Improvement of poly(gamma-glutamic acid) biosynthesis and redistribution of metabolic flux with the presence of different additives in Bacillus subtilis CGMCC 0833.
Tween-80, dimethyl sulfoxide (DMSO), and glycerol could be used as novel materials to regulate the central carbon metabolic pathway and improve gamma-PGA biosynthesis by Bacillus subtilis CGMCC 0833. With glycerol in the medium, the activity of 2-oxoglutarate dehydrogenase complex at the key node of 2-oxoglutarate was depressed, more carbon flux distribution was directed to synthesize glutamate, the substrate of gamma-PGA, which led to overproducing of gamma-PGA, reached 31.7 g/l, compared to the original value of 26.7 g/l. When Tween-80 or DMSO was in the medium, the activity of isocitrate dehydrogenase was stimulated, the branch flux from 2-oxoglutarate to glutamate was also enhanced due to the increasing of total flux from iso-citrate to 2-oxoglutarate, then a large amount of glutamate was produced, and formation of gamma-PGA was also improved, which was a different process compared with that of glycerol. Moreover, with the addition of Tween-80 or DMSO, cell membrane permeability was increased, which facilitated the uptake of extracellular substrates and the secretion of gamma-PGA by this strain; therefore, gamma-PGA production was further stimulated, and 34.4 and 32.7 g/l gamma-PGA were obtained, respectively. This work firstly employed additives to improve the biosynthesis of gamma-PGA and would be helpful in understanding the biosynthesis mechanism of gamma-PGA by Bacillus species deeply.


Renal Squamous Cell Carcinoma Presented With Bone Metastasis and Coexistence With Xanthogranulomatous Pyelonephritis: A Case Report.
Squamous cell carcinoma (SCC) of the kidney is a rare entity. Coexistence of SCC with xanthogranulomatous pyelonephrits is exceedingly rare with only few reports in the literature. We report a case of a 45-year-old male patient with xanthogranulomatous pyelonephritis coexistence with renal SCC in one kidney, which proved radiologically and histopathologically. The patient presented to the medical care with bone metastasis. Full radiology workup is also provided which includes computed tomography, magnetic resonance imaging, and positron emission tomography-computed tomography.


Field Evaluation of Cholkit Rapid Diagnostic Test for Vibrio Cholerae O1 During a Cholera Outbreak in Malawi, 2018.
Rapid diagnostic tests (RDTs) for cholera are an important emerging tool for surveillance, yet the currently available tests have several limitations. We assess the performance of a new RDT, Cholkit, during a cholera outbreak in Malawi compared with culture and find a sensitivity of 93.0% (95% CI, 83.0%-98.1%) and a specificity of 95.7% (95% CI, 78.1%-100.0%).


Effects of genetic polymorphisms of CYP2J2, CYP2C9, CYP2C19, CYP4F2, CYP4F3 and CYP4A11 enzymes in preeclampsia and gestational hypertension.
The aim of this study was to investigate the effects of cytochrome P450 (CYP) 2J2, CYP2C9, CYP2C19 and CYP4F2, CYP4F3 and CYP4A11 genetic polymorphisms in preeclampsia and gestational hypertension (GHT) patients in a sample of Turkish population. Patients (n = 168; 110 GHT and 58 preeclampsia) and healthy pregnant women (n = 155, controls) participated in the study. For genotyping, polymerase chain reaction (PCR) and restriction analysis (RFLP) were used. Substance levels were measured using LC-MS. Plasma DHET levels in GHT and preeclampsia patients were significantly lower than those in the control group (62.7%, 66.3% vs.100.0%, respectively, p < 0.0001). An increase in CYP2J2*7 allele frequency was observed in the preeclampsia group, as compared to GHT group (12.1% vs. 4.5%; odds ratio, O.R. = 2.88, p < 0.01). The frequencies of CYP2C19*2 and*17 alleles were higher in GHT group as compared to the control group (17.7% vs. 11.6%, O.R. = 1.99, p < 0.01; and 28.6% vs.18.4%, O.R. = 2.03, p < 0.01, respectively). An increased frequency of CYP4F3 rs3794987 G allele was found in GHT group as compared to the control group (48.0% vs. 38.0%; O.R. = 1.53, p < 0.01). DHET plasma levels were significantly reduced in hypertensive pregnant groups as compared to the control group. The allele frequency distributions for CYP2J2*7, CYP2C19 *2, *17 and CYP4F3 rs3794987 were significantly different in hypertensive pregnant patients as compared to the healthy control subjects. Our results may suggest that investigated genetic polymorphisms may be useful in diagnosis and clinical management of GHT and preeclampsia patients.


Initial Clinical Experience Treating Patients with Breast Cancer on a 6-MV Flattening-Filter-Free O-Ring Linear Accelerator.
Three-dimensional (3D) conformal radiation therapy is the standard technique used for adjuvant breast radiation. We report the clinical use of a novel 6-MV flattening-filter-free O-ring linear accelerator (6X-FFF ORL) for breast cancer that may improve upon 3D conformal radiation therapy with its higher dose rate and faster rotation and leaf speed than traditional C-arm gantries. We retrospectively identified consecutive women with breast cancer who underwent surgery followed by radiation therapy to the breast or chest wall on Halcyon (Varian Medical Systems, Palo Alto, CA), a novel 6X-FFF ORL. We report their clinicopathologic information, radiation therapy details, acute toxicities, dose-volume histogram data, couch corrections, and treatment times. Thirty-four women were treated for breast cancer on a 6X-FFF ORL between February 2018 and September 2018. All patients underwent lumpectomy (92%) or mastectomy (8%). Tumors were left sided in 44% and bilateral in 9%, and 9% included comprehensive nodal radiation therapy. Twelve percent of patients were treated prone and 29% with deep-inspiration breath hold. Standard target and normal-tissue constraints were met in nearly all plans. The 3D vector couch correction average was 0.77 ± 0.05 cm. The mean beam-on time was 2.0 ± 0.3 minutes, and mean treatment time from start of imaging to beam-off was 4.4 ± 0.4 minutes. Grade 2 dermatitis, fatigue, and breast pain occurred in 18%, 9%, and 3% of patients, respectively. In this first clinical report of breast radiation therapy with a 6X-FFF ORL, treatment was versatile and fast for complex setups and techniques, with acceptable toxicity and organ-at-risk doses. Thus, a 6X-FFF ORL can increase throughput or reduce length of day compared with a conventional C-arm linear accelerator in departments with a busy breast service.


Long-term patency of arteriovenous fistulas for hemodialysis: A decade's experience in a transplant unit.
The heterogeneous quality of studies on arteriovenous fistulas outcome, with variable clinical settings and large variations in definitions of patency and failure rates, leads to frequent misinterpretations and overestimation of arteriovenous fistula patency. Hence, this study aimed to provide realistic and clinically relevant long-term arteriovenous fistula outcomes. We retrospectively analyzed all autologous arteriovenous fistulas at our center over a 10-year period (2012-2022). Primary and secondary patency analysis was conducted using the Kaplan-Meier method; multivariate analysis of variance was used to detect outcome predictors. Vascular access-specific endpoints were defined according to the European guidelines on vascular access formation. Of 312 arteriovenous fistulas, 57.5% (n = 181) were radio-cephalic (RC_AVF), 35.2% (n = 111) brachio-cephalic (BC_AVF), and 6.3% (n = 20) brachio-basilic (BB_AVF). 6, 12, and 24 months follow-up was available in 290 (92.1%), 282 (89.5%), and 259 (82.2%) patients, respectively. Primary patency rates at 6, 12, and 24 months were 39.5%, 34.8%, and 27.2% for RC_AVF, 58.3%, 44.4%, and 27.8% for BC_AVF, and 40.0%, 42.1%, and 22.2% for BB_AVF (p = 0.15). Secondary patency rates at 6, 12, and 24 months were 65.7%, 63.8%, and 59.0% for RC_AVF, 77.7%, 72.0%, and 59.6% for BC_AVF, and 65.0%, 68.4%, and 61.1% for BB_AVF (p = 0.29). Factors associated with lower primary and secondary patency were hemodialysis at time of arteriovenous fistula formation (p = 0.037 and p = 0.024, respectively) and higher Charlson Comorbidity Index (p = 0.036 and p < 0.001, respectively). Previous kidney transplant showed inferior primary patency (p = 0.005); higher age inferior secondary patency (p < 0.001). Vascular access care remains challenging and salvage interventions are often needed to achieve maturation or maintain patency. Strict adherence to standardized outcome reporting in vascular access surgery paints a more realistic picture of arteriovenous fistula patency and enables reliable intercenter comparison.


A Retrospective Chart Review of Patients With Reactive Rapid Plasma Reagin Tests in the Emergency Department of an Urban Safety Net Hospital.
Emergency departments are a critical access point to care in safety net health systems. A retrospective chart review of patients with reactive rapid plasma reagins collected in the emergency department of an Atlanta safety net hospital from May 1, 2020, to October 31, 2020, found suboptimal rates of syphilis treatment completion and linkage to human immunodeficiency virus pre-exposure prophylaxis services.


Design, synthesis and biological evaluation of benzylisoquinoline derivatives as multifunctional agents against Alzheimer's disease.
A novel series of benzylisoquinoline derivatives were designed, synthesized, and evaluated as multifunctional agents against Alzheimer's disease (AD). The screening results showed that most of the compounds significantly inhibited cholinesterases (ChEs), human cholinesterases (h-ChEs) and self-induced β-amyloid (Aβ) aggregation. In particular, compound 9k showed the strongest acetylcholinesterase (AChE) inhibitory activity, being 1000-fold and 3-fold more potent than its precursor benzylisoquinoline (10) and the positive control galanthamine, respectively. In addition, 9k was a moderately potent inhibitor for h-ChEs. Compared with precursor benzylisoquinoline (36.0% at 20μМ), 9k (78.4% at 20μМ) could further inhibit Aβ aggregation. Moreover, 9k showed low cell toxicity in human SH-SY5Y neuroblastoma cells. Therefore, compound 9k might be a promising lead compound for AD treatment.


Dereplication by HPLC-DAD-ESI-MS/MS and Screening for Biological Activities of Byrsonima Species (Malpighiaceae).
Byrsonima species have been used in the treatment of gastrointestinal and gynecological inflammations, skin infections and snakebites. Based on their biological activities, it is important to study other organisms from this genus and to identify their metabolites. To determine the metabolic fingerprinting of methanol and ethyl acetate extracts of four Byrsonima species (B. intermedia, B. coccolobifolia, B. verbascifolia and B. sericea) by HPLC-DAD-ESI-MS/MS and evaluate their in vitro antioxidant, anti-glycation, anti-inflammatory and cytotoxic activities. Antioxidant activity was determined by DPPH˙, ABTS˙+ and ROO˙ scavenging assays. Anti-glycation activity was evaluated by the ability to inhibit the formation of advanced glycation endproducts (AGEs). Anti-inflammatory activity was evaluated using a murine macrophage cell line (RAW 264-7) in the presence of lipopolysaccharide (LPS). Tumour necrosis factor alpha (TNF-α) and nitrite (NO2- ) production were measured by ELISA and the Griess reaction, respectively. The compounds present in the extracts were tentatively identified by HPLC-DAD-ESI-MS/MS. The evaluation of the biological activities showed the potential of the extracts. The activities were assigned to the presence of glycoside flavonoids mainly derived from quercetin, quinic acid derivatives, gallic acid derivatives, galloylquinic acids and proanthocyanidins. Two isomers of sinapic acid-O-hexoside were described for the first time in a Byrsonima species. This research contributes to the study of the genus, it is the first report of the chemical composition of B. sericea and demonstrates the importance of the dereplication process, allowing the identification of known compounds without time-consuming procedures. Copyright © 2017 John Wiley & Sons, Ltd.


High-performance liquid chromatography with electrochemical detection for the simultaneous determination of the methoxylated amines, normetanephrine, metanephrine and 3-methoxytyramine, in urine.
A simple method for the simultaneous analysis of normetanephrine, metanephrine and 3-methoxytyramine (both free and conjugated) in human urine by reversed-phase ion-pair high-performance liquid chromatography with electrochemical detection has been developed. Existing methods have been optimized for extraction by study of analytical parameters. The hydrolysed urines are purified and concentrated by successive passages on two ion-exchange resins and ammoniacal elution to eliminate interference from pigments or related chemical compounds. The methoxyamines are separated by high-performance liquid chromatography on a reversed-phase column. Detection and quantitation are achieved with an electrochemical detector using a vitreous carbon electrode. Samples can be injected at 25-min intervals. Reference values of adults and children are given.


[Research on the annual periodicity of carabid beetles in the Cyrenaika (North Africa)].
In a 30 km deep coastal area of the Cyrenaika (Libya) a total of 28 species of carabids was collected on regular excursions in 1967 and 1968. In midsummer almost no carabids were caught. They appeared at the beginning of autumn and were caught in winter regularly in equal numbers. During March, April and May the number of catches increased considerably. Using ground traps no more than two species (Carterus silvestrii, Pristonychus picicornis) were caught and that only from December to May. From the pattern of their occurrence we can conclude that in the Cyrenaika carabid beetles propagate mainly during the winter.By examining the gonads, catching larvae and breeding, it was possible to prove that the speciesBroscus laevigatus, Harpalus litigiosus ssp.cyrenaicus andPristonychus picicornis propagate in winter. Examination of the gonads also indicates a winter propagation of the speciesPterostichus barbarus andLaemostenus complanatus ssp.cyrenaicus. It is not certain, however, thatCarterus silvestrii propagates during the winter.The speciesCalathus fuscipes andCalathus mollis from the temperate northern regions, which are known as autumn breeders, were caught in Libya too, which indicates a relation between the autumn breeders of Middle and Northern Europe and the 'winter breeders' of the South Mediterranean countries. The presence ofPogonus chalceus in Libya strongly suggests that, in certain biotopes, spring breeders may develop besides the dominant autumn breeders.


Transcatheter Aortic Valve Replacement as a Rescue Treatment for Prosthetic Valve Endocarditis.
A 74-year-old man with a history of bioprosthetic aortic valve replacement presented with acute severe aortic insufficiency and cardiogenic shock secondary to Diphtheroides and Enterococcus endocarditis. The patient was deemed to be not a surgical candidate by the multidisciplinary team and underwent rescue transcatheter aortic valve-in-valve replacement, resulting in complete recovery. (Level of Difficulty: Advanced.).


Mid-infrared III-V semiconductor lasers epitaxially grown on Si substrates.
There is currently much activity toward the integration of mid-infrared semiconductor lasers on Si substrates for developing a variety of smart, compact, sensors based on Si-photonics integrated circuits. We review this rapidly-evolving research field, focusing on the epitaxial integration of antimonide lasers, the only technology covering the whole mid-to-far-infrared spectral range. We explain how a dedicated molecular-beam epitaxy strategy allows for achieving high-performance GaSb-based diode lasers, InAs/AlSb quantum cascade lasers, and InAs/GaInSb interband cascade lasers by direct growth on on-axis (001)Si substrates, whereas GaAs-on-Si or GaSb-on-Si layers grown by metal-organic vapor phase epitaxy in large capability epitaxy tools are suitable templates for antimonide laser overgrowth. We also show that etching the facets of antimonide lasers grown on Si is a viable approach in view of photonic integrated circuits. Remarkably, this review shows that while diode lasers are sensitive to residual crystal defects, the quantum cascade and interband cascade lasers grown on Si exhibit performances comparable to those of similar devices grown on their native substrates, due to their particular band structures and radiative recombination channels. Long device lifetimes have been extrapolated for interband cascade lasers. Finally, routes to be further explored are also presented.


Analyzing the effects of hydrophobic mismatch on transmembrane α-helices using tryptophan fluorescence spectroscopy.
Hydrophobic matching between transmembrane protein segments and the lipid bilayer in which they are embedded is a significant factor in the behavior and orientation of such transmembrane segments. The condition of hydrophobic mismatch occurs when the hydrophobic thickness of a lipid bilayer is significantly different than the length of the membrane spanning segment of a protein, resulting in a mismatch. This mismatch can result in altered function of proteins as well as nonnative structural arrangements including effects on transmembrane α-helix tilt angles, oligomerization state, and/or the formation of non-transmembrane topographies. Here, a fluorescence-based protocol is described for testing model transmembrane α-helices and their sensitivity to hydrophobic mismatch by measuring the propensity of these helices to form non-transmembrane structures. Overall, good hydrophobic matching between the bilayer and transmembrane segments is an important factor that must be considered when designing membrane proteins or peptides.


Relation between lower limb comfort and performance in elite footballers.
Successes of the professional athlete as individuals and team, across codes of football are contingent upon performance measures. The aim of the study was to examine associations between comfort and performance. Prospective measures of lower limb comfort and coach rating performance criterion. Comfort and performance associations were described using Pearson's correlation coefficient (r) or the R square value from the regression estimate. Two professional football codes in Australia. 79 professional footballers were followed for one football season, a total of 1724 player weeks. Study hypothesis was poor lower limb comfort is negatively correlated with good match day rated performance. Aspects of validity and responsiveness to change tested the hypothesis that lower limb comfort, affects rated performance. A validated lower limb comfort index (LLCI) was used to test comfort. Rated performance was evaluated by subjective rating criterion of experienced team coaches. Poor lower limb comfort and good match day ratings were not well correlated (R(2) = 0.25, P ≤ 0.001) and usual-high comfort was correlated with usual-good performance (R(2) = 0.69, P ≤ 0.001). Lower limb comfort may be a sensitive measure of rated performance in football. The LLCI is the first measurement tool to show association between comfort and rated performance.


Unusual leiomyoblastoma of the stomach.
Only about 250 cases of benign gastric leiomyoblastoma of the stomach have been reported. Most occur in the smooth muscle of the antrum, and may be intramural, intraluminal, or exogastric in position. A pedunculated gastric leiomyoblastoma occurs less frequently, and those containing radiographically identifiable calcifications are extremely rare.


Rapid method demonstration project at four New Jersey marine beaches using real time quantitative Polymerase Chain Reaction (qPCR).
Real time quantitative Polymerase Chain Reaction (qPCR) was used at four marine bathing beaches in New Jersey as part of a demonstration project to evaluate the potential for use of qPCR as part of a routine beach monitoring program. Split sample analyses for Enterococcus spp. using membrane filtration (MF) and qPCR were performed for 11weeks during the summer of 2011 using swimming advisories based on qPCR results. Comparison of qPCR and MF results from split samples indicated that there was an 82% overall agreement rate between the two methods. Results from the qPCR tests were available by noon the same day of sample collection and swimming advisories were posted on a dedicated website. The qPCR method can be more labor intensive and requires a higher level of training to perform, however, qPCR was able to assess beach water quality in a timelier manner compared to conventional MF techniques.


Development of a DNA-binding TEMPO derivative for evaluation of nuclear oxidative stress and its application in living cells.
Oxidative stress in nuclei is known to induce either oxidative modification of DNA bases or single/double-strand breaks, which may lead to carcinogenesis. To evaluate the redox status in nuclei in living cells, we designed a novel nucleus-localizing redox spin probe, F-DisT, which contains a fluorescein fluorophore linked to a DNA minor-groove-binding moiety. Nuclear distribution of the probe was easily confirmed by colocalization with a nuclear stain, Hoechst 33342, in confocal microscopy. Measurement of oxidative stress with F-DisT in a murine macrophage cell line exposed to endotoxin (lipopolysaccharide) showed a remarkable increase in the ESR signal decay rate. This increase was significantly inhibited by N(ω)-nitro-l-arginine (nitric oxide synthase inhibitor) and diphenyleneiodonium chloride (NADPH oxidase inhibitor). These results indicate that nitric oxide and superoxide contribute to oxidative stress in nuclei. Similar studies in membrane or mitochondria using respective organelle-specific spin probes indicated that the redox microenvironments in these organelles are markedly different from that in nuclei. Thus, subcellular redox microenvironments show marked variability in endotoxin-stimulated living cells.


Clinicopathologic differentiation of atrophy of the pancreatic body and tail aplasia.
Congenital aplasia of the body and tail of the pancreas is derived from a defect of the dorsal pancreatic anlage and should not be considered a type of acquired atrophy of these structures. Congenital aplasia of the body and tail of the pancreas radiologically mimics acquired atrophy of the pancreatic body and tail. Two patients with radiologically identified aplasia of the body and tail of the pancreas were studied clinicopathologically. An 82-yr-old man was diagnosed radiologically as having both carcinoma of the head of the pancreas and aplasia of the body and tail of the pancreas and underwent pancreatoduodenectomy. Pathologically the carcinoma was distributed in the anterosuperior part of the head of the pancreas, and spread into the duct of Santorini and intraductally to a portion of the main pancreatic duct beyond the junction of the ducts of Santorini and Wirsung. Consequently, obstructive pancreatitis of the body and tail of the pancreas developed, resulting in marked atrophy that mimicked aplasia of the body and tail of the pancreas. A 74-yr-old woman was diagnosed radiologically as having two carcinomas, one of the gallbladder and one of the stomach, and aplasia of the body and tail of the pancreas. During surgery, suspected parenchymal disappearance and fatty replacement in the body and tail of the pancreas were noted. Histologic examination of biopsy specimens from the body portion revealed atrophic pancreatic tissue surrounded by fat. Therefore, these patients had atrophy of the pancreatic body and tail.


The effects of mono-(2-ethylhexyl)-phthalate on rat Sertoli cell-enriched primary cultures.
There is considerable evidence from in vivo studies that the Sertoli cell is an initial target cell for the actions of phthalates in the rodent testis. Because this metabolically active cell type plays a central role in spermatogenesis, we examined the effects of a toxic phthalate, mono-(2-ethylhexyl)-phthalate (MEHP), on the secretory and synthetic activities of primary testicular cell cultures isolated from 18-day-old rats. These cultures were 78-84% Sertoli cells. Exposure to MEHP decreased cellular ATP by ca. 20%, decreased production of radiolabeled 14CO2 from acetate, and decreased media levels of pyruvate, while it increased media levels of lactate and intracellular lipid. Protein synthesis, evaluated by radiolabeled leucine incorporation, was not affected by MEHP. Mitochondrial succinate dehydrogenase activity was decreased in the presence of MEHP. Michaelis-Menton kinetic analysis indicated this was a mixed inhibition. There was no apparent change in mitochondrial Rhodamine 123 uptake. These data are consistent with the concept that mitochondria are one target for the actions of MEHP in the Sertoli cell.


Motivations, aims and communication around advance directives: a mixed-methods study into the perspective of their owners and the influence of a current illness.
What are motivations of owners of an advance directive (AD) to draft an AD, what do they aim for with their AD and do they communicate about their AD? Written questionnaires were sent to a cohort of people owning different types of ADs (n=5768). A purposive sample of people suffering from an illness was selected from the cohort for an in-depth interview (n=29). About half of our population had no direct motivation to draft their AD. Most mentioned motivation for the other half was an illness of a family member or friend. Many different and specific aims for drafting an AD were mentioned. An often mentioned more general aim in people with different ADs was to prevent unnecessary lengthening of life or treatment (14-16%). Most respondents communicated about having an AD with close-ones (63-88%) and with their GP (65-79%). In the interviews people gave vivid examples of experiences of what they hoped to prevent at the end of life. Some mentioned difficulties foreseeing the future and gave examples of response shift. ADs can give directions to caregivers about what people want at the end of life. ADs have to be discussed in detail by their owners and caregivers, since owners often have specific aims with their AD.


Transtympanic neurectomy: a solution to drooling problems.
Thirty patients underwent transtympanic neurectomy because of severe drooling problems. Most of the group had cerebral palsy and/or mental retardation and the remainder had suffered brain damage. Their ages ranged from four to 56 years. There was improvement in drooling in 87 per cent of the group 14 months post-operatively, and there was no serious, irreversible complications. In this series of patients, transtympanic neurectomy has proved to be a safe and effective procedure for the control of drooling.


Application of a cognitive neuroscience perspective of cognitive control to late-life anxiety.
Recent evidence supports a negative association between anxiety and cognitive control. Given age-related reductions in some cognitive abilities and the relation of late life anxiety to cognitive impairment, this negative association may be particularly relevant to older adults. This critical review conceptualizes anxiety and cognitive control from cognitive neuroscience and cognitive aging theoretical perspectives and evaluates the methodological approaches and measures used to assess cognitive control. Consistent with behavioral investigations of young adults, the studies reviewed implicate specific and potentially negative effects of anxiety on cognitive control processes in older adults. Hypotheses regarding the role of both aging and anxiety on cognitive control, the bi-directionality between anxiety and cognitive control, and the potential for specific symptoms of anxiety (particularly worry) to mediate this association, are specified and discussed.


Gut microbiota Turicibacter strains differentially modify bile acids and host lipids.
Bacteria from the Turicibacter genus are prominent members of the mammalian gut microbiota and correlate with alterations in dietary fat and body weight, but the specific connections between these symbionts and host physiology are poorly understood. To address this knowledge gap, we characterize a diverse set of mouse- and human-derived Turicibacter isolates, and find they group into clades that differ in their transformations of specific bile acids. We identify Turicibacter bile salt hydrolases that confer strain-specific differences in bile deconjugation. Using male and female gnotobiotic mice, we find colonization with individual Turicibacter strains leads to changes in host bile acid profiles, generally aligning with those produced in vitro. Further, colonizing mice with another bacterium exogenously expressing bile-modifying genes from Turicibacter strains decreases serum cholesterol, triglycerides, and adipose tissue mass. This identifies genes that enable Turicibacter strains to modify host bile acids and lipid metabolism, and positions Turicibacter bacteria as modulators of host fat biology.


Efficacy and safety of pembrolizumab monotherapy in EGFR-mutant squamous cell lung cancer with PD-L1 over-expression: A case report.
Epidermal growth factor receptor (EGFR)-mutant nonsmall cell lung cancer (NSCLC) patients are less likely to be programmed death-ligand 1 (PD-L1)-positive compared with wild-type EGFR mutant tumors. Given the rarity of actionable driver genes in squamous cell lung cancer (SQCC), the frequency of SQCC patients simultaneously carrying EGFR driver gene mutation and having PD-L1 over-expression is extremely low. Studies on the effectiveness and safety of EGFR-TKIs or immune-checkpoint inhibitors (ICIs) in this subset of patients are lacking. The patient suffered from coughing and chest pain for 1 month. A chest CT revealed a mass with a cavity in the right lung, enlarged mediastinal lymph nodes, diffuse pleural thickening in the right pleura, and pleural effusion of the right chest. A pleural biopsy was performed using a video-assisted thoracoscope. The pathological examination revealed a poorly differentiated squamous cell carcinoma of lung. Further genetic testing identified exon 19 deletion mutation in EGFR with abundance of 0.27%. Meanwhile, immunohistochemical PD-L1 analysis showed a TPS of 90%. The patient was initially resistant to EGFR-TKIs but exhibited a rapid and marked response to pembrolizumab. After 5 cycles of pembrolizumab monotherapy, the patient developed Grade 3 immune-related dermatitis, and ICI therapy was suspended. ICI monotherapy could be an effective therapy in SQCC patients with low-abundance of EGFR mutations and PD-L1 over-expression. However, close attention should be paid to immune-related adverse events.


Relationship between accumulation, storage and overflow of noradrenaline in the rat salivary gland after chronic treatment with guanethidine.
1 The effect of guanethidine on the endogenous noradrenaline (NA) content, accumulation and overflow of [3H]-noradrenaline ([3H]-NA) in the rat salivary gland was examined at various times after drug administration. 2 Twenty-four h after a single injection of guanethidine (1 or 10 mg/kg s.c.), the respective values for the endogenous NA content and for the accumulation and overflow of [3H]-NA were approximately 55, 85, 30%, and 15, 55, 10% of the controls. 3 Although [3H]-NA accumulation had returned to control levels within 48 h after the dose of 10 mg/kg guanethidine, the overflow of [3H]-NA evoked by electrical stimulation or excess potassium (K+) remained depressed. 4 After the low or the high dose of guanethidine, the NA content of the salivary gland was restored to about 50% of the normal value between 4 to 24 and 48 to 72 h, respectively. 5 The accumulation of [3H]-NA was inhibited by about 75% by cocaine. The same degree of inhibition was obtained 4 h after 10 mg/kg guanethidine. In these experiments phenoxybenzamine did not reduce the residual (25%) uptake. 6 The reasons for differential rates of recovery of the endogenous NA content and the storage of [3H]-NA after guanethidine are discussed.


Cortical Bone Trajectory-Based Dynamic Stabilization.
Cortical bone trajectory (CBT) screws have demonstrated similar biomechanical strength and fusion rates as conventional pedicle screws for short-segment lumbar fusion. However, very few studies have verified the viability of CBT screws in dynamic stabilization. In the present study, we compared the clinical outcomes of CBT-based Dynesys dynamic stabilization (CBT-DDS) with standard minimally invasive transforaminal lumbar interbody fusion (MI-TLIF). Consecutive patients who had undergone CBT-DDS or MI-TLIF at L4-L5 or L3-L5 were retrospectively reviewed. All the radiological and clinical outcomes were compared between the 2 groups. The patient-reported outcomes included the visual analog scale scores for back and leg pain, Japanese Orthopaedic Association scores, and Oswestry disability index. The pre- and postoperative radiological evaluations were also reviewed. A total of 60 patients (20 patients in the CBT-DDS and 40 in the MI-TLIF groups) were analyzed. The MI-TLIF group had required a longer operation time (P = 0.010) than the CBT-DDS group but had had a similar estimated blood loss (P = 0.484). The perioperative complications, including screw loosening, wound infection, and radiculopathy, were similar in both groups (P > 0.05 for all). The degree of decreased range of motion after surgery was similar between the 2 groups (P = 0.781), and no pseudarthrosis developed in the MI-TLIF group. Because the clinical and radiological outcomes of CBT-DDS were similar to those of MI-TLIF in patients with L4-L5 or L3-L5 spondylosis and spondylolisthesis, CBT-DDS appears to be a viable and effective alternative to MI-TLIF, with a shorter operation time and similarly limited segmental motility.


Trends of Sex Differences in Outcomes of Cardiac Electronic Device Implantations in the United States.
The disparity in outcomes of cardiac electronic device implantations between sexes has been previously demonstrated in device-specific cohorts (eg, implantable cardioverter-defibrillators [ICDs]). However, it is unclear whether sex differences are present with all types of cardiac implantable electronic devices (CIEDs) and, if so, what the trends of such differences have been in recent years. With the use of the National Inpatient Sample, all hospitalizations from 2004 to 2014 for de novo implantation of permanent pacemakers, cardiac resynchronization therapy with or without a defibrillator, and ICDs were analyzed to examine the association between sex and in-hospital acute complications of CIED implantation. Out of 2,815,613 hospitalizations for de novo CIED implantation, 41.9% were performed on women. Women were associated with increased adjusted odds (95% confidence interval) of adverse procedural complications (major adverse cardiovascular complications: 1.17 [1.16-1.19]; bleeding: 1.13 [1.12-1.15],-thoracic: 1.42 [1.40-1.44]; cardiac: 1.44 [1.38-1.50]), whereas the adjusted odds of in-hospital all-cause mortality compared with men was 0.96 (0.94-1.00). The odds of adverse complications in the overall CIED cohort were persistently raised in women throughout the study period, whereas similar odds of all-cause mortality across the sexes were observed throughout the study period. In a national cohort of CIED implantations we demonstrate that women are at an overall higher risk of procedure-related adverse events compared with men, but not at increased risk of all-cause mortality. Further studies are required to identify procedural techniques that would improve outcomes among women undergoing such procedures.


Digital workflow for image-guided immediate implant placement by using the socket-shield technique and custom abutment in the esthetic area.
Marginal bone loss around immediately placed implants may compromise esthetic outcomes in the anterior maxilla. The relatively recent socket-shield technique of partial extraction therapy has been described as being useful to maintain the structure of peri-implant tissues. The present technique report introduced a digital workflow to perform image-guided implant placement after partial extraction therapy. In this technique, digital 3-dimensional images are used to plan and orient the flapless surgical procedure and to digitally design a custom abutment based on the natural emergence profile as segmented from the tooth to be extracted.


The neurophysiological basis of reward effects on backward inhibition processes.
The ability to flexibly switch between tasks is an important faculty in daily life. One process that has been suggested to be an important aspect of flexible task switching is the inhibition of a recently performed task. This is called backward inhibition. Several studies suggest that task switching performance can be enhanced by rewards. However, it is less clear in how far backward inhibition mechanisms are also affected by rewards, especially when it comes to the neuronal mechanisms underlying reward-related modulations of backward inhibition. We therefore investigated this using a system neurophysiological approach combining EEG recordings with source localization techniques. We demonstrate that rewards reduce the strength of backward inhibition processes. The neurophysiological data shows that these reward-related effects emerge from response and/or conflict monitoring processes within medial frontal cortical structures. Upstream processes of perceptual gating and attentional selection, as well as downstream processes of context updating and stimulus-response mapping are not modulated by reward, even though they also play a role in backward inhibition effects.


Circulating cell-specific extracellular vesicles as biomarkers for the diagnosis and monitoring of chronic liver diseases.
Chronic liver diseases represent a burgeoning health problem affecting billions of people worldwide. The insufficient performance of current minimally invasive tools is recognised as a significant barrier to the clinical management of these conditions. Extracellular vesicles (EVs) have emerged as a rich source of circulating biomarkers closely linked to pathological processes in originating tissues. Here, we summarise the contribution of EVs to normal liver function and to chronic liver pathologies; and explore the use of circulating EV biomarkers, with a particular focus on techniques to isolate and analyse cell- or tissue-specific EVs. Such approaches present a novel strategy to inform disease status and monitor changes in response to treatment in a minimally invasive manner. Emerging technologies that support the selective isolation and analysis of circulating EVs derived only from hepatic cells, have driven recent advancements in EV-based biomarker platforms for chronic liver diseases and show promise to bring these techniques to clinical settings.


Amino acid sequences of the human kidney cathepsins H and L.
The complete amino acid sequences of human kidney cathepsin H (EC 18.104.22.168) and human kidney cathepsin L (EC 22.214.171.124) were determined. Cathepsin H contains 230 residues and has an Mr of 25116. The sequence was obtained by sequencing the light, heavy and mini chain and the peptides produced by cyanogen bromide cleavage of the single-chain form of the enzyme. The glycosylated mini chain is a proteolytic fragment of the propeptide of cathepsin H. Human cathepsin L has 217 amino acid residues and an Mr of 23720. Its amino acid sequence was deduced from N-terminal sequences of the heavy and light chains and from the sequences of cyanogen bromide fragments of the heavy chain. The fragments were aligned by comparison with known sequences of cathepsins H and L from other species. Cathepsins H and L exhibit a high degree of sequence homology to cathepsin B (EC 188.8.131.52) and other cysteine proteinases of the papain superfamily.


Supportive care in neurooncology.
The quality of life of patients treated for brain tumor is, in all cases, deeply altered by the tumor and the treatments. Optimizing the symptomatic management is a key objective for all care givers. We present in this paper a very pragmatic focus concerning the management of intracranial hypertension (and/or neurological deficits), venous thromboembolism, confusion, epilepsy and symptoms more directly associated with the end of life.


T-cell repertoire complexity after allogeneic bone marrow transplantation.
We analyzed the T-cell repertoire in patients transplanted with bone marrow from an HLA identical sibling by determining the TCR diversity through Vbeta-CDR3-size spectratyping with Vbeta/Cbeta- and Vbeta/Jbeta-specific primers. Using the Vbeta/Cbeta primers, we observed limited TCR diversity only in recipients of a T-cell-depleted graft, whereas the TCR diversity of patients transplanted with an unmanipulated graft seemed to be indistinguishable from the one of a normal individual. However, with Vbeta/Jbeta-specific primers, increase of the resolution by approximately 10-fold also allowed the detection of imbalances in the TCR repertoire of recipients of an unmanipulated graft. This demonstrates that when high numbers of T cells are cotransfused with marrow, the TCR repertoire is more complete but still not as complete as in normal individuals, thereby emphasizing the important role of coinfused mature T cells in the restoration of the T-cell compartment after bone marrow transplantation.


Dermatology leadership and a top-down approach to increasing diversity.
The reasons underlying the lack of diversity within dermatology can be broadly categorized into lack of mentorship, decreased awareness of the specialty during medical school, socioeconomic barriers associated with the application process, and implicit bias during resident selection. This contribution examines the need for diversity in medicine and provides insight into the reasons behind the low number of underrepresented minority residents in dermatology. Leadership strategies that may help increase diversity in the field are also reviewed.


Serum responsiveness of the rat PCNA promoter involves the proximal ATF and AP-1 sites.
We have previously shown that the rat PCNA (proliferating cell nuclear antigen) gene promoter is responsive to serum stimulation. In this study, the sequence of the promoter responsive to serum stimulation has been localized in the region between nucleotides -70 and +125 relative to the transcription initiation site. This region contains an ATF site (nucleotides -51 to -44) and an AP-1 site (nucleotides -64 to -58). Mutation at either the ATF or the AP-1 site reduced the serum responsiveness of the promoter. In gel mobility shift assays, nuclear extracts from serum stimulated cells, compared to those from quiescent cells, exhibit an increasing binding activity toward a promoter related oligonucleotide (-70 to -42) which includes the ATF site and the AP-1 site. Formation of the DNA:protein complexes requires the simultaneous involvement of ATF and AP-1 sites as either element can abrogate the complexes in the competition experiment. Both the distance and sequence are essential to complex formation. Moreover, ATF-1 but not ATF-2 (or CREB) has been identified as a major component of the complexes in the antibody supershift or interference experiment. The results of this study suggest that ATF-1 in association with other factors is involved in regulating the serum stimulation of the rat PCNA promoter activity via the proximal ATF and AP-1 sites.


Sustained inhibition of antigen-induced histamine release from human lung by the beta 2-adrenoceptor agonist terbutaline.
Antigen-induced histamine release from passively sensitized human lung tissue was inhibited in the presence of the beta 2-adrenoceptor agonist, terbutaline. A sustained and statistically significant suppression was detected in the concentration interval 3 X 10(-8)-1 X 10(-6) M. Fifty per cent inhibition IC50, was obtained at an interpolated concentration of 5.3 X 10(-8) +/- 0.4 X 10(-8) M (n = 13), when the histamine secretion was elicited with optimum concentration of antigen. Histamine release induced with a suboptimum concentration of antigen was inhibited to a greater extent than release initiated with optimum concentration. The data in the present investigation support the concept that terbutaline-induced inhibition of mediator release from human lung tissue can contribute to the clinical effectiveness of the drug during treatment of allergic asthma.


Mitochondrial DNA A3243G mutation involved in familial diabetes, chronic intestinal pseudo-obstruction and recurrent pancreatitis.
To report on a family with five members who carry the A3243G mutation in mitochondrial tRNA for leucine 1 (MTTL1) and present with diabetes, chronic intestinal pseudo-obstruction (CIPO) and recurrent pancreatitis, and to screen for this mutation in a cohort of 36 unrelated patients with recurrent pancreatitis. The mutation was quantified in several tissue samples from patients. Respiratory chain activity was studied in muscle biopsies and fibroblast cultures. In addition, the thymidine phosphorylase gene (TP) involved in mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) and three genes involved in chronic pancreatitis - PRSS1, SPINK1 and CFTR - were sequenced in affected patients. Finally, the MTTL1 gene was examined in 36 unrelated patients who had recurrent pancreatitis, but no mutations in the PRSS1 and SPINK1 genes. Heteroplasmy for the mtDNA A3243G mutation was found in all tissue samples from these patients, but no mutations were found in the genes coding for thymidine phosphorylase, PRSS1, SPINK1 and CFTR. Also, none of the 36 unrelated patients with recurrent pancreatitis were carrying any MTTL1 mutations. The mtDNA A3243G mutation associated with the gastrointestinal manifestations observed in the affected family should be regarded as a possible cause of CIPO and unexplained recurrent pancreatitis. However, the mutation is probably only weakly involved in cases of isolated recurrent pancreatitis.


The impact of sensory impairment on cognitive performance, quality of life, depression, and loneliness in older adults.
The objective of this study was to evaluate the relationship between sensory impairment (hearing loss only, vision loss only, and dual sensory impairment [DSI]) and depression, loneliness, quality of life, and cognitive performance in older adults. A total of 229 community-dwelling older adults aged 60 years or older participated in this study. Variables were measured using the Geriatric Depression Scale (GDS-15), Revised University of California at Los Angeles Loneliness Scale (R-UCLA), Satisfaction with Life Scale (SWLS), and Mini-Mental State Examination (MMSE). There was an independent association between DSI and quality of life (P < .05) and between DSI and hearing loss alone and cognitive function (P < .05) in older adults. In addition, higher education was associated with better quality of life and cognitive function. DSI is a significant factor affecting the quality of life and cognitive function in older adults. Sociodemographic factors such as education play an important role in improving quality of life and cognitive function. Thus, increasing the awareness of this disability is important to ensure that older adults receive the necessary support services and rehabilitation to improve their level of independence.


Probiotic-fortified fruit juices: Health benefits, challenges, and future perspective.
Consumers' growing interest in using foods that improve health has motivated researchers and the food industry to develop new functional products, such as foods containing probiotics or live microbes. Probiotics have functional attributes that could satisfy most basic nutritional and therapeutic supplementation requirements. These microbes positively respond to clinical therapies against diseases and illnesses such as rotavirus-associated diarrhea, irritable bowel syndrome, and food allergies. Moreover, the role of probiotics in the prevention and treatment of obesity, diabetes, cancer, and diseases related to pathogenic microbes is an exciting and rapidly advancing research arena. Probiotic supplementation usually involves dairy products. However, because of the growing number of individuals affected by lactose intolerance and/or vegans, other food matrices like fruits, vegetables, cereals, and so on, have been studied as potential carriers for these microorganisms, presenting an alternative and better source in the process of assessing novel probiotic strains. The present review discusses the various factors affecting the survival of probiotics during storage in fruit juices, the possible effect of probiotics on sensory attributes and the overall acceptance of the products, and future technologies to improve the viability of probiotics.


Origins of emaciation in cancer patients.
Anorexia and elevated energy expenditure contributes to weight loss in cancer patients. Increased energy expenditure may explain 0.5-1.0 kg weight loss per month with regard to resting energy expenditure and even more with regard to total energy expenditure. The explanation behind elevated energy expenditure does not seem to be due to the tumour metabolism itself but rather to an increased host metabolic demand aimed at providing life-supporting protein synthesis and perhaps the immune system with energy. The trigger behind the activation of the synthesis of certain proteins is unknown. Only sparse information is available on the altered regulation of appetite in cancer disease. Changed metabolism and activity of neurotransmitters seem to be involved.


Biochemical characterization of novel glucokinase mutations isolated from Spanish maturity-onset diabetes of the young (MODY2) patients.
Mature-onset diabetes of the young, type 2 (MODY2) is associated with mutations in the glucokinase (GCK) gene that result in impaired glucokinase activity. Although more than 200 inactivating GCK mutations have been reported, only less than 20% of these mutations have been functionally characterized. In this work, we describe the biochemical characterization of six missense glucokinase mutations associated with MODY2 from Spanish patients, namely, Y61S, V182L, C233R, E265K, A379V, and K420E. All these mutations produced enzymes that presented reduced enzymatic activity in various degrees, from a mild affectation (K420E) to a more severe effect (C233R). Mutation severity correlated with the importance of the structural changes introduced by the mutations. For example, C233R affected a critical residue of the active center of the enzyme and rendered a protein with undetectable enzymatic activity. These data add new information on the structure-function relationship of human glucokinase.


Cation-exchange displacement chromatography for the purification of recombinant protein therapeutics from variants.
Removal of low level impurities that are closely related to the bioproduct is a commonly encountered challenge in the purification of biopharmaceuticals. These separations are typically carried out by using shallow linear salt gradients at relatively low column loadings, significantly limiting the throughput of the purification process. In this manuscript we examine the utility of displacement chromatography for the purification of recombinant human brain-derived neurotrophic factor, rHuBDNF. The utility of displacement chromatography is compared to gradient elution for the removal of variants of the rHuBDNF. The results demonstrate that displacement chromatography is capable of achieving high yields and purity at high column loadings. Displacements developed on 20 microns and 50 microns cation-exchange resins are shown to provide 8-fold and 4.5-fold increases in production rates, respectively, when compared to an existing linear gradient elution operation. These results demonstrate the efficacy of displacement chromatography for the purification of therapeutic proteins from complex feed streams.


Effects of antidepressants in the reproduction of aquatic organisms: a meta-analysis.
Pharmaceuticals are widespread contaminants across the aquatic systems and despite the role that these compounds play in society today, little is known about their effects in aquatic organisms once they are released into the environment. This study aims to provide quantitative insight on the effects of antidepressants on the reproduction of fish and crustaceans with a multilevel meta-analysis. A systematic literature search identified 19 studies investigating effects of antidepressant exposure in fish or crustaceans' reproduction, according to specific selection criteria. Heterogeneity analysis was performed and the moderating effect of the range of exposure concentrations, exposure time, organism group and toxicant was tested. Additionally, publication bias was also addressed. The results showed that, overall, there is no significant association between antidepressant exposure and the reproduction of fish and crustaceans, however, moderator analysis revealed that the range of concentrations is a significant moderator for fish and crustacean fecundity, showing contrary results between the two groups. Antidepressant concentration had a small, yet positive effect on fish fecundity, meaning that increased concentrations resulted in increased fish fecundity, whilst a negative effect on crustaceans' fecundity was apparent with increasing concentrations. This difference could be related to data artifacts, or, more likely, evidencing a hormetic dose-response curve, with different ranges of exposure concentrations considered in studies on fish and crustaceans. Antidepressants have shown effects on reproductive outcomes in aquatic organisms, based on individual studies and narrative reviews. However, our results show that other factors can have an important role. Additionally, data available for a quantitative assessment is scarce, focusing mainly on a few freshwater species, low concentration ranges and one SSRI compound, fluoxetine. Thus, more research on the subject is needed since meta-analysis are only as statistically powerful as the number of, good quality, studies they include.


Hydroxyl radical-induced cross-linking of thymine and lysine: identification of the primary structure and mechanism.
Hydroxyl radical-induced formation of a cross-link of thymine (Thy) and lysine (Lys) in the gamma-radiolysis of N2O-saturated aqueous solution was studied. A Thy-Lys cross-link (I) of the formal structure that OH radical and 4-carbon-centered Lys radical added respectively to C(5) and C(6) positions of Thy was isolated by a preparative HPLC and identified by a FAB-HRMS. The primary cross-link I was dehydrated by treatment with HCl at 120 degrees C to yield the secondary structure (II) possessing a C(5)-C(6) double bond in the Thy moiety: the latter structure II was reported previously (Dizdaroglu, M.; Gajewski, E. Cancer Res. 1989, 49, 3463-3467). A pulse radiolysis study with a redox titration method indicated that 4-carbon centered Lys radical intermediate was of neutral redox reactivity in contrast to reducing reactivity of 5-hydroxy-5,6-dihydrothymin-6-yl radical intermediate. The cross-link I could be formed by a conventional radical recombination mechanism, but not by an ionic recombination mechanism involving a redox reaction between the radical intermediates.


Long-Term Effect of Flow Diversion on Large and Giant Aneurysms: MRI-DSA Clinical Correlation Study.
The long-term effect of flow diversion (FD) on aneurysms has not been well studied. We aimed to assess the effect of the Pipeline embolization device (Covidien, Irvine, California, USA) on large and giant intracranial aneurysms with magnetic resonance imaging (MRI) and digital subtraction angiography (DSA) and then correlate with clinical follow-up. We conducted a retrospective analysis of aneurysms treated solely with Pipeline without adjunctive therapy. The largest aneurysm diameters were compared with pretreatment and post treatment MRI and correlated with DSA and clinical symptoms. Twelve patients harboring large and giant saccular unruptured aneurysms, treated with Pipeline, were included. Mean follow-up times were as follows: MRI 22.8 months (range 1-57), DSA 18.2 months (range 1-33), and clinical 29 months (range 1-48). Ten (83%) of 12 treated aneurysms decreased in size on MRI, which was first seen at an average of 17 months (range 4-57). In 70% of patients the change was noted on MRI ≤1 year after treatment, at an average of 9.1 months (range 4-12). MRI reduction was more likely in angiographically occluded aneurysms. Five aneurysms with complete occlusion on early angiogram showed size reduction on MRI only at ≥12 months. Mean aneurysm size reduction was 57% (range 19.6-94.2), which correlated well with improvement of cranial neuropathies in 75% and headaches in 100% of patients. Most aneurysms treated with Pipeline decreased in size, correlating with clinical improvement. Some aneurysms remained unchanged on MRI until a later time point despite early DSA occlusion. It may be reasonable to eliminate early postprocedural imaging and start follow-up only as late as 1 year after FD treatment in clinically stable, asymptomatic patients.


Long-term outcomes of patients receiving right internal thoracic artery or radial artery as a second arterial conduit. A propensity score matching study.
This study aimed to compare the long-term outcomes in propensity matched patients receiving right internal thoracic artery(RITA) or radial artery(RA) as second arterial conduit during coronary artery bypass grafting(CABG) with internal thoracic artery to the left anterior descending artery(LAD). In this retrospective study, propensity score matching was performed including 1198 patients from 3 centers resulting in 389-pairs who received either RITA or RA. In the matched cohort, median follow-up time was 7.53 years(interquartile range, 4.35-11.81). Survival probabilities at 5, 10 and 15-years were 93.8% versus 94.5%, 81.2% versus 76.2% and 63.2% vs 62.5% in the RITA and RA groups, respectively(HR: 1.11; 95%CI;0.80-1.53; P = 0.533) Freedom from MACCE in the matched cohort at 5, 10 and 15-years were 92.0% versus 93.7%, 75.0% versus 73.8%, 72.2% and 46.9% vs 47.2% in the RITA and RA groups, respectively(HR: 0.96; 95%CI;0.74-1.26; P = 0.774). Subgroup analyses of the matched cohort showed comparable long-term outcomes in terms of MACCE at follow-up in patients with age older than 65-years, obese patients, diabetics, female patients and with impaired EF. As for target vessel revascularization, RITA and RA had comparable outcomes in terms of MACCE when the conduit was used to graft either the left coronary system or the right coronary system. The use of RITA or RA as second arterial conduit during CABG with internal thoracic artery to the LAD is safe and associated with comparable long-term clinical outcomes. The choice of optimal second arterial conduit should be guided mainly by patients' characteristics and surgeons' preferences.


Two unusual cases of first trimester prenatal diagnosis of cystic fibrosis using DNA probes.
There are now several DNA probes which localize the cystic fibrosis mutation (CF) to chromosome 7q2.2-q3.1. The most tightly linked probes, pJ3.11 and met, are useful for first trimester prenatal diagnosis for many families provided that there is at least one living child affected by CF (Farrall et al., 1986). We describe here two families seeking prenatal diagnosis for CF which present unusual counselling problems. The first is an extended family in which there is no living affected member with CF; the second, a consanguinous marriage at risk both for cystic fibrosis and beta-thalassaemia. In both cases first trimester chorionic villus sampling and DNA haplotype analysis predicted that the fetus is a carrier for CF, and in the doubly affected family a carrier for beta-thalassaemia as well. Both pregnancies resulted in live births and subsequent immunoreactive trypsin estimations were both in the normal range.


Harpin Hpa1 Interacts with Aquaporin PIP1;4 to Promote the Substrate Transport and Photosynthesis in Arabidopsis.
Harpin proteins produced by plant-pathogenic Gram-negative bacteria are the venerable player in regulating bacterial virulence and inducing plant growth and defenses. A major gap in these effects is plant sensing linked to cellular responses, and plant sensor for harpin Hpa1 from rice bacterial blight pathogen points to plasma membrane intrinsic protein (PIP). Here we show that Arabidopsis AtPIP1;4 is a plasma membrane sensor of Hpa1 and plays a dual role in plasma membrane permeability of CO2 and H2O. In particular, AtPIP1;4 mediates CO2 transport with a substantial contribute to photosynthesis and further increases this function upon interacting with Hpa1 at the plasma membrane. As a result, leaf photosynthesis rates are increased and the plant growth is enhanced in contrast to the normal process without Hpa1-AtPIP1;4 interaction. Our findings demonstrate the first case that plant sensing of a bacterial harpin protein is connected with photosynthetic physiology to regulate plant growth.


Early estimation of pandemic influenza Antiviral and Vaccine Effectiveness (EAVE): use of a unique community and laboratory national data-linked cohort study.
After the introduction of any new pandemic influenza, population-level surveillance and rapid assessment of the effectiveness of a new vaccination will be required to ensure that it is targeted to those at increased risk of serious illness or death from influenza. We aimed to build a pandemic influenza reporting platform that will determine, once a new pandemic is under way: the uptake and effectiveness of any new pandemic vaccine or any protective effect conferred by antiviral drugs once available; the clinical attack rate of pandemic influenza; and the existence of protection provided by previous exposure to, and vaccination from, A/H1N1 pandemic or seasonal influenza/identification of susceptible groups. An observational cohort and test-negative study design will be used (post pandemic). A national linkage of patient-level general practice data from 41 Practice Team Information general practices, hospitalisation and death certification, virological swab and serology-linked data. We will study a nationally representative sample of the Scottish population comprising 300,000 patients. Confirmation of influenza using reverse transcription polymerase chain reaction and, in a subset of the population, serology. Future available pandemic influenza vaccination and antivirals will be evaluated. To build a reporting platform tailored towards the evaluation of pandemic influenza vaccination. This system will rapidly measure vaccine effectiveness (VE), adjusting for confounders, estimated by determining laboratory-confirmed influenza; influenza-related morbidity and mortality, including general practice influenza-like illnesses (ILIs); and hospitalisation and death from influenza and pneumonia. Once a validated haemagglutination inhibition assay has been developed (and prior to the introduction of any vaccination), cross-reactivity with previous exposure to A/H1N1 or A/H1N1 vaccination, other pandemic influenza or other seasonal influenza vaccination or exposure will be measured. A new sentinel system, capable of rapidly determining the estimated incidence of pandemic influenza, and pandemic influenza vaccine and antiviral uptake and effectiveness in preventing influenza and influenza-related clinical outcomes, has been created. We have all of the required regulatory approvals to allow rapid activation of the sentinel systems in the event of a pandemic. Of the 41 practices expressing an interest in participating, 40 have completed all of the necessary paperwork to take part in the reporting platform. The data extraction tool has been installed in these practices. Data extraction and deterministic linkage systems have been tested. Four biochemistry laboratories have been recruited, and systems for serology collection and linkage of samples to general practice data have been put in place. The reporting platform has been set up and is ready to be activated in the event of any pandemic of influenza. Building on this infrastructure, there is now the opportunity to extend the network of general practices to allow important subgroup analyses of VE (e.g. for patients with comorbidities, at risk of serious ILI) and to link to other data sources, in particular to test for maternal outcomes in pregnant patients. This study is registered as ISRCTN55398410. The National Institute for Health Research Health Technology Assessment programme.


The spatial-temporal evolution mechanism of PM2.5 concentration based on China's climate zoning.
Increasing attention has been given to the impact of PM2.5 concentration on human health. Exploring the influential factors of PM2.5 is conducive to improving air quality. Most existing studies explore the factors that influence the PM2.5 concentration from the perspective of cities or urban agglomerations, while few studies are conducted from the perspective of climate zones. We used the standard deviation ellipse and spatial autocorrelation analysis to explore the spatial-temporal evolution of the PM2.5 concentration in different climate zones in China during 2000-2018. We used differentiated EKC to construct panel regression models to explore the differences in the influential factors of the PM2.5 concentration in three climate zones. The number of cities with PM2.5 concentration less than 35 μg/m3 increased in the different climate zones. The center of gravity of the PM2.5 concentration has remained at the junction of the temperate and subtropical monsoon climate zones. The PM2.5 concentration had a high positive spatial autocorrelation in the different climate zones. The high-high clustering areas were located in the south of the temperate monsoon climate zone and the north of the subtropical monsoon climate zone. There was an inverted "U-shaped" curve between the PM2.5 concentration and economic development in China that varied in different climate zones. Identifying the differences in the influential factors of PM2.5 concentration in different climate zones will help to accelerate the implementation of the EKC inflection point.


Cholera. Lessons from the epidemic in Peru.
It is not known why the cholera epidemic, which affected all other continents, has not affected Latin America in the past 30 years. In addition, it is unclear why cholera appeared in Peru in 1991. Improvements in scientific knowledge and technology have occurred in Peru during the last 30 months. While it is impossible to summarize in only one article all these concepts, this article presents a few of the most important issues and recent developments in the treatment and prevention of cholera.


Exonization of AluYa5 in the human ACE gene requires mutations in both 3' and 5' splice sites and is facilitated by a conserved splicing enhancer.
Ancient Alu elements have been shown to be included in mature transcripts by point mutations that improve their 5' or 3' splice sites. We have examined requirements for exonization of a younger, disease-associated AluYa5 in intron 16 of the human ACE gene. A single G>C transversion in position -3 of the new Alu exon was insufficient for Alu exonization and a significant inclusion in mRNA was only observed when improving several potential splice donor sites in the presence of 3' CAG. Since complete Alu exonization was not achieved by optimizing traditional splicing signals, including the branch site, we tested whether auxiliary elements in AluYa5 were required for constitutive inclusion. Exonization was promoted by a SELEX-predicted heptamer in Alu consensus sequence 222-228 and point mutations in highly conserved nucleotides of this heptamer decreased Alu inclusion. In addition, we show that Alu exonization was facilitated by a subset of serine/arginine-rich (SR) proteins through activation of the optimized 3' splice site. Finally, the haplotype- and allele-specific ACE minigenes generated similar splicing patterns in both ACE-expressing and non-expressing cells, suggesting that previously reported allelic association with plasma ACE activity and cardiovascular disease is not attributable to differential splicing of introns 16 and 17.


[The patient with unclear palpitations: how extensive should the diagnostic approach be?].
The symptom "palpitation" may be caused by a cardiac arrhythmia; however, in a significant number of events no concomitant arrhythmia can be documented. In addition, a patient complaining about palpitations can be threatened by an event leading to significant morbidity or even increased mortality (e.g., atrial fibrillation in a patient with risk factors for stroke) or, in contrast, the symptom palpitation may have no implications in terms of morbidity or prognosis (e.g., premature atrial contractions in a healthy person). This unspecific character of the symptom "palpitation" makes clear that the diagnostic approach has to be individualized: the patient with severe concomitant symptoms, hemodynamic deterioration and/or risk factors for cardiogenic thromboembolism should be thoroughly investigated to detect an underlying disease and to document an ECG during the palpitations. Those with only mild symptoms and no evidence for a cardiac disease in a non-invasive work-up can be simply followed-up.


Fetal tissue/organ transplant in HLA-randomized adult vascular subcutaneous axillary folds: preliminary report of 14 patients.
In the year 1902, the first successful experimental organ transplant, i.e., an autotransplant of a dog's kidney from its normal position to the vessels of the neck, which resulted in some urine flow, was performed in the Vienna Physiology Institute under the direction of Hofrath Exner by Dr. Emerich Ullman (1861-1937). Since then, the art of transplant surgery has come a long way in establishing itself as an important discipline with the support disciplines of immunology, molecular biology, etc., for the restoration of a failing organ. Today there is a major discrepancy in the demand and supply of organ grafts. The aim of the present study is to see whether fetal organ and tissue, with its intrinsic advantages of hypo-antigenicity, can survive in a HLA and sex-randomized host in a surgically prepared vascular subcutaneous axillary fold, without any immunosuppressive support. We have earlier reported two cases of fetal thymic transplant, collected from consenting mothers undergoing hysterotomy and ligation. Fourteen cases were recruited for the present study after thorough informed consent and approval by the Ethical Committee of the Project. Of these, five patients were suffering from advanced cancer, three from diabetic gangrene, three from ischaemic heart disease and three from rheumatoid arthritis, liver abscess and disc prolapse. The ages of the patients varied from 39 to 82 years. Six fetal thymuses, three fetal liver tissues, three fetal cardiac tissues, one fetal pancreas and one fetal lung tissue were transplanted. All the fetuses were dissected and the selected tissues/organs were transplanted within one to three minutes after collecting them from the consenting mothers undergoing hysterotomy and ligation. The fetal tissue graft was placed in a surgically prepared subcutaneous vascular axillary fold, 2x1 cm, under local anaesthesia in the consenting adult recipient. Sequential Hb, Tc, Dc, ESR were done to see the impact of the transplant on the host system. After one month, the transplanted fetal tissue was taken out by an elliptical incision and the tissue was processed for histological staining. All the 14 patients tolerated the transplant procedure well. There was no fever, intractable pain or any other specific serious side-effect justifying removal of the transplant earlier. There was no discharge from the incision site and the healing and scar were by and large normal. There was no unusual leucocytosis or lymphocytosis. The serial histological study did not suggest features of transplant rejection. Pregnancy and neoplasm are two outstanding examples of natural tolerance to homograft. In both cases, blocking antibody has an important role in the phenomenon of immunotolerance. From our experiments mentioned above transplantation and our earlier reported studies, we believe that the hypo-antigenic fetal tissue has distinct advantages over adult tissue for transplant purposes.


Nucleotide sequence of the second psbG gene in Synechocystis 6803. Possible implications for psbG function as a NAD(P)H dehydrogenase subunit gene.
Nucleotide sequencing of the second Synechocystis 6803 psbG gene, psbG2 shows the predicted polypeptide to be 219 amino acids long. It is less similar to chloroplast psbG genes than is the Synechocystis psbG1 copy. Alignment with seven other psbG protein sequences, including that from the Paramecium mitochondrial genome reveals a central highly conserved region common to each. This is discussed as evidence supporting the proposal that the psbG polypeptide is a NAD(P)H dehydrogenase (complex I) subunit in cyanobacteria, chloroplasts and mitochondria.


HBV-positive and HIV-positive organs in transplantation: A clinical guide for the hepatologist.
Organ transplantation is a life-saving treatment for patients with end-stage organ disease, a severe condition associated with a high risk of waitlist mortality. It is primarily limited by a shortage of available organs. Maximising available donors can increase access to transplantation. Transplantation from donors positive for HBV and HIV has increased in many countries. However, antiviral therapies need to be readily available for recipients after transplantation to prevent possible reactivation of the virus following the administration of immunosuppressive therapies. Furthermore, the intentional transmission of a virus has practical, ethical, and clinical implications. In this review, we summarise the current research, focusing on grafts from donors positive for the HBV surface antigen, antibodies against the HBV core antigen, and HIV, to help hepatologists and physicians interested in transplantation to select the best antiviral and/or prophylactic regimens for after transplantation.


Use of an inflatable balloon on a catheter to prevent subdural fluid collections after intraventricular tumor surgery: technical note.
Ventricular tumor surgeries pose significant challenges to the neurosurgeon. Apart from the operative challenges and typical complications of surgery, subdural fluid collection is common after these surgeries. Various methods have been used to prevent this complication. The authors' technique of implanting an inflatable balloon attached to a catheter after resection of a huge intraventricular tumor was highly effective in preventing the development of a subdural fluid collection.


Importance of orthodontic intervention of the Class III malocclusion in mixed dentition.
Supervising the development of occlusion, managing problems during the transition from mixed to permanent dentition, as well as controlling environmental factors that contribute to establishing malocclusion, are important actions to achieve a Class I occlusion with facial balance. Among these problems, the malocclusions associated with dysfunctions such as mouth breathing or obstructive sleep apnea syndrome (OSAS), atypical swallowing and abnormal tongue position, open bites, crossbites and maxillomandibular discrepancies, and especially the Class III malocclusion can be listed. The purpose of this article is to present and discuss the main aspects relevant to the benefits of performing the treatment of Class III malocclusion in patients with growth.


Helical (spiral) CT of the pancreas and biliary tract.
Helical scanning offers many advantages for the evaluation of benign and malignant pancreatobiliary disease. This article presents these advantages, recommended screening protocols, and guidelines for the use of three-dimensional rendering of the peripancreatic vessels and bile ducts.


Financing long-term care. An insurance-based approach.
A joint public-private insurance program is the best approach to resolving the problem of financing long-term care. In this report, we describe one possible approach in detail. A modest expansion of the current (ie, after repeal of the Medicare Catastrophic Coverage Law of 1988) Medicare benefit for persons needing relatively short-term nursing home and home care services would be a first step. For those with extended long-term service needs, a non-means tested, publicly funded program with joint federal-state financing and administration would provide coverage after a substantial elimination period and with an income-related copayment. Private long-term care insurance purchased through employers before retirement or in the periretirement period, through use of income or equity accumulated in life insurance, pension funds, or home ownership, would be used to fund the exclusionary period or copayments of the public program by those who wish to have greater protection for income or assets. The role of Medicaid would be limited to paying for the deductible, copayments, and initial long-stay expenses of those with low incomes and limited assets.


Imputation of Human Primary Osteoblast Single Cell RNA-Seq Data Identified Three Novel Osteoblastic Subtypes.
Recently, single-cell RNA sequencing (scRNA-seq) technology was increasingly used to study transcriptomics at a single-cell resolution, scRNA-seq analysis was complicated by the "dropout", where the data only captures a small fraction of the transcriptome. This phenomenon can lead to the fact that the actual expressed transcript may not be detected. We previously performed osteoblast subtypes classification and dissection on freshly isolated human osteoblasts. Here, we used the scImpute method to impute the missing values of dropout genes from a scRNA-seq dataset generated on freshly isolated human osteoblasts. Based on the imputed gene expression patterns, we discovered three new osteoblast subtypes. Specifically, these newfound osteoblast subtypes are osteoblast progenitors, and two undetermined osteoblasts. Osteoblast progenitors showed significantly high expression of proliferation related genes (FOS, JUN, JUNB and JUND). Analysis of each subtype showed that in addition to bone formation, these undetermined osteoblasts may involve osteoclast and adipocyte differentiation and have the potential function of regulate immune activation. Our findings provided a new perspective for studying the osteoblast heterogeneity and potential biological functions of these freshly isolated human osteoblasts at the single-cell level, which provides further insight into osteoblasts subtypes under various (pathological) physiological conditions.


Sodium alginate-carboxymethyl chitosan hydrogels loaded with difenoconazole for pH-responsive release to control wheat crown rot.
Increasing concern about environmental pollution has driven the development of controlled release formulations for agrochemicals. Due to the advantages of degradability and responsiveness to environmental stimuli, polysaccharide-based hydrogel is an ideal carrier for agrochemicals controlled release. In this study, a method-easy polysaccharide hydrogel for controlled release of difenoconazole (DZ) was prepared with sodium alginate (SA) and carboxymethyl chitosan (CMCS). Due to its three-dimensional crosslinked mesh structure, the prepared hydrogels (CSDZ) showed an agrochemical load capacity of 9.03 % and an encapsulation efficiency of 68.64 %. The release rate is faster in alkaline solution, followed by neutral solution, and slowest in an acid environment, which is consistent with the swelling behavior. Furthermore, leaching studies showed that CSDZ hydrogels have excellent protective properties for encapsulated agrochemicals. Compared with technical DZ, the results of in vitro and pot antifungal testing showed that CSDZ had a better control effect against wheat crown rot (Fusarium pseudograminearum). Safety assessment studies indicated that CSDZ hydrogels exhibit good biocompatibility on nontargeted organisms (Daphnia magna, zebrafish and Eisenia fetida) and wheat. This study aims to provide a potentially promising approach for the preparation and application of biocompatible polysaccharide-based hydrogels for agrochemical-controlled release in sustainable disease management.


Reactivity of cytochromes c and f with mutant forms of spinach plastocyanin.
The reduction of plastocyanin by cytochromes c and f has been investigated with mutants of spinach plastocyanin in which individual, highly conserved surface residues have been modified. These include Leu-12 and Phe-35 in the 'northern' hydrophobic patch and Tyr-83 and Asp-42 in the 'eastern' acidic patch. The differences observed all involved binding rather than the intrinsic rates of electron transfer. The Glu-12 and Ala-12 mutants showed small but significant decreases in binding constant with cytochrome c, even though the cytochrome is not expected to make contact with the northern face of plastocyanin. These results, and small changes in the EPR parameters, suggested that these mutations cause small conformational changes in surface residues on the eastern face of plastocyanin, transmitted through the copper centre. In the case of cytochrome f, the Glu-12 and Ala-12 mutants also bound less strongly, but Leu12Asn showed a marked increase in binding constant, suggesting that cytochrome f can hydrogen bond directly to Asn-12 in the reaction complex. A surprising result was that the kinetics of reduction of Asp42Asn were not significantly different from wild type, despite the loss of a negative charge.


CMA/DAPI Banding of Plant Chromosomes.
Chromosome banding based on base-specific fluorochromes, mainly double staining with chromomycin A3 (CMA) and 4'-6-diamidino-2-phenylindole (DAPI), has been widely used since the 1970s. This technique allows the differential staining of distinct types of heterochromatin. Afterward, the fluorochromes can be easily removed and leave the preparation ready for sequential procedures such as FISH or immunodetection. Interpretations of similar bands obtained with different techniques, however, merit certain caution. Here we present a detailed protocol for CMA/DAPI staining optimized for plant cytogenetics and call attention to the most common sources of misinterpretation of DAPI bands.


Motor neuron degeneration in spinal and Bulbar Muscular Atrophy is a skeletal muscle-driven process: Relevance to therapy development and implications for related motor neuron diseases.
Non-cell autonomous degeneration has arisen as an important mechanism in neurodegenerative disorders. Using a novel line of BAC androgen receptor (AR) transgenic mice with a floxed transgene (BAC fxAR121), we uncovered a key role for skeletal muscle in X-linked Spinal and Bulbar Muscular Atrophy (SBMA), a motor neuronopathy caused by a polyglutamine expansion in exon 1 of the AR gene. By excising the mutant AR transgene from muscle only, we achieved complete rescue of neuromuscular phenotypes in these mice, despite retaining strong CNS expression. Furthermore, we delivered an antisense oligonucleotide (ASO) directed against the human AR transgene by peripheral injection, and documented that peripheral ASO delivery could rescue muscle weakness and premature death in BAC fxAR121 mice. Our results reveal a crucial role for skeletal muscle in SBMA disease pathogenesis, and offer an appealing avenue for therapy development for SBMA and perhaps also for related motor neuron diseases.


Patterns of sedentary time and ambulatory physical activity in a Danish population of girls and women with Rett syndrome.
Rett syndrome (RTT) is a rare neurodevelopmental disorder leading to multiple disabilities and high dependency on caregivers. This study aimed to: (1) describe the patterns of sedentary time and daily steps and (2) identify the association of individual and environmental characteristics with sedentary time. All Danish females with RTT older than 5 years of age and with a MECP2 mutation were invited to participate. The activPAL and StepWatch Activity Monitor (SAM) were worn by participants for at least four days. Sedentary time and step counts were plotted by time to examine daily activity patterns. Associations between sedentary time and individual and environmental covariates were assessed with linear regression models. The median (interquartile range) age of participants was 22.0 (14.3-36.5) years. On average 83.3% (standard deviation 13.9%) of waking hours were spent in sedentary behaviours (n = 48) and the median (interquartile range) daily step count was 5128 (2829-7704) (n = 28). Females older than 33.5 years, and those unable to walk independently were more sedentary. This study demonstrated high levels of sedentary time and low daily step counts in a Danish population of females with RTT. Advancing age and lower walking skills were associated with higher levels of sedentary time. Implications for Rehabilitation Sedentary lifestyles in individuals with disabilities have a negative impact on health and quality of life. High levels of sedentary time and low daily step counts were demonstrated in a Danish population of females with Rett syndrome. Advancing age and inability to walk independently were strongly associated with higher levels of sedentary time in females with Rett syndrome. Understanding patterns of sedentary behaviour and physical activity can aid health care professionals in developing health-promoting physical activity interventions.


Selection of the T-cell repertoire during ontogeny.
This report examines conflicting hypotheses concerning T-cell repertoire selection in terms of H-2 restriction during ontogeny. The experiments described in this report were incompatible with the hypothesis that bias in the repertoire is solely a consequence of "more or less intentional priming" by foreign antigen. Rather, results indicate that the repertoire is selected by self major histocompatibility complex (MHC) antigens in the absence of foreign antigens. Allorestricted T cells, the existence of which was previously thought to be incompatible with the concept of complete repertoire selection by self MHC antigens, were shown to significantly cross-react on targets expressing self MHC antigens. Thus, it is possible that allorestricted T cells are simply cross-reactive T-cell clones restricted by self MHC antigens; indeed, all experimental data were compatible with the idea of complete selection of the T-cell repertoire in terms of H-2 restriction by self MHC antigens.


Evaluation of the reentry vulnerability index to predict ventricular tachycardia circuits using high-density contact mapping.
Identifying arrhythmogenic sites to improve ventricular tachycardia (VT) ablation outcomes remains unresolved. The reentry vulnerability index (RVI) combines activation and repolarization timings to identify sites critical for reentrant arrhythmia initiation without inducing VT. The purpose of this study was to provide the first assessment of RVI's capability to identify VT sites of origin using high-density contact mapping and comparison with other activation-repolarization markers of functional substrate. Eighteen VT ablation patients (16 male; 72% ischemic) were studied. Unipolar electrograms were recorded during ventricular pacing and analyzed offline. Activation time (AT), activation-recovery interval (ARI), and repolarization time (RT) were measured. Vulnerability to reentry was mapped based on RVI and spatial distribution of AT, ARI, and RT. The distance from sites identified as vulnerable to reentry to the VT site of origin was measured, with distances <10 mm and >20 mm indicating accurate and inaccurate localization, respectively. The origins of 18 VTs (6 entrainment, 12 pace-mapping) were identified. RVI maps included 1012 (408-2098) (median, 1st-3rd quartiles) points per patient. RVI accurately localized 72.2% VT sites of origin, with median distance of 5.1 (3.2-10.1) mm. Inaccurate localization was significantly less frequent for RVI than AT (5.6% vs 33.3%; odds ratio 0.12; P = .035). Compared to RVI, distance to VT sites of origin was significantly larger for sites showing prolonged RT and ARI and were nonsignificantly larger for sites showing highest AT and ARI gradients. RVI identifies vulnerable regions closest to VT sites of origin. Activation-repolarization metrics may improve VT substrate delineation and inform novel ablation strategies.


Effect of aluminium on migration of oestrogen unresponsive MDA-MB-231 human breast cancer cells in culture.
Aluminium (Al) has been measured in human breast tissue, and may be a contributory factor in breast cancer development. At the 10th Keele meeting, we reported that long-term exposure to Al could increase migratory properties of oestrogen-responsive MCF-7 human breast cancer cells suggesting a role for Al in the metastatic process. We now report that long-term exposure (20-25 weeks) to Al chloride or Al chlorohydrate at 10(-4) M or 10(-5) M concentrations can also increase the migration of oestrogen unresponsive MDA-MB-231 human breast cancer cells as measured using time-lapse microscopy and xCELLigence technology. In parallel, Al exposure was found to give rise to increased secretion of active matrix metalloproteinase MMP9 as measured by zymography, and increased intracellular levels of activated MMP14 as measured by western immunoblotting. These results demonstrate that Al can increase migration of human breast cancer cells irrespective of their oestrogen responsiveness, and implicate alterations to MMPs as a potential mechanism worthy of further study.


Regulation of L-proline transport in Leishmania donovani by extracellular pH.
We have previously shown that Leishmania donovani promastigotes adapted to long-term culture at acidic pH can serve as a model to study parasite development in a lysosomal-like environment. In this study we investigated the effect of growth pH on L. donovani L-proline transport systems. Reducing the pH of the growth medium causes an up to 7-fold decrease in the extent of L-proline transport. Transport resumes after switching the culture from pH 4.5 to pH 7 for 48 h by a protein synthesis-dependent process. The pH optimum for transport changes from 7.5 in promastigotes grown at pH 7 to 5.5 in cells grown at pH 4.5. In addition, kinetic analysis of L-proline transport showed that Vmax in pH 4.5-grown L. donovani promastigotes is one-tenth that of cells grown at pH 7 (4.5 and 44.7 nmol min-1 (10(8) cells)-1, respectively). The apparent Km for L-proline in pH 4.5 promastigotes is one-half of the Km in pH 7 cells (0.30 and 0.65 mM, respectively). In contrast to L-proline transport, D-glucose transport demonstrates a growth pH-independent activity: Km and Vmax as well as optimum pH of transport are similar in promastigotes grown at either pH 7 or pH 4.5. Taken together, the results indicate that in L. donovani, expression and activity of L-proline transport is regulated by culture pH. The pH-dependent expression of L-proline transporters may be of physiological significance during the promastigote-amastigote transition.


Perinatal choline treatment modifies the effects of a visuo-spatial attractive cue upon spatial memory in naive adult rats.
The improvement in memory functions by choline supplementation is hypothesized to be due to increased synthesis and release of acetylcholine in the brain. We have found previously that combined pre- and postnatal choline supplementation results in long-lasting facilitation of spatial memory in juvenile rats when training was conducted in presence of a local salient cue. The present work aims to analyze the effects of peri- and postnatal choline supplementation on spatial abilities of naive adult rats. Treated rats were trained in various cued procedures of the Morris navigation task of 5 months of age. The treatment had a specific effect of reducing the escape latency of the rats when the platform was at a fixed location in space and indicated by a suspended cue. This effect was associated with an improved spatial memory when the cue and the platform were removed. In this condition, the control rats showed impaired spatial discrimination following the removal of the target cue, most likely due to an overshadowing of the distant environmental cues. This impairment was not observed in the treated rats. Further training with the suspended cue at unpredictable places in the pool revealed longer escape latencies in the control than in the treated rats suggesting that this procedure induced a selective perturbation of the normal but not of the treated rats. A special probe trial with the cue at an irrelevant location and no escape platform revealed a significant bias of the control rats towards the cue, but in treated rats towards the uncued spatial escape position. This behavioral dissociation suggests that a salient cue associated with the target induces an alternative "non spatial" guidance strategy in normal rats, with the risk of overshadowing attention towards more distant spatial cues. As a consequence, the improved escape in the presence of the cue in the treated rats is associated with a stronger memory of the spatial position following disappearance of the cue. This and previous observations suggest that a specific spatial attentional process relies on the buffering of highly salient visual cues to facilitate integration of their relative position in the environment.


Effect of Ramadan fasting in patients with type 2 diabetes mellitus treated with sodium-glucose cotransporter 2 inhibitors: A systematic review and meta-analysis.
Sodium-glucose cotransporter 2 inhibitors (SGLT-2i) improve glycemic control and weight, but might be associated with dehydration, hypotension and ketoacidosis, especially in patients with type 2 diabetes mellitus who fast during Ramadan. This meta-analysis evaluates the effects of Ramadan fasting on patients with type 2 diabetes mellitus treated with SGLT-2i. A literature search was carried out in PubMed, Embase and the Cochrane Library. Quality assessment was carried out using the ROBINS-I and Cochrane tools for risk of bias, and analyses were carried out using RevMan version 5.3. A total of five studies were included in this meta-analysis. During Ramadan, there was a significant reduction in glycated hemoglobin (P < 0.00001) and diastolic blood pressure (P = 0.006), with a non-significant trend for a reduction in weight (P = 0.44) and systolic blood pressure (P = 0.67). The number and severity of hypoglycemic episodes was lower in patients with type 2 diabetes mellitus treated with SGLT-2i compared with sulfonylureas. There was no significant change in estimated glomerular filtration rate, β-hydroxybutyrate, bicarbonate or anion gap. However, we identified considerable heterogeneity among studies, and a lack of head-to-head studies with structured outcome reporting on the risks and benefits of SGLT-2i during Ramadan. This systematic review and meta-analysis shows that patients with type 2 diabetes treated with SGLT2i's during Ramadan have an improvement in HbA1c, less hypoglycemia and no major adverse effects.


In vivo real-time confocal laser scanning endomicroscopic colonoscopy for the detection and characterization of colorectal neoplasia.
Conventional colonoscopy has a significant false-negative rate for intraepithelial neoplasia. Chromoendoscopy increases sensitivity but lacks specificity. The aim was to assess prospectively the clinical applicability and predictive power of the EC3870CIFK confocal laser endomicroscope (CLE) for the in vivo diagnosis of intraepithelial neoplasia during colonoscopy. Lesions were identified using chromoscopy followed by CLE imaging and graded according to vascular and cellular changes. CLE imaging of circumscribed lesions and four segmental 'normal' colorectal quadrants was performed. Targeted biopsy specimens were then compared with histopathological results. Forty patients completed the protocol (22 men and 18 women; median age 62 (range 39-82) years). The median duration of ileal intubation and total procedure time were 12 (range 5-26) and 55 (range 28-92) min respectively. Chromoscopic colonoscopy revealed 162 lesions in 39 patients. CLE imaging was obtained on all 162 lesions. Some 5422 confocal images were compared with 802 targeted biopsy specimens. Intraepithelial neoplasia was predicted with an accuracy of 99.1 per cent (sensitivity 97.4 per cent and specificity 99.3 per cent) (P = 0.711). Confocal laser endomicroscopy permits high-quality cellular, subsurface vascular and stromal imaging, enabling prediction of intraepithelial neoplasia with a high level of accuracy.


Diagnosis and clinical features of borderline personality disorder in the east and west: a preliminary report.
In the first clinical study of the borderline personality disorder (BPD) in Asia, the Diagnostic Interview for Borderlines (DIB) was performed on 85 female outpatients aged 18 to 30 years in Japan. BPD was diagnosed in 32 (38%) of these patients. The International Classification of Diseases-9 (ICD-9) diagnoses made at the initial examination of the DIB-identified BPD patients were, in order of frequency, neurotic disorders, anorexia nervosa and other eating disorders, and manic-depressive psychoses. The DSM-III axis I diagnoses of the DIB-identified BPD patients were affective disorders in 63%, eating disorders in 34%, and substance use disorders in 22%. This study suggests that there are indeed BPD patients in Japan. An exception is made of the low incidence of substance use disorders, and of the fact that most Japanese patients continue to maintain stormy one-to-one or masochistic relationships with their parents because they live at home--their clinical picture is not different from that of American patients.