Photorespiratory ammonia does not inhibit photosynthesis in glutamate synthase mutants of Arabidopsis.
Exposure of ferredoxin-dependent glutamate synthase (EC 22.214.171.124) mutants of Arabidopsis thaliana to photorespiratory conditions resulted in the accumulation of NH(4) (+) and the inhibition of photosynthesis. However, upon transfer from 2% O(2), 350 microliters per liter CO(2), to 21% O(2), 350 microliters per liter CO(2), net photosynthesis declined at a slower rate in methionine sulfoximine treated leaf discs relative to controls. The recovery of photosynthesis was also more rapid in MSO-treated leaf discs although ammonia levels were more than threefold higher. Photosynthesis in leaf discs treated with azaserine was inhibited more than controls when transferred to 21% O(2) and recovered less than controls when returned to 2% O(2) although NH(4) (+) levels were not significantly different. The results obtained are consistent with the view that the rapid inhibition of photosynthesis in the glutamate synthase mutants in photorespiratory conditions is not due to the accumulation of NH(4) (+) but rather to the depletion of amino donors for glyoxylate and the consequent effects of glyoxylate on the lack of return of carbon to the chloroplast.


Collagen-Based Bioinductive Implant for Treatment of Partial Thickness Rotator Cuff Tears.
Partial thicknessrotator cuff tears(PTRCT) have low healing potential and tend to progress over time if not addressed surgically. There is a relative paucity of literature discussing optimal treatment for symptomatic PTRCT as compared to full thicknessrotator cuff tears. The available data supports a treatment course of debridement with or without acromioplasty for symptomatic patients with tears less than 50% in thickness combined with the use of repair (conversion to full thickness or transtendinous) for symptomatic patients with tears greater than 50% in thickness. The aim of this study was to evaluate functional and radiographic outcomes following surgical implantation of a collagen-based bioinductive implant for PTRCT. Patients with PTRCT who underwent implantation of a collagen-based bioinductive implant over the bursal surface of the rotator cuff were identified. Patients who had an implant placed to augment a standard full thickness repair were excluded from analysis. We administered questionnaires to patients with a minimum of 6 months since surgery and collected patient data including demographics, preoperative and postoperative American Shoulder and Elbow Society (ASES) scores, preoperative and postoperative visual analog scale (VAS) pain scores, complications, and satisfaction level. Magnetic resonance imaging (MRI) was obtained as needed, at 6 months, or at 12 months postoperatively, and tendon thickness was measured on coronal views. Thirty patients were identified who met the inclusion and exclusion criteria. Twenty-four patients (80% response rate) were available for follow-up; 19 were male and five were female. Mean age and body mass index were 54.5 ± 11.6 years and 28.6 ± 5.9 kg/m2 , respectively. Mean tear thickness as measured intraoperatively was 56.6%. There were 16 (66.7%) articular-sided, five (20.8%) bursal-sided, and three (12.5%) intrasubstance tears. Mean survey follow-up time was 19.1 months. Mean ASES scores increased significantly from 45.6 preoperatively to 68.1 postoperatively (p = 0.001). Mean VAS pain scores decreased significantly from 8.3 preoperatively to 3.8 postoperatively (p < 0.001). Mean patient satisfaction level was 7.5. Ten patients had both a preoperative and postoperative MRI available for comparison. Tendon thickness at the tear site increased significantly from 5.7 mm preoperatively to 6.5 mm at mean 9.9 months follow-up (p = 0.007). There were no implant-related complications. One patient suffered a traumatic re-tear 4 months postoperatively. Highly porous collagen-based bioinductive implants are safe and effective forreducing pain and improving shoulderfunction in patientswith PTRCT of approximately 50%, with radiographic evidence of new tissue formation. Randomized controlled trials are needed to assess efficacy relative to debridement and standard repair techniques.


A Copper-Catalyzed Decarboxylative Amination/Hydroamination Sequence: Switchable Synthesis of Functionalized Indoles.
A copper-catalyzed decarboxylative amination/hydroamination sequence of propargylic carbamates with various nucleophiles is described for the first time. It features an earth-abundant metal catalyst, mild reaction conditions, and high efficiency. Further treatments of the resultant key intermediates using an acid or a base in one pot enable the controllable and divergent synthesis of two types of functionalized indoles. Moreover, experiments to demonstrate the synthetic potential of this methodology are performed.


Characteristics of a photoresist hologram and its replica.
The characteristics of the surface relief hologram which is produced by a photoresist and its replica by an embossing method are experimentally studied. It becomes clear that the maximum diffraction efficiency of the photoresist hologram and that of the replica hologram are almost the same value-21-25%-and the maximum SNR of the original hologram and its replica are fairly good.


Avian influenza A(H5N1) in humans: lessons from Egypt.
Highly pathogenic avian influenza A(H5N1) has ravaged the Egyptian poultry population. Ninety human cases, including 27 fatalities have been recorded by 30 December, 2009. However, epidemiological information on the infection in humans in Egypt is scarce. We analysed the first three years of highly pathogenic avian influenza A(H5N1) in Egypt between 20 March 2006 and 31 August 2009) and found that more cases occurred in females than males, especially in 2006 and 2007. Women in the age group 20-39 years had the greatest tendency to be infected. It took an average of one day and 18 hours to seek medical assistance in patients who recovered and of six days in fatal cases. Children sought treatment much earlier than adults. On average, a patient died 11 days after the onset of symptoms. Exposure to infected poultry remained the most important risk factor.


The impact of unemployment insurance benefits on the probability of migration of the unemployed.
"By ignoring individual unemployment compensation benefits and conditions of job termination, past migration research has concluded that personal unemployment doubles the likelihood of interstate labor-force migration. Findings from the present study indicate that aggregating the unemployed, without adjusting for these two factors, overstates the probability of migration for the involuntarily unemployed benefit recipient and understates the likelihood of migration for the voluntarily unemployed benefit recipient. The results suggest that federal discretionary unemployment-compensation programs, which are implemented during recessionary periods, likely serve to retard out-migration of those who are involuntarily unemployed." Data are from a 1982 sample of unemployed U.S. workers.


[Exercise-induced laryngomalacia].
Exercise-induced laryngomalacia is a rare differential diagnosis of exercise-induced asthma. We report the case of a previously fit 14-year-old female patient who presented with dyspnoea on intense exertion and whose condition had not improved with treatment prescribed for a misdiagnosed exercise-induced asthma. A diagnosis of exercise-induced laryngomalacia was eventually made when a variable extra-thoracic airway obstruction on the flow-volume loops was obtained after an incremental exercise test. Flexible fiberoptic rhinolaryngoscopy confirmed this abnormality and demonstrated an anterior prolapse of the arytenoid region, partially obstructing the airway. The patient therefore underwent laser aryepiglottoplasty which produced a satisfactory anatomical outcome in the larynx but which only resulted in a partial functional improvement. Laryngomalacia is rare and its pathophysiology is not well understood. However, it should be considered in patients presenting with exertional dyspnoea, particularly if there is inspiratory embarrassment, who fail to improve with beta-2-adrenergic agonists. The diagnosis can be made from the flow-volume loop and direct laryngoscopy, before and after exercise.


Influence of day of postpartum breeding on pregnancy rate, pregnancy loss rate, and foaling rate in Thoroughbred mares.
Records (years 2005-2007) were analyzed from a Thoroughbred stud farm in central Kentucky. Data from all breeding cycles of foaling mares were tabulated (3184 cycles of 2003 foaling mares bred between 7 and 163 days postpartum). A multiple logistic regression model employing Bayesian statistics was used to adjust for factors that significantly affected outcome; odds ratios (ORs) for pregnancy rate, pregnancy loss rate, and foaling rate were determined to examine the influence of day of postpartum breeding on these parameters. Mares bred before Day 22 (Day 0 = day of foaling) postpartum had a decreased OR for becoming pregnant (P < 0.05); the median OR for becoming pregnant (1.00) was not reached until Day 46 postpartum. Mares bred before Day 13 postpartum had an increased OR for pregnancy loss (P < 0.05). The median OR for pregnancy loss did not decline below 1.00 until Day 78 postpartum. Mares bred before Day 20 postpartum had a decreased OR for producing a foal (P < 0.05). The median OR for producing a foal (1.00) was not reached until Day 75 postpartum. We concluded that fertility (in terms of a higher OR for becoming pregnant and a lower OR for pregnancy loss, resulting in a higher OR for producing a foal) continued to improve in Thoroughbred mares for approximately 2.5 mo postpartum. These findings are of importance to management strategies directed at early postpartum breeding, and explain some of the reported drift in subsequent foaling dates of Thoroughbred mares, despite management practices that employ early postpartum breeding.


SARS-CoV-2 placentitis as a cause of intrauterine death of a fetus in a patient with covid-19 infection and ongoing HELLP syndrome.
We hereby present a case of intrauterine death of a fetus due to placental damage by placentitis caused by SARS-CoV-2 infection in ongoing covid-19 disease and HELLP syndrome. In this case report, we describe a patient who experienced intrauterine death of a fetus and the development of HELLP syndrome in ongoing covid-19 infection. Placentitis caused by SARS-CoV-2 infection was identified as the cause of intrauterine death of a fetus. After the end of pregnancy, a patient experienced an improvement in the symptoms of covid-19 and also a gradual improvement and adjustment of laboratory and coagulation parameters. SARS-CoV-2 infection in pregnancy can be complicated by the development of SARS-CoV-2 placenitis, which can cause intrauterine death of a fetus. Covid-19 infection can even have similar signs to HELLP syndrome and therefore careful monitoring of pregnant women with covid-19 is recommended.


Blood Thiol Redox State in Chronic Kidney Disease.
Thiols (sulfhydryl groups) are effective antioxidants that can preserve the correct structure of proteins, and can protect cells and tissues from damage induced by oxidative stress. Abnormal levels of thiols have been measured in the blood of patients with moderate-to-severe chronic kidney disease (CKD) compared to healthy subjects, as well as in end-stage renal disease (ESRD) patients on haemodialysis or peritoneal dialysis. The levels of protein thiols (a measure of the endogenous antioxidant capacity inversely related to protein oxidation) and S-thiolated proteins (mixed disulphides of protein thiols and low molecular mass thiols), and the protein thiolation index (the molar ratio of the S-thiolated proteins to free protein thiols in plasma) have been investigated in the plasma or red blood cells of CKD and ESRD patients as possible biomarkers of oxidative stress. This type of minimally invasive analysis provides valuable information on the redox status of the less-easily accessible tissues and organs, and of the whole organism. This review provides an overview of reversible modifications in protein thiols in the setting of CKD and renal replacement therapy. The evidence suggests that protein thiols, S-thiolated proteins, and the protein thiolation index are promising biomarkers of reversible oxidative stress that could be included in the routine monitoring of CKD and ESRD patients.


Changes in energetic metabolism of Biomphalaria glabrata (Mollusca, Planorbidae) in response to exogenous calcium.
Calcium is considered an essential element for the metabolism of aquatic snail Biomphalaria glabrata (Say, 1818), intermediate host of Schistosoma mansoni Sambon, 1907 in Brazil, and represents a limiting factor to its distribution and adaptation to the environment. This study investigated the effect of different concentrations of exogenous CaCO3 on the energetic metabolism of B. glabrata for better understanding the physiological interference of chemical elements dissolved in the environment with the physiology of this species. Sixty-day-old snails were distributed into six groups, five exposed to different concentrations of CaCO3 (20, 40, 60, 80 and 100 mg/L) and a control group. The exposure to CaCO3 was assessed over time, with analysis of 15 snails of each group in the following intervals: 1, 14, 21 or 30 days for hemolymph extraction. Concentrations of calcium and glucose in the hemolymph were determined by commercial kits, and organic acids were extracted using an ion exchange column and analyzed by high-performance liquid chromatography. Concentration of calcium in the hemolymph showed no significant difference (p>0.05) from the control group and between the concentrations tested. Concentration of glucose decreased (p<0.05) in the treatments of exposure to 20 and 40 mg/L and increased when exposed to 80 and 100 mg/L CaCO3 compared to control and to other concentrations tested over 30 days. The organic acids pyruvate, oxaloacetate, citrate, succinate, fumarate, beta-hydroxybutyrate and lactate presented increased concentrations, while propionate and acetoacetate, decreased concentrations, when exposed to CaCO3 compared to control. Considering the influence of different periods of exposure to CaCO3, on the 14th day, there were stronger alterations in the metabolism of B. glabrata. In conclusion, exposure to CaCO3 reduced the concentration of glucose, which is metabolized into pyruvate, the final product of glycolysis, and also influenced the energetic metabolism pathways, indicating an aerobic or partially anaerobic functioning.


Transcriptomic analysis of Saccharomyces cerevisiae physiology in the context of galactose assimilation perturbations.
Despite being extensively studied in various organisms due to scientific, industrial and medical interest, the galactose assimilation function and regulation, and especially its interaction with other parts of cellular physiology, have not been fully elucidated yet. The post-genomic era holistic techniques ("omics") could assist towards this goal. In this paper, we holistically analyzed full-genome Saccharomyces cerevisiae transcriptional profiling data concerning its glucose- and galactose-grown wild-type (WT) physiology and its glucose-grown gal7-deficient (GAL7Delta) physiology, as these were obtained in the experiment described in Lai and Elsas (Biochem. Biophys. Res. Commun. 2000, 271, 392-400). The gal7 gene encodes for the second enzyme of the galactose assimilation, Leloir, pathway, and its deficiency in humans causes a potentially lethal disease, named "classic galactosemia". Analysis of the galactose-grown compared to the glucose-grown WT physiology indicated a significant increase in the transcriptional expression of the ribosomal machinery and decrease in the transcriptional activity of the fatty acids' beta-oxidation at the peroxisomes. Comparison of GAL7Delta to WT physiology in glucose indicated a significant transcriptional increase in the mitochondrial activity and the rate of catabolism of disaccharides, including sucrose, mannose and trehalose, towards amplified biosynthesis of the main cell wall components. Comparison with other physiological conditions indicated strong correlation between the glucose-grown GAL7Delta transcriptional physiology and the WT growth under glucose derepression conditions. Finally, the acquired results and the large number of still unknown genes that were related to the galactose assimilation regulation indicated the need for further, specifically designed, perturbations and integrated analyses of multiple levels of cellular function.


Respiratory function of the placenta as determined with carbon monoxide in sheep and dogs.
A technique is described for studying the respiratory function of the placenta using carbon monoxide, a gas whose exchange across the placenta between the maternal and fetal circulations is limited by diffusion rather than blood flow. During the steady state before the introduction of CO, the normal concentration of carboxyhemoglobin in the ewe, [COHb](M), is approximately 0.90%, and that in the fetus is 2.9%, the ratio [COHb](F)/[COHb](M) being 3.2. In dogs the corresponding values are 1.9%, 4.8%, and 2.4%. After the introduction of CO into the mother animal, CO diffused across the placenta slowly with an equilibration half-time of approximately 2 hours. The average carbon monoxide diffusing capacity (D(Pco)) of the placenta during maternal to fetal exchange was 0.54 ml per (minute x mm Hg x kg fetal weight) (SD +/- 0.13) in sheep and 0.57 ml per (minute x mm Hg x kg) (SD +/- 0.18) in dogs. The fetal to maternal placental diffusing capacity in two sheep was 0.54 ml per (minute x mm Hg x kg). Calculations considering the relative rates of reaction of O(2) and CO with red cell hemoglobin and the relative rates of diffusion of the two gases suggest that the true D(Po2) should be about 1.2 to 2 times greater than the D(Pco) or 0.65 to 1.1 per (minute x mm Hg x kg). This is about 5 times greater than the reported value of D(Po2) calculated from measurements of P(O2) in the mixed uterine and umbilical venous blood. With a diffusing capacity of this magnitude the maternal and fetal placental end capillary P(O2) would approach equilibrium, becoming too small to measure, and the calculation of D(Po2) would be unreliable. We suggest that the apparent end capillary P(o2) gradients of 15 to 20 mm Hg, obtained from sampling uterine and umbilical venous blood, result from a combination of uneven distribution of maternal and fetal placental blood flow and from placental oxygen consumption.


Multiple sclerosis: hyperintense dentate nucleus on unenhanced T1-weighted MR images is associated with the secondary progressive subtype.
To describe the occurrence of abnormal hyperintensity in the dentate nucleus on T1-weighted magnetic resonance (MR) images in patients with multiple sclerosis (MS) as a neuroradiologic sign of gray matter involvement. Presence of the finding was evaluated for association with disability, clinical MS subtype, total lesion volume on T1- and T2-weighted MR images (lesion load), and brain atrophy. Written informed consent was waived by the Ethics Committee because of the retrospective nature of this single-center Institutional Review Board-approved study. MR examinations of 185 patients with MS were reviewed, and 119 patients were included for analysis. Two neuroimagers, who were blinded to clinical data, assessed the presence of a hyperintense dentate nucleus on T1-weighted MR images. The presence of this radiologic alteration was then evaluated in relation to MS subtype, clinical disability, T1 and T2 lesion load, and whole-brain atrophy measurements. Fisher exact, chi(2), and Mann-Whitney U tests were used to evaluate differences in clinical and imaging features between patients with and those without a T1 hyperintense dentate nucleus. Twenty-three (19.3%) of the 119 patients had a hyperintense dentate nucleus on unenhanced T1-weighted MR images. This finding was related to the secondary progressive subtype of the disease, a higher score on the Expanded Disability Status Scale, a higher brain lesion load, and tissue loss. None of the patients with primary progressive MS had a hypterintense dentate nucleus. Hyperintensity of the dentate nucleus may be present on unenhanced T1-weighted MR images of patients with MS and is associated with the secondary progressive disease subtype and with increased clinical disability, lesion load, and brain atrophy.


Decreased expression of miR-125b and miR-100 in oral cancer cells contributes to malignancy.
Altered microRNA (miRNA) expression profiles have been observed in numerous malignancies, including oral squamous cell carcinoma (OSCC). However, their role in disease is not entirely clear. Several genetic aberrations are characteristic of OSCC, with amplification of chromosomal band 11q13 and loss of distal 11q being among the most prevalent. It is not known if the expression levels of miRNAs in these regions are altered or whether they play a role in disease. We hypothesize that the expression of miRNAs mapping to 11q are altered in OSCC because of loss or amplification of chromosomal material, and that this contributes to the development and progression of OSCC. We found that miR-125b and miR-100 are down-regulated in OSCC tumor and cell lines, and that transfecting cells with exogenous miR-125b and miR-100 significantly reduced cell proliferation and modified the expression of target and nontarget genes, including some that are overexpressed in radioresistant OSCC cells. In conclusion, the down-regulation of miR-125b and miR-100 in OSCC appears to play an important role in the development and/or progression of disease and may contribute to the loss of sensitivity to ionizing radiation.


Interaction of torsional and longitudinal guided waves in weakly nonlinear circular cylinders.
The nonlinear forcing terms for the wave equation in general curvilinear coordinates are derived based on an isotropic homogeneous weakly nonlinear elastic material. The expressions for the nonlinear part of the first Piola-Kirchhoff stress are specialized for axisymmetric torsional and longitudinal fundamental waves in a circular cylinder. The matrix characteristics of the nonlinear forcing terms and secondary mode wave structures are manipulated to analyze the higher harmonic generation due to the guided wave mode self-interactions and mutual interactions. It is proved that both torsional and longitudinal secondary wave fields can be cumulative by a specific type of guided wave mode interactions. A method for the selection of preferred fundamental excitations that generate strong cumulative higher harmonics is formulated, and described in detail for second harmonic generation. Nonlinear finite element simulations demonstrate second harmonic generation by T(0,3) and L(0,4) modes at the internal resonance points. A linear increase of the normalized modal amplitude ratio A2/A1(2) over the propagation distance is observed for both cases, which indicates that mode L(0,5) is effectively generated as a cumulative second harmonic. Counter numerical examples demonstrate that synchronism and sufficient power flux from the fundamental mode to the secondary mode must occur for the secondary wave field to be strongly cumulative.


Disability assessment in population surveys: results of the O.E.C.D. Common Development Effort.
In 1976, the OECD sponsored the "Healthfulness of Life" Common Development Effort under the direction of the Social Indicators Programme. With eight member countries and WHO participating, the group set out to develop and statistically define indicators of short and long-term disability for inclusion in population surveys. This paper concentrates on the development and validation testing of the long-term disability items. The technique adopted was one of measuring an individual's ability to perform tasks essential to daily living. The conceptual background and definitions are outlined along with the final recommandations for survey questions and indicator construction put forward to the OECD countries. In addition to the specific testing programmes carried out by the participating countries, the recommended items have been included in other large scale surveys such as the OECD Comprehensive Survey which has been carried out in several countries.


[Update on current care guidelines: Hip fracture].
In Finland approximately 7,000 hip fractures occurred annually during 1996-2008. Risk of hip fracture can be diminished through efforts to prevent falls and osteoporosis. A hip fracture is treated operatively, with the aim of early mobilisation and full weight bearing. Postoperative care and rehabilitation requires multidisciplinary and multifaceted management, focusing on improvement of the patient's physical condition, appropriate pain management, the prevention of delirium and other possible complications. Rehabilitation should be centralised in specialised rehabilitation centres for the elderly. Secondary prevention of future fractures should include management of osteoporosis and fall prevention.


Transition to the adult nephrologist does not induce acute renal transplant rejection.
In spite of the overall increased renal graft survival, long-term allograft survival has remained least successful in adolescent recipients. A major change in their care is the transition from the paediatric to the adult nephrology unit. To analyse the effect of transition on the acute rejection frequency and graft survival, we performed a historical cohort study in all patients transplanted at the paediatric unit between 1980 and 2004. Data were obtained by reviewing medical charts in two of the four Dutch pediatric renal transplantation centers from time of transplantation until 3 years after transition. For analysis, we used a Cox proportional hazards model. The cohort consisted of 162 patients: 133 native Dutch and 29 immigrant patients. Transition occurred at a mean age of 18 years (range 14-22). At transition, 72% had a functioning allograft. Acute rejections occurred in 92/162 patients before (median follow-up 4.8 years, range 0.2-12.8) and in 15/116 patients after transition (median follow-up 3.0 years, range 1.6-3.0). Most rejections (62%) occurred within the first year after transplantation. The relative risk of acute rejections after transition in comparison to before transition was 0.10 [95% confidence interval (95% CI) 0.04-0.28] in Dutch patients and 0.69 (95% CI 0.33-1.40) in immigrant patients. In the 3 years before transition, 28/154 patients (18%) experienced graft failure compared to 19/116 patients (16%) in the 3 years after transition. The risk for acute rejection decreases after transition to the adult unit. There is less risk reduction in immigrant patients. Nephrologists should pay special attention to these patients.


Is high concentration of parvalbumin a requirement for superfast relaxation?
It is generally thought that the rapid relaxation of fast muscles is facilitated by the Ca(2+) binding protein parvalbumin (Parv). Indeed superfast swimbladder (SWB) muscle of toadfish contains the largest concentration of this protein ever observed (up to 1.5 mM). At 15 degrees C toadfish perform a 100 Hz call, 400 ms in duration, followed by a long (5-15 s) intercall interval. It has been proposed that Parv helps sequester the Ca(2+) during the call, and then Ca(2+) unbinds and is pumped back into the sarcoplasmic reticulum during the long intercall interval. Midshipman (Porichthys notatus) is another fish which calls at a high frequency; 80-100 Hz at a temperature of 12-15 degrees C. However, unlike toadfish, midshipman call with a 100% duty cycle. Without an intercall interval, Parv would seem of little use as it would become saturated early in calling. Here we show that the midshipman SWB has only about 1/8th of the Parv in toadfish. Moreover, total Parv content in calling male midshipman SWB was not different from that in the non-calling female and the much slower locomotory muscles. These data suggest that Parv does not play a large role in the calling of midshipman, which is accomplished without a high concentration of this protein. Native gel-electrophoresis also revealed presence of three major (PA-I, PA-II and PA-III) and two minor (PA-Ia and PA-IIIa, <5% of total content) Parv isoforms in adult toadfish SWB. Midshipman SWB contained about equal amounts of PA-I and PA-II and also a small (approximately 10%) amount of PA-III. By amino acid composition, toadfish PA-Ia and PA-I isoforms were different from PA-II and PA-III isoforms (by 24 and 14 residues, respectively).


Electrostatic force microscope for probing surface charges in aqueous solutions.
A scanning force microscope was converted to an electrostatic force microscope by charging the usually neutral cantilever with phospholipids. The electrostatic force microscope was used to study surface electrostatic charges of samples in aqueous solutions. Lysozymes, DEAE-Sephadex beads, 3-propyltriethoxysilane-treated glass and mica were imaged in water or phosphate buffer with electrostatic force microscopy. The adhesion force measured when a charged probe and oppositely charged specimen interacted was up to 500 times greater than when a bare probe was used. This dramatic increase in measured adhesion force can be attributed to the energy required to break the salt bridges formed between the charged probe and the specimen. The use of phospholipids to functionalize the cantilever tip allows the incorporation of other biomolecules and ligands that can be used as biologically specific tips (e.g., receptors, drugs) for the study of intermolecular interactions.


Characterization of the cooperative function of inhibitory sequences in Ets-1.
DNA binding by the eukaryotic transcription factor Ets-1 is negatively regulated by an intramolecular mechanism. Quantitative binding assays compared the DNA-binding activities of native Ets-1, three deletion mutants, and three tryptic fragments. Ets-1 and activated Ets-1 polypeptides differed in DNA-binding affinity as much as 23-fold. Inhibition was mediated by two regions flanking the minimal DNA-binding domain. Both regions regulated affinity by enhancing dissociation of the protein-DNA complex. Three lines of evidence indicated that inhibition requires cooperative interaction between the two regions: first, the two inhibitory regions acted through a common mechanism; second, neither region functioned independently of the other; finally, mutation of the C-terminal inhibitory region altered the conformation of the N-terminal inhibitory region. In addition, partial proteolysis detected an identical altered conformation in the N-terminal inhibitory region of Ets-1 bound to DNA. This finding suggested that repression is transiently disrupted during DNA binding. These results provide evidence that the two inhibitory regions of Ets-1 are structurally, as well as functionally, coupled. In addition, conformational change is shown to be a critical component of the inhibition mechanism. A cooperative, allosteric model of autoinhibition is described. Autoinhibition of Ets-1 could be relieved by either protein partner(s) or posttranslational modifications.


Growth hormone-releasing factor analogue (hGRF1-29NH2): immunoreactive-GRF plasma levels after intravenous and subcutaneous administration.
A homologous radioimmunoassay (RIA) system for human growth hormone-releasing factor 1-29NH2 (hGRF 1-29-NH2) was developed and applied to the measurement of immunoreactive (IR) concentrations of the peptide in anaesthetized rats to determine some of its pharmaco-kinetics after i.v. and s.c. administration. Analysis of the biphasic disappearance curve of IR-hGRF-1-29NH2 from plasma after i.v. injection (10 micrograms) gave values for the half-lives of the initial distribution phase (alpha) and for the elimination phase (beta) of 1.9 +/- 0.2 min and 10.4 +/- 0.2 min respectively. There was rapid uptake of the peptide from the site of s.c. injection but comparison of areas under the plasma IR-hGRF1-29NH2/time curves showed that the estimated total amount in the circulation after s.c. injection was only 4% of that obtained after i.v. administration. A large degree of degradation of the peptide is indicated either at the site of injection or during transfer to plasma; this susceptibility to rapid breakdown is reflected in the short half-life of the peptide in the circulation. Therefore the measurement of the above parameters is a prerequisite when assessing potency of GRF analogues in-vivo and when implementing suitable dosage regimens for clinical purposes.


Association of Otolaryngology Resident Duty Hour Restrictions With Procedure-Specific Outcomes in Head and Neck Endocrine Surgery.
Graduate medical education has undergone a transformation from traditional long work hours to a restricted plan to allow adequate rest for residents. The initial goal of this restriction is to improve patient outcomes. To determine whether duty hour restrictions had any impact on surgery-specific outcomes by analyzing complications following thyroid and parathyroid procedures performed before and after duty hour reform. Retrospective cross-sectional analysis of the National Inpatient Sample (NIS).The NIS was queried for procedure codes associated with thyroid and parathyroid procedures for the years 2000 to 2002 and 2006 to 2008. Hospitals were divided based on teaching status into 3 groups: nonteaching hospitals (NTHs), teaching hospitals without otolaryngology programs (THs), and teaching hospitals with otolaryngology programs (THs-OTO). Procedure-specific complication rates, length of stay, and mortality rates were collected. SAS statistical software (version 9.4) was used for analysis with adjustment using Charlson comorbidity index. Total numbers of head and neck endocrine procedures were 34 685 and 39 770 (a 14.7% increase), for 2000 to 2002 and 2006 to 2008, respectively. THs-OTO contributed a greater share of procedures in 2006 to 2008 (from 18% to 25%). With the earlier period serving as the reference, length of stay remained constant (2.1 days); however, total hospital charges increased (from $12 978 to $23 708; P < .001). Rates of postoperative hematoma (odds ratio [OR], 1.21; 95% CI, 1.06-1.38), hypoparathyroidism (OR, 1.27; 95% CI, 1.06-1.52), and unintentional vessel lacerations (OR, 1.36; 95% CI, 1.02-1.83) increased overall with NTHs (OR, 1.26; 95% CI, 1.04-1.52), THs (OR, 1.65; 95% CI, 1.15-2.37), and THs-OTO (OR, 1.98; 95% CI, 1.09-3.61) accounting for these differences, respectively. Overall mortality decreased (OR, 0.66; 95% CI, 0.47-0.94) following a decrease in the TH-OTO mortality rate (OR, 0.34; 95% CI, 0.12-0.93). While recurrent laryngeal nerve injury, hematoma formation, and hypoparathyroidism did not change, length of stay and mortality improved within THs-OTO following head and neck endocrine procedures after implementation of duty hour regulations. This finding refutes the concern that duty hour restrictions result in poorer overall outcomes. Less time available to develop technical competence may play a factor in some outcomes in lieu of recurrent laryngeal nerve injury increasing within THs and accidental injury to vessels, organs, or nerves and hypocalcemia increasing within THs-OTO. Furthermore, head and neck endocrine cases increased at THs with otolaryngology programs.


Relationship of intrinsic connections to forelimb movement representations in monkey motor cortex: a correlative anatomic and physiological study.
1. Intracortical microstimulation (ICMS) and horseradish peroxidase (HRP) histochemistry were combined to examine the relationship between intrinsic connections and intracortical microstimulation sites eliciting evoked movements in the forelimb representation of adult macaque monkey motor cortex. 2. The distribution of sites from which stimulation-evoked movements about individual forelimb joints were elicited under anesthesia varied considerably among animals. Identical movements could often be elicited from multiple, noncontiguous sites. 3. After single, small extracellular HRP injections at sites from which thumb movement was evoked, small groups of retrogradely labeled cells and dense patches of axon terminations were found scattered across a wide area of the forelimb representation. Terminal patches were discontinuous and arose from horizontal, intracortical axons. 4. Correlating the HRP labeling with the physiologically defined movement maps revealed a profuse set of intrinsic, bidirectional connections that connect digit representations and representations of movements about the wrist, elbow, and shoulder. 5. HRP injections placed in the forelimb representation close to the physiologically defined face representation resulted in virtually no retrogradely labeled cells or terminal fiber labeling that crossed into the face representation. A patch of anterolaterally placed label that was present may be the dissociated rostrolateral arm area of other authors. 6. Taken together, these data suggest that extensive, horizontally oriented, intrinsic axon collaterals provide inputs to many different forelimb movement representations and may be recruited during complex movements to coordinate the activity of motor cortical zones whose predominant output is to forelimb muscle groups acting synchronously.


[Effect of products of germ-line and somatic gene mutations on chromatin remodeling].
Chromatin remodeling is engaged in basic cell functions as DNA replication, recombination, DNA repair and transcription of genes. Chromatin is remodeled by ATP-dependent chromatin remodeling complexes and protein complexes covalently modifying histones. Germ-line mutations of the genes coding for proteins which participate in chromatin remodeling cause severe developmental diseases and they increase a risk of cancer development. Somatic mutations and translocations of such genes are associated with cancer development of certain tumors. Chromatin remodeling mechanisms can thus be targeted by special strategy of cancer therapy.


Flat Panel Detector c-Arms Are Associated with Dramatically Reduced Radiation Exposure During Ureteroscopy and Produce Superior Images.
Background: We wished to determine whether newly available flat panel detector (FPD) c-arms were (1) associated with lower radiation dose during ureteroscopy (URS) than conventional image intensifier (CII) c-arms and (2) to compare fluoroscopic image quality between the units. Materials and Methods: We retrospectively reviewed 44 consecutive patients undergoing URS at a pediatric hospital, with c-arms assigned by availability in the operating room. We performed dosimetry experiments using the same c-arms on standard phantoms. Results: Patient and case characteristics did not differ significantly between the two groups of patients. The median dose in the FPD group was less than a quarter of the dose in the CII group, 0.48 [0.42, 0.97] mGy vs 2.2 [1.1, 3.8] mGy, p < 0.0001. The FPD dose remained at less than one-third of the CII dose accounting for any difference in fluoroscopy time, and remained significant in a multivariate model including fluoroscopy time and patient weight (β = 2.4, p = 0.007). Phantom studies showed higher image quality for FPDs at all simulated patient sizes, even at lower radiation doses. Conclusions: This is the first report comparing radiation dose from c-arms of image intensifiers and FPDs in adults or children. Use of an FPD during URS was associated with a substantially decreased absorbed dose for patients while simultaneously improving image quality.


Impact of occupational dust exposure on dermatologic symptoms among Korean workers.
Occupation-related dust exposure is common, especially with increased industrialization. While occupational dust-related health issues caused by inhalation or ingestion have been well studied, only a few studies have examined the dermatologic effects of occupational dust exposure. This study aimed to investigate the association between occupational dust exposure and dermatologic symptoms in Korean workers. Among the large-scale representative data from the fifth Korean Working Conditions Survey, 45,700 workers were selected for study. Occupational dust exposure level was categorized as none, moderate, and severe, and dermatologic symptoms were assessed using a questionnaire on health problems. We analyzed the association between occupational dust exposure and dermatologic symptoms using multivariate logistic regression. Risks of skin problems and work-related skin problems were significantly associated with the level of occupational dust exposure in a dose-dependent manner (odds ratio (95% confidence interval): moderate, 1.51 (1.14-2.01); severe, 2.39 (1.74-3.29) in general skin problems; moderate, 1.72 (1.22-2.42); severe, 3.06 (2.11-4.44) in work-related skin problems). We, thus, demonstrate an association between occupational dust exposure and skin problems. As skin absorption of dust is a major route of dust exposure at the workplace, it is necessary to determine the efficacy of continuous management of occupational dust exposure.


Identification of a unique gene expression signature that differentiates hepatocellular adenoma from well-differentiated hepatocellular carcinoma.
It is often difficult to distinguish hepatocellular adenoma (HCA) from well-differentiated hepatocellular carcinoma (WDHCC) when limited tissue from a needle biopsy is evaluated. The aim of this study was to identify gene expression patterns that can distinguish HCA from WDHCC, with the ultimate goal of discovering novel diagnostic markers. Gene expression profile analysis was performed using Affymetrix U133Plus2 GeneChip microarrays on RNA isolated from frozen tissue of 6 HCA and 8 WDHCC specimens. Statistical analysis of microarray data identified 63 genes whose expression levels were significantly different between HCA and WDHCC. These included 57 genes overexpressed by HCA and 6 overexpressed by WDHCC. Eight genes were chosen for further analysis by quantitative RT-PCR on RNA derived from archived, paraffin-embedded tissue blocks of an independent validation set comprising 9 HCAs and 9 HCCs. Seven of the 8 genes demonstrated average expression differences between HCA and HCC that were concordant with the microarray findings, and their expression pattern correctly classified the 18 tumors into HCA and HCC using unsupervised clustering analysis. Furthermore, immunohistochemical staining performed on a third, independent set of 27 HCAs and 33 HCCs confirmed the expression differences at protein levels for 5 of the genes. Taken together, our data demonstrate significant molecular differences between HCA and WDHCC, despite their morphologic similarity. More importantly, we have identified a unique set of genes whose expression pattern can discriminate between these two types of hepatocellular neoplasms, suggesting the possibility of future development of ancillary molecular and immunohistochemical diagnostic methods.


Role of intracellular cation abnormalities in development of left ventricular hypertrophy.
To elucidate factors determining the development of left ventricular hypertrophy, we performed measurements of intraerythrocyte sodium and intraplatelet calcium concentrations, measurements of plasma and urinary catecholamine levels and plasma renin activity, calculation of left ventricular mass using echocardiography, and 24-h ambulatory blood pressure (BP) monitoring. The results indicate that abnormalities of sodium and calcium mobilization may play an important role in the adaptation of left ventricular muscle to a persistent increase in BP.


[Beta-lactam antibiotic sensitivity in Aeromonas spp. of clinical, animal, and environmental origin].
Susceptibility to beta-lactam antibiotics was investigated in Aeromonas spp. Microorganisms were isolated from both, clinical and water creek samples, as well as from processed raw chicken carcasses. Aeromonas like colonies were identified by means of Aerokey II and API 20 E System (Bio-Merieux). A. hydrophila prevailed both of human origin (44%) and water creek samples (41%), while A. caviae ranked first among raw chicken samples (65%). Dilution testing by Agar Method was performed to determine minimum inhibitory concentration (MIC), following NCCLS standards. All tested microorganisms were susceptible to third generation cephalosporin, cefepime, imipenem, aztreonam, and resistant to ampicillin. Only with cefepime and aztreonam exceptions, strains of human origin showed higher values of MIC90 than environmental ones. These results suggest that antibiotic resistance is mainly due to a steady environmental pressure, on account of the widely used above mentioned compounds.


Pathological lesions in vascular dementia.
According to current diagnostic criteria, a definite diagnosis of vascular dementia (VaD) can be reached on pathological grounds by showing the presence of vascular lesions and the absence of degenerative changes exceeding those expected for age. However, while it is commonly accepted that VaD is a group of heterogeneous entities rather than a process with a unique pathological substrate, the spectrum of vessel and parenchyma changes etiologically associated with the clinical syndrome remains basically unidentified. The review of some recent clinical-pathological series shows that different studies have assessed the presence of dissimilar vascular lesions and that, in many cases, no pathological definition was given. This has hindered the clarification of clinical-pathological correlations in the field of VaD. In this scenario, the use of animal models of cerebrovascular diseases may help to elucidate the type of lesions possibly linked with cognitive impairment in humans and might provide insight into some of the pathophysiological mechanisms of vascular cognitive impairment. A consensus is today needed in order to harmonize the pathological examination of vascular lesions in cases of dementia. An ongoing survey aimed at collecting information about the procedures used in different pathological laboratories in the assessment of lesions possibly associated with dementia is finally presented.


Distinguishing transmembrane helices from peripheral helices by circular dichrosim.
The interpretation of the circular dichroism (c.d.) spectra of proteins to date requires additional secondary structural information of the proteins to be analysed (e.g., X-ray or n.m.r. data). Therefore these methods are inappropriate for a c.d. database whose secondary structures are unknown, as in the case of the membrane proteins. The Convex Constraint Analysis algorithm [Perczel, Hollósi, Tusnady, and Fasman (1991) Convex constraint analysis: a natural deconvolution of circular dichroism curves of proteins. Protein Eng. 4, 669-679] operates on a collection of c.d. spectral data to extract the common spectral components with their spectral weights. The linear combinations of these derived 'pure' c.d. curves can reconstruct the original data set with great accuracy. For a membrane protein data set, the five-component spectra so obtained from the deconvolution consisted of two different types of alpha-helices (the alpha-helix in the soluble domain and the alpha1-helix, for the transmembrane alpha-helix), a beta-pleated sheet, a class-C-like spectrum related to beta-turns and a spectrum correlated with the unordered conformation. The deconvoluted c.d. spectrum for the alpha1-helix was characterized by a positive red-shifted band in the range 195-200 nm (+95,000 degrees.cm2-dmol-1), with the intensity of the negative band at 208 nm being slightly less negative than that of the 222 nm band (-50,000 and -60,000 degrees.cm2.dmol-1 respectively) in comparison with the regular alpha-helix, with a positive band at 190 nm and two negative bands at 208 and 222 nm with magnitudes of +70,000, -30,000 and -30,000 degrees.cm2.dmol-1 respectively.


Measuring Perceived Level of Integration During the Process of Primary Care Behavioral Health Implementation.
Provider- and staff-perceived levels of integration were measured during implementation of a primary care behavioral health clinic; these data were used to tailor and evaluate quality improvement strategies. Providers and staff at an urban, academic, adult primary care clinic completed the 32-item Level of Integration Measure (LIM) at baseline and 7 months. The LIM assesses 6 domains of integrated care. Overall and domain scores were calibrated from 0 to 100, with ≥80 representing a highly integrated clinic. Response rate was 79% (N = 46/58) at baseline and 83% (N = 52/63) at follow-up. Overall, LIM score increased from 64.5 to 70.1, P = .001. The lowest scoring domains at baseline were targeted for quality improvement and increased significantly: integrated clinical practice, 60.0 versus 68.4, P < .001; systems integration, 57.0 versus 63.8, P = .001; and training, 56.7 versus 65.3, P = .001. Ongoing quality improvement, including organizational and financial strategies, is needed to achieve higher levels of integration.


Structural and functional characterization of gene products encoded in the human genome by homology detection.
Availability of the human genome data has enabled the exploration of a huge amount of biological information encoded in it. There are extensive ongoing experimental efforts to understand the biological functions of the gene products encoded in the human genome. However, computational analysis can aid immensely in the interpretation of biological function by associating known functional/structural domains to the human proteins. In this article we have discussed the implications of such associations. The association of structural domains to human proteins could help in prioritizing the targets for structure determination in the structural genomics initiatives. The protein kinase family is one of the most frequently occurring protein domain families in the human proteome while P-loop hydrolase, which comprises many GTPases and ATPases, is a highly represented superfamily. Using the superfamily relationships between families of unknown and known structures we could increase structural information content of the human genome by about 5%. We could also make new associations of domain families to 33 human proteins that are potentially linked to genetically inherited diseases.


[Distribution of recombinant human immunodeficiency virus type-1 CRF01_AE strains in China and its sequence variations in the env V3-C3 region].
To characterize CRF01_AE strains of recombinant human immunodeficiency virus type-1 (HIV-1) found in the Second National Molecular Epidemiology Study on HIV in China and to analyze its sequence variation in the env V3-C3 region during the First National Molecular Epidemiology Study (NMES1, 1996 - 1998) to the Second National Molecular Epidemiology Study (NMES2, 2001 - 2002). DNA was extracted from peripheal blood mononuclear cells of the subjects with HIV infection. The env C2-V4 region of HIV-1 was amplified with nested polymerase chain reaction (n-PCR). PCR products were directly sequenced using ABI 377 DNA sequencer, then the gene-based phylogenetic tree was constructed and its variation of amino acids was analyzed with GCG software. Totally, 169 strains of recombinant HIV-1 CRF01_AE were identified from blood samples collected from different high risk groups in 17 of 31 provinces, municipalities and autonomous regions all over China by the end of 2002. Although sexual transmission still dominated during NMES1 (62.2%, 23/37) and NMES2 (55.3%, 73/132), prevalence of HIV-1 CRF01_AE in intravenous drug users (IDUs) increased to 41.6% (57/137) during NMES2 from 27% (10/37) during NMES1. Phylogenetic tree analysis showed that HIV-1 CRF01_AE strains prevalent in IDUs during NMES2 did not cluster with those prevalent in the subjects infected by sexual transmission during NMES2 and those in IDUs during NMES1. The amino acid residues of V3 region of HIV-1 CRF01_AE in IDUs were relatively conservative, but the sixth, eighth, ninth, tenth, twelfth, fifteenth, sixteenth amino acid residues of C3 region displayed regular changes. HIV-1 CRF01_AE strain has been introduced into inland provinces from southeastern coast areas and southwestern border areas, with an increasing prevalence in IDUs. The sequence of env V3-C3 region of recombinant HIV-1 CRF01_AE strains prevalent in IDUs during NMES2 was obviously different from that during NMES1, suggesting that HIV-1 CRF01_AE strains prevalent in IDUs during NMES2 might come from a new source and have a potential to spread.


In vitro dimensions and curvatures of human lenses.
The purpose of this study was to determine dimensions and curvatures of excised human lenses using the technique of shadowphotogrammetry. A modified optical comparator and digital camera were used to photograph magnified sagittal and coronal lens profiles. Equatorial diameter, anterior and posterior sagittal thickness, anterior and posterior curvatures, and shape factors were obtained from these images. The data were used to calculate lens volumes, which were compared with the lens weights. Measurements were made on 37 human lenses ranging in age from 20 to 99 years. These showed that lens dimensions and the anterior radius of curvature increase linearly throughout adult life while posterior curvature remains constant. The relative shape (or aspect ratio) of the posterior lens is unchanged through adult life since both equatorial diameter and posterior thickness increase at the same rate. The ratio of anterior thickness to posterior thickness is constant at 0.70. It is suggested that in vivo forces alter the apparent location of the lens equator, that the in vitro lens shape corresponds to the maximally accommodated shape in vivo and that the shapes of the accommodated and unaccommodated lens progressively converge toward each other due to lens growth with age, with a convergence point located near the age of total loss of accommodation (55-60 years). Together, these observations provide additional support for the Helmholtz theory of accommodation.


Magnetic sensing via ultrasonic excitation.
We present ultrasonic techniques for magnetic measurements. Acoustically modulated magnetization is investigated with sensitive rf detection by narrowband loop antennas. Magnetization on the surface of ferromagnetic metals is temporally modulated with the rf frequency of the irradiated ultrasonic waves, and the near-field components emitted from the focal point of the ultrasonic beam are detected. Based on the principle of the acoustically stimulated electromagnetic (ASEM) response, magnetic sensing and tomography are demonstrated by ultrasonic scanning. We show that ASEM imaging combines good acoustic resolution with magnetic contrast. The sensitivity of this method is estimated to be about 6 G/Hz(1∕2) in our current setup.


Allelic silencing at the tumor-suppressor locus 13q14.3 suggests an epigenetic tumor-suppressor mechanism.
Genomic material from chromosome band 13q14.3 distal to the retinoblastoma locus is recurrently lost in a variety of human neoplasms, indicating an as-yet-unidentified tumor-suppressor mechanism. No pathogenic mutations have been found in the minimally deleted region until now. However, in B cell chronic lymphocytic leukemia tumors with loss of one copy of the critical region, respective candidate tumor-suppressor genes are down-regulated by a factor >2, which would be expected by a normal gene-dosage effect. This finding points to an epigenetic pathomechanism. We find that the two copies of the critical region replicate asynchronously, suggesting differential chromatin packaging of the two copies of 13q14.3. Although we also detect monoallelic silencing of genes localized in the critical region, monoallelic expression originates from either the maternal or paternal copy, excluding an imprinting mechanism. DNA methylation analyses revealed one CpG island of the region to be methylated. DNA demethylation of this CpG island and global histone hyperacetylation induced biallelic expression, whereas replication timing was not affected. We propose that differential replication timing represents an early epigenetic mark that distinguishes the two copies of 13q14.3, resulting in differential chromatin packaging and monoallelic expression. Accordingly, deletion of the single active copy of 13q14.3 results in significant down-regulation of the candidate genes and loss of function, providing a model for the interaction of genetic lesions and epigenetic silencing at 13q14.3 in B cell chronic lymphocytic leukemia.


Crisaborole Ointment 2%: A Review in Mild to Moderate Atopic Dermatitis.
Crisaborole ointment 2% (Eucrisa™) is a novel, anti-inflammatory inhibitor of phosphodiesterase 4 (PDE4) that is available in the USA for the topical treatment of mild to moderate atopic dermatitis in patients aged ≥ 2 years. In two short-term (28 days), identically designed, multicentre, phase III studies in this patient population, topical therapy with crisaborole ointment 2% reduced disease severity and pruritus severity compared with vehicle, with the effect established early and sustained over the course of treatment. Improvements in the other signs of atopic dermatitis (erythema, exudation, excoriation, induration/papulation, and lichenification) were also seen. Crisaborole ointment 2% was generally well tolerated in the short-term studies, with its favorable safety profile maintained over the longer term (up to 52 weeks) in a multicentre, extension study. Most treatment-emergent adverse events (TEAEs) were of mild to moderate severity and considered unrelated to the study medication. Moreover, the incidence of application-site pain following short- and longer-term topical therapy with crisaborole ointment 2% was low. In conclusion, crisaborole ointment 2% is an effective and generally well tolerated new topical option for the management of mild to moderate atopic dermatitis in patients aged ≥ 2 years, with the potential to effectively treat this patient population over the longer term without the safety concerns associated with other current topical anti-inflammatory agents.


Relationships between hypothalamic neuropeptide Y and food intake in the lactating rat.
NPYergic neurons in the hypothalamic arcuate nucleus (ARC) that project to the paraventricular nucleus (PVN) are postulated to regulate food intake and energy balance. This projection is overactive in lactation and is thought to drive hyperphagia in this condition. We have explored further the relationship between hypothalamic NPY and food intake in lactation and tested the hypothesis that hypoinsulinemia is the stimulus to NPY neuronal activity. Compared with nonlactating controls (n = 10), freely fed lactating rats (n = 9) showed significantly increased (p < 0.05) NPY levels in the ARC and medial preoptic area (MPO), but there was no significant increase in whole hypothalamic NPY mRNA levels. Lactating rats (n = 8) that were restricted to control rats' food intake for 3 days showed generally higher hypothalamic NPY levels, with significantly higher concentrations than controls (p < 0.05) in the ARC, MPO, PVN, and lateral hypothalamic area (LHA); NPY mRNA levels were also significantly increased (p < 0.05). Across all three experimental groups, there was a significant inverse correlation between plasma insulin concentration and hypothalamic NPY mRNA levels (r = -0.39, p < 0.01). We conclude that the ARC-PVN projection is overactive in lactation and that this is not a consequence of hyperphagia. Hypoinsulinemia may stimulate these neurons, as it is thought to do in other conditions of energy deficit.


Enhancing passive sampling tools for detecting marine bioinvasions.
Early detection is important for successful management of invasive species, but optimising monitoring systems to detect multiple species from different taxonomic groups remains a major challenge. Settlement plates are often used to monitor non-indigenous marine species (NIMS) associated with vessel biofouling, but there have been few assessments of their fitness-for-purpose. We deployed arrays of settlement plates ("settlement arrays") containing combinations of treatments that reflected conditions associated with the vessel transport pathway (i.e., copper based antifouling coatings, shaded habitat) to determine the treatment combinations that maximised NIMS diversity. Horizontal (shaded) treatments preferentially sampled higher NIS diversity than vertical plates. Although plates with copper-based biocides had larger proportions of NIS to indigenous species, they sampled only a subset of NIS diversity. Overall diversity was greatly enhanced through use of multiple treatments, demonstrating benefits of multi-faceted sampling arrays for maximising the potential taxonomic and species richness.


Hepatocellular carcinoma with mediastinal and pericardial invasion: report of two cases.
Mediastinal invasion with pericardial involvement in hepatocellular carcinoma (HCC) is rarely described. We report two patients with hepatitis-C-related HCC, who, after several courses of transcatheter arterial chemoembolization (TACE), developed mediastinal and pericardial neoplastic growth. Both patients presented with clinical manifestations of exertional dyspnea, chest pain and orthopnea. The diagnosis of HCC with pericardial involvement, through direct invasion of the anterior mediastinum, was established by computerized tomography and magnetic resonance imaging. These patients' symptoms were relieved after they received radiotherapy with a total dosage of 3,000 and 4,000 cGy over a three-week and four-week period, respectively. We suggest that direct mediastinal invasion with pericardial involvement should be considered when evaluating patients with advanced HCC who developed precordial distress following palliative TACE. Early recognition of this unusual complication is important in the management of HCC.


Start-up of a pilot-scale anaerobic fixed film reactor at low temperature treating slaughterhouse wastewater.
A pilot-scale anaerobic fixed film reactor (AFFR) with vertically arranged PVC tubes as biomass carrier, treating poultry slaughterhouse wastewater was started-up in 74 days at temperatures between 20-24 degrees C. The start-up process consisted of a long acclimatization phase followed by a low loaded growth phase, a gradual increase of OLR upto 9.2 kg COD/m3d, and a final maturation phase at moderated loads of 2.7 kg COD/m3d at which total COD removal efficiencies of 57% were achieved. Alkalinity ratio IA:PA was found to be the best control parameter to avoid VFA accumulation. OLR increase based on pH control was not satisfactory because changes in CO2 solubility caused daily by temperature and flow variations led to pH oscillations of 0.2 units. The low wastewater alkalinity, 260 mg/l CaCO3 was insufficient to buffer the pH system, therefore the pH decrease associated with the VFA accumulation was not easily detected and could not be used as a way of OLR control. Organic matter removal took place by accumulation and biodegradation processes. Limitation in the reactor hydrodynamics and particulate fraction hydrolysis was detected at high flow rates.


Stevens-Johnson syndrome-toxic epidermal necrolysis (SJS-TEN) overlap associated with carbamazepine use.
A benign pruritic rash occurs in 10%-15% of persons treated with carbamazepine. A small fraction of them may experience life-threatening dermatological syndromes such as exfoliative dermatitis, erythema multiforme, Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). The case of an 18-year-old female suffering from bipolar affective disorder (mania) who was being treated with carbamazepine, lithium, chlorpromazine and benzhexol is presented. After 10 days of treatment, she developed high-grade fever and mucocutaneous manifestations of SJS-TEN overlap. She was treated in hospital with systemic corticosteroids, antibiotics, intravenous fluids and other supportive measures, and recovered after 3 weeks.


[Characteristics and prognosis of visual deficit caused by parasellar meningiomas].
Meningiomas are the most common slow growing tumors of the central nervous system which arise from the arachnoid cells. One certain group of them--tuberculum sellae (TS), cavernous sinus (CS), anterior clinoid (AC), and planum sphenoidale (PS) meningiomas, a group often called "parasellar", present with monocular or binocular visual deterioration, which is often unrecognized by patients until visual loss is severe and the tumor has reached a significant size. To describes the type of visual deficit, severity of the symptoms and surgical treatment of tumors with the above mentioned localization. During the period 2003-2007 year, 93 patients (31 man, 62 women) diagnosed with meningiomas of the TS (31 cases), CS (16 cases), AC (21 cases), PS (17 cases), (see text) - 8, were diagnosed and operated on in the Department of Neurosurgery, University Hospital "St. Ivan Rilsky", Sofia, Bulgaria. Retrospective analysis was done. Age at diagnosis ranged from 12 to 74 years, but the majority of cases were in the 5th and 6th decade of life. Localization of was: TS 33%, CS 17%, AC 24%, PS 18% and with more than one area affected in 8 %. The most common symptom was slowly progressing loss of monocular visual acuity (81% of cases). 68% of patients presented with already severely deteriorated vision (0.1-0). Neuro-ophthalmological examination (see text) the patients. Computed tomography and magnetic resonance imaging were the main diagnostic methods. "Gross" total resection was achieved in 62%, subtotal in 12%, partial in 26% of the cases. There was early improvement in 57% of the patients. Death due to cardio-respiratory insufficiency occurred in one case. the progressive monocular visual deficit is often the first clinical symptom of the parasellar menigiomas. The early diagnosis is important for the success of surgery. Close collaboration between different specialists (ophthalmologists, neurologist and neurosurgeons) is essential for treatment of these tumors and prevention of the visual deterioration.


Training at the optimum power zone produces similar performance improvements to traditional strength training.
The purpose of this study was to test if substituting a regular maximum strength-oriented training regimen by a power-oriented one at the optimal power load in the first phase of a traditional periodization produces similar performance improvements later on into the training period. Forty five soldiers of the Brazilian brigade of special operations with at least one year of army training experience were divided into a control group (CG - n = 15, 20.18 ± 0.72 yrs, 1.74 ± 0.06 m, 66.7 ± 9.8 kg, and 1RM/weight ratio = 1.14 ± 0.12), a traditional periodization group (TG - n = 15, 20.11 ± 0.7 yrs, 1.72 ± 0.045 m, 63.1 ± 3.6 kg, and 1RM/weight ratio = 1.21 ± 0.16); and a maximum-power group (MPG - n = 15, 20.5 ± 0.6 yrs, 1.73 ± 0.049m, 67.3 ± 9.8 kg, 1RM/weight ratio = 1.20 ± 0.14). Maximum strength (26.2% and 24.6%), CMJ height (30.8% and 39.1%) and sprint speed (11.6% and 14.5%) increased significantly (p < 0.05) and similarly for the MPG and TG, respectively, from pre- to post-assessments. Our data suggests that a power training regimen may be used in the initial phase of the training cycle without impairing performance later on into the training period. Key pointsTraining at the optimal power zone during two mesocycles of a traditional periodization did not hamper strength, speed and power performance improvements.Additional research is required in order to find out if longer periods of training at optimal power zone are capable of producing similar performance improvements to traditional strength training regimen.


Spatial differentiation of metabolism in prostate cancer tissue by MALDI-TOF MSI.
Prostate cancer tissues are inherently heterogeneous, which presents a challenge for metabolic profiling using traditional bulk analysis methods that produce an averaged profile. The aim of this study was therefore to spatially detect metabolites and lipids on prostate tissue sections by using mass spectrometry imaging (MSI), a method that facilitates molecular imaging of heterogeneous tissue sections, which can subsequently be related to the histology of the same section. Here, we simultaneously obtained metabolic and lipidomic profiles in different prostate tissue types using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MSI. Both positive and negative ion mode were applied to analyze consecutive sections from 45 fresh-frozen human prostate tissue samples (N = 15 patients). Mass identification was performed with tandem MS. Pairwise comparisons of cancer, non-cancer epithelium, and stroma revealed several metabolic differences between the tissue types. We detected increased levels of metabolites crucial for lipid metabolism in cancer, including metabolites involved in the carnitine shuttle, which facilitates fatty acid oxidation, and building blocks needed for lipid synthesis. Metabolites associated with healthy prostate functions, including citrate, aspartate, zinc, and spermine had lower levels in cancer compared to non-cancer epithelium. Profiling of stroma revealed higher levels of important energy metabolites, such as ADP, ATP, and glucose, and higher levels of the antioxidant taurine compared to cancer and non-cancer epithelium. This study shows that specific tissue compartments within prostate cancer samples have distinct metabolic profiles and pinpoint the advantage of methodology providing spatial information compared to bulk analysis. We identified several differential metabolites and lipids that have potential to be developed further as diagnostic and prognostic biomarkers for prostate cancer. Spatial and rapid detection of cancer-related analytes showcases MALDI-TOF MSI as a promising and innovative diagnostic tool for the clinic.


Tannin-Based Copolymer Resins: Synthesis and Characterization by Solid State 13C NMR and FT-IR Spectroscopy.
In recent years, the interest for bio-sources is rising exponentially and tannins extracts are one of the most interesting, easily-available, phenolic building blocks. The condensed tannins or proanthocyanidins are already known for their polymerization chemistry, which is the basis for several natural-based materials (e.g., adhesives, foams). In the present work we aim to observe the behavior of the extract of Acacia Mimosa (Acacia mearnsii) when reacted with several possible co-monomers at different relative amount, pH and temperature conditions. The more insoluble copolymers obtained with formaldehyde, hexamine, glyoxal, maleic anhydride, furfural and furfuryl alcohol were analyzed through solid state 13C NMR (Nuclear magnetic resonance) and FT-IR (Fourier Transform-Infrared) spectroscopy. The 13C NMR afforded the opportunity to detect: (i) aromatic substitutions and consequent poly-condensations for the majority of the hardeners studied; (ii) acylation for the maleic anhydride and also some; (iii) Diels⁻Alder arrangements for the furanic co-monomers; the FT-IR spectroscopy suggested that the formaldehyde and hexamine copolymers present a higher cross-linking degree.


Enantioselective Biosynthesis of L-Phenyllactic Acid From Phenylpyruvic Acid In Vitro by L-Lactate Dehydrogenase Coupling With Glucose Dehydrogenase.
As a valuable versatile building block, L-phenyllactic acid (L-PLA) has numerous applications in the fields of agriculture, pharmaceuticals, and biodegradable plastics. However, both normally chemically synthesized and naturally occurring PLA are racemic, and the production titer of L-PLA is not satisfactory. To improve L-PLA production and reduce the high cost of NADH, an in vitro coenzyme regeneration system of NADH was achieved using the glucose dehydrogenase variant LsGDHD255C and introduced into the L-PLA production process. Here an NADH-dependent L-lactate dehydrogenase-encoding variant gene (L-Lcldh1Q88A/I229A) was expressed in Pichia pastoris GS115. The specific activity of L-LcLDH1Q88A/I229A (Pp) was as high as 447.6 U/mg at the optimum temperature and pH of 40°C and 5.0, which was 38.26-fold higher than that of wild-type L-LcLDH1 (Pp). The catalytic efficiency (k cat/K m) of L-LcLDH1Q88A/I229A (Pp) was 94.3 mM-1 s-1, which was 67.4- and 25.5-fold higher than that of L-LcLDH1(Pp) and L-LcLDH1Q88A/I229A (Ec) expressed in Escherichia coli, respectively. Optimum reactions of L-PLA production by dual-enzyme catalysis were at 40°C and pH 5.0 with 10.0 U/ml L-LcLDH1Q88A/I229A (Pp) and 4.0 U/ml LsGDHD255C. Using 0.1 mM NAD+, 400 mM (65.66 g/L) phenylpyruvic acid was completely hydrolyzed by fed-batch process within 6 h, affording L-PLA with 90.0% yield and over 99.9% ee p. This work would be a promising technical strategy for the preparation of L-PLA at an industrial scale.


Molecular cloning, expression, and characterization of myo-inositol oxygenase from mouse, rat, and human kidney.
myo-Inositol oxygenase (MIOX) is a non-heme iron enzyme, which catalyzes the conversion of myo-inositol to d-glucuronic acid, the first committed step in myo-inositol catabolism. Full-length cDNAs of 858bp each coding for 33kDa protein were cloned from kidney cDNA libraries of mouse, rat, and human. The individual clones were expressed in Escherichia coli and recombinant MIOX proteins were purified to electrophoretic homogeneity. A hydrophobic interaction chromatography step yielded multiple conformers, with mouse and human MIOX showing three peaks and rat enzyme revealing two peaks. Individual MIOX peaks exhibited distinct V(max) and K(m) values. Interestingly, upon storage, the 33kDa protein was degraded to a approximately 30kDa truncated protein in each species, and formed small amounts of dimers of identical subunits. While MIOX is a highly conserved enzyme in all mammalian species, the labile nature and tendency to degrade in solution may be the source of significant differences in size previously reported in the literature. Regardless of the source, our results strongly dispel previous conflicting literature reports on the size of the protein and confirm that MIOX is a 33kDa protein.


Electrical Stimulation Therapy and Wound Healing: Where Are We Now?
Healing chronic wounds is an ongoing challenge for clinicians and poses a serious public health burden. Electrical stimulation (ES), broadly defined as the application of electrical current via electrodes placed on the skin adjacent to or directly within the wound, has been proposed as a therapeutic modality over a century ago, and recent advances in understanding the biology of electrical phenomena in the skin have rekindled an interest in this modality. Despite evidence that has shown ES to be effective for wound healing, it has been slow to gain acceptance in the United States. Also, there has been no consensus in terms of standardization of parameters to devise a systematic protocol for implementation of this technology. The epidermis maintains a "skin battery" that generates an endogenous electric field and current flow when wounded. Experimental models have demonstrated that most of the cell types within the wound can sense an electric field in the range of that endogenously generated in the wound, and respond with a variety of biological and functional responses that can contribute to healing. Multiple animal wound models have demonstrated enhancement of a number of parameters of healing when ES is exogenously supplied. Clinical trials have investigated the efficacy of multiple forms of ES for improving healing in a wide variety of human chronic wounds. In 2002 the Centers for Medicare and Medicaid Services approved reimbursement for use of ES in a clinical setting for certain chronic wounds. THERE REMAIN MANY VOIDS IN OUR KNOWLEDGE BASE: clinical evidence is limited by deficiencies in the design of many of the trials, a multiplicity of ES application modes and waveforms used in trials prevent selection of an optimal modality, and lack of uniformity in reporting ES dosages leave us not much advanced from our clinical knowledge base a decade ago.


Genetic Variability of the Functional Domains of Chromodomains Helicase DNA-Binding (CHD) Proteins.
In the past few years, there has been an increasing neuroscientific interest in understanding the function of mammalian chromodomains helicase DNA-binding (CHD) proteins due to their association with severe developmental syndromes. Mammalian CHDs include nine members (CHD1 to CHD9), grouped into subfamilies according to the presence of specific functional domains, generally highly conserved in evolutionary terms. Mutations affecting these domains hold great potential to disrupt protein function, leading to meaningful pathogenic scenarios, such as embryonic defects incompatible with life. Here, we analysed the evolution of CHD proteins by performing a comparative study of the functional domains of CHD proteins between orthologous and paralogous protein sequences. Our findings show that the highest degree of inter-species conservation was observed at Group II (CHD3, CHD4, and CHD5) and that most of the pathological variations documented in humans involve amino acid residues that are conserved not only between species but also between paralogs. The parallel analysis of both orthologous and paralogous proteins, in cases where gene duplications have occurred, provided extra information showing patterns of flexibility as well as interchangeability between amino acid positions. This added complexity needs to be considered when the impact of novel mutations is assessed in terms of evolutionary conservation.


[The vascular aspect of kidney transplantation].
The vascular aspect of renal transplantation is a factor of major importance throughout the various phases of surgery. It includes nephrectomy from living donors, or intact cadaveric kidney with a sufficiently long renal artery and vein (with or without patch), organic ex situ surgery, and implantation surgical technique. In 1994 fifteen renal transplantations are performed in the Clinical Center of Urology. In thirteen patients the kidneys transplanted are obtained from living donors, and in two-from cadavers. The vascular surgery techniques used, and the problems faced during their application are surveyed. Vascular surgery performed successfully is among the factors guaranteeing good survival of the organ transplanted.


Disturbance of response to acute thermal pain in naturally occurring cholecystokinin-a receptor gene knockout Otsuka Long-Evans Tokushima Fatty (OLETF) rats.
Otsuka Long-Evans Tokushima Fatty (OLETF) rats lack cholecystokinin-A receptor (CCK-AR) because of a genetic abnormality. We observed that body temperature homeostasis in response to changes in ambient temperature was deteriorated in OLETF rats, while the functions of the signal outputs from the hypothalamus to effectors were not impaired. Deteriorated homeostasis was also seen in CCK-AR deficient (-/-) mice. In the present study, we examined whether the sensory pathway involved in transmitting signals about temperature from the skin to the brain was impaired in OLETF rats. To elucidate the involvement of CCK-AR function, we conducted the same experiment in CCK-AR(-/-) mice. Responses to thermal pain were assessed using the Hargreaves' plantar test apparatus. Shortening of withdrawal latency was observed in OLETF rats compared to control rats, indicating thermal hyperalgesia. Behavioral responses following paw withdrawal were disturbed in OLETF rats. The 5-hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid contents in the hippocampus and frontal cortex of OLETF rats were significantly higher than in those of the controls. CCK-AR(-/-) mice did not show any differences from wild-type mice. In conclusion, OLETF rats showed thermal hyperalgesia and disturbed responses to thermal pain, and an alteration of 5-HT function might have a role in this disturbance.


Human embryonic stem cell-extracts inhibit the differentiation and function of monocyte-derived dendritic cells.
Embryonic stem cells (ESC) possess inherent properties of immune privilege with the capacity to evade allogeneic immune responses. Moreover, ESCs have been shown to prevent immune activation in response to third party antigen presenting cells in vitro and have the capacity to promote allograft survival in vivo. However, clinical use of human ESCs to treat immunological disorders may risk teratoma or ectopic tissue formation. Here, we show that cellular extracts from both human and mouse ESCs retain the immune modulatory properties of intact cells. ESC-extracts that contained 12-24 μg of total protein effectively prevented T cell proliferation in allogeneic mixed lymphocyte reactions (MLR), whereas control fibroblast extracts did not affect proliferation. Cellular mechanisms underlying hESC extract-mediated immune modulation involve the maturation of monocyte derived dendritic cells (mDC). hESC extract-treated mDCs had reduced surface expression of co-stimulatory and maturation markers CD80, HLA-DR and CD83 and secreted lower levels of IL12p40. Accordingly, hESC extract-treated DCs were found to be poor stimulators of purified allogeneic T cells compared to those DCs treated with vehicle or fibroblast extracts. Our results demonstrate that ESC extracts retain the immune modulatory properties of ESCs and for the first time demonstrates that ESC derived factors can inhibit human mDC maturation and function.


Elevated soluble Fas/APO-1 (CD95) levels in silicosis patients without clinical symptoms of autoimmune diseases or malignant tumours.
Soluble Fas (sFas) is produced as translation products of alternative mRNA splicing, and antagonizes the membranous Fas molecule in Fas/Fas ligand interactions. We investigated the serum sFas levels in 64 Japanese silicosis patients with no clinical symptoms of autoimmune diseases or malignant tumours, using ELISA for sFas. The serum sFas levels in the silicosis patients were significantly higher than those in healthy volunteers. Elevated serum sFas levels were also detected in patients with systemic lupus erythematosus but, unexpectedly, no difference was observed in sFas levels between progressive systemic sclerosis patients and healthy volunteers. On the other hand, there was no significant difference in the expression of Fas on peripheral blood lymphocytes between the patients with silicosis and age-matched healthy volunteers. These observations provided the first evidence that serum sFas levels are elevated in silicosis patients without clinical symptoms of autoimmune diseases or malignant tumours. It remains to be clarified whether patients with elevated sFas levels have a tendency to develop autoimmune diseases later, or whether some other distinct factor(s) is necessary to initiate the progression of autoimmune diseases.


Efficacy of an Enhanced Recovery After Surgery (ERAS) Pathway in Elderly Patients Undergoing Spine and Peripheral Nerve Surgery.
Elderly patients are a vulnerable patient population in elective spinal surgery. Older patients have more medical comorbidities and are also more sensitive to opiate medications. Despite this, spine and peripheral nerve surgery is still feasible in these patients, and an Enhanced Recovery After Surgery (ERAS) regimen can further enhance the safety profile. This is a before and after cohort study at a single institution on elderly patients who underwent elective spine and peripheral nerve surgery. Patients were prospectively enrolled in a novel ERAS protocol from April 2017 to December 2018. The control group was a historical cohort of patients who underwent surgery from September 2016 to December 2016. The primary outcome was self-reported opioid use at 1- and 3-months postoperatively. The secondary outcome was compliance with the ERAS protocol across several measures including patient-controlled (PCA) use, patient-reported pain scores, mobilization and ambulation status, and Foley catheter use. Among 504 patients aged 65 and older compared to historic controls there was a significant reduction in the use of post-operative opioids at one month (36.2% vs. 71.7%, p < 0.001) and 3 months after surgery (33.0% vs. 80.0%, p < 0.001). 504 consecutive elderly patients were included in the ERAS protocol compared to a control group of 60. The two groups had similar surgical procedures and baseline demographics, with similar mean ages (ERAS 73.2 years vs. control 73.5 years, p = 0.67). The ERAS group showed improved mobilization and ambulation on POD 0 in compliance with our protocol compared to the control group (mobilization: 60.0% vs. 10.0%, p < 0.001; ambulation: 36.1% vs. 10.0%, p < 0.001), with no inpatient falls reported for either group. ERAS facilitates reduction in opiate use at 1- and 3-month intervals postoperatively in patients greater than 65 years old undergoing elective spine and peripheral nerve surgery. Early mobilization and ambulation are safe and feasible in this population.


[Effect of gamma-aminobutyric acid and its derivatives on convulsions induced by L-kynurenine and quinolinic acid].
Derivatives of GABA which are ineffective against seizures produced by typical convulsants (strychnine, pentylenetetrazole, etc.), phenibut and sodium hydroxybutyrate in doses of 200 mg/kg and higher possess marked anticonvulsant action against seizures induced by intraventricular injection of L-kynurenine and quinolinic acid in mice. Phepiron, a cyclic form of phenibut, had dissimilar action on the effects of L-kynurenine and a quinolinic acid, enhancing that produced by the former agent and diminishing tonic extension and lethality after the latter one. GABA and piracetam appeared to be ineffective. Efficacy of GABA derivatives against seizures produced by L-kynurenine and quinolinic acid suggests that the system of GABA plays a significant role in the genesis of seizures produced by kynurenines.


Sequential-Anaerobic and Sequential-Aerobic Bioleaching of Metals (Ni, Mo, Al and V) from Spent Petroleum Catalyst in Stirred Tank Batch Reactor: A Comparative Study.
Spent petroleum catalyst as a repository of several toxic metals is recommended for metal removal before safe disposal. To evaluate an effective biotechnological approach for metal removal, a comparative study between sequential-aerobic and sequential-anaerobic bioleaching processes was conducted for the removal of metals from crushed-acetone-pretreated spent petroleum catalyst. The SEM-EDX and XPS analysis confirmed the presence of Ni, Al, Mo and V in their oxidic and sulphidic forms in spent catalyst. The bioleaching experiments were performed in stirred tank batch reactors (2.5 L), temperature 30 °C, pH 1.4 and stirring speed 250 rpm for the period of 160 h. Sulfuric acid acted as lechant for both sequential-aerobic (Acidithiobacillus ferrooxidans oxidised sulfur to sulfuric acid aerobically) and sequential-anaerobic (Acidithiobacillus ferrooxidans oxidised sulphur to sulfuric acid coupled with the ferric reduction to ferrous anaerobically) bioleaching studies. The higher Ni and V extractions compared to Al and Mo for all the studies were due to increased solubility of Ni and V, and supported by XPS which showed marginal signs of Ni and V peaks in leach residues compared to feed spent catalyst. At the end (320 h), sequential-aerobic bioleaching was resulted to 99% Ni, 65% Al, 90% Mo and 99% V extraction quite more effective than sequential-anaerobic bioleaching (88% Ni, 28% Al, 33% Mo and 77% V) and sequential-control leaching (94% Ni, 20% Al, 40% Mo and 57% V). Although anaerobic bioleaching a possible approach, aerobic condition was found to be more suitable for sulfuric acid generation by A. ferrooxidans and high yield. So aerobic bioleaching is recommended to be favourable approach compared to anaerobic counterpart for future study and extrapolation. The online version contains supplementary material available at 10.1007/s12088-021-00978-8.


Probing the origins of human acetylcholinesterase inhibition via QSAR modeling and molecular docking.
Alzheimer's disease (AD) is a chronic neurodegenerative disease which leads to the gradual loss of neuronal cells. Several hypotheses for AD exists (e.g., cholinergic, amyloid, tau hypotheses, etc.). As per the cholinergic hypothesis, the deficiency of choline is responsible for AD; therefore, the inhibition of AChE is a lucrative therapeutic strategy for the treatment of AD. Acetylcholinesterase (AChE) is an enzyme that catalyzes the breakdown of the neurotransmitter acetylcholine that is essential for cognition and memory. A large non-redundant data set of 2,570 compounds with reported IC50 values against AChE was obtained from ChEMBL and employed in quantitative structure-activity relationship (QSAR) study so as to gain insights on their origin of bioactivity. AChE inhibitors were described by a set of 12 fingerprint descriptors and predictive models were constructed from 100 different data splits using random forest. Generated models afforded R (2), [Formula: see text] and [Formula: see text] values in ranges of 0.66-0.93, 0.55-0.79 and 0.56-0.81 for the training set, 10-fold cross-validated set and external set, respectively. The best model built using the substructure count was selected according to the OECD guidelines and it afforded R (2), [Formula: see text] and [Formula: see text] values of 0.92 ± 0.01, 0.78 ± 0.06 and 0.78 ± 0.05, respectively. Furthermore, Y-scrambling was applied to evaluate the possibility of chance correlation of the predictive model. Subsequently, a thorough analysis of the substructure fingerprint count was conducted to provide informative insights on the inhibitory activity of AChE inhibitors. Moreover, Kennard-Stone sampling of the actives were applied to select 30 diverse compounds for further molecular docking studies in order to gain structural insights on the origin of AChE inhibition. Site-moiety mapping of compounds from the diversity set revealed three binding anchors encompassing both hydrogen bonding and van der Waals interaction. Molecular docking revealed that compounds 13, 5 and 28 exhibited the lowest binding energies of -12.2, -12.0 and -12.0 kcal/mol, respectively, against human AChE, which is modulated by hydrogen bonding, π-π stacking and hydrophobic interaction inside the binding pocket. These information may be used as guidelines for the design of novel and robust AChE inhibitors.


Patient-reported outcomes of topical therapies in actinic keratosis: A systematic review.
Patients' perspectives on actinic keratosis treatments may have an impact on treatment adherence and, therefore, therapeutic outcomes. We performed a systematic review to assess patients' perspectives of topical, field-directed treatments for actinic keratoses. A literature search was conducted, and 14 studies were identified encompassing 4433 patients. Only four studies were focused on face and/or scalp, which are the locations that typically impact patients' quality of life. Four studies were clinical trials. One study utilized a validated patient-reported outcomes (PRO) instrument specifically developed for actinic keratosis. In general, treatment adherence and patient satisfaction were better with shorter-duration treatment regimens such as ingenol mebutate gel. Imiquimod improved quality of life in one study but not in another. No data was available on topical piroxicam. The findings underscore the need for effective and well-tolerated, short-duration topical treatment for actinic keratosis.


In vivo analysis of Chlamydomonas chloroplast petD gene expression using stable transformation of beta-glucuronidase translational fusions.
We have used the Escherichia coli beta-glucuronidase (uidA) gene as a reporter gene to localize the promoter and analyze the function of the 5' untranslated region (UTR) of the Chlamydomonas chloroplast petD gene. Using particle bombardment, petD-uidA transcriptional and translational fusion genes were introduced into the chloroplast genome in the large inverted repeat flanking the atpB gene. In transformants carrying a petD-uidA transcriptional fusion, uidA mRNA accumulated but was not translated. However, in a translational fusion that included the entire petD 5' UTR, uidA mRNA accumulated and a high level of beta-glucuronidase activity was detected. When approximately 70% of the petD 5' UTR was deleted from the translational fusion, uidA mRNA accumulation and beta-glucuronidase activity decreased 4- to 6-fold and 8-fold, respectively. Run-on transcription assays demonstrated that all strains transcribe the uidA gene at equivalent rates. Our results show that sequences essential for translation reside in the petD 5' UTR and also that sequences within the 5' UTR directly or indirectly affect mRNA stability. The expression of beta-glucuronidase under the control of chloroplast transcriptional and translational signals will facilitate further studies of chloroplast gene regulatory mechanisms.


Carbon Transfer and Partitioning between Vegetative and Reproductive Organs in Pisum sativum L.
Assimilate partitioning was studied in the common pea (Pisum sativum L.) by feeding (14)CO(2) to whole plants and measuring radioactivity in different organs 48 hours after labeling. Two experimental protocols were used. For the first, one reproductive node was darkened with an aluminum foil, to prevent photosynthesis during labeling. The aim was to study assimilate translocation among nodes. The second was carried out to assess any priority among sinks. Whole plants were shaded, during labeling, to reduce carbon assimilation. Various developmental stages between the onset of flowering and the final stage in seed abortion of the last pod were chosen for labeling. When all photosynthetic structures at the first reproductive node were darkened at any stage of development after the formation of the first flower, the first pod was supplied with assimilates from other nodes. In contrast, later developed pods, when photosynthetic structures at their node were darkened, received assimilates from other nodes only when they were beyond their final stage in seed abortion. Reducing illumination to 30% did not change distribution of assimilated carbon between vegetative and reproductive structures, nor among pods. It appears that the relative proportion of (14)C allocated to any one pod, compared to other pods, depends on the dry weight of that pod as a proportion of the total reproductive dry weight. When the plant was growing actively, following the start of the reproductive phase until a few days before the end of flowering, the top of the plant (i.e., all the organs above the last opened flower) had a higher sink strength and a higher relative specific activity than pods, suggesting that it was a more competitive sink for assimilates. The pattern of assimilate distribution described here provides an explanation for pod and seed abortion.


[Stabilimetry and cranio-cervico-mandibular disorders].
Cranio-cervico-mandibular disorders can cause disturbances in posture. Stabilimetry measures spatio-temporal variations in the center of body pressure and evaluates the mechanisms of maintenance of balance. The study used a stabilimetric platform to evaluate posture changes in patients with cranio-cervico-mandibular disorders before and after treatment for malocclusion. Between February 1998 and December 2000, 60 patients with cranio-cervico-mandibular disorders were recruited from the Dentistry Clinic of the University of Sassari. Each patient underwent two stabilimetric examinations (closed mouth with cotton wads inserted between the dental arches). The tests were conducted on a stabilimetric platform. Measurement of body posture load and sway were analyzed with a specific software program that correlated the vestibular, somatosensory and visual systems, and determined the role of each in postural control. The stabilimetric analysis showed that the adoption of the plaque allowed rebalance of the postural system, without affecting the visual system. After treatment, 64% of patients experienced remission of pain symptoms with orthotic therapy. The relationship between malocclusion and posture should be seen from a holistic standpoint in other to gain a global therapeutic outcome.


Electrospray ionization mass spectrometry as a tool to analyze hydrogen/deuterium exchange kinetics of transmembrane peptides in lipid bilayers.
A method is described to study the precise positioning of transmembrane peptides in a phospholipid bilayer combining hydrogen/deuterium (H/D) exchange and nanoelectrospray ionization mass spectrometry. The method was tested by using model systems consisting of designed alpha-helical transmembrane peptides [acetylGW(2)(LA)(5)W(2)Aethanolamine (WALP16) and acetyl-(GA)(3)W(2)(LA)(5)W(2)(AG)(3)ethanolamine (WALP16(+10))] incorporated in large unilamellar vesicles of 1, 2-dimyristoyl-sn-glycero-3-phosphocholine. Both peptides consist of an alternating leucine/alanine hydrophobic core sequence flanked by tryptophan residues as interfacial anchor residues. In the case of WALP16(+10), this sequence is extended at both ends by 5-aa glycine/alanine tails extending into the aqueous phase surrounding the bilayer. H/D exchange of labile hydrogens in these peptides was monitored in time after dilution of the vesicles in buffered deuterium oxide. It was found that the peptides can be measured by direct introduction of the proteoliposome suspension into the mass spectrometer. Several distinct H/D exchange rates were observed (corresponding to half-life values varying from </=2 to approximately 2 x 10(4) min). Fast exchange rates were assigned to the water-exposed tails of WALP16(+10). For both WALP16 and WALP16(+10), intermediate exchange rates were assigned to the residues close to the membrane/water interface, and the slow exchange rates to the membrane-embedded hydrophobic core. These assignments were confirmed by results from collision-induced dissociation tandem mass spectrometry experiments, which allowed analysis of exchange of individual peptide amide linkages. This proteoliposome nanoelectrospray ionization mass spectrometry technique is shown to be an extremely sensitive and powerful tool for revealing site-specific information on peptide-membrane interactions.