based on clinical symptoms after excluding other causes of pruritus and presence of autoimmune diseases. Total bile acids were not obtained in this cohort. ICP pregnancies were assessed with a functional echocardiography at diagnosis including PR-interval, isovolumetric contraction time (ICT), ejection time (ET), and isovolumetric relaxation time (IRT) for electrical, systolic, and diastolic function, respectively. Controls were assessed at recruitment. Perinatal outcomes were obtained from delivery books. The main outcome was the presence of PR-interval prolongation or first-degree fetal heart block, and echographic signs of diastolic and systolic dysfunction. Compared to controls, ICP were above upper limit in conjugated bilirubin (2.0% vs. 20.4%; p = 0.008), aspartate aminotransferase (2.0% vs. 24.5%; p = 0.002), and alanine aminotransferase (4.1% vs. 28.6%; p = 0.002). ICP was associated with a higher PR-interval (130 ± 12 ms vs. 121 ± 6 ms; p < 0.0001) with five first-degree fetal heart blocks. IRT was significantly higher in ICP (42 ± 6 ms vs. 37 ± 5 ms; p = 0.0001), with no differences in ICT and ET. PR-interval trend was only positively correlated with IRT in ICP pregnancies (p = 0.04 and p = 0.34, in ICP and controls, respectively). Our study demonstrates that fetuses affected by maternal ICP are associated with electrical and diastolic myocardial dysfunction. More studies focused on antenatal and postnatal functional echocardiography are necessary to validate our results and consider these markers in the clinical management of ICP pregnancies.


Lessons learned from catheter ablation of ventricular arrhythmias in patients with a fully magnetically levitated left ventricular assist device.
Data on catheter ablation of ventricular arrhythmias (VA) are scarce in patients with left ventricular assist devices (LVADs) and current evidence predominantly consists of case reports with outdated LVAD. This prospective observational study reports our experience in terms of catheter ablation of VAs in patients with novel 3rd generation LVADs. Between 2018 and 2020, nine consecutive patients undergoing a total number of ten ablation procedures for VAs were analyzed. The mean duration between LVAD implantation and catheter ablation was 23 ± 16 months. Acute procedural success was achieved in all patients. VA substrates were not related to the LVAD scarring (cannula) site in the majority of patients. All procedures were conducted without any relevant procedure-related complications. In terms of follow-up, only one patient presented with a repeat episode of electrical storm requiring ICD-shocks 16 months after the initial ablation procedure. Four patients suffered of singular VA effectively treated with antitachycardia pacing via their ICD. The remainder were free of any VA relapse (n = 4). Two non-procedure-related deaths occurred during follow-up. Catheter ablation of VAs in patients with 3rd generation LVAD is feasible and leads to satisfying clinical results in terms of freedom from VA recurrence and quality of life. The majority of arrhythmia substrates in these patients are not directly related to the LVAD cannulation site and may represent a progress of heart failure.


PerSubs: A Graph-Based Algorithm for the Identification of Perturbed Subpathways Caused by Complex Diseases.
In the era of Systems Biology and growing flow of omics experimental data from high throughput techniques, experimentalists are in need of more precise pathway-based tools to unravel the inherent complexity of diseases and biological processes. Subpathway-based approaches are the emerging generation of pathway-based analysis elucidating the biological mechanisms under the perspective of local topologies onto a complex pathway network. Towards this orientation, we developed PerSub, a graph-based algorithm which detects subpathways perturbed by a complex disease. The perturbations are imprinted through differentially expressed and co-expressed subpathways as recorded by RNA-seq experiments. Our novel algorithm is applied on data obtained from a real experimental study and the identified subpathways provide biological evidence for the brain aging.


[Ewing's sarcoma, fibrogenic tumors, giant cell tumor, hemangioma of bone : Radiology and pathology].
Radiological imaging only reflects the anatomy and its pathological abnormalities. Therefore, the radiologist should be able to recognize the basic features of the pathological anatomy of bone tumors. This can only be learned working closely with a pathologist who is experienced in this field. On the other hand, the pathologist needs from the radiologist their diagnostic assessment with information on size, location, aggressiveness and the existence of a bone tumor's matrix, of the whole lesion, because he usually only receives a small part for examination in the form of a biopsy. In this article, the features and fundamentals (standards) of radiological-pathological cooperation as the mainstay for a precise diagnosis in bone tumors are outlined. The radiological appearance and the histopathological features behind it are presented for Ewing's sarcoma, fibrogenic tumors, giant cell tumor, and hemangioma of the bone.


Zinc deficiency activates S100A8 inflammation in the absence of COX-2 and promotes murine oral-esophageal tumor progression.
Zinc (Zn)-deficiency (ZD) is implicated in the pathogenesis of human oral-esophageal cancers. Previously, we showed that in ZD mice genetic deletion of cyclooxygenase-2 (Cox-2) enhances N-nitrosomethylbenzylamine-induced forestomach carcinogenesis. By contrast, Cox-2 deletion offers protection in Zn-sufficient (ZS) mice. We hypothesize that ZD activates pathways insensitive to COX-2 inhibition, thereby promoting carcinogenesis. This hypothesis is tested in a Cox-2(-/-) mouse tongue cancer model that mimics pharmacologic blockade of COX-2 by firstly examining transcriptome profiles of forestomach mucosa from Cox-2(-/-) and wild-type mice on a ZD vs. ZS diet, and secondly investigating the roles of identified markers in mouse forestomach/tongue preneoplasia and carcinomas. In Cox-2(-/-) mice exposed to the tongue carcinogen 4-nitroquinoline 1-oxide, dietary ZD elicited tongue/esophagus/forestomach carcinomas that were prevented by ZS. The precancerous ZD:Cox-2(-/-) vs. ZS:Cox-2(-/-) forestomach had an inflammatory signature with upregulation of the proinflammation genes S100a8 and S100a9. Bioinformatics analysis revealed overrepresentation of inflammation processes comprising S100a8/a9 and an nuclear factor (NF)-κB network with connectivity to S100A8. Immunohistochemistry revealed co-overexpression of S100A8, its heterodimeric partner S100A9, the receptor for advanced glycation end-products (RAGE), NF-κB p65, and cyclin D1, in ZD:Cox-2(-/-) forestomach/tongue preneoplasia and carcinomas, evidence for the activation of a RAGE-S100A8/A9 inflammatory pathway. Accumulation of p53 in these carcinomas indicated activation of additional inflammatory pathways. Zn-replenishment in ZD:Cox-2(-/-) mice reversed the inflammation and inhibited carcinogenesis. Thus, ZD activates alternative inflammation-associated cancer pathways that fuel tumor progression and bypass the antitumor effect of Cox-2 ablation. These findings have important clinical implications, as combination cancer therapy that includes Zn may improve efficacy.


Viruses PLAY DEAD to TAMe interferon responses.
TAM receptors promote apoptotic cell uptake and function as inflammation suppressors. Many viruses mimic apoptotic cells, thus exploiting TAM receptors for attachment and entry. In this issue of Cell Host & Microbe, Bhattacharyya et al. show that TAM binding by enveloped viruses also induces receptor signaling to suppress cellular interferon responses.


Kinetic analysis of [18F] altanserin bolus injection in the canine brain using PET imaging.
Currently, [18F] altanserin is the most frequently used PET-radioligand for serotonin2A (5-HT2A) receptor imaging in the human brain but has never been validated in dogs. In vivo imaging of this receptor in the canine brain could improve diagnosis and therapy of several behavioural disorders in dogs. Furthermore, since dogs are considered as a valuable animal model for human psychiatric disorders, the ability to image this receptor in dogs could help to increase our understanding of the pathophysiology of these diseases. Therefore, five healthy laboratory beagles underwent a 90-min dynamic PET scan with arterial blood sampling after [18F] altanserin bolus injection. Compartmental modelling using metabolite corrected arterial input functions was compared with reference tissue modelling with the cerebellum as reference region. The distribution of [18F] altanserin in the canine brain corresponded well to the distribution of 5-HT2A receptors in human and rodent studies. The kinetics could be best described by a 2-Tissue compartment (2-TC) model. All reference tissue models were highly correlated with the 2-TC model, indicating compartmental modelling can be replaced by reference tissue models to avoid arterial blood sampling. This study demonstrates that [18F] altanserin PET is a reliable tool to visualize and quantify the 5-HT2A receptor in the canine brain.


Effects of an androgenic growth promoter 17beta-trenbolone on masculinization of Mosquitofish (Gambusia affinis affinis).
Endocrine disrupting chemicals can affect normal hormone dependent processes through numerous mechanisms, including ligand mimicky. 17beta-Trenbolone (TB), a pharmaceutical, androgenic, anabolic steroid, is a potent agonist of androgen receptors, and has been extensively used as a growth promoter for beef cattle in the US. The effects of TB on adult and newborn mosquitofish (Gambusia affinis affinis) were examined. Two forms of mosquitofish androgen receptor (AR), ARalpha and ARbeta, were cloned. The mRNA expression levels of ARalpha and ARbeta were transiently increased in the anal fin of adult females at day 3 following exposure to TB (1-10 microg/L) or methyltestosterone (MT) (0.1-10 microg/L), a pharmaceutical androgen used as a positive control. Gonopodium differentiation from the adult female anal fin was induced after 28 days of exposure to TB (1-10 microg/L) or MT (0.1-10 microg/L). Gonopodium differentiation also was induced in all mosquitofish fry exposed for 28 days to 0.3, 1 or 10 microg/L TB. Furthermore, spermatozoa were observed histologically in the testes of male fry exposed for 28 days to 1 or 10 microg/L TB; spermatozoa are normally observed only in the testes of mature males. Surprisingly, all female fry exposed for 28 days to 1 or 10 microg/L TB displayed the formation of an ovotestis, as spermatozoa were found in the ovary. Thus, TB, like MT, induced masculinization of the anal fin accompanied by a transient up-regulation of ARalpha and ARbeta in adult females. TB also induced differentiation of the anal fin into a gonopodium in fry of both sexes, stimulated precocious spermatogenesis in the testes of males and the formation of ovotestes in females.


High-throughput immunoaffinity enrichment and N-glycan analysis of human plasma haptoglobin.
Haptoglobin (Hp) is a positive acute phase protein, synthesized in the liver, with four N-glycosylation sites carrying mainly complex type N-glycans. Its glycosylation is altered in different types of diseases but still has not been extensively studied mainly due to analytical challenges, especially the lack of a fast, efficient, and robust high-throughput Hp isolation procedure. Here, we describe the development of a high-throughput method for Hp enrichment from human plasma, based on monolithic chromatographic support in immunoaffinity mode and downstream Hp N-glycome analysis by hydrophilic interaction ultrahigh-performance liquid chromatography with fluorescent detection (HILIC-UHPLC-FLR). Chromatographic monolithic supports in a 96-well format enable fast, efficient, and robust Hp enrichment directly from diluted plasma samples. The N-glycome analysis demonstrated that a degree of Hp deglycosylation differs depending on the conditions used for N-glycan release and on the specific glycosylation site, with Asn 241 being the most resistant to deglycosylation under tested conditions. HILIC-UHPLC-FLR analysis enables robust quantification of 28 individual chromatographic peaks, in which N-glycan compositions were determined by UHPLC coupled to electrospray ionization quadrupole time of flight mass spectrometry. The developed analytical approach enables fast evaluation of total Hp N-glycosylation and is applicable in large-scale studies.


Identification and cDNA cloning of a new nodule-specific gene, Nms-25 (nodulin-25) of Medicago sativa.
A new nodule-specific gene, Nms-25 (nodulin-25), was identified in cDNA clones isolated from a nodule-specific cDNA library of Medicago sativa. The first transcript of this gene appeared 9 days after inoculation of the roots with Rhizobium meliloti. The time of expression and the quantity of the transcripts of the Nms-25 gene was similar to that of leghemoglobin genes suggesting a similar regulation. A protein of 246 amino acids could be deduced from a full-length cDNA clone. The first 24 amino acids at the N-terminal end of this protein formed a signal sequence which might direct membrane transport into the peribacteroid space. Using different predictive methods the signal sequence cleaved protein was tentatively predicted to be a water-soluble enzyme, but not hydrolase.


Effects of Energy Drink Acute Assumption in Gastrointestinal Tract of Rats.
Energy drinks (EDs) are non-alcoholic beverages containing high amounts of caffeine and other psychoactive substances. EDs also contain herbal extract whose concentration is usually unknown. EDs can have several adverse effects on different organs and systems, but their effects on the gastrointestinal (GI) tract have been poorly investigated. To determine the acute effects of EDs on the GI tract, we administered EDs, coffee, soda cola, or water to Sprague-Dawley rats (n = 7 per group, randomly assigned) for up to five days, and analyzed the histopathological changes in the GI tract. Data were compared among groups by Kruskal-Wallis or Mann-Whitney tests. We found that, while EDs did not cause any evident acute lesion to the GI tract, they triggered eosinophilic infiltration in the intestinal mucosa; treatment with caffeine alone at the same doses found in EDs leads to the same effects, suggesting that it is caffeine and not other substances present in the EDs that causes this infiltration. The interruption of caffeine administration leads to the complete resolution of eosinophilic infiltration. As no systemic changes in pro-inflammatory or immunomodulating molecules were observed, our data suggest that caffeine present in ED can cause a local, transient inflammatory status that recruits eosinophils.


Heat fatalities in Pima County, Arizona.
The most common cause of heat fatalities is environmental exposure during heat waves. Deserts of the southwestern USA are known for temperatures that exceed 32 degrees C for 30 days or more; yet, heat-related fatalities are rare among residents of the region. We compiled data from the National Weather Service and the Office of the Medical Examiner in order to determine the relationship between temperature and occurrence of heat fatalities in Pima County, AZ. Logistic regression indicated that for each degree of increase in temperature (degrees C), there was a 35% increase in the odds of a heat fatality occurring (p<0.001).


Mutants of Chinese hamster ovary cells with altered glucose-6-phosphate dehydrogenase activity.
Two mutant clones of a Chinese hamster ovary cell line deficient in glucose-6-phosphate dehydrogenase (G6PD) activity have been characterized. In each case, there is evidence that a structural gene mutation has taken place. The first mutant produces 11% specific enzyme activity compared to wild-type parental cells, but this residual activity is much more heat sensitive than that of the wild type. The second mutant contains no residual activity, but a revertant was isolated that exhibits a partial restoration of G6PD activity with, again, an increased heat sensitivity. The selection of G6PD+ cells from G6PD- populations can be effected by exploiting the increased sensitivity of the latter to diamide, a compound that depletes the cell of reduced glutathione.


Radical scavenging activities of Tyr-, Trp-, Cys- and Met-Gly and their protective effects against AAPH-induced oxidative damage in human erythrocytes.
Radical scavenging activities of Tyr-, Trp-, Cys- and Met-Gly and their protective effects against AAPH-induced oxidative damage in erythrocytes were evaluated in this study. This damage includes hemolysis, oxidation of hemoglobin, formation of MDA and the depletion of glutathione (GSH) and catalase (CAT). Results showed that Tyr- and Trp-Gly could quench the radicals effectively in ABTS and ORAC assays with TE (Trolox equivalent) values of more than 1.0 μmol TE/μmol, followed by Cys- and Met-Gly. All these dipeptides could protect erythrocytes against AAPH-induced hemolysis in a dose-dependent manner. They could also significantly (p<0.05) retard the oxidation of hemoglobin and depletion of GSH in erythrocytes. The protective effects of these dipeptides decreased in the following order: Trp-Gly>Tyr-Gly>Met-Gly>Cys-Gly, which were consistent with their peroxyl radical scavenging activities. It suggested that these dipeptides might protect erythrocytes against AAPH-induced oxidative damage, mainly by acting as the direct radical scavengers.


Dosimetric comparison of VMAT standard optimization (SO) and multi-criteria optimization (MCO) treatment plans with standard mode delivery (STD) or sliding window (SW) for head and neck cancer.
A new development on the RayStation treatment planning system (TPS) allows a plan to be planned by imposing a constraint on the leaf sequencing: all leaves move in the same direction before moving again in the opposite direction to create a succession of sliding windows (SWs). The study aims to investigate this new leaf sequencing, coupled with standard optimization (SO) and multi-criteria optimization (MCO) and to compare it with the standard sequencing (STD). Sixty plans were replanned for 10 head and neck cancer patients (two dose levels simultaneously SIB, 56 and 70 Gy in 35 fractions). All plans were compared, and a Wilcoxon signed-rank test was performed. Pre-processing QA and metrics of multileaf collimator (MLC) complexity were studied. All methodologies met the dose requirements for the planning target volumes (PTVs) and organs at risk (OARs). SO demonstrates significantly best results for homogeneity index (HI), conformity index (CI), and target coverage (TC). SO-SW gives best results for PTVs (D98% and D2% ) but the differences between techniques are less than 1%. Only the D2%,PTV-56 Gy is higher with both MCO methods. MCO-STD offer the best sparing OARs (parotids, spinal cord, larynx, oral cavity). The gamma passing rates (GPRs) with 3%/3 mm criteria between the measured and calculated dose distributions are higher than 95%, slightly lowest with SW. The number of monitor units (MUs) and MLC metrics are higher in SW show a higher modulation. All plans are feasible for the treatment. A clear advantage of SO-SW is that the treatment plan is more straightforward to planning by the user due to the more advanced modulation. MCO stands out for its ease of use and will allow a less experienced user to offer a better plan than in SO. In addition, MCO-STD will reduce the dose to the OARs while maintaining good TC.


Long-term recording on multi-electrode array reveals degraded inhibitory connection in neuronal network development.
Spontaneous neuronal activity plays an important role in development. However, the mechanism that underlies the long-term spontaneous developmental change of cultured neuronal networks in vitro is not well understood. To investigate the contribution of inhibitory and excitatory connections to the development of neuronal networks, dissociated neurons from an embryonic rat hippocampal formation were cultured on a multi-electrode array plate and spontaneous activities were recorded by multi-channel system. These spontaneous activities were compared to bicuculline-induced firings, which were recorded by 60 electrodes simultaneously from 1 to 14 weeks in vitro (WIV). The phenomena showed that the spontaneous firing activities changed from an initial pattern of synchronized bursts to a later pattern of high frequency random spikes. The bicuculline-induced firing activities transformed from a pattern of synchronized bursts throughout all active sites in 3 WIV, to a pattern of local synchronized or random spikes appearing in the intervals of synchronized bursts after 11 WIV, while the firing rate hardly changed. Kynurenic acid, a broad-spectrum glutamate receptor antagonist, blocked all activities while CNQX inhibited only the local synchronized or random spikes. These suggest that the inhibitory connection was age-dependent degraded in vitro and the developmental spontaneous firing pattern was built by the homeostatic balance of the excitatory-inhibitory connection networks. Long-term cultures on MEA provided a useful tool to measure the relationship between spontaneous developmental change and pharmacological influence in vitro.


Quantification of ternary microplastic mixtures through an ultra-compact near-infrared spectrometer coupled with chemometric tools.
The ubiquitous distribution of plastics and microplastics (MPs) and their resistance to biological and chemical decay is adversely affecting the environment. MPs are considered as emerging contaminants of concern in all the compartments, including terrestrial, aquatic, and atmospheric environments. Efficient monitoring, detection, and removal technologies require reliable methods for a qualitative and quantitative analysis of MPs, considering point-of-need testing a new evolution and a great trend at the market level. In the last years, portable spectrometers have gained popularity thanks to the excellent capability for fast and on-site measurements. Ultra-compact spectrometers coupled with chemometric tools have shown great potential in the polymer analysis, showing promising applications in the environmental field. Nevertheless, systematic studies are still required, in particular for the identification and quantification of fragments at the microscale. This study demonstrates the proof-of-concept of a Miniaturized Near-Infrared (MicroNIR) spectrometer coupled with chemometrics for the quantitative analysis of ternary mixtures of MPs. Polymers were chosen representing the three most common polymers found in the environment (polypropylene, polyethene, and polystyrene). Daily used plastic items were mechanically fragmented at laboratory scale mimicking the environmental breakdown process and creating "true-to-life" MPs for the assessment of analytical methods for MPs identification and quantification. The chemical nature of samples before and after fragmentation was checked by Raman spectroscopy. Sixty three different mixtures were prepared: 42 for the training set and 21 for the test set. Blends were investigated by the MicroNIR spectrometer, and the dataset was analysed using Principal Component Analysis (PCA) and Partial Least Square (PLS) Regression. PCA score plot showed a samples distribution consistent with their composition. Quantitative analysis by PLS showed the great capability prediction of the polymer's percentage in the mixtures, with R2 greater than 0.9 for the three analytes and a low and comparable Root-Mean Square Error. In addition, the developed model was challenged with environmental weathered materials to validate the system with real plastic pollution. The findings show the feasibility of employing a portable tool in conjunction with chemometrics to quantify the most abundant forms of MPs found in the environment.


Influence of fatty chain length and starch composition on structure and properties of fully substituted fatty acid starch esters.
A series of almost fully substituted Fatty Acid Starch Esters (FASEs) has been obtained in a homogeneous LiCl/DMAc medium by grafting octanoyl (C8), lauroyl (C12) and palmitoyl (C16) chlorides onto 3 starch species: Amylo-Maize, Potato and Waxy Maize. Structure-property relationships of FASEs are investigated as a function of both fatty acid chain length and amylose/amylopectin ratio of the starch. The structural study has revealed a layered type organization in which starch chain planes are separated by fatty chains. The latter are interpenetrated and/or tilted for FASE-C16 whatever the origin of the starch is, and fatty chains partially crystallizes into a structure with hexagonal symmetry. FASEs with C8 and C12 side chains are totally amorphous. The mechanical behavior of FASEs is shown to depend on both side chain length and amylose/amylopectin ratio, and an increase in material ductility is observed at increasing amylose content for C8 and C12 side chains.


Radioimmunoassay of cholecystokinin in human tissue and plasma.
A highly sensitive radioimmunoassay for cholecystokinin (CCK) without any cross-reactivity with gastrin is described. The antibody was raised in a rabbit by immunisation with 30% CCK and bound to all COOH-terminal CCK-peptides containing at least 14 amino acid residues. The affinity constant of the antibody was 59.4 X 10(10) l/mol. CCK 33 conjugated to [125I]hydroxyphenylpropionic acid-succinimide ester was used as label. The binding between label and antibody was inhibited by 50% (ID50) at a concentration of 2.8 pmol/l cholecystokinin 33. The detection limit of the assay was between 0.5 and 1.0 pmol/l plasma. Concentrations of CCK in aqueous acid extracts of human upper small intestine were 36.5 +/- 9.8 pmol/g and of human cerebral cortex 28.2 +/- 2.5 pmol/g tissue. Plasma samples were extracted in 96% ethanol prior to assay. No advantage was obtained by adding aprotinin to the tubes. When frozen at -20 degrees C plasma CCK was stable for at least 6 months. Basal plasma CCK concentrations in 30 normal subjects were very low, 0.9 +/- 0.1 pmol/l, range 0.5 to 3.1 pmol/l. Intraduodenal administration of fat induced significant increases in plasma CCK from 1.1 +/- 0.1 to 8.2 +/- 1.3 pmol/l (p = 0.01). Infusion of exogenous CCK, resulting in plasma CCK levels slightly lower than those measured during administration of fat, induced pancreatic enzyme secretion and gallbladder contraction. The reliability of this radioimmunoassay for measurements of CCK in human plasma was extensively evaluated.


Processive Activity of Replicative DNA Polymerases in the Replisome of Live Eukaryotic Cells.
DNA replication is carried out by a multi-protein machine called the replisome. In Saccharomyces cerevisiae, the replisome is composed of over 30 different proteins arranged into multiple subassemblies, each performing distinct activities. Synchrony of these activities is required for efficient replication and preservation of genomic integrity. How this is achieved is particularly puzzling at the lagging strand, where current models of the replisome architecture propose turnover of the canonical lagging strand polymerase, Pol δ, at every cycle of Okazaki fragment synthesis. Here, we established single-molecule fluorescence microscopy protocols to study the binding kinetics of individual replisome subunits in live S. cerevisiae. Our results show long residence times for most subunits at the active replisome, supporting a model where all subassemblies bind tightly and work in a coordinated manner for extended periods, including Pol δ, redefining the architecture of the active eukaryotic replisome.


Synthesis of proto-oncogene proteins and cyclins depends on intact microfilaments.
It is well established that microfilament disintegration by cytochalasin D (CD) as well as latrunculin (LAT)-A and LAT-B causes an inhibition of S phase entry of various nontransformed cell lines. Our experiments extended these observations to human embryonal diploid fibroblasts (Wi-38). To investigate the question whether this stop of DNA synthesis is due to a decline of the synthesis of proteins that are necessary for G1 progression and S phase entry, we examined the expression of two proto-oncogenes (c-fos, c-jun) and three cyclins (D1, E, A) after altering the microfilament system. Disintegration of microfilaments by CD, LAT-A, or LAT-B of asynchronously growing fibroblasts caused a strong dose-dependent and time-dependent inhibition of total protein synthesis. Expression of c-jun, cyclins D1, E, and A decreased by about the same percentage as total protein synthesis. The strong induction of total protein synthesis after reactivating serum-starved fibroblasts by adding fetal calf serum was suppressed, when CD or LAT-A were added to the culture medium during this reactivation process. While expression of cyclin E as well as cyclin A decreased by about the same percentage as total protein synthesis, cyclin D1 was more suppressed after microfilament disintegration. After reactivating growth-arrested Wi-38 fibroblasts, cultured in suspension for 12 h, by transferring them to a rigid substratum they could adhere to, total protein synthesis was strongly induced. Again alteration of microfilaments by CD suppressed that increase. The expression of cyclin D1 was slightly more suppressed than total protein synthesis after addition of CD during that reactivation process. Our results suggest that alteration of microfilaments causes a strong decline of total protein synthesis accompanied by a decrease of the expression of proteins that are required for G1 progression and S phase entry. The diminished presence of proteins that are important for cell cycle progression could explain the inhibition of DNA synthesis after microfilament disintegration by various drugs.


An established kidney cell line from humpback grouper (Cromileptes altivelis) and its susceptibility to bacteria and heavy metals.
Humpback grouper (Cromileptes altivelis), one kind of commercial fish with considerable economic value, has been recognized as a promising candidate for mariculture. In the wake of the development of aquaculture industry, the breeding density of C. altivelis has increased gradually, which gave rise to the occurrence of various pathogenic diseases. In our research, we established a new kidney cell line (designated as CAK) from humpback grouper and evaluated its susceptibility to bacteria and heavy metals. The results of our study showed that the optimal growth temperature was 26 °C, and optimal medium was L-15 supplemented with 20% fetal bovine serum (FBS). The sequencing of 18S rRNA gene indicated that CAK cell line was derived from C. altivelis. Chromosome analysis showed that the number of chromosome in CAK was 48. After being transfected of pEGFP-N3 plasmid, high transfection efficiency of CAK was observed, suggesting the potential to be used for the study of foreign functional genes. Moreover, the bacterial susceptibility results revealed that CAK cells were sensitive to Vibrio harveyi and Edwardsiella tarda, especially V. harveyi. Meanwhile, three heavy metals (Hg, Cu, and Cd) had toxic effects on the CAK cells with a dose-dependent manner. To sum up, the CAK cell line might be an ideal tool in vitro for analyzing the function of exogenous genes, bacterial susceptibility, and toxicity assay of heavy metals.