A light microscopic and ultrastructural examination of calcified dental tissues of horses: 3. Dentine.
Ultrastructural examinations of equine dentine found that dentinal tubules extended from the amelodentinal junction towards the pulp forming primary curvatures. The number of dentinal tubules/unit area and their diameters increased significantly from the amelodentinal junction towards the pulp cavities, particularly in regular secondary dentine, but irregular secondary (tertiary) dentine contained no dentinal tubules. Dentinal tubules contained odontoblast processes that appeared to extend as far as the amelodentinal junction, but due to iatrogenic loss during specimen preparation, odontoblasts were seldom found in regular secondary dentine. In primary dentine, the dentinal tubules were surrounded by large amounts of peritubular dentine that increased in diameter from the amelodentinal junction towards the junction of primary and secondary dentine. The site of the dentinal tubule within the peritubular dentine varied at different dentinal sites. Peritubular dentine was present in primary dentine only and was surrounded by a thin layer of intertubular dentine. When acid etched, peritubular dentine gave dentine a honeycomb appearance adjacent to the junction of primary and secondary dentine.


Type 2 Diabetes and the Use of Real-Time Continuous Glucose Monitoring.
The role of continuous glucose monitoring (CGM) in type 1 diabetes (T1D) is well established in improving glycemic control and reducing hypoglycemia. Type 2 diabetes (T2D) is more prevalent than T1D and management of T2D is more heterogeneous, requiring treatment ranging from lifestyle modification to oral medications to intensive insulin therapy. Recent randomized controlled trials in intensively insulin-treated T2D demonstrated the efficacy and safety of real-time CGM (rtCGM) in reducing glycated hemoglobin without increasing hypoglycemia. Although evidence is limited, early studies have indicated a role for rtCGM in selected patients with non-insulin requiring T2D to improve glycemic control and/or reduce hypoglycemia. Based on literature review, we summarized current data on the use of rtCGM in T2D management and provided future research direction to generate more evidence on the utility of CGM in this population.


Intraocular Injection of StivantⓇ (A Biosimilar to Bevacizumab): A Case Series.
To report the results of intravitreal injection of a bevacizumab biosimilar called StivantⓇ. This prospective interventional case series was conducted on eyes with neovascular age-related macular degeneration (nAMD), retinal vein occlusion (RVO), and diabetic macular edema (DME). StivantⓇ was injected in three consecutive months and changes in best-corrected visual acuity (BCVA) and central macular thickness (CMT) were measured at baseline and monthly up to one month after the third injection. Three hundred and eighty-five eyes with DME (234 eyes, 61%), nAMD (87 eyes, 22%), and macular edema secondary to RVO (64 eyes, 17%) were enrolled. The mean ± standard deviation age of the patients was 61.7 ± 7.20 years. The mean BCVA and CMT changed from 0.63 ± 0.3 to 0.51 ± 0.3 LogMAR (P = 0.12 ) and from 420.4 ± 47.3μm at baseline to 316.7 ± 50.6 μm (P < 0.001) in the DME group; from 0.79 ± 0.3 to 0.68 ± 0.3 LogMAR (P = 0.19) and from 376.1 ± 31.7 μm to 303 ± 31.3 μm (P = 0.019) in the nAMD group; and from 0.81 ± 0.4 to 0.63 ± 0.4 LogMAR (P = 0.05) and from 424.21 ± 18 μm to 303.4 ± 18.8 μm (P < 0.001) in the RVO group, respectively. Our limited experience showed that the intravitreal injection of StivantⓇ was well tolerated. Although the results of this case series showed relative improvement in CMT one month after the last injection of StivantⓇ, BCVA improvement was statistically significant only in the RVO group. This would be essential to design a randomized clinical trial to evaluate the non-inferiority of StivantⓇ in comparison to bevacizumab.


SOCS3 Gene Polymorphism and Hypertension Susceptibility in Chinese Population: A Two-Center Case-Control Study.
Endothelial inflammation and vascular damage are essential risk factors contributing to hypertension. Suppressor of cytokine signaling 3 (SOCS3) is involved in the regulation of multiple inflammatory pathways. A large number of studies have shown that the anti-inflammatory effect of SOCS3 in hypertension, obesity, and allergic reactions has brought more insights into the inhibition of inflammation. Therefore, we selected a tagSNP of SOCS3 (rs8064821) to investigate whether they are contributing to the risk of hypertension in the Chinese population. In total, 532 patients with hypertension and 569 healthy controls were enrolled for two central of China. SOCS3 rs8064821 C>A polymorphism was genotyped using TaqMan assay. SOCS3 rs8064821 CA genotype was associated with an increased risk of hypertension (OR = 1.821, 95%CI = 1.276-2.600, P = 0.001). Rs8064821 A allele was associated with higher SOCS3 mRNA level in PBMCs from healthy donors. SOCS3 rs8064821 C>A polymorphism may contribute to the risk of hypertension in the Chinese population by regulating the expression of SOCS3.


The residues 4 to 6 at the N-terminus in particular modulate fibril propagation of β-microglobulin.
The ΔN6 truncation is the main posttranslational modification of β-microglobulin (βM) found in dialysis-related amyloid. Investigation of the interaction of wild-type (WT) βM with N-terminally truncated variants is therefore of medical relevance. However, it is unclear which residues among the six residues at the N-terminus are crucial to the interactions and the modulation of amyloid fibril propagation of βM. We herein analyzed homo- and heterotypic seeding of amyloid fibrils of WT human βM and its N-terminally-truncated variants ΔN1 to ΔN6, lacking up to six residues at the N-terminus. At acidic pH 2.5, we produced amyloid fibrils from recombinant, WT βM and its six truncated variants, and found that ΔN6 βM fibrils exhibit a significantly lower conformational stability than WT βM fibrils. Importantly, under more physiological conditions (pH 6.2), we assembled amyloid fibrils only from recombinant, ΔN4, ΔN5, and ΔN6 βM but not from WT βM and its three truncated variants ΔN1 to ΔN3. Notably, the removal of the six, five or four residues at the N-terminus leads to enhanced fibril formation, and homo- and heterotypic seeding of ΔN6 fibrils strongly promotes amyloid fibril formation of WT βM and its six truncated variants, including at more physiological pH 6.2. Collectively, these results demonstrated that the residues 4 to 6 at the N-terminus particularly modulate amyloid fibril propagation of βM and the interactions of WT βM with N-terminally truncated variants, potentially indicating the direct relevance to the involvement of the protein's aggregation in dialysis-related amyloidosis.


Effects of autogenic and reciprocal inhibition muscle energy techniques on isometric muscle strength in neck pain: A randomized controlled trial.
Neck is one of the most common sites of musculoskeletal symptoms, and muscle shortening and weakness is observed to be a common cause of neck pain and disability. To compare the immediate and short term effects of static stretching (SS), autogenic inhibition (AI) and reciprocal inhibition (RI) muscle energy techniques (MET) on isometric muscle strength in the management of mechanical neck pain. A randomized controlled trial was conducted on 78 participants with neck pain randomly allocated to SS, AI-MET and RI-MET groups. All the participants received Trans Cutaneous Electrical Nerve Stimulation (TENS), hot pack and unilateral postero-anterior glide, followed by 3-5 repetitions of either SS, AI-MET or RI-MET for five consecutive sessions. Numeric pain rating scale (NPRS) and Modified Sphygmomanometer Dynamometry (MSD) were used as outcome measurement tools. One way ANOVA and repeated measures ANOVA were used for inter-group and intra-group comparison. In terms of MSD scores, a significant difference (p< 0.05) was observed between the groups. Both AI-MET and RI-MET were found to be comparatively more effective than SS, however AI-MET was found to be the most effective. AI-MET is more effective than SS and RI-MET in terms of improving isometric muscle strength in patients with mechanical neck pain.


Protective effect of valproic acid in streptozotocin-induced sporadic Alzheimer's disease mouse model: possible involvement of the cholinergic system.
Sporadic Alzheimer's disease (SAD) is a slowly progressive neurological disorder that is the most common form of dementia. Cholinergic system dysfunction and amyloid beta formation are the two main underlying pathological mechanisms for the disease development. In recent studies, insulin receptor desensitization and disturbances in the downstream effects of insulin receptor signaling were observed in the brains of Alzheimer's patients. Currently, intracereberoventricular (ICV) injection of streptozotocin (STZ) is found to induce behavioral, neurochemical, and structural alterations in animals resembling those found in SAD patients. Valproic acid (VPA), a histone deacetylase inhibitor (HDACi), was recently shown to regulate the transcription of several genes in both in vivo and in vitro models of Alzheimer's disease. The aim of the current study is to investigate the potential effect of different doses of valproic acid, in an ICV-STZ-induced animal model of SAD. Streptozotocin-injected mice showed cognitive and spatial memory dysfunction in the Y-maze, object recognition test, and Morris water maze (MWM) neurobehavioral tests. The mice also exhibited a decrease in acetylcholine (ACh) and neprilysin (NEP) levels accompanied by an increase in acetylcholinesterase (AChE) activity. For the first time to our knowledge, our findings have shown that VPA is capable of restoring ACh levels in ICV-STZ-injected mice, as well as normalizing both NEP levels and AChE activity. Via this mechanism, an enhancement of cognitive functions is observed. Thus, VPA is suggested to be a promising therapeutic approach against SAD.


Repetitive Transcranial Magnetic Stimulation for Improving Cognitive Function in Patients With Mild Cognitive Impairment: A Systematic Review.
Background: Mild cognitive impairment (MCI) is an early stage of Alzheimer's disease. Repetitive transcranial magnetic stimulation (rTMS) has been widely employed in MCI research. However, there is no reliable systematic evidence regarding the effects of rTMS on MCI. The aim of this review was to evaluate the efficacy and safety of rTMS in the treatment of MCI. Methods: A comprehensive literature search of nine electronic databases was performed to identify articles published in English or Chinese before June 20, 2019. The identified articles were screened, data were extracted, and the methodological quality of the included trials was assessed. The meta-analysis was performed using the RevMan 5.3 software. We used the GRADE approach to rate the quality of the evidence. Results: Nine studies comprising 369 patients were included. The meta-analysis showed that rTMS may significantly improve global cognitive function (standardized mean difference [SMD] 2.09, 95% confidence interval [CI] 0.94 to 3.24, p = 0.0004, seven studies, n = 296; low-quality evidence) and memory (SMD 0.44, 95% CI 0.16 to 0.72, p = 0.002, six studies, n = 204; moderate-quality evidence). However, there was no significant improvement in executive function and attention (p > 0.05). Subgroup analyses revealed the following: (1) rTMS targeting the left hemisphere significantly enhanced global cognitive function, while rTMS targeting the bilateral hemispheres significantly enhanced global cognitive function and memory; (2) high-frequency rTMS significantly enhanced global cognitive function and memory; and (3) a high number of treatments ≥20 times could improve global cognitive function and memory. There was no significant difference in dropout rate (p > 0.05) between the rTMS and control groups. However, patients who received rTMS had a higher rate of mild adverse effects (risk ratio 2.03, 95% CI 1.16 to 3.52, p = 0.01, seven studies, n = 317; moderate-quality evidence). Conclusions: rTMS appears to improve global cognitive function and memory in patients with MCI and may have good acceptability and mild adverse effects. Nevertheless, these results should be interpreted cautiously due to the relatively small number of trials, particularly for low-frequency rTMS.


The Changing Landscape in Supporting Preterm Infants at Birth.
Noninvasive ventilation for preterm infants at birth has been recommended and universally adopted. The umbilical cord is often clamped immediately in order to provide the support the infant needs for stabilization. However, recent scientific data from experimental studies that involve animals in transition and human studies using physiological measurements at birth have increased awareness as to how little we know about how these interventions interact and integrate with the infant's changing physiology. It has become clear that in apneic infants the larynx is closed immediately after birth, which can completely negate the effect of noninvasive ventilation of the lung. For this reason, stimulating and supporting spontaneous breathing could enhance the success of noninvasive ventilation. Animal data also demonstrated that the large swings in blood pressure, blood flow, and oxygenation caused by immediate cord clamping can be avoided by postponing cord clamping until lung aeration has been established. In this review we will focus on these "game changers" that have the potential to completely change the approach used in stabilizing preterm infants at birth.


On the feasibility of absolute 3D dosimetry using LiF thermoluminescence detectors and polymer gels on a 0.35T MR-LINAC.
As shown in our previous study, highly accurate absolute dosimetry in 3D is feasible by combining polymer gels (PG) with thermoluminescence dosimetry (TLD). In this setup, the thermoluminescence (TL)-based point dose information is used to renormalize the PG. This new PG-TLD reference system is now extended to measurements in magnetic fields. Experiments were carried out on a conventional 6 MV linear accelerator (LINAC) and a 6 MV 0.35 T magnetic resonance (MR)-LINAC. Signal stability of TLD600 and TLD700 was examined without and with magnetic field. Afterwards, the combination of PAGAT PG and TL detectors was employed within a cylindrical phantom in presence of the magnetic field. Two scenarios were tested: (I) an air-filled phantom and (II) a water-filled phantom. For each scenario, two plans were irradiated: (a) opposed beams with a field size of 10 × 10 cm2 and (b) a 3D conformal plan assuring homogeneous target coverage using three equally distributed coplanar beams. Mean relative uncertainty of TL calibration reproducibility for TLD600/TLD700 was 0.49%/0.85% at the MR-LINAC and 0.48%/0.83% for the conventional LINAC. Individual TL calibration coefficients of TLD600 and TLD700 behaved differently in the presence of the magnetic field. An average difference of (3.29 ± 0.89)% occurred for all TLD600, whereas the result for TLD700 is not quite as clear with (1.09 ± 0.89)% after excluding some outliers. Using the TL dose information for PG renormalization, high 3D gamma passing rates were achieved using the 3%/2 mm criteria: 91.0% (Ia), 92.6% (Ib), 94.3% (IIa), 97.4% (IIb). This study shows that TL signal reproducibility is not affected by a low magnetic field. Nevertheless, absolute calibration coefficients of the individual detectors indicate a dependency on the magnetic field. Hence, a calibration at the appropriate LINAC type is recommended. Furthermore, the previously established renormalization method for PG was applied to measurements at a MR-LINAC and was verified as suitable for evaluations of homogeneous dose distribution in the target volume.


Effect of methyl jasmonate in combination with ethanol treatment on postharvest decay and antioxidant capacity in Chinese bayberries.
The effects of methyl jasmonate (MeJA) in combination with ethanol (EtOH) treatment on green mold rot caused by Penicillium citrinum , natural decay, and antioxidant capacity in harvested Chinese bayberries were investigated. MeJA at 10 micromol L(-1) in combination with EtOH at 22.32 micromol L(-1) was most effective in reducing green mold rot and natural decay. This combined treatment also significantly inhibited spore germination and germ tube elongation of the pathogen in vitro compared with MeJA alone or control. Meanwhile, the combination-treated bayberries exhibited highest reducing power, scavenging activities against superoxide, hydroxyl, and 1,1-diphenyl-2-picrylhydrazyl radicals, and contents of total phenolics, flavonoids, and anthocyanins as well as individual phenolic compounds. Fruit quality parameters were not significantly affected by these treatments. These results suggest that the combination of MeJA and EtOH had an additive effect in reducing postharvest decay and improving antioxidant capacity in Chinese bayberries.


The key role of ergothioneine in label-free surface-enhanced Raman scattering spectra of biofluids: a retrospective re-assessment of the literature.
Label-free surface-enhanced Raman scattering (SERS) has recently gained attention in the field of liquid biopsy as a rapid and relatively inexpensive technique that could significantly ease clinical diagnosis and prognosis by investigating a biofluid sample with a laser. Indeed, SERS spectra provide information about a set of metabolites present in the analysed biofluid, thereby offering biochemical insight into specific health conditions. Ergothioneine plays a key role since it is one of the few metabolites in biofluids that are detectable by label-free SERS. In the past decade, many studies characterizing biofluids or other biological samples have unknowingly linked this amino acid with crucial metabolic processes, including inflammation, in a plethora of diseases. However, since the SERS spectrum of ergothioneine has been reported only recently, most past studies inadvertently assigned what are now recognized as the spectral features of this compound to other molecules. The purpose of the present review is to summarize and re-evaluate these studies in the light of the recent SERS characterization of ergothioneine so as to better recognize the role of ergothioneine in many clinical conditions.


Characterization of the principal metabolite of quinine in human urine by 1H-n.m.r. spectroscopy.
1. The major metabolite of quinine in human urine, which is also the sole metabolite in human plasma and saliva, has been identified and characterized by chemical ionization mass spectrometry and 1H-n.m.r. spectrometry. 2. The mass spectrum showed that an oxygen atom is incorporated in the quinuclidine nucleus, and the exact position of the oxidation was established from the n.m.r. spectrum to be at the C-3 position.


The Synthesis of Backbone Thermo and pH Responsive Hyperbranched Poly(Bis(N,N-Propyl Acryl Amide))s by RAFT.
Hyperbranched poly(methylene-bis-acrylamide), poly(bis(N,N-propyl acryl amide)) (HPNPAM) and poly(bis(N,N-butyl acryl amide)) were synthesized by reversible addition-fragmentation chain transfer polymerization. HPNPAMs showed lower critical solution temperature (LCST) due to an appropriate ratio between hydrophilic and hydrophobic groups. The effects of reaction conditions on polymerization were investigated in detail. The structure of HPNPAM was characterized by ¹H NMR, FT-IR, Muti detector-size exclusion chromatography (MDSEC) and Ultravioletvisble (UV-Vis). The α value reached 0.20 and DB was 90%, indicating HPNPAMs with compact topology structure were successfully prepared. LCSTs were tuned by Mw and the pH value of the solution. The change of molecular size was assayed by dynamic light scattering and scanning electron microscope. These results indicated that the stable uniform nanomicelles were destroyed and macromolecules aggregated together, forming large particles as temperature exceeded LCST. In addition, after the cells were incubated for 24 h, the cell viability reached 80%, which confirmed this new dual responsive HPNPAM had low cytotoxicity.


Retinal hemorrhage associated with perinatal distress in newborns.
To describe the ophthalmoscopic features and natural history in a case series of eyes that developed intraocular hemorrhages associated with perinatal distress and to evaluate their clinical courses. A retrospective chart review of 289 neonates with a medical history of perinatal distress was conducted. Among these 289 patients (578 eyes), 29 eyes of 17 neonates were found to have had retinal hemorrhages or vitreous hemorrhages (VH). A comprehensive chart review, including details of fundoscopic findings and perinatal history, was conducted. Intraocular hemorrhage was present in 5.5% of the patients. Most hemorrhages (82.7%) were intraretinal. In our population, 17% (n = 5) of hemorrhages resolved within two weeks, but 31% (n = 9) did not resolve even after four weeks. Most hemorrhages spontaneously resolved without any specific sequelae; however, one infant's dense unilateral VH persisted up to three months after birth. When the patient was seen again at 3.5 years of age, she had developed axial myopia and severe amblyopia of the involved eye. In asphyxiated newborns, the possibility of intraocular hemorrhages should be considered. Long-standing, dense hemorrhages obscuring the macula may lead to severe vision deprivation amblyopia. Therefore, ophthalmic examination should be considered in neonates with perinatal distress, and close observation is necessary for hemorrhages that do not resolve in this amblyogenic age group.


Unique and redundant functional domains of APETALA1 and CAULIFLOWER, two recently duplicated Arabidopsis thaliana floral MADS-box genes.
APETALA1 (AP1) and CAULIFLOWER (CAL) are closely related MADS box genes that are partially redundant during Arabidopsis thaliana floral meristem determination. AP1 is able to fully substitute for CAL functions, but not vice versa, and AP1 has unique sepal and petal identity specification functions. In this study, the unique and redundant functions of these two genes has been mapped to the four protein domains that characterize type-II MADS-domain proteins by expressing all 15 chimeric combinations of AP1 and CAL cDNA regions under control of the AP1 promoter in ap1-1 loss-of-function plants. The "in vivo" function of these chimeric genes was analysed in Arabidopsis plants by expressing the chimeras. Rescue of flower meristem and sepal/petal identities was scored in single and multiple insert homozygous transgenic lines. Using these chimeric lines, it was found that distinct residues of the AP1 K domain not shared by the same CAL domain are necessary and sufficient for complete recovery of floral meristem identity, in the context of the CAL protein sequence, while both AP1 COOH and K domains are indispensable for complete rescue of sepal identity. By contrast, either one of these two AP1 domains is necessary and sufficient for complete petal identity recovery. It was also found that there were positive and negative synergies among protein domains and their combinations, and that multiple-insert lines showed relatively better rescue than equivalent single-insert lines. Finally, several lines had flowers with extra sepals and petals suggesting that chimeric proteins yield abnormal transcriptional complexes that may alter the expression or regulation of genes that control floral organ number under normal conditions.


Prevalence of microorganisms of hygienic interest in an organized abattoir in Mumbai, India.
The magnitude of food-borne illnesses in India is unknown because of lack of surveillance networks. Monitoring the prevalence of food-borne pathogens and indicators of contamination in primary production at abattoirs is imperative for creating a data bank and for effective control of such pathogens before they enter the food chain. Microorganisms of hygienic interest were screened for their prevalence at Deonar Abattoir, Mumbai. Swab samples from 96 sheep/goat carcass sites were collected and analyzed for Staphylococcus spp., Bacillaceae, Clostridiaceae and Enterobacteriaceae. Average Staphylococcus aureus and Staphylococcus epidermidis counts were 3.15 +/- 0.18 and 3.46 +/- 0.17 log10 CFU/cm(2), respectively. Bacillus cereus, Bacillus subtilis and Clostridium spp. counts were 3.10 +/- 0.08, 3.41 +/- 0.19 and 0.76 +/- 0.06 log10 CFU/cm(2), respectively. The Escherichia coli count was 3.54 +/- 0.06 and the Klebsiella aerogenes count was 3.22 +/- 0.22 log10 CFU/cm(2). Counts for Proteus vulgaris and Proteus mirabilis were 3.44 +/- 0.14 log10 CFU/cm(2) and 3.71 +/- 0.12 log10 CFU/cm(2), respectively. S. epidermidis had the highest percentage prevalence at (41.6%), followed by K. aerogenes (31.9%), B. subtilis (28.2%) and P. vulgaris (23.6%). Salmonella spp. were not isolated. The data demonstrate high prevalence and diversity of micro flora on carcasses in the primary Indian production facility, which might be attributed to either human handling or improper dressing especially during evisceration process. Appropriate training for personal and production hygiene is essential for workers in Indian meat production facilities.


ZnPP reduces autophagy and induces apoptosis, thus aggravating liver ischemia/reperfusion injury in vitro.
There is growing evidence indicating that autophagy plays a protective role in liver ischemia/reperfusion (IR) injury. Heme oxygenase-1 (HO-1) can also prevent liver IR injury by limiting inflammation and inducing an anti-apoptotic response. Autophagy also plays a crucial role in liver IR injury. The aim of the present study was to investigate the role of HO-1 in liver IR injury and the association between HO-1, autophagy and apoptotic pathways. IR simulation was performed using buffalo rat liver (BRL) cells, and HO-1 activity was either induced by hemin (HIR group) or inhibited by zinc protoporphyrin (ZnPP) (ZIR group). In the HIR and ZIR group, the expression of HO-1 and autophagy-related genes [light chain 3-Ⅱ (LC3-Ⅱ)] was assessed by RT-qPCR and the protein expression of caspases, autophagy-related genes and genes associated with apoptotic pathways (Bax) was detected by western blot anlaysis. The results of RT-PCR revealed the genetically decreased expression of HO-1 and autophagy-related genes in the ZIR group. Similar results were obtained by western blot analysis and immunofluorescence. An ultrastructural analysis revealed a lower number of autophagosomes in the ZIR group; in the HIR group, the number of autophagosomes was increased. The expression of Bax and cytosolic cytochrome c was increased, while that of Bcl-2 was decreased following treatment of the cells with ZnPP prior to IR simulation; the oppostie occurred in the HIR group. Cleaved caspase-3, caspase-9 and poly(ADP-ribose) polymerase (PARP) protein were activated in the IR and ZIR groups. The disruption of mitochondrial membrane potential was also observed in the ZIR group. In general, the downregulation of HO-1 reduced autophagy and activated the mitochondrial apoptotic pathway.


Asymmetric α-Chlorination of β-Keto Esters Using Hypervalent Iodine-Based Cl-Transfer Reagents in Combination with Cinchona Alkaloid Catalysts.
We herein report an unprecedented strategy for the asymmetric α-chlorination of β-keto esters with hypervalent iodine-based Cl-transfer reagents using simple Cinchona alkaloid catalysts. Our investigations support an α-chlorination mechanism where the Cinchona species serves as a nucleophilic catalyst by reacting with the chlorinating agent to generate a chiral electrophilic Cl-transfer reagent in situ. Using at least 20 mol-% of the alkaloid catalyst allows for good yields and enantioselectivities for a variety of different β-keto esters under operationally simple conditions.


Utility of signs and symptoms of chronic cough in predicting specific cause in children.
Paediatricians rely on cough descriptors to direct them to the level of investigations needed for a child presenting with chronic cough, yet there is a lack of published data to support this approach. A study was undertaken to evaluate (1) whether historical cough pointers can predict which children have a specific cause for their cough and (2) the usefulness of chest radiography and spirometry as standard investigations in children with chronic cough. This was a prospective cohort study of children referred to a tertiary hospital with a cough lasting >3 weeks between June 2002 and July 2004. All included children completed a detailed history and examination using a standardised data collection sheet and followed a pathway of investigation until a diagnosis was made. In 100 consecutively recruited children of median age 2.8 years, the best predictor of specific cough observed was a moist cough at the time of consultation with an odds ratio (OR) of 9.34 (95% CI 3.49 to 25.03). Chest examination or chest radiographic abnormalities were also predictive with OR 3.60 (95% CI 1.31 to 9.90) and 3.16 (95% CI 1.32 to 7.62), respectively. The most significant historical pointer for predicting a specific cause of the cough was a parental history of moist cough (sensitivity 96%, specificity 26%, positive predictive value 74%). The most useful clinical marker in predicting specific cough is the presence of a daily moist cough. Both chest examination and chest radiographic abnormalities are also useful in predicting whether children have a specific cause of their cough.


[Opiate substitution treatment's cycles in a five-year followed-up cohort in ambulatory practice].
To describe opiate substitution treatment (OST) cycles in a five year followed-up cohort of patients initiating OST during 5 years in ambulatory practice. Data were extracted from Health Insurance System Database from January 1(st) 2005 to December 31(st) 2010 for "Haute Garonne" (South Western France area, 1 million inhabitants). Patients starting OST were included. Drop-outs were identified throughout the follow-up. Five hundred patients were included, 72% of men, median age of 33 years. Median number of OST cycles was 2 [1; 3] on 5 years. Median duration of first OST cycle was 214 [77; 689] days. Median delay between first and second OST cycle was 80 [23; 241] days. Retention rate at one year was 38.2%. More than 50% of patients had at least 2 cycles of OST.


Polyphosphate accumulation among denitrifying bacteria in activated sludge.
Bacterial polyphosphate accumulation and denitrification are important processes in biological removal of nutrients from wastewater. It has been suggested that phosphorus accumulators are able to denitrify. However, the bacteria known as the most important phosphorus accumulators, belonging to the genus Acinetobacter are generally not known to denitrify. To clarify how commonly both physiological traits are present in the same organism, we screened 165 isolates from activated sludge and wastewater for their ability to denitrify, and the ability of the denitrifying isolates to accumulate polyphosphate. Of the 165 isolates, 149 were from acetate mineral medium (87 of these identified as Acinetobacter by the API 20 NE identification system) and 16 were from nutrient broth and nitrate medium. Only 15 of 165 isolates tested showed true respiratory denitrification activity. In the presence of acetylene they converted more than 80% of 5mM NO3- to N2O in 6 days. None of the Acinetobacter isolates were among the 15 respiratory denitrifiers. The denitrifying isolates were identified as species of Pseudomonas, Agrobacterium, Pasteurella, Sphingomonas or could not be identified by the API 20 NE identification system. According to the BIOLOG identification system the denitrifiers were species of Pseudomonas, Hydrogenophaga, Citrobacter, Xanthomonas or they could not be identified. The ability of confirmed denitrifiers to accumulate phosphate was measured in experiments where cells pregrown under phosphorus limitation were exposed to phosphate (8 mg P/L) under aerobic conditions. The rates of excess phosphate uptake varied from 0.3 to more than 23 mg P/g dry matter/h. Rates for four isolates were higher than those reported for Acinetobacter strains. These results show that polyphosphate accumulation and denitrification in activated sludge can be carried out by the same organisms.


[Clinical study of lamivudine and interferon combinate administration to inhibit hepatitis B virus replication].
To explore a new strategy for effective and economical anti-virus therapy for HBV infection, we conducted a sequence administration of lamivudine and interferon alpha 1b to evaluate its effects on HBV replication and rebound as well as YMDD mutation induced by lamivudine. 150 HBV patients having at least 6 months history of infection were assigned randomly into 5 groups. Each group of these patients was either treated with lamivudine, interferon alpha 1b, lamivudine combined with interferon, sequence administration of lamivudine and interferon (sequence group) or no anti-virus therapy (control group) for 12 months. The serum samples were collected at 0, 3, 6, 9, 12 and 18th months and were assayed for ALT, AST, HBeAg, HBV DNA (quantitive PCR) as well as YMDD mutation types by microarray. The anti-virus replication effects were shown as early as the 3rd month in the sequence group but not in the IFN and control groups. The significant and persistent inhibition effect of it on HBV replication and improvement of liver function was shown. It was more effective than lamivudine or IFN treatments at the end of the drug administration and 6 months later after the drug was withdrawn. We also found that this sequence administration pattern can significantly shorten the period of treatment of lamivudine as well as reduce the rate of YMDD mutation and rebound of HBV replication after lamivudine withdrawal. It is also more economical than a combined therapy of lamivudine with IFN. This sequence administration of lamivudine and IFN pattern can significantly improve the anti-virus effect on HBV replication, shorten the period of treatment with lamivudine, reduce the mutation rate of YMDD and prevent the rebound of HBV after drug withdrawal.


Sex differences in psychological response to pain in patients with fibromyalgia syndrome.
To examine whether men and women with fibromyalgia syndrome (FMS) differ with respect to pain severity and functioning, pain-related beliefs, or pain-related coping. We hypothesized no significant sex differences in measures of pain and functioning, but that we would observe differences between men and women in how they view and how they cope with FMS-related pain. A total of 747 women and 48 men with FMS who attended a multidisciplinary treatment program completed the study measures. Analyses of covariance were used to examine sex differences in the study measures, with a P-value of ≤0.01 and at least a moderate effect size (Cohen d≥0.5) required for a difference to be deemed statistically significant. Men and women did not differ on demographic measures except for their age, with the men in our sample being significantly younger than the women. Consistent with the study hypothesis, the results revealed no sex differences in the measures of pain and functioning. For pain-related beliefs, men were more likely to view pain as reflecting harm, and they were also more likely than women to use activity avoidance as a pain-coping strategy. The study findings suggest that women and men with FMS may think about and cope with pain somewhat differently, and may therefore benefit from different types of psychosocial pain intervention.


CRISPR/Cas9-mediated targeted mutagenesis of GmSPL9 genes alters plant architecture in soybean.
The plant architecture has significant effects on grain yield of various crops, including soybean (Glycine max), but the knowledge on optimization of plant architecture in order to increase yield potential is still limited. Recently, CRISPR/Cas9 system has revolutionized genome editing, and has been widely utilized to edit the genomes of a diverse range of crop plants. In the present study, we employed the CRISPR/Cas9 system to mutate four genes encoding SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors of the SPL9 family in soybean. These four GmSPL9 genes are negatively regulated by GmmiR156b, a target for the improvement of soybean plant architecture and yields. The soybean Williams 82 was transformed with the binary CRISPR/Cas9 plasmid, assembled with four sgRNA expression cassettes driven by the Arabidopsis thaliana U3 or U6 promoter, targeting different sites of these four SPL9 genes via Agrobacterium tumefaciens-mediated transformation. A 1-bp deletion was detected in one target site of the GmSPL9a and one target site of the GmSPL9b, respectively, by DNA sequencing analysis of two T0-generation plants. T2-generation spl9a and spl9b homozygous single mutants exhibited no obvious phenotype changes; but the T2 double homozygous mutant spl9a/spl9b possessed shorter plastochron length. In T4 generation, higher-order mutant plants carrying various combinations of mutations showed increased node number on the main stem and branch number, consequently increased total node number per plants at different levels. In addition, the expression levels of the examined GmSPL9 genes were higher in the spl9b-1 single mutant than wild-type plants, which might suggest a feedback regulation on the expression of the investigated GmSPL9 genes in soybean. Our results showed that CRISPR/Cas9-mediated targeted mutagenesis of four GmSPL9 genes in different combinations altered plant architecture in soybean. The findings demonstrated that GmSPL9a, GmSPL9b, GmSPL9c and GmSPL9 function as redundant transcription factors in regulating plant architecture in soybean.


Perceptions of Clinical Dental Students Toward Online Education During the COVID-19 Crisis: An Egyptian Multicenter Cross-Sectional Survey.
Objectives: To evaluate the perceptions of clinical dental students on the role of online education in providing dental education during the COVID-19 crisis. Materials and Methods: A cross-sectional online survey was sent to four Egyptian dental schools from the 20th of January 2021 to the 3rd of February 2021. Survey questions included the demographics, uses, experiences, perceived benefits, and barriers of distance learning in dentistry during the COVID-19 pandemic. Responses were collected from the clinical dental school students. Categorical data were presented as frequencies (n) and percentages (%) and were analyzed using Fisher's exact test. Results: Three hundred thirty-seven clinical dental students across four Egyptian dental schools responded. Most students used either Google Classroom or Microsoft Teams to access the online content. The data showed that the COVID-19 pandemic affected the academic performance of most participants (97.4%) with varying degrees. On average, students were neutral when asked to rate the online lectures, but did not find online practical education as effective (81.3%) as online theoretical teaching. The commonly described barriers to online teaching included loss of interaction with educators, inappropriateness in gaining clinical skills, and the instability of the internet connection. Conclusion: Despite the reported benefits, clinical dental students in Egypt preferred the hybrid approach in dental education as distance learning represented a prime challenge to gain adequate clinical dental skills.


Investigation of mechanism of desmopressin binding in vasopressin V2 receptor versus vasopressin V1a and oxytocin receptors: molecular dynamics simulation of the agonist-bound state in the membrane-aqueous system.
The vasopressin V2 receptor (V2R) belongs to the Class A G protein-coupled receptors (GPCRs). V2R is expressed in the renal collecting duct (CD), where it mediates the antidiuretic action of the neurohypophyseal hormone arginine vasopressin (CYFQNCPRG-NH2, AVP). Desmopressin ([1-deamino, 8-D]AVP, dDAVP) is strong selective V2R agonist with negligible pressor and uterotonic activity. In this paper, the interactions responsible for binding of dDAVP to vasopressin V2 receptor versus vasopressin V1a and oxytocin receptors has been examined. Three-dimensional activated models of the receptors were constructed using the multiple sequence alignment and the complex of activated rhodopsin with Gt(alpha) C-terminal peptide of transducin MII-Gt(alpha) (338-350) prototype (Slusarz, R.; Ciarkowski, J. Acta Biochim Pol 2004 51, 129-136) as a template. The 1-ns unconstrained molecular dynamics (MD) of receptor-dDAVP complexes immersed in the fully hydrated 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) membrane model was conducted in an Amber 7.0 force field. Highly conserved transmembrane residues have been proposed as being responsible for V2R activation and G protein coupling. Molecular mechanism of the dDAVP binding has been suggested. The internal water molecules involved in an intricate network of the hydrogen bonds inside the receptor cavity have been identified and their role in the stabilization of the agonist-bound state proposed.


Initial immunochemical characterization of MX35 ovarian cancer antigen.
Monoclonal antibody (mAb) MX35 reacts with approximately 90% of ovarian epithelial cancers and has been studied in localization and biodistribution trials in ovarian cancer patients. This study shows that mAb MX35 recognizes a cell surface antigen of about 95,000 D on OVCAR-3 ovarian cancer cells. The antigen could be immunoprecipitated from lysates of cells metabolically labeled with [3H]glucosamine and it bound to concanavalin A and wheat germ agglutinin lectins, showing that it is a glycoprotein. MX35 antigen can also be detected in detergent lysates of OVCAR-3 cells by Western blotting. Using this technique the MX35 epitope(s) was shown to be heat stable but susceptible to reduction by 2-mercaptoethanol. Protease digestion of the antigen resulted in smaller fragments (42-52 kDa) that still reacted with antibody. We conclude that MX35 antigen is a 95 kDa glycoprotein, stabilized by disulfide bonds, with a large protease-resistant region that carries the MX35 epitopes.


[Observation on therapeutic effect of acupuncture and moxibustion combined with cupping on senile psychophysiologic disorders].
To probe into the feasibility of acupuncture and moxibustion combined with rapid and repeated cupping at points on the back for treatment of senile psychophysiologic disorders. Sixty cases of clinically commonly-seen senile psychophysiologic disorders were randomly divided into the treatment group treated by acupuncture and moxibustion combined with rapid and repeated cupping at points on the back and the control group treated by oral administration of small dose of medicine for expectant treatment. Their therapeutic effects were compared. The therapeutic effect of the treatment group was better than that of the control group (P < 0.01). Acupuncture and moxibustion combined with rapid and repeated cupping at points on the back can more rapidly alleviate the psychotic symptoms of thinking more and misgivings, gloominess, desire to cry, restlessness due to deficiency, anger, impatient for senile psychophysiologic disorders.


[Successful child delivery in treatment-free remission of chronic myeloid leukemia after discontinuation of dasatinib].
A 21-year-old woman was diagnosed with chronic myeloid leukemia in March 2014. The patient and her family did not wish to freeze eggs before dasatinib initiation. After 66 months of oral dasatinib administration and 40 months of MR4.5 maintenance, the patient requested to discontinue dasatinib due to a desire to conceive. MR4.5 maintenance was continued, and she achieved spontaneous pregnancy 6 months after dasatinib discontinuation. The patient gave birth to a normal baby 13 months later and was on MR4.5 maintenance 21 months after dasatinib discontinuation.


Dietary native resistant starch but not retrograded resistant starch raises magnesium and calcium absorption in rats.
The effects on calcium and magnesium absorption of dietary native and retrograded cornstarch were studied in rats. Uncooked high amylose starch granules (35% of total glucose equivalents as enzyme-resistant starch) and cooked and cooled (-20 degrees C) high amylose starch (24% of total glucose equivalents as retrograded resistant starch) were used as test starches, and cooked normal starch (3% of total glucose equivalents as resistant starch) was used as control starch. Native vs. control starch raised the amount of polymerized glucose in ileum, but not in feces. Retrograded starch produced more polymerized glucose than control starch in both ileum and feces. When compared with control starch, ileal pH was significantly lowered by native starch and tended to be raised by retrograded starch. Cecal pH was lowered by the two preparations rich in resistant starch. Apparent absorption of calcium and magnesium was raised by native starch but not by retrograded resistant starch. Calcium concentrations in the liquid phase of the ileum tended to be elevated by native starch but were significantly lowered by retrograded starch relative to control starch. Magnesium and calcium concentrations in liquid cecal contents tended to be raised with native starch; they were unchanged with retrograded starch. It is suggested that native resistant starch raised calcium and magnesium absorption because it tended to enhance the solubility of these minerals in ileal and cecal digesta.


Evaluation of current prognostic and predictive markers in breast cancer: a validation study of tissue microarrays.
Tissue microarrays (TMAs) are an attractive alternative to analysis of whole sections (WS). For breast carcinomas, the recent recommendations for cut-offs (i.e. Ki67, H-score) have necessitated the re-evaluation of TMAs. TMA results of immunohistochemistry (IHC) and Fluorescence in situ hybridization (FISH) testing for Estrogen receptors (ER), Progesterone receptors (PgR), Ki67 and HER2 were compared against the results of WS for 88 breast carcinomas. We found excellent agreement between the two methods for ER and PgR IHC evaluation, using the H-score (Kappa coefficient 0.972 and 0.9, respectively). There was also excellent correlation for HER2 IHC (Kappa coefficient 1) and amplification (Kappa coefficient 0.933). Furthermore, scoring of Ki67 was highly-correlated between TMAs and WS (Kappa coefficient 0.954). The latter excellent correlation has not, to our knowledge, been previously reported. For breast cancer, TMAs are an efficient and reliable alternative to the use of WS, using the currently recommended markers, evaluation protocols and cut-off values.


Primary liposarcoma of the orbit.
Liposarcoma is the most common sarcoma in adults, but is an extremely rare tumor of the orbit. We report 4 cases of primary orbital liposarcoma treated at our center. All cases had tumor localized to the orbit and underwent exenteration followed by radiotherapy in one patient. None of the patients had a recurrence until a follow-up of 1-5 years (mean: 4 years).


Modern Techniques for the Isolation of Extracellular Vesicles and Viruses.
Extracellular signaling is pivotal to maintain organismal homeostasis. A quickly emerging field of interest within extracellular signaling is the study of extracellular vesicles (EV), which act as messaging vehicles for nucleic acids, proteins, metabolites, lipids, etc. from donor cells to recipient cells. This transfer of biologically active material within a vesicular body is similar to the infection of a cell through a virus particle, which transfers genetic material from one cell to another to preserve an infection state, and viruses are known to modulate EV. Although considerable heterogeneity exists within EV and viruses, this review focuses on those that are small (< 200 nm in diameter) and of relatively low density (< 1.3 g/mL). A multitude of isolation methods for EV and virus particles exist. In this review, we present an update on methods for their isolation, purification, and phenotypic characterization. We hope that the information we provide will be of use to basic science and clinical investigators, as well as biotechnologists in this emerging field. Graphical Abstract.


Technical know-how in stereotactic ablative radiotherapy (SABR).
Stereotactic ablative radiotherapy (SABR) is an addition to the armamentarium against cancer. The technical requirements for SABR are very stringent, given its very narrow therapeutic window. However, when the principles are strictly followed, it is possible to deliver SABR to extracranial tumours safely and effectively.


When are tricoordinated PdII species accessible? Stability trends and mechanistic consequences.
The ease of access to Pd(II) tricoordinated species (whether intermediates or transition states) in organometallic and catalytic reactions has been assessed with DFT methods to analyze the relative stability of tricoordinated [PdArXL] complexes versus their tetracoordinated derivatives formed by two most common processes of filling the fourth coordination site: solvent coordination (with tetrahydrofuran), or dimerization to give [Pd2Ar2(micro-X)2L2]. The effect of each ligand (L=PH3, PMe3, PPh3, PtBu3, 1-AdPtBu2; Ar=C6F5, C6H5, C6H4OH, C6H4OCH3, C6H4NH2, C6H2(NH2)3; X=F, Cl, Br, I, OH, SH, NH2, PH2, CH3) on these two processes has been systematically considered, and the results have been compared with the experimental information available. The trends observed, match the experimental results and suggest that: 1) the formation of bridged dimeric complexes is strongly preferred; 2) electronic effects are in general less important compared to steric effects; 3) when steric effects prevent formation of bridges and coordination of a fourth external ligand, intramolecular agostic interactions are established with C--H groups of one ligand; 4) as an exception, for X=NR2 true tricoordinated complexes, not showing agostic interactions, become stable. In the later case NR2 seems to act as pi-donor with its lone pair to the empty orbital at the fourth coordination site of palladium, thus avoiding a true 14e configuration for the tricoordinated PdII complex.


QCD-Electroweak First-Order Phase Transition in a Supercooled Universe.
If the electroweak sector of the standard model is described by classically conformal dynamics, the early Universe evolution can be substantially altered. It is already known that-contrarily to the standard model case-a first-order electroweak phase transition may occur. Here we show that, depending on the model parameters, a dramatically different scenario may happen: A first-order, six massless quark QCD phase transition occurs first, which then triggers the electroweak symmetry breaking. We derive the necessary conditions for this dynamics to occur, using the specific example of the classically conformal B-L model. In particular, relatively light weakly coupled particles are predicted, with implications for collider searches. This scenario is also potentially rich in cosmological consequences, such as renewed possibilities for electroweak baryogenesis, altered dark matter production, and gravitational wave production, as we briefly comment upon.


Contractile responses to sumatriptan in isolated bovine pulmonary artery rings: relationship to tone and cyclic nucleotide levels.
We examined responses to the 5-hydroxytryptamine 1D (5-HT1D)-receptor agonist sumatriptan in bovine pulmonary artery rings (2-3 mm ID). The effects of agonist-induced tone and agents that alter intracellular cyclic AMP [cyclic AMP]i or [cyclic GMP]i on responses to sumatriptan were investigated. At resting tension, responses to sumatriptan were slight or not evident. In the presence of tone induced by U46619, responses to sumatriptan (1 nM-30 mM) were greatly potentiated, as were responses to the alpha2-adrenoceptor agonist UK14304. Responses to the alpha 1-adrenoceptor agonist phenylephrine (PE) were potentiated only slightly. In the presence of U46619, addition of the adenylyl cyclase activator, forskolin (1 nM-0.1 microM or isoprenaline (ISO 1 microM) induced relaxations and increases in [cyclic AMP]i and resulted in further potentiation of the contractile response to sumatriptan. Addition of 0.1 microM sodium nitroprusside (SNP) inhibited sumatriptan-induced contractions. Whereas sumatriptan alone did not significantly affect [cyclic AMP]i, in the presence of U46619 it decreased [cyclic AMP]i. This effect of sumatriptan was further enhanced in the presence of forskolin. Sumatriptan increased [cyclic GMP]i. Using a nitric oxide (NO) synthase inhibitor and vessels denuded of endothelium, we showed that the increased [cyclic GMP]i in response to sumatriptan was endothelium-dependent and mediated by NO. This increase in [cyclic GMP]i was not observed in the presence of U46619. By measuring cyclic AMP and cyclic GMP phosphodiesterase (PDE) levels, we demonstrated that the point of "cross-talk" between cyclic nucleotides may not be at the level of total PDE activity. These results highlight the important role of [cyclic AMP], [cyclic GMP]i, and endothelium function in the control of 5-HT1D receptor-mediated vasoconstriction, which is dependent on a decrease in [cyclic AMP]i in the absence of an increase in [cyclic GMP]i.


[Apoptosis in radiosensitive malignant tumors].
This study reported the radiation induced apoptosis in 6 cases with malignancies that were observed rapid tumor regression with radiotherapy up to 10 Gy. Microscopic examinations of 6 cases revealed the tumor cell number reduction and the appearance of apoptotic cells in the irradiated cases. It is suggested that the radiation-induced apoptosis plays an important role in rapid tumor regression by radiotherapy and occurs not only in lymphoma which has been recently reported in vitro but also in carcinoma.


Escherichia Coli Increases its ATP Concentration in Weakly Acidic Environments Principally through the Glycolytic Pathway.
Acid resistance is an intrinsic characteristic of intestinal bacteria in order to survive passage through the stomach. Adenosine triphosphate (ATP), the ubiquitous chemical used to power metabolic reactions, activate signaling cascades, and form precursors of nucleic acids, was also found to be associated with the survival of Escherichia coli (E. coli) in acidic environments. The metabolic pathway responsible for elevating the level of ATP inside these bacteria during acid adaptation has been unclear. E. coli uses several mechanisms of ATP production, including oxidative phosphorylation, glycolysis and the oxidation of organic compounds. To uncover which is primarily used during adaptation to acidic conditions, we broadly analyzed the levels of gene transcription of multiple E. coli metabolic pathway components. Our findings confirmed that the primary producers of ATP in E. coli undergoing mild acidic stress are the glycolytic enzymes Glk, PykF and Pgk, which are also essential for survival under markedly acidic conditions. By contrast, the transcription of genes related to oxidative phosphorylation was downregulated, despite it being the major producer of ATP in neutral pH environments.


Spatial modeling of long-term air temperatures for sustainability: evolutionary fuzzy approach and neuro-fuzzy methods.
This paper investigates the capabilities of the evolutionary fuzzy genetic (FG) approach and compares it with three neuro-fuzzy methods-neuro-fuzzy with grid partitioning (ANFIS-GP), neuro-fuzzy with subtractive clustering (ANFIS-SC), and neuro-fuzzy with fuzzy c-means clustering (ANFIS-FCM)-in terms of modeling long-term air temperatures for sustainability based on geographical information. In this regard, to estimate long-term air temperatures for a 40-year (1970-2011) period, the models were developed using data for the month of the year, latitude, longitude, and altitude obtained from 71 stations in Turkey. The models were evaluated with respect to mean absolute error (MAE), root mean square error (RMSE), Nash-Sutcliffe efficiency (NSE), and the determination coefficient (R2). All data were divided into three parts and every model was tested on each. The FG approach outperformed the other models, enhancing the MAE, RMSE, NSE, and R2 of the ANFIS-GP model, which yielded the highest accuracy among the neuro-fuzzy models by 20%, 30%, and 4%, respectively. A geographical information system was used to obtain temperature maps using estimates of the optimal models, and the results of the model were assessed using it.


[Differences in fraction of exhaled nitric oxide values measured by two offline methods or NIOXmino in adult Japanese asthmatics].
The fraction of exhaled nitric oxide (FeNO) is a useful marker of asthma control. The FeNO measurement with two offline methods and NIOXmino may be more affordable, no studies have examined the differences in FeNO values measured with various methods in adult asthmatics. The study population comprised 39 stable asthmatics treated with inhaled steroids at our outpatient clinic. FeNO values were measured by two offline methods (Sievers and CEIS), NIOXmino. FeNO(NIOXmino) values were significantly correlated with those of FeNO(Sievers) (r=0.935, P< 0.001) and FeNOCEIS (r=0.908, P< 0.001). However, FeNO(NIOXmino) values were low compared with FeNO(Sievers) (FeNO(NIOXmino)=0.848 x FeNO(Sievers)) and FeNO(CEIS) (FeNO(NIOXmino)=0.672 x FeNO(CEIS)). Differences exist in the values of FeNO measured by various methods: conversion equations are needed to compare the FeNO values among these three methods.


Contact toxicity, feeding reduction, and repellency of essential oils from three plants from the ginger family (Zingiberaceae) and their major components against Sitophilus zeamais and Tribolium castaneum.
The essential oils from rhizomes of Alpinia conchigera Griff, Zingiber zerumbet Smitt, Curcuma zedoaria (Berg.) Roscoe; their major compounds (camphene, camphor, 1,8-cineole, alpha-humulene, isoborneol, alpha-pinene, beta-pinene and terpinen-4-ol); and synthetic essential oils comprised of mixtures of major pure compounds in the same ratios as the extracted essential oils were tested for contact, feeding reduction, and repellency against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst) adults. Via topical applications, the three extracted oils had similar toxicity against S. zeamais (LD50 fiducial limits: 18-24 microg oil/mg insect). T. castaneum had similar sensitivity to all three oils (35-58 microg/mg), and it was less sensitive than S. zeamais. The LD50 values of synthetic A. conchigera and synthetic Z. zerumbet oils were similar to those of their corresponding extracted essential oils. The synthetic C. zedoaria oils showed lower contact toxicity than the extracted C. zedoaria oils to both insects. Sitophilus zeamais and T. castaneum were sensitive to terpinen-4-ol and isoborneol in contact toxicity tests. In antifeedant tests, the three extracted oils were able to decrease the consumption of flour disks, especially Z. zerumbet oils, whereas both insect species could feed on the flour disks treated with three synthetic essential oils. Only terpinen-4-ol deterred feeding in both insects. In repellency tests, A. conchigera oils at highest concentration repelled S. zeamais and T. castaneum. None of the synthetic essential oils repelled S. zeamais (315 microl/cm2) and T. castaneum (31 microl/cm2) Only terpinen-4-ol showed repellent activity against both insects.


Abdominal Wall Masses: CT Findings and Clues to Differential Diagnosis.
The abdominal wall does not comprise a distinct organ, and is often cursorily evaluated on CT. However, it is affected by many different pathological processes. These may be categorized according to their underlying etiology-trauma, infection or inflammation, iatrogenic and neoplastic process-or according to the abdominal wall layer they affect. We chose instead to group these lesions into 6 distinct categories based on their CT characteristic density: solid, infiltrative, hypervascular, fluid, fat, and bone density lesions. We highlight throughout the article the importance of integrating pertinent clinical history to narrow the differential diagnosis.


Recombinant human cytidine deaminase: expression, purification, and characterization.
The complementary DNA (cDNA) coding for human cytidine deaminase (CDA) was obtained using two specific primers to screen RNA from peripheral blood polymorphonuclear leukocytes by reverse transcriptase PCR. The cDNA fragment was ligated into the expression vector pTrc99-A and expressed in Escherichia coli following induction with isopropyl-1-thio-beta-D-galactopyranoside (IPTG). The nucleotide sequence of the cDNA corresponded to that published by Laliberté and Momparler (Cancer Res. 54, 5401-5407, 1994). It contained a 438-bp open reading frame encoding a polypeptide of 146 amino acids with a predicted molecular mass of 16.2 kDa. The protein expressed in E. coli showed high cytidine deaminase activity and its molecular mass was estimated to be 57 kDa by gel filtration and 16 kDa by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Both values are in agreement with those already published and suggest that human CDA contains three or four identical subunits. Cross-linking experiment indicated that the enzyme is a tetramer. The recombinant CDA has been purified to homogeneity by a rapid procedure consisting of heat inactivation followed by affinity chromatography. The final enzyme preparation showed a specific activity of 105 U/mg, corresponding to about 88-fold purification with respect to the crude extract and was judged to be >98% pure by SDS-PAGE. Inductively coupled plasma-optical emission spectroscopy (ICP-OES) analysis revealed the presence of 1 atom of Zn per subunit. Since CDA causes the deamination of several antitumoral cytidine-analog drugs, the recombinant enzyme was characterized kinetically and several pyrimidine nucleoside analogs were tested as potential substrates and inhibitors. The results obtained agreed closely with those previously reported for the purified human placenta CDA.


Speech priming: an examination of rate and syntactic persistence in preschoolers.
Interactional coordination is important for conversational competence. For example, the syntactic form and rate of perceived speech can influence future productions in adults. Previous work has shown that children are similarly primed by syntax. This experiment demonstrates that syntactic priming and rate priming exist simultaneously in children. Participants (4- and 5-years-old) alternated between listening to priming sentences that described visual scenes and producing their own descriptions of similar scenes. The priming sentences varied in rate (fast and slow) and syntactic structure (active and passive). Children's sentences reflected the timing and syntactic structure of the primes, and there were developmental differences in their rate persistence.


Entrapment neuropathies: 2. Lower limb.
Entrapment neuropathy is a form of compression neuropathy (see previous article). It is rare in the lower limb. Fibular tunnel syndrome, meralgia paraesthetica and tarsal tunnel syndrome are reviewed here.


Fabrication of self-supporting porous silicon membranes and tuning transport properties by surface functionalization.
This study presents a simple approach to perform selective mass transport through freestanding porous silicon (pSi) membranes. pSi membranes were fabricated by the electrochemical etching of silicon to produce membranes with controlled structure and pore sizes close to molecular dimensions (approximately 12 nm in diameter). While these membranes are capable of size-exclusion based separations, chemically specific filtration remains a great challenge especially in the biomedical field. Herein, we investigate the transport properties of chemically functionalized pSi membranes. The membranes were functionalized using silanes (heptadecafluoro-1,1,2,2-tetrahydrodecyl)dimethylchlorosilane (PFDS) and N-(triethoxysilylpropyl)-o-polyethylene oxide urethane (PEGS) to give membranes hydrophobic (PFDS) and hydrophilic (PEGS) properties. The transport of probe dyes tris(2,2'-bipyridyl)dichlororuthenium(ii) hexahydrate (Rubpy) and Rose Bengal (RB) through these functionalized membranes was examined to determine the effect surface functionalization has on the selectivity and separation ability of pSi membranes. This study provides the basis for further investigation into more sophisticated surface functionalization and coupled with the biocompatibility of pSi will lead to new advances in membrane based bio-separations.


A carbon dioxide gas sensor by combination of multivalent cation and anion conductors with a water-insoluble oxycarbonate-based auxiliary electrode.
A compact and inexpensive carbon dioxide gas sensor was successfully realized by the combination of a divalent magnesium ionic conductor of Mg0.7(Zr0.85Nb0.15)4P6O24 and a divalent oxide anion conducting ZrO2-Y2O3 solid electrolyte with the water-insoluble Li- and Ba-codoped Nd2O2CO3 solid solution as the auxiliary electrode. The sensor response was continuous and reproducible, and the present sensor also demonstrated a theoretical Nernst response in the atmosphere where water vapor, nitrogen oxides, ammonia, etc., coexist. The exposure of the present sensor to water dew and variation in oxygen concentration does not interfere with the sensor response, which will be a great advantage in applying the in situ practical CO2 detection in combustion exhaust gas atmospheres.


Does methylphenidate normalize the classroom performance of children with attention deficit disorder?
The present study examined the degree to which methylphenidate (MPH) normalized the classroom behavior and academic functioning of 31 children with attention deficit disorder (ADD). Subjects with ADD participated in a double-blind, placebo-controlled trial in which children received each of four doses (5, 10, 15, and 20 mg) of MPH and a placebo following baseline measures. Children with ADD were compared with a group of 25 normal control children on teacher ratings of social conduct, direct observations of on-task behavior, and academic efficiency. At a group level of analysis, MPH exerted a significant effect on classroom measures of attention and academic efficiency to a point where they were no longer statistically deviant from scores obtained by normal control children. Nevertheless, when examined at the individual level, 25% of the sample failed to show normalized levels of classroom performance, thus implying the need for ancillary school-based interventions.


Up-regulation of oxytocin receptors in rabbit amnion by glucocorticoids: potentiation by cyclic adenosine 3',5'-monophosphate.
Oxytocin (OT) receptors (OTR) in rabbit amnion increase more than 200-fold at the end of gestation. In the present report, we studied the basis of this up-regulation. Incubation of amnion cells with cortisol (20 nM) for 24 h increased the amount of 125I-labeled OT antagonist bound by 16- to 18-fold. The effects of cortisol were dose and steroid dependent. Administration of glucocorticoid to pregnant does also increased the concentration of OTRs in amnion. The effects of cortisol in vitro were potentiated by the addition of forskolin (50 microM), so that OTR number increased by as much as 182 times. The effects of cortisol and forskolin, either separately or in combination, were inhibited by activation of protein kinase-C or coincubation with transforming growth factor-alpha (10 nM). Cyclosporin-A (5 microM) selectively inhibited cortisol-induced rises in the OTR concentration. The addition of cortisol to amnion cells increased OT-stimulated prostaglandin E2 (PGE2) release almost 100-fold; the combination of forskolin and cortisol increased the PGE2 response to OT about 5600 times. Judging from the greater effects on PGE2 release, these results suggest that forskolin and cortisol up-regulate the signal response mechanism to OT as well as the OTR concentration. The findings show that changes occurring in the amnion in vivo can be mimicked in vitro, and they elucidate the mechanism of up-regulation of OTR concentrations.


Effects of resveratrol and flavonols on cardiovascular function: Physiological mechanisms.
Resveratrol and flavonols are commonly found together in fruits and vegetables and, therefore, consumed in the diet. These two polyphenols share both vasorelaxant and antioxidant activity and may act together to improve cardiovascular function. This review examines the mechanisms by which resveratrol and flavonols influence cardiovascular function and perhaps offer a new approach for the development of therapeutic agents for the prevention and/or treatment of coronary artery disease.


Effect of microenvironment and cell-line type on carbohydrate-binding proteins of macrophage-like cells.
The pattern of sugar inhibition of rosette formation, a model for intercellular interaction between cultured cells and glutaraldehyde-fixed, trypsinated rabbit erythrocytes, served to infer the presence of carbohydrate-binding proteins. This profile from cell extracts for the two murine macrophage-like cell lines, P388D1 and J774A.1, was comparatively analyzed by affinity chromatography on supports with immobilized carbohydrates (lactose, L-fucose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and maltose) or with the immobilized mannose-rich yeast glycoprotein mannan or fetuin-derived glycopeptides containing sialic acid residues. After elution with specific sugar in the absence of Ca2+ ions, the proteins were separated by sodium dodecyl sulfate - polyacrylamide slab gel electrophoresis. The composition of carbohydrate-binding proteins of the two lines clearly exhibited quantitative and qualitative differences. Moreover, the pattern of P388D1 cells was also demonstrated to change significantly in response to alterations in the conditions of the physiological environment. These alterations were imposed by in vitro growth, by subsequent in vivo growth in nude mice, and by re-adaptation of cells to culture after in vivo passage. Collectively, our observations and other physiological and biochemical reports on macrophage lectins indicate that the presence of sugar receptors with different specificities may be an indicator of macrophage differentiation, being reversibly modulated to a considerable extent by external factors, e.g., microenvironment. Extensive but selective alterations in this respect could play an important role in the control of recognition and effector mechanisms within diverse functions of macrophage subpopulations.


Is the Sustainable Choice a Healthy Choice?-Water Footprint Consequence of Changing Dietary Patterns.
It is evident that the modification of dietary patterns is a necessary precondition of disease prevention and health improvement. Changing nutritional habits also has deep-rooted consequences on the environmental burden. The majority of similar previous studies have analyzed the change in greenhouse gas emissions against theoretical modifications in current food consumption. The analysis on the effect of diet on the water footprint is also gaining in importance, since water supply is a critical global issue. Based on current nutritional patterns of a Central European country-Hungary-as well as dietary recommendations and scientific literature, we generated six dietary scenarios and determined the consequences of these on green (originally from precipitation) and blue (sourced from surface or groundwater) water consumption and dietary quality. Compared to the baseline scenario (current local nutritional pattern) of both genders, based on the integrated aspect of water footprint and dietary quality, the most disadvantageous scenario was the ketogenic (ca. -2% in dietary quality, +18% in blue water footprint, and +16% in green water footprint) and the most advantageous was the sustainable scenario (ca. +9% in dietary quality, -42% in green water footprint, and -29% in blue water footprint). As a summary it can be stated, that (1) there is no clear linear relationship between the "healthiness" and water footprint of different diets, but (2) a more balanced diet, which integrates nutritional and environmental considerations could decrease the environmental burden in an efficient way.


Characterization of IgG2 Disulfide Bonds with LC/MS/MS and Postcolumn Online Reduction.
The complication of IgG2 disulfide connections demands advances in techniques for disulfide bond determination. We have developed a new LC/MS/MS method for improved disulfide analysis. With postcolumn introduction of dithiothreitol (DTT) and ammonium hydroxide, each disulfide-containing peptide eluted out of LC in an acidic mobile phase can be rapidly reduced prior to MS analysis. The reduction can be driven to near completion. The reagents are MS-friendly, and the reaction occurs at no cost of separation (little is added to the postcolumn dead volume of the LC system). Comparing LC/MS data with and without online reduction, a direct correlation can be established between a disulfide peptide and its composing peptides using retention time. With disulfide online removal, high-quality MS/MS fragmentation data can be acquired and allows for definitive determination of the disulfide peptide. This technique is especially valuable in determining the disulfide bond linkage of complicated molecules such as the hinge-containing disulfide peptides produced from IgG2 disulfide isoforms. Due to over/under enzymatic cleavages, multiple hinge-containing disulfide peptides are produced from each isoform. Twenty-two hinge-containing disulfide peptides in total have been confidently identified with this technique. Without the method, successful identification to many of these peptides would have become extremely difficult.


Role of PSMA PET-guided metastases-directed therapy in oligometastatic recurrent prostate cancer.
Oligometastatic prostate cancer (OMPC) has been proposed as an intermediary state between localised disease and widespread metastases, with varying definitions including 1, 3, or ≤5 visceral or bone metastasis. Traditional definitions of OMPC are based on staging with conventional imaging, such as computerised tomography (CT) and whole-body bone scan (WBBS). Novel imaging modalities such as prostate-specific membrane antigen positron emission tomography (PSMA PET) have improved diagnostic utility in detecting early metastatic prostate cancer (PC) metastases compared with conventional imaging. Specifically, meta-analytical data suggest that PSMA PET is sensitive in detecting oligometastatic disease in patients with biochemical recurrence (BCR) post-radical treatment of PC. Recent trials have evaluated PSMA PET-guided metastases-directed therapy (MDT) in oligometastatic recurrent disease, typically with salvage surgery or radiotherapy (RT). To date, these preliminary studies demonstrate promising results, potentially delaying the need for systemic therapy. We aim to report a comprehensive, multidisciplinary review of PSMA-guided MDT in OMPC. In this review, we highlight the utility of PMSA PET in biochemically recurrent disease and impact of PSMA PET on the definition of oligometastatic disease and outline data pertaining to PSMA-guided MDT.


Evaluation of optical coherence tomography findings in age-related macular degeneration: a reproducibility study of two independent reading centres.
To determine the reproducibility among readers of two independent certified centres, the Vienna Reading Center (VRC) and the University of Wisconsin-Madison Reading Center (UW-FPRC) for optical coherence tomography (OCT) images in age-related macular degeneration (AMD). Fast macular thickness scans and 6 mm cross hair scans were obtained from 100 eyes with all subtypes of AMD using Stratus OCT. Consensus readings were performed by two certified OCT readers of each reading center using their grading protocol. Common variables of both grading protocols, such as presence of cystoid spaces, subretinal fluid, vitreomacular traction and retinal pigment epithelial detachment, were compared using κ statistics. In addition, the intraclass correlation coefficient (ICC) was calculated for centre point thickness (CPT) of values re-measured manually in the presence of alignment errors. The reproducibility was dependent on the variable measured with a κ value of 0.81 for the presence of cystoid spaces, 0.78 for the presence of subretinal fluid and 0.795 for the presence of vitreomacular traction. The lowest reproducibility was found for the presence of retinal pigment epithelial detachment with a κ value of 0.51. The CPT was re-measured in 29 out of 100 scans at both sites with an ICC of the re-measured thicknesses of 0.92. OCT scan data are crucial in monitoring treatment efficacy in AMD clinical trials. For comparison of results obtained by different reading centers, the inter-reading center reproducibility is essential. Although the reproducibility is generally high, the reliability depends on the selected morphological parameters.


Structural and magnetic behaviour of soft magnetic Finemet-type ribbons.
Different kinds of magnetic anisotropies have been induced during the nanocrystallization process of Co- and Ni-rich amorphous ferromagnetic (Finemet) ribbons by the application of a constant stress or an axial magnetic field during the annealing process. Magnetization measurements have evidenced the presence od macroscopic anisotropy in the treated samples. The main goal of this work has been, after a careful DSC study, the structural analysis of the treated ribbons using X-ray Diffraction and Atomic Force Microscopy (AFM), detecting substantial differences in the crystallization state and grain size of the samples depending on the thermal treatment that was carried out. Moreover, AFM measurements revealed in all the treated samples a strong nanocrystallisation of the surface without evidences of amorphous matrix, which contrast with XRD measurements that have shown a high content of amorphous phase in the bulk of the ribbons. Magneto-optical Kerr effect measurements have been performed with the aim to elucidate the complex magnetic behaviour that is expected for the surface of the ribbons, measuring surface hysteresis loops that showed much higher coercive field values than that obtained in the bulk material.


Biosynthesis, cleavage, and degradation of the human immunodeficiency virus 1 envelope glycoprotein gp160.
The synthesis and processing of the human immunodeficiency virus 1 (HIV-1) envelope precursor glycoprotein gp 160 was studied in an infected CD4+ lymphocytic cell line. Surprisingly, only a small percentage (5-15%) of gp160 is cleaved to produce the mature gp120 component. Intracellular sorting results in the transfer of most uncleaved gp160 to lysosomes, where it is degraded, while gp120 is transported to the cell surface and subsequently secreted. Cleavage of gp160 to generate gp120 occurs intracellularly and can be inhibited by NH4Cl. Taken together, these results indicate that intracellular cleavage of gp160 determines the intracellular transport and survival of the envelope glycoproteins necessary to produce infectious virus.


Avascular necrosis and the Aberdeen splint in developmental dysplasia of the hip.
Between January 1987 and December 1988 there were 7575 births in the Swansea maternity unit. Of these 823 (10.9%) were considered to be at 'high risk' for developmental dysplasia of the hip (DDH). Static ultrasound examination was performed in each case and the results classified on the basis of the method of Graf. A total of 117 type III-IV hips in 83 infants was splinted using the Aberdeen splint. Radiographs of these hips were taken at six and 12 months. Hilgenreiner's measurements of the acetabular angle were made in all cases and the development of the femoral capital epiphysis was assessed by measuring the epiphyseal area. The effect of splintage on the acetabular angle and the epiphyseal area between the normal and abnormal splinted hips was compared. Radiographs of 16 normal infants (32 normal unsplinted hips) were used as a control group. This cohort has now been followed up for a minimum of nine years. There have been no complications as a result of splintage. The failure rate was 1.7% or 0.25 per 1000 live births. No statistical difference was found when comparing the effect of splintage on the acetabular angle and epiphyseal area between normal and abnormal splinted hips and normal unsplinted hips. Our study has shown that while the Aberdeen splint had a definite but small failure rate, it was safe in that it did not produce avascular necrosis. The current conventional view that a low rate of splintage is always best is therefore brought into question if the Aberdeen splint is chosen for the management of neonatal DDH.


Comparative Enzymology of the Glyceraldehyde 3-Phosphate Dehydrogenases from Pisum sativum.
Glyceraldehyde 3-phosphate dehydrogenases (EC 126.96.36.199 and 18.104.22.168) have been purified from the seed, root, etiolated, and green shoot of peas (Pisum sativum). These enzymes are tetramers of 140,000 daltons, with subunits of 35,000 daltons. The enzymes differ in isoelectric point. The seed enzyme has a pI of 5.1, and the root enzyme has a pI of 4.5. The cytoplasmic enzyme from etiolated shoots is slightly acidic with a pI of 5.7 to 6.1 and is found in two separable forms. The chloroplast enzyme (from green shoots) is most basic with a pI of 8.0.In immunodiffusion experiments, the seed, root, and cytoplasmic enzymes of the etiolated shoot share antigenic homology, while the chloroplast enzyme does not cross react antigenically with the extra-chloroplast enzymes. The antiserum to the pea chloroplast enzyme did, however, cross react with glyceraldehyde 3-phosphate dehydrogenase purified from the spinach chloroplast. Therefore, the chloroplast enzyme is significantly different from the extra-chloroplast enzymes with respect to primary sequence.The NADP analog phosphoadenosine diphosphoribose showed competitive inhibition to the chloroplast enzyme with either pyridine nucleotide. The NAD analog pyridine 3-aldehyde NAD was competitive with respect to the NAD activity but was hyperbolic competitive in the presence of NADP, indicating a complexity in the binding of pyridine nucleotide to the chloroplast enzyme.


[Determination of danshensu in urine and its pharmacokinetics in human].
To determine Danshensu in urine and study its pharmacokinetics in human. A solid phase extraction-HPLC method was used for determination of Danshensu in urine of human. HPLC separation is performed on a Shim-pack CLC-ODS column (150 mm x 6.0 mm ID, 5 microns) with a mobile phase composed of acetonitrile -0.01 mol.L-1 KH2PO4 (adjusted to pH 2.8 with phosphoric acid). The flow rate was 1.0 mL.min-1 and the UV detector was set at 280 nm. The linear range of Danshensu was 0.2-50 mg.L-1 (r = 0.9999), and its limit of detection was 1.5 ng. The mean recovery was 99.4% (RSD = 2.9%). The pharmacokinetics of Danshensu after p.o. administration of two kinds of pharmaceutical preparations containing Danshen (with 20 mg of Danshensu) were investigated in 6 healthy human volunteers by determining the Danshensu in urine samples. The elimination half lives (T1/2) of Danshensu after p.o. administration of compound granule preparation A and decoction of Danshen were (0.92 +/- 0.16) h and (0.94 +/- 0.21) h, respectively. Their excretions of Danshensu in urine were (6.2 +/- 2.8)% and (14 +/- 4)% of the dose in 8 hours, respectively. Under normal doses, Danshensu can be eliminated from kidney. There is no evident difference on elimination half lives of Danshensu after p.o. administration of the two doses, but the excretions of Danshensu by urine after p.o. administration of compound granule preparation A were lower than that of decoction of Danshen.


Ginsenoside Rb3 alleviates smoke-induced lung injury via the H19/miR-29b-3p/HGMB1/TLR4 signalling pathway.
The over-activation of inflammation is involved in the pathogenesis of smoke-induced lung injury (SILI), while Rb3 treatment may alleviate smoke-induced lung injury by down-regulating the expression of H19, a regulator of miR-29b expression. Moreover, HMGB1 is an important mediator of inflammation. Therefore, in this study, we set up an animal model of SILI and treated it with Rb3 to study the effect of Rb3 on the treatment of SILI and the involvement of H19/miR-29b/HMGB1/TLR4 signalling. SILI mice treated with Rb3 before H&E staining and TUNEL assay were conducted to observe the pathological damages and status of apoptosis in each group. Real-time PCR, Western blot, computational analysis and luciferase assays were utilized to establish the signalling pathway involved in the pathogenesis of SILI and the action of Rb3 treatment. Rb3 treatment alleviated pathological changes in the lungs while decreasing the levels of W/D ratio and cell apoptotic index. H19 was validated to sponge miR-29b-3p, while HMGB1 mRNA was validated to be a target gene of miR-29b-3. As a result, a signalling pathway of H19/miR-29b-3p/HMGB1 was established. Cell viability was evidently reduced after 72 hours of treatment with CSE, but the treatment of Rb3 elevated the expression of H19 and HMBG1 in the presence of CSE. Also, CSE-induced inhibition of miR-29b-3p expression was restored by Rb3. The findings of this study collectively demonstrated that Rb3 exhibited its therapeutic effect during the treatment of SILI via modulating the H19/miR-29b-3p/HMBG1 signalling pathway.


Conjugate of N4-(4-carboxybutyryl)-ara-C and ethylenediamine-introduced dextran. Drug release profiles and further in vivo study of its antitumor effects.
In vitro and further in vivo work with a conjugate formed from the cytotoxic drug 1-beta-arabinofuranosylcytosine (ara-C) and dextran 2000 are described. In the preparation of this conjugate, functionalisation of ara-C was via N4-(4-carboxybutyryl)-ara-C (glu-ara-C), permitting conjugation with amino groups introduced by prior reaction of the oxidised dextran with ethylenediamine; by varying the proportions of the reaction components, 5.4 to 7.7% w/w loadings of ara-C were obtained. At physiological pH, in vitro, drug release from a 5.4% loaded conjugate was gradual and was dominantly ara-C; at lysosomal pH (pH 5) the release rate was much slower and more ara-U was formed. Antitumour effects were evaluated in L1210 leukaemic mice following single (1 day after inoculation) or double (2 and 6 days after inoculation) intraperitoneal injection of ara-C, glu-ara-C, or a 7.7% loaded conjugate at three dose levels. In all cases, the increase in lifespan was greatest following use of the conjugate, but the differences in the effects of ara-C and the conjugate were only significant at the lowest dose level. Glu-ara-C was virtually inactive under all conditions.


A microfluidic concentration generator for dose-response assays on ion channel pharmacology.
We present a microfluidic device to generate either statically spatial or dynamically temporal logarithmic concentrations. The temporal logarithmic concentration generator was also integrated with planar patch-clamp chips for dose-response assays on ion channels. Proposed serial dilution principle controls the flow pattern at each branching point via designing the flow resistance of microchannels. Simple and linear ratios of the flow resistance results in desired logarithmic concentration at outlets, where the concentrations can be dynamically altered by different combination of valve actuations, were demonstrated. Single-cell pharmacology on ion channels was implemented by sequentially applying logarithmic drug concentrations to patched cells. Inhibitory activity of potassium channels of human embryonic kidney cells was examined by tetraethylammonium solutions. Resulted IC(50) and Hill slope reveal excellent agreement with assays from manually prepared drug concentrations showing the practicability and preciseness of the present approach. Applications include cellular analysis under various drugs and/or logarithmic concentrations at the single-cell level.


Amorphous materials for elementary-gas-involved electrocatalysis: an overview.
Given the critical demands on energy conversion, storage, and transportation, tremendous interest has been devoted to the field of material development related to energy harvesting, recently. As the only route towards energy utilization, the carriers with the characteristics of low carbon are regarded as the future choice, e.g., hydrogen and ammonia. To this end, electrocatalysis provides a green way to access these substances. However, the unfulfilled conversion efficiency is the bottleneck for practical application. In this review, the promising characteristics of amorphous materials and the amorphous-induced electrocatalytic enhancement (AIEE) were emphasized. In the beginning, the characteristics of amorphous materials are briefly summarized. The basic mechanism of heterogeneous electrocatalytic reactions is illustrated, including the hydrogen/oxygen evolution and oxygen/nitrogen reduction. In the third part, the electrocatalytic performance of amorphous materials is discussed in detail, and the mechanism of AIEE is highlighted. In the last section of this review, the challenges and outlook for the development of amorphous enhanced electrocatalysis are presented.


Crystallization and preliminary X-ray diffraction analysis of two pH-dependent forms of a di-haem cytochrome c peroxidase from Pseudomonas nautica.
Two crystal forms of cytochrome c peroxidase from Pseudomonas nautica were obtained, one at pH 4.0 using sodium citrate as precipitant and another at pH 5.3 using ammonium phosphate and sodium citrate as precipitants. The two forms belong to different space groups P3(1)21 (pH 4.0) and P6(4)22 (pH 5.3), with unit-cell parameters a = b = 114.5, c = 90.7 A and a = b = 151.0, c = 155.9 A, respectively. Several complete data sets were collected using synchrotron radiation at ESRF and Cu K(alpha) X-ray radiation from a rotating-anode generator. These results will contribute to clarifying the haem transitions occurring during peroxidatic reaction and the required electron-transfer processes and to elucidating the catalytic mechanism of the enzyme and the role of calcium in the activation process.


First field application of a thermal desorption resonance-enhanced multiphoton-ionisation single particle time-of-flight mass spectrometer for the on-line detection of particle-bound polycyclic aromatic hydrocarbons.
The on-line analysis of single aerosol particles with mass spectrometrical methods is an important tool for the investigation of aerosols. Often, a single laser pulse is used for one-step laser desorption/ionisation of aerosol particles. Resulting ions are detected with time-of-flight mass spectrometry. With this method, the detection of inorganic compounds is possible. The detection of more fragile organic compounds and carbon clusters can be accomplished by separating the desorption and the ionisation in two steps, e.g. by using two laser pulses. A further method is, using a heated metal surface for thermal desorption of aerosol particles. If an ultraviolet laser is used for ionisation, a selective ionisation of polycyclic aromatic hydrocarbons (PAH) and alkylated PAH is possible via a resonance-enhanced multiphoton-ionisation process. Laser velocimetry allows individual laser triggering for single particles and additionally delivers information on aerodynamic particle diameters. It was shown that particles deriving from different combustion sources can be differentiated according to their PAH patterns. For example, retene, a C(4)-alkylated phenanthrene derivative, is a marker for the combustion of coniferous wood. In this paper, the first field application of a thermal desorption resonance-enhanced multiphoton-ionisation single particle time-of-flight mass spectrometer during a measurement campaign in Augsburg, Germany in winter 2010 is presented. Larger PAH-containing particles (i.e. with aerodynamic diameters larger than 1 μm), which are suspected to be originated by re-suspension processes of agglomerated material, were in the focus of the investigation. Due to the low concentration of these particles, an on-line virtual impactor enrichment system was used. The detection of particle-bound PAH in ambient particles in this larger size region was possible and in addition, retene could be detected on several particles, which allows to identify wood combustion as generic source of these particles. The observed diurnal distribution of these larger particles, however, support the origin by traffic induced re-suspension of sedimented/agglomerated material.


Measuring and monitoring radiation dose during fluoroscopically guided procedures.
The principal problem in measuring patient radiation dose during fluoroscopically guided procedures is that dose is not administered uniformly throughout the patient's body. Four dose metrics have been developed to quantify patient radiation dose for fluoroscopically guided procedures: fluoroscopy time, peak skin dose, reference dose, and kerma-area-product. Each metric must be understood to be used appropriately. Fluoroscopy time correlates poorly with other dose metrics. It should not be used as the sole method to estimate, monitor, or record patient radiation dose unless no alternative is available. Kerma-area-product is a good metric for estimating stochastic risk. Reference dose is a conservative method to estimate peak skin dose and deterministic risk and is recommended for this purpose. Every fluoroscope sold in the USA since mid 2006 is able to measure, display, and record reference dose. Radiation dose should be monitored during fluoroscopically guided procedures, either by the operator or by a designated individual in the procedure room, such as a technologist or nurse. Patient radiation dose should be recorded appropriately in the medical record. Patients who have received a sufficiently large radiation dose should have follow-up at 10-14 days and at 1 month after the procedure for possible deterministic effects.


[Molecular and cytogenetic identification of Triticum aestivum-Leymus racemosus translocation line T6DL·7LrS].
Leymus racemosus had a high resistant capacity to wheat scab (Fusarum head blight). The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the germplasm of wheat resistance. To obtain Triticum aestivum-Leymus racemosus translocation line with scab resistance, we irradiated the pollen of T. aestivum-L. racemosus disomic addition line DA7Lr by ⁶⁰Co-γ-rays 1 200 R (100 R/min) prior to pollinating to emasculation T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M1 by GISH. The plant with one translocation chromosome was self-pollinated, and at meiotic metaphase I its progenies with two translocation chromosomes were analyzed for chromosome pairing behavior in their pollen mother cells (PMCs). One rod bivalent was observed at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. Sequential GISH-FISH analysis, using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probe, translocation line was confirmed as T6DL·7LrS. The translocation line had higher resistance to wheat scab and feasibility to be used as a new source in wheat breeding resistant to scab disease.


Inhibition of blood-brain barrier disruption in experimental allergic encephalomyelitis by short-term therapy with dexamethasone or cyclosporin A.
Double radioisotope measurement of neurovascular integrity in Lewis rats inoculated for experimental allergic encephalomyelitis (EAE) showed abnormal elevation of albumin extravasation in the cerebellum, medulla-pons and cervical spinal cord at the time of clinical manifestation. Therapeutically administered dexamethasone (Dex) (0.1-1 mg/kg body weight) or cyclosporin A (CsA) (25-75 mg/kg body weight) dose-dependently reduced albumin movement across the blood-brain barrier (BBB). Dex at a dose of 1 mg/kg completely suppressed abnormal BBB permeability in all tissues (P < or = 0.001), while CsA at the highest dose of 75 mg/kg achieved highly significant (P < or = 0.001), but not complete, suppression of aberrant barrier leakage in the areas studied. The implications of these findings to possible drug action at the immunocompromised cerebrovasculature are discussed.


Cell Wall Free Space of Cucumis Hypocotyls Contains NAD and a Blue Light-Regulated Peroxidase Activity.
Solutions were obtained from the cell wall free space of red light-grown cucumber (Cucumis sativus L.) hypocotyl sections by a low-speed centrifugation technique. The centrifugate contained NAD and peroxidase but no detectable cytoplasmic contamination, as indicated by the absence of the activity of glucose-6-phosphate dehydrogenase from the cell wall solution. Peroxidase activity centrifuged from the cell wall of red light-grown cucumber hypocotyl section could be resolved into at least three cathodic isoforms and two anodic isoforms by isoelectric focusing. Treatment of red light-grown cucumber seedlings with a 10-minute pulse of high-intensity blue light increased the level of cell wall peroxidase by about 60% and caused a qualitative change in the anodic isoforms of this enzyme. The increase in peroxidase activity was detectable within 25 minutes after the start of the blue light pulse, was maximal at 35 minutes, and declined to control levels by 45 minutes of irradiation. The inhibitory effect of blue light on hypocotyl elongation was more rapid than the effect of blue light on total wall peroxidase activity, leading to the conclusion that growth and peroxidase activity are not causally related.


Female long-term survivors after allogeneic hematopoietic stem cell transplantation: evaluation and management.
Female long-term survivors of allogeneic hematopoietic stem cell transplantation (allo-HSCT) incur a significant burden of late effects. Genital graft-versus-host disease (GVHD), human papillomavirus (HPV) reactivation, ovarian failure and infertility, sexual dysfunction, and osteoporosis are concerns that can significantly impact quality of life. This review examines the risk, pathogenesis, clinical presentation, and implications of these common complications. Recommendations are provided for evaluation and management of these late effects and other obstetric and gynecologic issues that may arise in this patient population.