Determinants of long act reversible contraceptive utilization among HIV positive reproductive age women attending ART clinic in South West Ethiopia.
Identifying the determinants of long-acting contraceptive utilization and managing the sexual and reproductive health of HIV-infected women is critical to reducing HIV transmission and maternal mortality. However, the determinants of long-acting contraceptive utilization have not been well understood in resource-limited settings like Ethiopia. The aim of this study was to identify determinants of long-acting reversible contraceptive utilization among HIV-positive women on ART in southwest Ethiopia. A facility-based, unmatched case control study was conducted from July 24 to August 28, 2021, in south-west Ethiopia. The study participants were HIV positive women, with a sample size of 109 cases and controls. An interviewer administered a questionnaire, and a check list was used for data collection. A systemic random sampling technique was used to collect data from cases and controls. Bivariate and multivariable logistic regressions were employed to determine the determinants of LARC utilization among HIV-positive women. To demonstrate the strength of the determinant, the odds ratio was calculated with 95% confidence intervals, and a P-value less than 0.05 was used to declare statistical significance. A total of 324 women (108 cases and 216 controls) of reproductive age who were HIV positive were interviewed, with a response rate of 99.0%. urban residence (AOR = 2.67, 95%CI: 1.23- 5.77), having formal education (AOR = 2.93, 95% CI:1.36, 6.34), being counseled by health care provider (AOR = 5.42,95% CI: 2.67-11.03), no future fertility intention (AOR = 2.87, 95% CI:1.44-5.70), having CD4 count less than 500 cell/mm3 (AOR = 4.18,95% CI:2.12-8.23), having information of HIV transmission from mother to child (AOR = 3.65,95% CI:1.49-8.95),not using condom during sexual intercourse (AOR = 4.86,95% CI:2.46-9.62),,having knowledge towards LARC (AOR = 2.38,95% CI:1.24-4.58) and attitude towards LARC (AOR = 6.41,95%CI:3.16-13.0) were independent determinants of LARC utilization among HIV positive women. Women being counseled by a health care provider, having no future fertility intention, and having a CD4 count less than 500 cells/mm3 were found to be determinants of long-acting contraceptive method utilization among HIV-positive reproductive-age women. Also, our study supports the WHO Strategic Concepts for Improving the Links between Family Planning and HIV/AIDS Policy, Programs, and Services. It is recommended that Health care providers should use these factors as base line during family planning counseling and service delivery.


Bloom's syndrome in two Dutch families.
The clinical and cytogenetic data are presented of four children with Bloom's syndrome, who belong to two unrelated Dutch families. The patients showed, in varying degrees, the clinical features most characteristic of the syndrome: stunted growth; telangiectatic facial erythema; sun-sensitivity of the skin; and decreased immuno-competence. In one child the skin lesions were only minor and the diagnosis would probably not have been made if her sib had not been recognized as having Bloom's syndrome. The cytogenetic characteristics of the syndrome were present in all patients. Each showed a high number of chromosomal aberrations and numerous sister-chromatid exchanges per cell.


Beta-carboline-carbohydrate hybrids: molecular design, chemical synthesis and evaluation of novel DNA photocleavers.
Beta-carboline present in beta-carboline alkaloids from marine organisms was found, for the first time, to cleave DNA at the guanine site upon irradiation with UV light with a long wavelength without any additive, and beta-carboline-carbohydrate hybrid system was effective for DNA cleavage.


Industrial medicine and acute musculoskeletal rehabilitation. 4. Evaluation and management of the injured worker with a lower-extremity injury.
This self-directed learning module highlights the importance of recognizing the relationships between multiple, concurrent joint injuries in the lower extremity of an injured worker. This chapter focuses on evaluating the lower extremity and devising a rehabilitation program that incorporates the entire kinetic chain. Further recommendations for return to work are discussed. (a) To accurately recognize and diagnose coexisting injuries in the lower extremity and (b) to devise an evaluation and treatment program enabling the worker to return to work.


Maillard reaction products-based encapsulant system formed between chitosan and corn syrup solids: Influence of solution pH on formation kinetic and antioxidant activity.
Maillard reaction products (MRPs) between chitosan and various sugars with enhanced antioxidant activity were previously produced. However, few reports address the chitosan and corn syrup solids system that has been successfully used to encapsulate nutraceutical oils. Maillard reaction is pH-responsive, the influence of solution pH on the formation kinetic and antioxidant activity of MRPs was therefore evaluated in this work. FT-IR and zeta-potential results confirmed the formation of MRPs between chitosan and corn syrup solids. Possible Amadori compounds signals were observed clearly in the 1H NMR spectrum. Brown color development depended on initial solution pH, following a zero-order kinetic regression. Antioxidant activity of reaction products was higher than the native system and increased with an increase in the initial pH of the solution. Developed MRPs with a dual function as antioxidant and encapsulant can possibly be used to protect emulsified oil from oxidation.


HMG-CoA synthase 2 drives brain metabolic reprogramming in cocaine exposure.
The brain is a high energy-consuming organ that typically utilizes glucose as the main energy source for cerebral activity. When glucose becomes scarce under conditions of stress, ketone bodies, such as β-hydroxybutyrate, acetoacetate and acetone, become extremely important. Alterations in brain energy metabolism have been observed in psychostimulant abusers; however, the mode of brain metabolic programming in cocaine dependence remains largely unknown. Here, we profiled the metabolites and metabolic enzymes from brain nucleus accumbens (NAc) of mice exposed to cocaine. We found that cocaine modified energy metabolism and markedly activated ketogenesis pathway in the NAc. The expression of HMG-CoA synthase 2 (HMGCS2), a critical rate-limiting ketogenesis enzyme, was markedly up-regulated. After switching metabolic pathways from ketogenesis to glycolysis through activation of glucokinase, cocaine-evoked metabolic reprogramming regained homeostasis, and the cocaine effect was attenuated. Importantly, both the pharmacological and genetic inhibition of HMGCS2 significantly suppressed cocaine-induced ketogenesis and behavior. In conclusion, cocaine induces a remarkable energy reprogramming in the NAc, which is characterized by HMGCS2-driven ketogenesis. Such effect may facilitate adaptations to cocaine-induced energy stress in the brain. Our findings establish an important link between drug-induced energy reprogramming and cocaine effect, and may have implication in the treatment of cocaine addiction.


The human papillomaviruses.
Recent biochemical and serological studies have shown the existence of at least four distinct types of human papillomaviruses (HPVs) causing benign skin lesions. These viruses show hardly no antigenic relationships; their DNAs differ by their sensitivity to restriction endonucleases, and show little, if any, sequence homology, as detected by molecular hybridization using complementary RNAs transcribed in vitro. Data on the pathogenicity of HPVs are still incomplete but indicate that some types of benign skin lesions (plantar warts, common warts, flat warts) may be preferentially associated with some types of HPV. Most interesting is that epidermodysplasia verruciformis has been found associated with two types of virus, and that malignant conversion of some lesions has been observed in all the patients infected with one of them. This suggests that at least a HPV may have a higher oncogenic potential, as do rabbit (Shope) papillomavirus and bovine alimentary tract papillomavirus. Much remains to be known on human papilloma-viruses and further studies may lead to the characterization of additional types of HPVs, especially in genital condylomata acuminata and laryngeal papillomas whose malignant conversion, although rare, may be observed. Progress in this field has been and remains hampered by the lack of cell culture systems allowing replication of these highly host and tissue specific viruses, and by the widely variable virus content of the different human lesions known to be associated with a papillomavirus. Further studies are warranted by the possible role of these widespread and epitheliotropic viruses in the origin of some carcinomas in man.


Antiarrhythmic therapy for asymptomatic ventricular arrhythmias.
The rationale for treatment of patients with nonsustained asymptomatic ventricular arrhythmias is the theoretical benefit of preventing more serious ventricular arrhythmias and sudden cardiac death. Because of the high costs involved and the serious side effects, such as proarrhythmia, associated with this therapy, the decision to treat this patient group for a potential protective effect must be weighed carefully. Risk factors identifying those patients most likely to have further complications include the presence of heart disease and left ventricular dysfunction, and the relative severity of these conditions. Those patients who fit into high-risk groups are the ones most likely to benefit, although this benefit is still unproved. If antiarrhythmic therapy is given, it is recommended that it be started in the hospital and that the efficacy of treatment be assessed by serial electrophysiologic-pharmacologic testing or noninvasive means. Empiric treatment, especially started out-of-hospital, is discouraged because it is least likely to benefit the patient and most likely to cause harm.


Genome-wide DNA methylation analysis of human peripheral blood reveals susceptibility loci of diabetes-related hearing loss.
Diabetes-related hearing loss (DRHL) is a complication of diabetes mellitus that is drawing more attention currently. DNA methylation has a critical role in the pathogenesis of type 2 diabetes mellitus (T2DM) and its complications. Therefore, we investigated the genome-wide DNA methylation of peripheral blood of T2DM patients with/without hearing loss in order to explore the susceptibility loci of DRHL. Between DRHL group and control group, 113 gene sites were identified to be differentially methylated regions (DMRs). Among 38 DMRs with whole samples, the classification accuracy is up to 90%. With alignment to T2DM susceptibility genes and deafness genes published, KCNJ11 was found to be the only overlapped gene. The DNA methylation level of KCNJ11 was associated with stroke (t = 2.595, p < 0.05), but not with diabetic nephropathy and diabetic retinopathy. The detective rate of distortion product otoacoustic emissions (DPOAE) from low to high frequencies (0.7-6 kHz) on the right ear was significantly correlated with the methylation level of KCNJ11. The auditory brainstem response (ABR) threshold on the right ear was also correlated (r = 0.678, p < 0.05). This DNA methylation profile indicates the susceptibility loci of DRHL. The potassium metabolism may have a critical role in the hearing loss caused by hyperglycemia.


Botulinum toxin as an adjunct to monocular recession-resection surgery for large-angle sensory strabismus.
To evaluate the result of using intraoperative botulinum toxin A (BTA) as an adjunct to monocular recession-resection surgery in the management of large-angle sensory strabismus. The medical records of patients diagnosed with sensory strabismus with constant large-angle strabismus were reviewed retrospectively to identify those who underwent monocular recession-resection surgery combined with 5 units of BTA injection into the recessed muscle of the nonfixating eye. Surgical outcome was considered successful if the final deviation was within the range of 0Δ-10Δ. A total of 13 patients (mean age, 31.04 ± 18.5 years) were included, 8 with exotropia and 5 with esotropia. The mean follow-up period was 52.77 ± 10.9 months. The mean preoperative deviation was 66Δ ± 16Δ in the esodeviation group and 56Δ ± 5Δ in the exodeviation group. The final postoperative mean deviation was 6Δ ± 7Δ in the esodeviation group and 6Δ ± 8Δ in the exodeviation group. There were 7 patients (87.5%) with final deviation of ≤10Δ in the exodeviaton group and 4 (80%) in the esodeviation group. Adjunctive usage of BTA with conventional surgery appears to enhance surgical outcomes by increasing the amount of expected correction. The combined use of BTA with monocular recession-resection rectus muscle surgery may be a good alternative in the treatment of large-angle sensory strabismus with the advantage of less risk of potential complications compared to supramaximal surgery or surgery in the "good" eye.


Regional differences in reactivity and in the influence of the epithelium on canine intrapulmonary bronchial smooth muscle responsiveness.
Differences in the reactivity and in the influence of the epithelium on responsiveness of canine 2nd and 3rd generation airway smooth muscle were examined. Epithelium-containing 3rd generation airways produced a greater maximum contraction and were more sensitive to methacholine and histamine, but not to KCl, than corresponding 2nd generation airways. Mechanical removal of the epithelium increased the sensitivity to methacholine and histamine in 2nd generation airways; there was also an increase in the maximum response elicited by histamine, but not by methacholine, in epithelium-free preparations. In contrast, there was no significant difference in the sensitivity to or the maximum response elicited by histamine or methacholine in epithelium-containing and epithelium-free 3rd generation airways. Epithelium removal had no effect on KCl-induced responses in either airway region. The inhibitory effects of verapamil (1 microM) against KCl- and methacholine-induced responses were identical in preparations containing and lacking the epithelium. The results support the postulate of an epithelium-derived inhibitory factor modulating airway smooth muscle reactivity. Furthermore, the influence of the epithelium exhibits regional differences, being greater in larger airways.


Mapping of TBARS distribution in frozen-thawed pork using NIR hyperspectral imaging.
In this study, NIR hyperspectral imaging technology was applied to determine the distribution of TBARS in frozen-thawed pork. A total of 240 fresh pork samples were assigned to 4 treatment groups (0, 1, 3, 5 frozen-thawed cycles). For each sample, a hyperspectral image (874-1734nm) was collected, followed by chemical TBARS analysis. Successive projection algorithm (SPA) was applied to choose effective wavelengths (EWs). The selected 13 EWs of the calibration set and relevant TBARS value were used as inputs of partial least squares regression (PLSR) model, yielding correlation coefficient of prediction of 0.81 and root mean square error of prediction of 0.33. The developed PLSR model were applied pixel-wise to produce chemical maps of TBARS for 24 selected samples in the prediction set. The results indicated that NIR hyperspectral imaging combined with image processing has the potential to visualize TBARS distribution in frozen-thawed pork. This technique could be useful in real-time quality monitoring in meat industry.


Sequential FISH allows the determination of the segregation outcome and the presence of numerical anomalies in spermatozoa from a t(1;8;2)(q42;p21;p15) carrier.
To find out the meiotic segregation behaviour of the t(1;8;2)(q42;p21;p15), to evaluate the occurrence of interchromosomal effects, and to determine whether there is an accumulation of unbalanced products in aneuploid/diploid gametes. A sequential FISH protocol based on two successive hybridization rounds over the same spermatozoa was performed to determine the segregation outcome of the rearranged chromosomes. The presence of numerical abnormalities for 13, 18, 21, X and Y was also evaluated by sperm FISH. Those aneuploid/diploid gametes were subsequently relocalized and analyzed for their segregation content through additional hybridization rounds. The segregation pattern observed reported a very low production of normal/balanced gametes (11.7 %). Significant increased frequencies of diploidies and disomies for chromosomes X/Y and 18 were detected (p < 0.001). Aneuploid and diploid spermatozoa displayed significant increases of 5:1, 6:0 and other unexpected disjunction modes (p < 0.001). The strategy developed in this study is a reliable new approach to establish the full segregation pattern of complex chromosome rearrangements (CCR). Results corroborate the low number of normal/balanced spermatozoa produced by CCR carriers and support previous findings regarding an altered segregation pattern in gametes with numerical abnormalities. Altogether this confirms the importance of PGD as a tool to prevent the transmission of chromosomal abnormalities to the offspring in CCR patients.


Sub-optimal CD4 reconstitution despite viral suppression in an urban cohort on antiretroviral therapy (ART) in sub-Saharan Africa: frequency and clinical significance.
A proportion of individuals who start antiretroviral therapy (ART) fail to achieve adequate CD4 cell reconstitution despite sustained viral suppression. We determined the frequency and clinical significance of suboptimal CD4 reconstitution despite viral suppression (SO-CD4) in an urban HIV research cohort in Kampala, Uganda. We analyzed data from a prospective research cohort of 559 patients initiating ART between 04/04-04/05. We described the patterns of SO-CD4 both in terms of:- I) magnitude of CD4 cell increase (a CD4 count increase < 50 CD4 cells/microl at 6 months, <100 cells/microl at 12 months; and <200 cells/microl at 24 months of ART) and II) failure to achieve a CD4 cell count above 200 cells/microl at 6,12 and 24 months of ART. Using criteria I) we used logistic regression to determine the predictors of SO-CD4. We compared the cumulative risk of clinical events (death and/or recurrent or new AIDS-defining illnesses) among patients with and without SO-CD4. Of 559 patients initiating ART, 386 (69%) were female. Median (IQR) age and baseline CD4 counts were 38 yrs (33-44) and 98 cells/microl (21-163) respectively; 414 (74%) started a d4T-based regimen (D4T+3TC+NVP) and 145 (26%) a ZDV-based regimen (ZDV+3TC+EFV). After 6, 12 and 24 months of ART, 380 (68%), 339 (61%) and 309 (55%) had attained and sustained HIV-RNA viral suppression. Of these, 78 (21%), 151 (45%) and 166 (54%) respectively had SO-CD4 based on criteria I), and 165(43%), 143(42%) and 58(19%) respectively based on criteria II). With both criteria combined, 56 (15%) and 129 (38%) had SO-CD4 at 6 and 12 months respectively. A high proportion (82% and 58%) of those that had SO-CD4 at 6 months (using criteria I) maintained SO-CD4 at 12 and 24 months respectively. There were no statistically significant differences in the incidence of clinical events among patients with [19/100PYO (12-29)] and without SO-CD4 [23/100PYO (19-28)]. Using criteria I), the frequency of SO-CD4 was 21% at 6 months. Majority of patients with SO-CD4 at 6 months maintained SO-CD4 up to 2 years. We recommend studies of CD4 T-cell functional recovery among patients with SO-CD4.


Mesopic contrast sensitivity function after excimer laser photorefractive keratectomy.
To evaluate contrast sensitivity under mesopic conditions in patients who had undergone uncomplicated excimer laser photorefractive keratectomy (PRK) for myopia. Monocular contrast sensitivity function was measured with the Stereo Optical F.A.C.T. chart in 26 patients who had received PRK using the Nidek EC-5000 excimer laser system. Mean preoperative refractive error was -6.23 +/- 1.69 D (range, -4.00 to -8.25 D); postoperatively, mean refractive error was -0.36 +/- 0.58 D (range, -0.75 to +0.50 D). Contrast sensitivity function was measured 6 months after surgery using four different chart luminances: 85, 5.0, 2.5, and 0.1 cd/m2, the first being a photopic level and the rest mesopic. A control group of eight emmetropic subjects was also studied to allow comparison of results for statistical purposes. Logarithmic values of contrast sensitivity at each spatial frequency were used for statistical analysis and normalized values were used for graphical representation. The results showed a statistically significant reduction (P < .01) in contrast sensitivity for the PRK patients in comparison with the control group under mesopic conditions for each spatial frequency tested (1.5, 3, 6, 12, and 18 c/deg), although no significant contrast sensitivity differences were observed between PRK and control groups at the photopic (85 cd/m2) level (P > .01 for all frequencies). Photorefractive keratectomy can induce significant reductions in contrast sensitivity under mesopic conditions, even though the photopic contrast sensitivity function is normal.


Study to promote innovation in rural integrated telepsychiatry (SPIRIT): Rationale and design of a randomized comparative effectiveness trial of managing complex psychiatric disorders in rural primary care clinics.
Managing complex psychiatric disorders like PTSD and bipolar disorder is challenging in Federally Qualified Health Centers (FQHCs) delivering care to U.S residents living in underserved rural areas. This protocol paper describes SPIRIT, a pragmatic comparative effectiveness trial designed to compare two approaches to managing PTSD and bipolar disorder in FQHCs. Treatment comparators are: 1) Telepsychiatry Collaborative Care, which integrates consulting telepsychiatrists into primary care teams, and 2) Telepsychiatry Enhanced Referral, where telepsychiatrists and telepsychologists treat patients directly. Because Telepsychiatry Enhanced Referral is an adaptive intervention, a Sequential, Multiple Assignment, Randomized Trial design is used. Twenty-four FQHC clinics without on-site psychiatrists or psychologists are participating in the trial. The sample is patients screening positive for PTSD and/or bipolar disorder who are not already engaged in pharmacotherapy with a mental health specialist. Intervention fidelity is measured but not controlled. Patient treatment engagement is measured but not required, and intent-to-treat analysis will be used. Survey questions measure treatment engagement and effectiveness. The Short-Form 12 Mental Health Component Summary (SF-12 MCS) is the primary outcome. A third (34%) of those enrolled (n = 1004) are racial/ethnic minorities, 81% are not fully employed, 68% are Medicaid enrollees, 7% are uninsured, and 62% live in poverty. Mental health related quality of life (SF-12 MCS) is 2.5 standard deviations below the national mean. We hypothesize that patients randomized to Telepsychiatry Collaborative Care will have better outcomes than those randomized to Telepsychiatry Enhanced Referral because a higher proportion will engage in evidence-based treatment.


Transfer of antibiotic resistance between commensal and pathogenic members of the Enterobacteriaceae under ileal conditions.
To determine the rate of antibiotic resistance transmission between commensal and pathogenic representatives of the Enterobacteriaceae. Through the use of a validated in vitro simulation of the porcine ileum, the transmission of antibiotic resistance was detected between commensal Escherichia coli, E. coli O157 and Salmonella spp. Countable transconjugant populations arose readily and, in one example, proved capable of indefinite persistence. Genetic material conferring antibiotic resistance is readily transmissible between members of the Enterobacteriaceae under ileal conditions. Recipient phenotype influences the persistence of multi-resistant transconjugants. The observation that the conjugal transmission of antibiotic resistance is commonplace under ileal conditions impacts primarily on the risk of food contamination by multi-resistant bacteria. The establishment of a multi-resistant transconjugant population as a dominant member of the microflora maintains a genetic reservoir of antimicrobial resistance.


Comparison of Two Analytical Methods for Busulfan Therapeutic Drug Monitoring.
Busulfan (Bu) is an old drug, but is still well recommended as an alkylating agent during conditioning therapy, before hematopoietic stem cell transplantation. Although its dose administration is standardized and based on patient weight, therapeutic drug monitoring is required in order to maintain its exposure [as area under the concentration-time curve (AUC) from 0 to infinity AUC0-∞] within a narrow therapeutic range and, if necessary, to adjust the dose with as short a lead time as possible. The aim of the study is to evaluate the agreement (as calculated AUC) between a gold standard analytical method and a new one that is faster and easier. We analyzed 221 plasma samples from 37 children (0.25-16 years; 4-62.5 kg) and 11 adults (21-59 years; 45-80 kg), corresponding to 52 AUC values (ng h/mL). The drug exposure was calculated, simultaneously, by two validated analytical methods. The reference method was a high-performance liquid chromatography (HPLC) assay combined with an ultraviolet detector (UV). The test method had a triple quadrupole mass spectrometer (MS) as detector; the clean-up procedures of the samples were different and faster. The agreement between the two methods (reference and test) was evaluated in terms of Bu exposure differences based on Lin's concordance correlation coefficient (CCC) and represented by the Bland-Altman plot. The CCC between the AUC of the two methods was excellent (0.868; 95% CI: 0.802-0.935). The precision of the measures (expressed by Pearson's italic "r") was 0.872, and the accuracy (accounted by the bias correction factor) was 0.996. We can conclude that the HPLC-MS/MS assay represents a very good alternative to the reference.


Enhanced ADAM17 expression is associated with cardiac remodeling in rats with acute myocardial infarction.
This study aimed to investigate the dynamic expression of A-disintegrin-and-metalloproteinase-17 (ADAM17) during cardiac remodeling after acute myocardial infarction (AMI). Forty male Wistar rats with a permanent ligation of the left anterior descending artery were equally divided into four groups based on predefined sacrifice time: MI1d, MI1w, MI4w and MI12w. As controls, 36 rats only with left thoracotomy were equally divided into four groups. Cardiac remodeling was assessed by echocardiography and hematoxylin and eosin (H&E) staining. ADAM17 mRNA was detected by real-time reverse transcription polymerase chain reaction, and protein expression of ADAM17, tissue inhibitor of metalloproteinases-3 (TIMP-3) and TNF-α was analyzed by western blotting. The systolic function was sharply worsened in the MI1w group (versus the Con1w group, P<0.05), but left ventricular weight index was significantly increased after 4weeks post-MI (P<0.05). H&E staining revealed that one week after AMI, myocardial tissue in the epicardial border zone of the infarcted heart was mixed with broken mitochondrial cristae and decreased matrix density. ADAM17 mRNA and protein expression was significantly increased, accompanied by decreased TIMP-3 and upregulated TNF-α expression in the MI1w group (versus the MI1d group, all P<0.05). Moreover, dynamic ADAM17 mRNA expression was positively correlated with enlarged LVEDd and LVESd (P=0.001, P=0.003) and negatively with LVEF (P=0.039) in AMI rats. Enhanced ADAM17 expression, along with decreased TIMP-3 and increased TNF-α expression, especially within one week after AMI, is associated with cardiac remodeling.


[The effects of flavonoids on rat liver during chronic palloidin intoxication/Morphological and biochemical studies (author's transl)].
Continuous intraperitoneal administration of phalloidin (0.5 mg/kg body weight/day) leads to an alteration of intracellular contractile acto-myosinfilaments in rat liver. The hepatocytes show an accumulation of fibrillar material with some loss of contractile function of the pericanalicular web. Biochemically an increase of serum transaminases and alkaline phosphatase occurs. Histochemically the liver exhibitis changes in the distribution of some hepatocellular enzymes. The influence of the flavonoid (+)-cyanidanol-3 on these phalloidin-induced lesions was studied by histochemical, immunofluorescence and biochemical methods. The results imply, that (+)-cyanidanol-3 is probably protecting the plasma membrane of hepatocytes and therefore reduces the entrance of phalloidin into the cytoplasm. In addition an increased activity of the reticuloendothelial system was observed, perhaps resulting from the flavonoid administration. Both effects could be discussed as mechanisms of flavonoid action in the liver.


Effects of altered extracellular and intracellular calcium concentration on hyperpolarizing responses of the hamster egg.
Upon fertilization the hamster egg shows transient, periodic hyperpolarizing responses (h.r.s) due to a Ca-activated K conductance; these are superimposed on a gradual, hyperpolarizing shift of the resting potential (h.s.) (Miyazaki & Igusa, 1981a, 1982a). The h.r.s and h.s. were further analysed by changing external divalent cations or by injection of Ca2+ into the egg, to study the mechanisms of the increase in the intracellular Ca2+ concentration ([Ca2+]i). The series of h.r.s was abolished by the removal of external Ca2+. The frequency of the h.r. was decreased by lowering the [Ca2+]o or by adding Mn2+ or Co2+, and it was increased by raising the [Ca2+]o in a time- and concentration-dependent manner. The h.r. frequency was decreased on sustained depolarization with steady current, and increased on hyperpolarization. In contrast to the h.r. frequency, the amplitude, conductance increase and reversal potential of each h.r. were little affected by [Ca2+]o, Mn2+ or Co2+. The h.s. was decreased by lowering the [Ca2+]o, by adding Mn2+ or Co2+, or by injection of EGTA. The h.s. may reflect continuous Ca influx stimulating a Ca-activated K conductance (GK). In unfertilized eggs a regenerative h.r. was induced by Ca injection with an apparent threshold. The relationship between GK and the injected Ca2+ showed a steep jump at the critical current, associated with a four-fold increase in GK. The regenerative h.r. was followed by a refractory period of 1-2 min. In inseminated eggs the periodic sperm-mediated h.r.s. (s.-h.r.s) were interrupted by interposed h.r.(s) induced by Ca injection(s): the periodicity of s.-h.r.s was reset by Ca-induced h.r. In inseminated eggs the regenerative h.r. was induced by Ca injection with a much smaller pulse than necessary in unfertilized eggs. The refractory period was shortened to 40-50 sec, comparable to the period of s.-h.r.s. In inseminated eggs periodic h.r.s similar to s.-h.r.s were produced by continuous, repetitive injections of Ca2+ with constant pulses. The frequency of these h.r.s was dependent on the injection current. It is concluded that each h.r. indicates an enhancement of the increase in [Ca2+]i, probably the result of Ca-induced Ca release from intracellular stores. A possible mechanism for periodic increase in [Ca2+]i reflected in s.-h.r.s is proposed, based on a linkage of the continuous Ca influx to Ca release.


Electrophysiological effects of the combination of imipramine and desipramine in guinea pig papillary muscles.
The electrophysiological effects of imipramine and its active metabolite desipramine, alone or in combination, were studied in isolated guinea pig papillary muscles. The maximum upstroke velocity (Vmax) of the action potential was used as an indirect index of the magnitude of the sodium inward current. In muscles driven at 0.02 Hz, imipramine alone (5 x 10(-6) M), desipramine alone (5 x 10(-6) M), or their combination had no effect on action potential characteristics. However, in the presence of imipramine or desipramine, trains of stimuli at rates between 0.5 and 3 Hz led to a frequency-dependent Vmax block that was augmented at higher stimulation rates. At each driving rate, the Vmax block produced by desipramine was significantly greater than that produced by imipramine. The combination of imipramine and desipramine increased the onset rate of the frequency-dependent Vmax block, but the total amount of Vmax block was similar to that produced by desipramine alone. In the presence of imipramine alone, the recovery of Vmax was a monoexponential process, the time constant (tau re) being 2.3 +/- 0.4 s, whereas in the presence of desipramine, it was better defined by a biexponential function (tau re = 1.5 +/- 0.4 and 15.8 +/- 2.8 s). In the presence of the combination, the recovery process was also defined by a biexponential function and the tau re values were similar to those found in the presence of desipramine alone. Thus, according to their onset and offset kinetics of the frequency-dependent Vmax block, imipramine and desipramine can be classified as sodium channel blockers with intermediate and slow kinetics, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Utilization of RNAi to validate antibodies for reverse phase protein arrays.
Reverse phase protein arrays (RPPAs) emerged as a very useful tool for high-throughput screening of protein expression in large numbers of small specimen. Similar to other protein chemistry methods, antibody specificity is also a major concern for RPPA. Currently, testing antibodies on Western blot for specificity and applying serial dilution curves to determine signal/concentration linearity of RPPA signals are most commonly employed to validate antibodies for RPPA applications. However, even the detection antibodies fulfilling both requirements do not always give the expected result. Chemically synthesized small interfering RNAs (siRNAs) are one of the most promising and time-efficient tools for loss-of-function studies by specifically targeting the gene of interest resulting in a reduction at the protein expression level, and are therefore used to dissect biological processes. Here, we report the utilization of siRNA-treated sample lysates for the quantification of a protein of interest as a useful and reliable tool to validate antibody specificity for RPPAs. As our results indicate, we recommend the use of antibodies which give the highest dynamic range between the control siRNA-treated samples and the target protein (here: EGFR) siRNA-treated ones on RPPAs, to be able to quantify even small differences of protein abundance with high confidence.


MR imaging of degenerative disc disease in the lumbar spine with ultrashort TE pulse sequences.
The objective of this study was to assess the feasibility of using ultrashort TE (UTE) pulse sequences to image the lumbar spine. Pulse sequences of TE=0.08 ms were used to image the lumbar spine in 5 normal subjects and 14 patients with degenerative disease. Contrast enhancement was administered in 11 cases. The sequences showed high signal in the anterior and posterior longitudinal ligaments, the cartilaginous end plate, the annulus fibrosus, the ligamentum flavum, interspinous ligaments and insertions of ligaments. Normal contrast enhancement was seen in these structures. Enhancement of hypertrophied ligaments and scar tissue was readily identified. Long T2 suppression techniques were useful in distinguishing enhancement of scar tissue from veins. Enhancement in discs was more obvious than with conventional sequences. In a case of thalassaemia bands of high signal were seen in the intervertebral discs parallel to the end plates. The UTE sequences offer new options for visualizing discs, scar tissue, ligaments and other structures of the lumbar spine in health and disease.


Maternal psychological stress moderates diurnal cortisol linkage in expectant fathers and mothers during late pregnancy.
Using data from a large international sample (N = 385) of first-time expectant parents, the current analysis investigated whether parents demonstrated diurnal cortisol linkage in late pregnancy and whether self-reported psychological stress moderated this linkage. At approximately 36 weeks gestation, mothers and fathers collected saliva samples in their home at three times on two consecutive days and reported on their psychological stress. Results from multilevel models indicated that there was significant positive within-couple diurnal cortisol linkage on average for the whole sample. However, this linkage was moderated by maternal self-reported psychological stress. Specifically, for couples with higher maternal psychological stress, cortisol linkage was strong. Conversely, for couples with lower maternal psychological stress, maternal and paternal cortisol were unrelated. These findings suggest that among higher-maternal-stress couples, lower paternal cortisol may buffer maternal cortisol, whereas higher paternal cortisol may amplify maternal cortisol. Our results support the idea that interpersonal psychological and physiological stress in close relationships is interdependent and mutually influenced. Further, our findings contribute to the field's understanding of interpersonal processes during pregnancy, which may have health-related implications in the prenatal and postnatal periods for both parents and the developing child.


Outcomes of colonic diverticulitis according to the reason of immunosuppression.
The aims of this study were to analyze the relationship between the different causes of immunosuppression (IMS) and diverticulitis. IMS patients admitted for colonic diverticulitis were included in the study. Patients were divided in 5 groups according to the reasons of IMS: group I, chronic corticosteroid therapy; group II, transplant patients; group III, malignant neoplasm disease; group IV, chronic renal failure; group V, others immunosuppressant treatment. Rate of emergency surgery, outcomes in terms of postoperative mortality, and recurrence rate after nonoperative management were analyzed according to the reason of IMS. Emergency surgery was performed in 76 patients (39.3%). It was needed more frequently in group I. Overall, postoperative mortality was of 31.6% and recurrence rate after successful nonoperative management occurred in 30 patients (27.8%). No differences were observed among the groups. The rate of emergency surgery in IMS patients at the first episode of acute colonic diverticulitis is high. Elective surgery in IMS patients should be individually indicated according to persistence of symptoms or early recurrences.


Programmable photonic neural networks combining WDM with coherent linear optics.
Neuromorphic photonics has relied so far either solely on coherent or Wavelength-Division-Multiplexing (WDM) designs for enabling dot-product or vector-by-matrix multiplication, which has led to an impressive variety of architectures. Here, we go a step further and employ WDM for enriching the layout with parallelization capabilities across fan-in and/or weighting stages instead of serving the computational purpose and present, for the first time, a neuron architecture that combines coherent optics with WDM towards a multifunctional programmable neural network platform. Our reconfigurable platform accommodates four different operational modes over the same photonic hardware, supporting multi-layer, convolutional, fully-connected and power-saving layers. We validate mathematically the successful performance along all four operational modes, taking into account crosstalk, channel spacing and spectral dependence of the critical optical elements, concluding to a reliable operation with MAC relative error [Formula: see text].


High-grade desmoplastic infantile astrocytoma in a 1-year-old child with Down's syndrome: a case report.
Down's syndrome is the most common chromosomal abnormality in humans. It has been associated with central nervous system tumors such as primary acute lymphoblastic leukemia and germinomas, but desmoplastic infantile astrocytoma has not yet been reported with Down's syndrome. Desmoplastic infantile astrocytoma is a rare intracranial tumor that mostly occurs in the first 2 years of life. It usually presents as a large, aggressive tumor with both solid and cystic components. Genetically, it has been linked to the BRAF V600E mutation. Despite the rapid growth pattern, it usually has a favorable prognosis after neurosurgical excision. The presence of this extremely rare, genetically linked tumor, and its combination with Down's syndrome, the most common human genetic defect, makes this a very novel clinical presentation. It also raises a very research-worthy question of an undiscovered link between these two genetic disorders. In this case, we report a 1-year-old Pakistani origin male child with Down's syndrome, who presented with progressive macrocephaly and developmental regression over the last 2 months. He was unable to sit by himself, and had lost his handgrip bilaterally. Down's Syndrome was diagnosed soon after birth, based on typical facial features and presence of palmar crease, and later confirmed karyotypically for Trisomy 21. Upon presentation, initial blood tests did not show any abnormality. Magnetic resonance imaging of the brain was done, and showed a mixed intensity cystic mass with solid dural component posteriorly in the right parieto temporo occipital region. Craniotomy was performed, and about 85% of the tumor mass was excised. Histological examination and immunochemistry confirmed the suspected radiological diagnosis of desmoplastic infantile astrocytoma. After surgical excision, our patient gradually reacquired his previously regressed developmental milestones. Unfortunately, the remaining mass, which could not be excised due to its attachment to the highly vascular dura mater, showed regrowth on repeat brain magnetic resonance imaging. As his parents did not consent to further surgery, chemotherapy was offered as the next treatment option to prevent tumor regrowth. This case report highlights the need for more case data and research to understand desmoplastic infantile astrocytoma, and their genetic correlation with Down's syndrome. From a clinical standpoint, since desmoplastic infantile astrocytoma has a good postresection prognosis in a majority of early-diagnosed clinical cases, pediatricians, radiologists, and pathologists should consider desmoplastic infantile astrocytoma in their initial differential diagnosis in Down's syndrome patients with macrocephaly and developmental regression during the first 2 years of life.


Nutritional support in the critical ill patient: Requirements, prescription and adherence.
In critically ill patients, nutritional support is a challenge in terms of both estimating their requirements and ensuring adherence to the prescribed treatment. To assess the association between requirements, prescription and adherence to energy and protein supplementation based on the phase of disease in critically ill patients. We conducted a prospective, observational and analytical study in patients aged 0-18 years admitted to the paediatric intensive or intermediate care unit in 2020-2021. We collected data on demographic and anthropometric characteristics and the phase of disease (acute phase [AP] vs. non-acute phase [nAP]), in addition to prescribing (P) (indication of nutritional support), basal metabolic rate (BMR, Schofield equation), adherence to nutritional support (A) and protein requirements (R), and calculated the following ratios: P/BMR, P/R, A/BMR, A/R, and A/P. The sample included 131 participants with a median age of 16 (4.5) months, of who 128 (97.7%) had comorbidities and 13 (9.9%) were in the AP. Comparing the phases of disease (AP vs. nAP), the median values for energy supplementation were P/BMR, 0.5 (IQR, 0.1-1.4) vs. 1.3 (IQR, 0.9-1.8) (P = 0.0054); A/BMR, 0.4 (IQR, 0-0.6) vs. 1.2 (IQR, 0.8-1.7) (P = 0.0005); A/P, 0.7 (IQR, 0-0.9) vs. 1 (IQR, 0.8-1) (P = 0.002), and for protein were P/R, 0.7 (IQR, 0-1.1) vs. 1.2 (0.9-1.6) (P = 0.0009); A/R 0.3 (IQR, 0-0.6) vs. 1.1 (IQR, 0.8-1.5) (P = 0.0002); A/P 0.7 (IQR, 0-1) vs. 1(IQR, 0.8-1) (P = 0.002). We found AP/nAP ratios greater than 110% for energy in the P/BMR (4 patients [30.8%]/72 patients [61%]; P = 0.007), A/BMR (3 [23%]/63 [53.4%]; P = 0.009) and A/P (1 [7%]/3 [2.5%]; P = 0.007). As for protein, more than 1.5 g/kg/day was prescribed in 3 patients (23.1%) in the AP and 71 (60.1%) in the nAP. We found adherence to the prescribed intake in 2 (15.4%) patients in the AP and 66 (56%) in the nAP. We found a correlation coefficient of 0.6 between the energy P/R and the protein P/R. Prescribed support was discontinued in 7 patients (53.8%) in the AP and 31 (26.3%) in the nAP (P = 0.002). The proportion of adherence to prescribed nutritional support was high in patients in the nAP of the disease. Overfeeding was frequent, more so in the nAP. We identified difficulties in adhering to prescribed support, chief of which was the discontinuation of feeding.


Dataset for deformation behavior of pure titanium grade 2 materials during continuous extrusion.
There is a huge application and demand for titanium alloys with excellent upgraded mechanical, metallurgical, and material properties in modern industries. To fulfill the demand of modern industries metal forming process is highly desirable. Among all metal forming processes, a special type of cold forming called the continuous extrusion process has been highly appropriate to fulfill the demands. The theoretical analysis has been carried out through Upper Bound Method. The numerical simulation has been carried out through the three-dimensional finite element tool DEFORM-3D. The experimental plan and design have been carried out using Taguchi (2^3) array methods on the MINITAB platform by considering extrusion wheel velocity and feedstock temperature as chief extrusion parameters. The experimental validation process was executed on 12.5 mm CP- Titanium grade 2 feedstock materials using a TBJ350 CONFORM machine setup. The optimization process of parameters for the optimum value of the response variable was carried out through Grey Relational Analysis.


Impact of an internet-based intervention on Finnish mothers' perceptions of parenting satisfaction, infant centrality and depressive symptoms during the postpartum year.
this study evaluates how an internet-based intervention affects mothers' parenting satisfaction, infant centrality and depressive symptoms. a quasi-experimental design and repeated measures were used. this study was conducted in two Finnish public maternity hospitals (intervention/control). a convenience sample (N=1300) of primiparous and multiparous Finnish-speaking mothers were invited to participate. Multiple-birth and early discharge mothers receiving home visits were excluded. The analysis included 760 mothers. the intervention offered online support for parenting, breast feeding and infant care beginning from the middle of pregnancy. It consisted of an information database, a peer discussion forum and expert advice. Outcomes were measured by the Evaluation and Infant Centrality subscales of the What Being the Parent of a New Baby is Like-Revised, and the Edinburgh Postnatal Depression Scale after childbirth, and six weeks, six months and 12 months post partum. Age, parity, parenting self-efficacy, and perception of infant and family functioning were used as the covariates. during the first postpartum year, mothers' parenting satisfaction increased significantly, whereas infant centrality and depressive symptoms decreased within groups. However, these changes were not linear. The mean difference between groups in parenting satisfaction and depressive symptoms was not significant within any of the four assessments. The mean difference in infant centrality between control and intervention mothers was significant only at six weeks post partum. Primiparas scored significantly higher in infant centrality and significantly lower in depressive symptoms than multiparas. an internet-based intervention did not significantly affect mothers' perceptions of parenting satisfaction and depressive symptoms, but intervention mothers experienced higher infant centrality compared with control mothers at six weeks. Parenting self-efficacy was a significant covariate for all measures. Parity needs to be taken into account when infant centrality and depressive symptoms are used as outcome variables in intervention studies. More research is needed to study the potential of information data bank, professional, and peer online support. This study also highlights the need to pay more attention to the selection of the target population, the selection of outcome measures, and implementation issues in intervention research.


Prognostic and predictive roles of microRNA‑411 and its target STK17A in evaluating radiotherapy efficacy and their effects on cell migration and invasion via the p53 signaling pathway in cervical cancer.
Cervical cancer is one of the most common gynecological malignancies worldwide. However, the pathogenesis of cervical cancer remains to be fully elucidated. Increasing evidence shows that microRNAs (miRNAs) may be involved in the pathogenesis of cervical cancer. The present study tested the hypothesis that the overexpression of miRNA (miR)‑411 may delay, whereas the overexpression of serine/threonine kinase 17a (STK17A) may contribute to, cervical cancer development and progression through the p53 pathway. Cervical cancer tissues and adjacent normal tissues were obtained from 141 patients with cervical cancer following radiotherapy, with efficacy evaluated. The receiver operating characteristic curve was plotted to show the value of miR‑411 and STK17A in predicting the efficacy of radiotherapy. Cox's proportional hazards regression model was utilized for multivariate analysis. A series of inhibitors, mimics or small interfering RNAs against STK17A were introduced to validate the regulatory mechanism of miR‑411 in governing STK17A, determined with a luciferase reporter gene assay. The expression of miR‑411 and STK17A, and the status of the p53 signaling pathway were evaluated. The colony forming ability, proliferation, migration, invasion and apoptosis of CaSki cells were assessed using a colony formation assay, 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay, Transwell assay and flow cytometry, respectively. miR‑411 was upregulated but STK17A was reciprocal in cervical tissues. The overexpression of miR‑411 and low expression of STK17A were correlated with high efficacy of radiotherapy. miR‑411 and STK17A had predictive value for the efficacy of radiotherapy; miR‑411 was the protective factor and STK17A was a risk factor for prognosis of cervical cancer. Increasing miR‑411 activated the p53 signaling pathway and promoted cell apoptosis, but inhibited cell proliferation, invasion and migration. STK17A, an miR‑411 target, increased following miR‑411 over‑expression, whereas the p53 signaling pathway was activated following STK17A inhibition. It was observed that the effect of miR‑411 inhibition was lost following STK17A silencing. These findings indicate that the miR‑411‑mediated direct suppression of STK17A induces apoptosis and suppresses the proliferation, migration and invasion of human cervical cancer cells via the p53 signaling pathway. Additionally, miR‑411 and STK17A have predictive value for the efficacy of radiotherapy.


Two Mms2 residues cooperatively interact with ubiquitin and are critical for Lys63 polyubiquitination in vitro and in vivo.
Recent structural analyses support a model whereby Mms2 interacts with and orientates Ub to promote Ubc13-mediated Lys63 chain formation. However, residues of the hMms2-Ub interface have not been addressed. We found two hMms2 residues to be critical for binding and polyUb conjugation. Surprisingly, while each single mutation reduces the binding affinity, the double mutation causes significant reduction of Ub binding and abolishes polyUb chain formation. Furthermore, the corresponding yeast mms2 double mutant exhibited an additive phenotype that caused a complete loss of MMS2 function. Taken together, this study identifies key residues of the Mms2-Ub interface and provides direct experimental evidence that Mms2 physical association with Ub is correlated with its ability to promote Lys63-linked Ub chain assembly.


Clinical Impact of Gastric Acid-Suppressing Medication Use on the Efficacy of Erlotinib and Gefitinib in Patients With Advanced Non-Small-Cell Lung Cancer Harboring EGFR Mutations.
Gastric acid-suppressing medications (AS), namely, proton pump inhibitors and histamine-2 receptor antagonists, increase gastric pH, which may reduce the absorption of the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors-erlotinib and gefitinib. From 2008 to 2011, 130 consecutive patients with advanced non-small-cell lung cancer (NSCLC) harboring EGFR mutations were treated with either erlotinib or gefitinib at our institution. The clinical characteristics of the patients were reviewed, and the efficacy and toxicity of erlotinib and gefitinib were compared for patients receiving and not receiving AS. Among the 130 patients, 47 received AS (AS users group), while the remaining 83 patients did not (AS non-users group). The overall response rate (ORR) and median progression-free survival (PFS) in the subject population was 60% and 10 months, respectively. In the AS users and non-users groups, the ORR was 64% and 63% (P = .92), while the median PFS was 8.7 and 10.7 months (P = .13), respectively. No significant difference in either ORR or PFS was observed between the 2 groups. With regard to the toxicity, the frequencies of rash (83% vs. 86%; P = .60) and diarrhea (34% vs. 29%; P = .55) were similar for both groups. A multivariate analysis identified that AS use was not a significant factor for either PFS or OS. Concurrent use of AS did not affect the efficacy or toxicity of erlotinib and gefitinib in patients with advanced NSCLC harboring EGFR mutations.


Pediatric health screening and referral in the ED.
Many studies have demonstrated the importance of performing preventive care in the ED. The primary objective of this study was to identify unmet health needs in the ED of the pediatric patient population. The secondary objective was to determine if the patient's parent or guardian would accept health referrals and bring the child to follow up with a doctor. Age- and sex-specific algorithms concerning preventive care were developed from the US Department of Public Health Clinicians' Book of Preventative Health. A convenience sample of patients and their families who presented to the ED were asked to participate in the study. The exclusion criteria consisted of patients who were institutionalized, unstable, and had parents who were unable to communicate or declined to participate. After 1 week, the parents were followed up by telephone to find out if they had made an appointment with a doctor as recommended. One month after the ED visit, the health care's computer system was queried to confirm that the appointment had been completed. Data were analyzed using SPSS (version 10.0; SPSS, Chicago, Ill), and tests of significance used were the Pearson chi2, frequency test, and crosstabs. This study was institutional review board-approved as exempt. Two hundred three pediatric patients were enrolled. Most of the patients had a primary care physician (87.1%, 176/203) and insurance (85.6%, 172/203). Only 25 (12.3%) of 203 needed any referrals, with an acceptance rate of 72.0% (18/25) and completion rate of 40% (10/25). The most frequent unmet need was for urine, lead, and anemia screening (19.4%, all 20/103). Few patients needed immunizations (1.0%, 2/203), alcohol screening (2.0%, 2/100), or blood pressure testing (3.0%, 3/100). Analysis of the correlation between getting 1 or more referrals and race was found to have a significant relationship (chi = 19.69, df = 6, P = .003) but not with sex, age group, insurance, or primary care physician (P > .05). In this study, 12.3% of the patients were found to have unmet health care needs. Insurance status had no bearing on the need for referrals. Assessment in the pediatric population for unmet health care needs was found to be a low-yielding, labor-intensive process.


GTP regeneration influences interactions of microtubules, neurofilaments, and microtubule-associated proteins in vitro.
Interactions of microtubules, neurofilaments, and microtubule-associated proteins were investigated by turbidity and falling-ball viscometry measurements. We found evidence of endogenous GTPase activity in neurofilaments and microtubule-associated proteins (MAPs) in preparations that do not include urea or heat treatment, respectively. The absence or presence of either adenyl-5'-yl imidodiphosphonic acid or a GTP-regenerating system markedly influenced observed polymerization and gelation characteristics. Most significantly, the apparent viscosity of neurofilament and microtubule samples did not display a biphasic optimal MAP concentration profile when a GTP-regenerating system was operant. Likewise, GTP regeneration promoted the recovery of gelation following mechanical disruption of neurofilament/MAP/microtubule mixtures. These and other observations require some reassessment of proposed roles for microtubule-associated proteins in modulating neurofilament-microtubule interactions in vitro.


A sensitive LC/MS/MS method using silica column and aqueous-organic mobile phase for the analysis of loratadine and descarboethoxy-loratadine in human plasma.
A sensitive method using liquid chromatography with tandem mass spectrometric detection (LC/MS/MS) was developed and validated for the simultaneous analysis of antihistamine drug loratadine (LOR) and its active metabolite descarboethoxy-loratadine (DCL) in human plasma. Deuterated analytes, i.e. LOR-d(3) and DCL-d(3) were used as the internal standards (I.S.). Analytes were extracted from alkalized human plasma by liquid/liquid extraction using hexane. The extract was evaporated to dryness under nitrogen, reconstituted with 0.1% (v/v) of trifluoroacetic acid (TFA) in acetonitrile, and injected onto a 50 x 3.0 mm I.D. 5 microm, silica column with an aqueous-organic mobile phase consisted of acetonitrile, water, and TFA (90:10:0.1, v/v/v). The chromatographic run time was 3.0 min per injection and flow rate was 0.5 ml/min. The retention time was 1.2 and 2.0 min for LOR and DCL, respectively. The tandem mass spectrometric detection was by monitoring singly charged precursor-->product ion transitions: 383-->337 (m/z) for LOR, 311-->259 (m/z) for DCL, 388-->342 (m/z) for LOR-d(3), and 316-->262 (m/z) for DCL-d(3). The low limit of quantitation (LLOQ) was 10 pg/ml for LOR and 25 pg/ml for DCL. The inter-day precision of the quality control (QC) samples was 3.5-9.4% relative standard deviation (R.S.D.). The inter-day accuracy of the QC samples was 99.0-107.9% of the nominal values.


Gene-specific dye bias in microarray reference designs.
The most widely used microarray experiment design includes the use of a reference standard. Comparisons of gene expression between samples are facilitated because each sample is directly measured against the reference standard, using two fluorescent dyes. Numerous reports indicate that some genes incorporate the two commonly used dyes with different efficiencies, contributing to inaccurate data. However, it is widely assumed that these effects will not corrupt results if the reference standard is labeled with the same dye on each microarray. We demonstrate that this assumption is not reliable and that dye orientation can significantly influence measured changes in gene expression.


Functionalising the wavefunction.
Functionalism is the view that being x is to play the role of x. This paper defends a functionalist account of three-dimensional entities in the context of Wave Function Realism (WFR), that can explain in details how we can recover three-dimensional entities out of the wavefunction. In particular, the essay advocates for a novel version of WFR in terms of a functional reductionist approach in the style of David Lewis. This account entails reduction of the upper entities to the bottom ones, when the latter behave appropriately. As applied to WFR, it shows how the wavefunction can turn out to be identical to three-dimensional objects, provided certain conditions. The first major goal of the paper is thus to put forward an improved and more rigorous version of WFR, which dissolves several extant issues about the theory, and can serve as a starting point for the future literature about the topic. Moreover, the second major goal of the article is to take WFR as a case study to demonstrate the pros of functional reductionism, especially in the form defended here, thereby helping to bring this view back in the philosophy of science debate. The positive upshots of this paper suggest a possible application of functional reductionism also to other contexts.


Oxidation of allylic alcohols to alpha,beta-unsaturated carbonyl compounds with aqueous hydrogen peroxide under organic solvent-free conditions.
Allylic alcohols are chemoselectively oxidized to alpha,beta-unsaturated carbonyl compounds in high yield with aqueous H(2)O(2) in the presence of Pt black catalyst under organic solvent-free conditions. The catalyst is easily recyclable and effective for at least 7 cycles.


Identification of self-incompatibility genotypes of apricot (Prunus armeniaca L.) by S-allele-specific PCR analysis.
A cDNA of 417 bp encoding an S-RNase gene, named PA S3, was isolated from apricot, Prunus aremeniaca. Nine S-alleles, S1-S9, were recognized by S-allele-specific PCR and confirmed by Southern blot analysis using PA S3 as probe. The S-genotypes of the six cultivars were determined and the results of self- and cross-pollination tests among the six cultivars were consistent with the predicted S-haplotypes by PCR analysis.


Self-disproportionation of Enantiomers of Enantiomerically Enriched Compounds.
This review describes self-disproportionation of enantiomers (SDE) of non-racemic mixtures, subjected to distillation, sublimation, or chromatography on achiral stationary phase using achiral eluent, which leads to the substantial enantiomeric enrichment and corresponding depletion in different fractions, as compared to the enantiomeric composition of the starting material. This phenomenon is of a very general nature as SDE has been reported for different classes of chiral organic compounds bearing various functional groups and possessing diverse elements of chirality. The literature data discussed in this review clearly suggests that SDE is typical for enantiomerically enriched chiral organic compounds and special care should always be taken in evaluation of the stereochemical outcome of enantioselective reactions as well as determination of enantiomeric ratios of non-racemic mixtures of natural products after any purification process. The role of molecular association of enantiomers on the magnitude and preparative efficiency of SDE, as a new, nonconventional method for enantiomerc purifications, is emphasized and discussed.


Importance of Multiple Reinforcing Comments and Areas for Change in Optimizing Dietary and Exercise Self-Monitoring Feedback in Behavioral Weight Loss Programs: Factorial Design.
Individualized dietary and physical activity self-monitoring feedback is a core element of behavioral weight loss interventions and is associated with clinically significant weight loss. To our knowledge, no studies have evaluated individuals' perspectives on the composition of feedback messages or the effect of feedback composition on the motivation to self-monitor. This study aims to assess the perceptions of feedback emails as a function of the number of comments that reinforce healthy behavior and the number of areas for change (ie, behavioral changes that the individual might make to have an impact on weight) identified. Emailed feedback followed a factorial design with 2 factors (ie, reinforcing comments and areas for change), each with 3 levels (ie, 1, 4, or 8 comments). A total of 250 adults with overweight or obesity who were interested in weight loss were recruited from the Qualtrics research panel. Participants read 9 emails presented in a random order. For each email, respondents answered 8 questions about the likelihood to self-monitor in the future, motivation for behavioral change, and perceptions of the counselor and the email. A mixed effects ordinal logistic model was used to compute conditional odds ratios and predictive margins (ie, average predicted probability) on a 5-point Likert response scale to investigate the optimal combination level of the 2 factors. Emails with more reinforcing comments or areas for change were better received, with small incremental benefits for 8 reinforcing comments or areas for change versus 4 reinforcing comments or areas for change. Interactions indicated that the best combination for 3 of 8 outcomes assessed (ie, motivation to make behavioral changes, counselor's concern for their welfare, and the perception that the counselor likes them) was the email with 8 reinforcing comments and 4 areas for change. Emails with 4 reinforcing comments and 4 areas for change resulted in the highest average probability of individuals who reported being very likely to self-monitor in the future. The study findings suggest how feedback might be optimized for efficacy. Future studies should explore whether the composition of feedback email affects actual self-monitoring performance.


Access block and overcrowding at the emergency department at Tupua Tamasese Meaole Hospital in Samoa.
Access block and overcrowding are known to adversely impact on patient outcomes, service delivery, and patient experiences within emergency departments (ED) worldwide. There are no studies on access block or overcrowding from the Pacific Islands. The aim of the present study is to provide preliminary data on access block and overcrowding in the ED of the national tertiary hospital of Samoa. Mixed methods study design. Data collection was performed in March 2020. The quantitative strand calculated (1) the point prevalence of patients impacted by access block in the ED, and (2) the ED bed occupancy rate to assess for overcrowding. The qualitative strand used thematic analysis of two focus group interviews exploring access block and overcrowding with ED medical and nursing staff members. On the day of data collection, a total of 60 patients presented through the ED triage system. Of the 20 patients who were admitted into ED, 80% were triaged as 'see without delay' (CAT1), 'emergency' (CAT2) or 'urgent' (CAT3). For patients requiring admission to hospital wards, 100% waited 4 + h in ED, and 100% waited 8 + h, suggesting the presence of access block. Overcrowding in the ED setting was also evident, with an ED bed occupancy rate of 0.95, and an adjusted bed occupancy rate of 1.43. The major themes emerging from the ED staff focus groups and individual in-depth interviews were (1) the adverse impacts of access block and overcrowding, i.e., violence towards ED staff members, (2) the preventable contributing factors, i.e., lack of physical beds in the ED, and (3) practical recommendations to improve patient flow through the ED, i.e., improved collaboration between ED, outpatient services, and the hospital wards. Preliminary evidence suggested the presence of access block and overcrowding in the ED of the national tertiary hospital of Samoa. ED staff interviews provided insight into the ED frontline challenges and offered practical recommendations for ED health service improvement.


Natural disease progression and novel survival prediction model for hepatocellular carcinoma with spinal metastases: a 10-year single-center study.
Individual prediction of life expectancy in patients with spinal metastases from hepatocellular carcinoma (HCC) is key for optimal treatment selection, especially when identifying potential candidates for surgery. Most reported prognostic tools provide categorical predictions, and only a few include HCC-related factors. This study aimed to investigate the natural progression of the disease and develop a prognostic tool that is capable of providing individualized predictions. Patients with HCC-derived metastatic spinal disease were identified from a retrospective cohort of patients with spinal metastases who were diagnosed at Chiang Mai University Hospital between 2006 and 2015. Kaplain-Meier methods and log-rank tests were used to statistically evaluate potential factors. Significant predictors from the univariable analysis were included in the flexible parametric survival regression for the development of a prognostic prediction model. Of the 1143 patients diagnosed with HCC, 69 (6%) had spinal metastases. The median survival time of patients with HCC after spinal metastases was 79 days. In the multivariable analysis, a total of 11 potential clinical predictors were included. After backward elimination, four final predictors remained: patients aged > 60 years, Karnofsky Performance Status, total bilirubin level, and multifocality of HCC. The model showed an acceptable discrimination at C-statistics 0.73 (95% confidence interval 0.68-0.79) and fair calibration. Four clinical parameters were used in the development of the individual survival prediction model for patients with HCC-derived spinal metastases of Chiang Mai University or HCC-SM CMU model. Prospective external validation studies in a larger population are required prior to the clinical implication of the model.


Physical and observational practices of unusual actions prime action verb processing.
Numerous studies have highlighted a strong relationship between language and sensorimotor processes, showing, for example, that perceiving an action influences subsequent language processing. Moreover, previous studies have demonstrated that the context in which actions are perceived is crucial to enable this action-language relationship. In particular, action verb processing is facilitated when an action is perceived in its usual context (e.g., someone watering a plant) but not in an unusual context (e.g., someone watering a computer). This difference could be explained in terms of experience; because people always practice actions in accordance with the context, they have no (visual or motor) experience related to the unusual context. The aim of the present study was to test this assumption by assessing and comparing the effect of physical practice and observational learning on the action-language relationship. The results of two experiments showed a facilitation effect of both training methods. Whereas usual actions systematically prime action verb processing, the link between action and language appears for unusual actions only after training by practicing (experiment 1, physical practice) or observing (experiment 2, observational learning). Overall, these findings support the role of experience in the activation of sensorimotor representations during action verb processing.


A simple method for quick evaluation of the anterior tooth ratio: an observational study.
An ideal relationship of anterior teeth is closely related to postoperative function, stability, and aesthetics. Therefore, it is necessary to estimate the proportion of anterior teeth when communicating with patients about possible treatment plans and outcomes. This study aimed to establish a simple method for assessing the proportion of anterior teeth and to identify the standard ratio value to provide references for clinical work. Five hundred fourteen patients were divided into derivation, standard, and validation datasets. We first deduced our novel simplified anterior tooth ratio (SATR) by finding the key teeth with the derivation datasets, then established standard values by measuring the standard models, and finally validated the diagnostic performance of SATR. Independent sample t-test was used to select key teeth. Pearson's correlation analysis and linear regression analysis was used to test and verify the correlation between SATR and the anterior Bolton ratio. Chi-square test and diagnostic test were used to verify the diagnostic results using SATR. P values of < 0.05 were considered statistically significant. Patients with an abnormal anterior Bolton ratio were more likely to have variations in the maxillary and mandibular lateral incisors. Therefore, the ratio of maxillary and mandibular lateral incisors was chosen as a simple way to assess the anterior tooth ratio and was defined as SATR (simplified anterior tooth ratio). A positive correlation was observed between SATR and anterior Bolton ratio (r = 0.702, p < 0.001), with the linear regression equation as follows: y = 0.503 + 0.328x, x = SATR, y = anterior Bolton ratio. The standard value of SATR was established (85.69% ± 3.57%) and proven reliable in clinical practice. The ratio of maxillary and mandibular lateral incisors can be used to estimate the anterior tooth ratio, which showed high reliability and efficiency.


Assessment of Human Papillomavirus Non-16/18, Type-Specific Risk for Cervical Intraepithelial Neoplasia Grade 3 or Worse Among Women With Cervical Atypical Glandular Cells.
To evaluate the risk for cervical intraepithelial neoplasia grade 3 (CIN 3) or worse (including adenocarcinoma in situ [AIS] and invasive cervical cancer) associated with non-16/18 human papillomavirus (HPV) types (other HPV) among women with atypical glandular cells (AGC) in cervical cytology. This population-based cohort study evaluates the risk of CIN 3 or worse associated with other HPV types. Human papillomavirus genotyping was performed on Pap tests collected in Sweden from 341 women with AGC that were positive for other HPV types from February 17, 2014, to December 31, 2018. The women were followed for histopathologic outcomes using comprehensive registry linkages until December 31, 2019. Cumulative incidence proportions of CIN 3 or worse by specific HPV type were calculated using 1-minus Kaplan-Meier function. Hazard ratios (HRs) for CIN 3 or worse were generated using multivariate Cox regression. Of 341 women, 134 (39.3%) had CIN 3-AIS, but there were only five (1.5%) women in the cohort with invasive cervical cancer. Human papillomavirus 45 preceded 80.0% of invasive cervical cancer cases. Among women positive for HPV33, 82.9% (95% CI 58.0-97.3%) had CIN 3 or worse during follow-up. Positivity for HPV31 conferred the highest HR for CIN 3 or worse relative to other types, both in primary cytology and primary HPV screening (HR 2.71, 95% CI 1.47-5.00 and HR 3.41, 95% CI 1.95-5.96, respectively). Among non-16/18 HPV types in AGC, HPV31 and 33 had the highest risk for CIN 3 or worse, whereas most of the women with invasive cancer were positive for HPV45. Extended HPV genotyping may be helpful for the management of AGC.


Diagnostic performance of an artificial intelligence-driven cardiac-structured reporting system for myocardial perfusion SPECT imaging.
To describe and validate an artificial intelligence (AI)-driven structured reporting system by direct comparison of automatically generated reports to results from actual clinical reports generated by nuclear cardiology experts. Quantitative parameters extracted from myocardial perfusion imaging (MPI) studies are used by our AI reporting system to generate automatically a guideline-compliant structured report (sR). A new nonparametric approach generates distribution functions of rest and stress, perfusion, and thickening, for each of 17 left ventricle segments that are then transformed to certainty factors (CFs) that a segment is hypoperfused, ischemic. These CFs are then input to our set of heuristic rules used to reach diagnostic findings and impressions propagated into a sR referred as an AI-driven structured report (AIsR). The diagnostic accuracy of the AIsR for detecting coronary artery disease (CAD) and ischemia was tested in 1,000 patients who had undergone rest/stress SPECT MPI. At the high-specificity (SP) level, in a subset of 100 patients, there were no statistical differences in the agreements between the AIsr, and nine experts' impressions of CAD (P = .33) or ischemia (P = .37). This high-SP level also yielded the highest accuracy across global and regional results in the 1,000 patients. These accuracies were statistically significantly better than the other two levels [sensitivity (SN)/SP tradeoff, high SN] across all comparisons. This AI reporting system automatically generates a structured natural language report with a diagnostic performance comparable to those of experts.


Seminal lipids as energy substrate for the spermatozoa.
The level of fructose, NEFA, triglycerides and lipase activity were determined in the semen of 17 normozoospermic, 10 oligozoospermic and 10 azoospermic men. This was done immediately and 2 hours after seminal liquefaction. The levels of fructose and NEFA were significantly decreased while triglycerides show significant increase. These phenomena were more marked in normozoospermic, less marked in oligozoospermic and absent in azoospermic men. These results may indicate that both fructose and free fatty acids are utilized by sperms to supply energy or incorporated by them into triglycerides. No lipase activity was detected by the method used in the present work.


Surgical Management of Gynecologic Cancers.
This article addresses the role of surgery in the management of gynecologic cancers with liver metastases. The authors review the short-term and long-term outcomes of aggressive resection through retrospective and randomized studies. Although the data supporting aggressive resection of liver metastasis are largely retrospective and case based, the randomized control data to address neoadjuvant versus chemotherapy have been widely criticized. Residual disease remains an important predictor for survival in ovarian cancer. If a patient cannot achieve near optimal cytoreduction, radical cytoreductive procedures, such as hepatic resection, should be considered for palliation only.


Management of Otic and Nasopharyngeal, and Nasal Polyps in Cats and Dogs.
Feline inflammatory polyps are the most common nonneoplastic lesion of ear and nasopharynx in cats. Minimally invasive techniques for polyp removal, such as traction avulsion combined with curettage of the tympanic cavity and per-endoscopic transtympanic traction, have been successful for long-term resolution. Feline nasal hamartomas are benign lesions of the nasopharynx, and most have a good prognosis after surgical removal. Canine aural and nasopharyngeal inflammatory polyps are rare and have a similar clinical presentation as cats with these lesions. In dogs, it is important to achieve an accurate histologic diagnosis of these masses before appropriate surgical treatment can be planned.


The coordinated functions of the E. coli MutS and MutL proteins in mismatch repair.
The Escherichia coli MutS and MutL proteins have been conserved throughout evolution, although their combined functions in mismatch repair (MMR) are poorly understood. We have used biochemical and genetic studies to ascertain a physiologically relevant mechanism for MMR. The MutS protein functions as a regional lesion sensor. ADP-bound MutS specifically recognizes a mismatch. Repetitive rounds of mismatch-provoked ADP-->ATP exchange results in the loading of multiple MutS hydrolysis-independent sliding clamps onto the adjoining duplex DNA. MutL can only associate with ATP-bound MutS sliding clamps. Interaction of the MutS-MutL sliding clamp complex with MutH triggers ATP binding by MutL that enhances the endonuclease activity of MutH. Additionally, MutL promotes ATP binding-independent turnover of idle MutS sliding clamps. These results support a model of MMR that relies on two dynamic and redundant ATP-regulated molecular switches.


Independent Association Between Use of Proton Pump Inhibitors and Muscle Wasting in Patients with Heart Failure: A Single-Center, Ambispective, Observational Study.
Although proton pump inhibitors (PPIs) play a pivotal role in the prevention and treatment of gastric acid-related diseases and gastrointestinal adverse events caused by antiplatelet therapies, the safety of long-term use of PPIs has been questioned. The aim of this study was to determine the effects of use of PPIs on muscle mass and bone mineral density in heart failure (HF) patients. This was a single-center, ambispective (combined retrospective and prospective), observational study. HF patients (n = 747; 72 years of age; males, 54%) who received a dual-energy x-ray absorptiometry scan were enrolled. Muscle wasting was defined as appendicular skeletal muscle mass index (ASMI) < 7.0 kg/m2 in males and <5.4 kg/m2 in females. Propensity scores for the use of PPIs were calculated using a multivariate logistic regression model to minimize selection bias. Before propensity score matching, ASMI was significantly lower in patients receiving PPIs than in patients not receiving PPIs, resulting in a higher prevalence of muscle wasting in the PPI group. Such a relationship between use of PPIs and muscle wasting remained after propensity score matching. In multivariate Cox regression analyses, use of PPIs was independently associated with presence of muscle wasting (hazard ratio 1.68, 95% confidence interval 1.05-2.69) after adjustment for established risk factors of sarcopenia. On the other hand, there were no differences in bone mineral density between the PPI group and the no-PPI group. Use of PPIs is associated with a high risk of muscle wasting in HF patients. Caution is warranted when long-term PPI treatment is performed in sarcopenic HF patients and HF patients with several risk factors for muscle wasting.


Temporal and spatial delineation of mouse Otx2 functions by conditional self-knockout.
To identify the independent spatial and temporal activities of the essential developmental gene the Otx2, the germline mutation of which is lethal at embryonic day 8.5, we floxed one allele and substituted the other with an inducible CreER recombinase gene. This makes 'trans' self-knockout possible at any developmental stage. The transient action of tamoxifen pulses allows time-course mutation. We demonstrate efficient temporal knockout and demarcate spatio-temporal windows in which Otx2 controls the head, brain structures and body development.


A review of the genus Maimuna Lehtinen, 1967 (Araneae, Agelenidae) in Turkey, with a description of a new species.
The records of the agelenid spider genus Maimuna Lehtinen, 1967 from Turkey are reviewed. A new species, Maimuna anatolica sp. n. is described from Mula Province, Anatolia. The other two Turkish species of the genus are illustrated and re-diagnosed. Some discrepancies in the different diagnoses of the genus Maimuna are pointed out and discussed.


The brain correlates of state proactive aggression.
Proactive aggression can result in many economic, social, and health problems. It has two levels: trait and state. Although the neural basis of trait proactive aggression is becoming clearer, the neural basis of state proactive aggression has been largely ignored and remains unclear. We explore the basic brain correlates of state proactive aggression using a reward-interference task (RIT), in which participants play a competitive reactive time task against an opponent. They can make decision to give the opponent noise interference for winning the game and gaining reward or not. Forty healthy participants from a university in China performed the RIT in the scanner and their functional images were obtained on a 3.0-T Siemens Tim Trio scanner. Finally, 38 participants (21 males, M age = 19.24, SD = 1.32) were included in analyses. The results show that relative to the baseline, the medial prefrontal cortex (MPFC), anterior cingulate cortex (ACC), temporoparietal junction (TPJ), and inferior frontal gyrus (IFG) were activated by aggression selection and nonaggression selection in the RIT. Furthermore, relative to nonaggression selection, the MPFC, ACC, TPJ, IFG, middle cingulate cortex (MCC), and middle temporal gyrus (MTG) were more strongly activated during aggression selection and correlated with the scores of some of the three basic proactive aggression motivations (instrumental motivation, moral inhibition motivation, and moral approval motivation). The results provide preliminary evidence of neural correlates for state proactive aggression and should be replicated by other methods or with other samples. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Seasonal variations in plasma luteinizing hormone and testosterone levels in the European badger Meles meles L.
Although active spermatogenesis occurs throughout the year in the male European badger Meles meles L., seasonal fluctuations of testicular activity were observed. The present study compared the pattern of luteinizing hormone (LH) and testosterone (T) secretions throughout the annual cycle. LH secretion was investigated by means of a heterologous radioimmunoassay using anti-ovine LH beta antiserum and canine reference standard. Seasonal and nycthemeral changes in LH and T secretions were observed. During the year a period of maximal values for both hormones is well correlated with the breeding. Moreover, episodic LH releases occurs during the period of minimal testosterone values, indicating resumption of hypothalamo-pituitary activity well before the breeding period.


Local connections in transplanted and normal cerebral cortex of rats.
Injections of the fluorescent tracer Fluoro-Gold were made in transplanted and normal cerebral cortex of rats in order to investigate and compare the local connectivities of both. In the normal somatosensory cortex, small injections in superficial layers (I to III) produced retrograde cell labeling below the injection site in two bands: in layer V and in the deep part of layer VI. Pieces of embryonic rat neocortical tissue were transplanted into a cavity made in the somatosensory cortex of young adult rats. After a survival period of 2-3 months, small injections of Fluoro-Gold were made in the superficial part of the grafts. These injections revealed multiple clusters of intratransplant-projecting cells. No callosal or thalamic neurons were labeled in these experiments. On occasion, a bilaminated pattern of retrograde cell labeling was observed inside the transplants. In both transplanted and normal cortices, pyramidal and non-pyramidal cells were retrograde-labeled. We conclude that in the neocortical transplants there is a pattern of local connectivity that is reminiscent of the pattern of intracortical connectivity in the normal neocortex in at least two aspects: first, the retrograde-labeled cells tended to form clusters or bands; second, both pyramidal and non-pyramidal cells were labeled.


[Infantile brucellosis (author's transl)].
Some aspects concerning brucellosis are revised. This is a disease with a high incidental rate among the children's population in Salamanca and bordering provinces. The local prevailing epidemiologic conditions are reviewed, as well as the most outstanding clinical features, which are in children, according to the author's personal experience, less protean than in adult's illness.


Activities of serine hydroxymethyltransferase in murine tissues and tumours.
The specific activity of the enzyme serine hydroxymethyltransferase (EC 22.214.171.124) was determined in various murine, rat and human tumour cell lines. The activities of the enzyme were also investigated in tissues of non-tumour bearing DBA/2 mice and BALB/c mice bearing the PC6 ascites tumour. The highest enzyme activity in the murine tissues was found in the liver and then the kidneys. The enzyme was present in all the tissues assayed. The activities of enzyme found in the tumours varied considerably, with the PC6 ascites, Walker 256 and Lewis lung cells, being the highest.


Quantitative 1H NMR method for the routine spectroscopic determination of enantiomeric purity of active pharmaceutical ingredients fenfluramine, sertraline, and paroxetine.
Determining the enantiomeric purity of chiral therapeutic agents is important in the development of active pharmaceutical ingredients (API). A strategy for determining the enantiomeric purity of three APIs was developed using nuclear magnetic resonance (NMR) and the chiral solvating agent (CSA) 1,1-bi-2-naphthyl (1). While chiral chromatography is widely used to evaluate enantiomeric purity, it can sometimes suffer from tedious sample preparation obviating rapid measurements that are sometimes needed during the manufacture of such agents. The techniques described herein provide comparable enantiomeric purity results with those obtained with traditional chiral HPLC and other published methods for these compounds. Chiral analysis of standard samples of methylbenzylamine enantiomeric mixtures using 1 were found to be quantitative to approximately 1% minor enantiomer. Enantiomeric purity determination by NMR utilizing chiral solvating agents do not require special instrumental techniques, chemical derivatization or standards and is therefore ideally suited for rapid routine analysis. As a result, the technique demonstrated is commonly used in our laboratory as a complementary or alternative method to chiral HPLC or optical rotation measurements for routine determination of enantiomeric purity.


Effects of STIP1 and GLCCI1 polymorphisms on the risk of childhood asthma and inhaled corticosteroid response in Chinese asthmatic children.
Asthma is a common chronic lung disease in children. We aimed to determine the associations between stress-induced phosphoprotein 1 (STIP1) and glucocorticoid-induced transcript 1 (GLCCI1) polymorphisms and susceptibility of childhood asthma and inhaled corticosteroid (ICS) response in children. A total of 263 Chinese Han asthmatic children were recruited from the Xiangya Hospital, Central South University. Pulmonary function tests were performed before the treatment and 3 months after the treatment. One hundred fifty non-asthmatic children were recruited. Each participant's DNA was extracted from the peripheral blood and Method of MassARRAY was used to genotype the single-nucleotide polymorphisms (SNPs). STIP1 rs2236647 wild-type homozygote (CC) was associated with increased asthma risk of children (OR = 1.858, 95% CI:1.205-2.864), but not associated with the ICS response. GLCCI1 rs37969, rs37972 and rs37973 polymorphisms were not associated with the risk of childhood asthma. However, rs37969 mutant genotypes (TT/GT) were significantly associated with less improvement in PD20 (p = 0.028). We also found significant associations between rs37969, rs37972 and rs37973 mutant genotypes and less improvement in maximal midexpiratory flow (MMEF) after ICS treatment for 3 months (p = 0.036, p = 0.010 and p = 0.003, respectively). STIP1 rs2236647 was associated with asthma risk of children and GLCCI1 rs37969 mutant genotypes were associated with less improvement in airway hyper-responsiveness. GLCCI1 rs37969, rs37972 and rs37973 polymorphisms might be associated with pulmonary function in childhood asthma patients after ICS treatment.


A new sequence for shaped voxel spectroscopy in the human brain using 2D spatially selective excitation and parallel transmission.
Spatially selective excitation in two dimensions (2D-SSE) utilizing parallel transmission was applied as a means to acquire signal from voxels adapted to the anatomy of interest for in vivo (1) H MR spectroscopy. A novel method to select spectroscopy voxels with arbitrary shapes in two dimensions was investigated. An on-off scheme with an adiabatic slice selective inversion pulse preceding a 2D-SSE pulse together with a segmented inward spiral excitation k-space trajectory enabled rapid free induction decay acquisitions. Performance of the sequence was evaluated in simulations, phantom experiments, and in vivo measurements at 3 T. High spatial fidelity of the excitation profile was achieved for different target shapes and with little off-resonance deterioration. Metabolite concentrations in human brain determined with the new sequence were quantified with Cramér-Rao lower bounds less than 20%. They were in the physiological range and did not deviate systematically from results acquired with a conventional SPECIAL sequence. In conclusion, a new approach for shaped voxel MRS in the human brain is presented, which complements existing sequences. Simulations show that 2D-SSE pulses yield reduced chemical shift artifact when compared with conventional localization methods. Copyright © 2016 John Wiley & Sons, Ltd.


Ovarian steroids modulate gonadotropin-releasing hormone-induced biphasic luteinizing hormone secretory responses and inositol phosphate accumulation in rat anterior pituitary cells and alpha T3-1 gonadotrophs.
The ovarian steroids estradiol and progesterone act as important modulators of GnRH-induced luteinizing hormone (LH) secretion from anterior pituitary cells. Recently, we demonstrated that the steroids are able to influence GnRH-stimulated Ca2+ mobilization from extra- and intracellular sources. Here we investigated the actions of estradiol and progesterone on GnRH-induced biphasic LH secretory responses in the model of perifused female rat pituitary cells. A 20 min GnRH stimulus elicited biphasic LH responses composed of an initial peak followed by a prolonged plateau phase. Both phases were equally enhanced by long-term (48 h) estradiol treatment. This action was facilitated by subsequent short-term progesterone treatment. In contrast, combined treatment with estradiol and progesterone for 48 h led to inhibited LH secretory profiles. To determine the steroid actions on the extracellular Ca2+ independent component of LH secretion we performed experiments using cells that were perifused with Ca2+ deficient medium. Under these conditions the cells responded exclusively with a single peak phase of LH secretion, which was augmented or inhibited by estradiol and progesterone treatment as described above. To test the hypothesis that an effect of estradiol and progesterone on GnRH-induced polyphophoinositide hydrolysis is responsible for their modulatory actions on Ca2+ signals and LH secretion we measured inositol phosphate (IP) accumulation after different steroid treatment paradigms in rat pituitary cells and alpha T3-1 immortalized gonadotrophs. GnRH-induced IP production was enhanced by long-term estradiol treatment. Short-term exposure of estradiol-primed cells to progesterone did not lead to significant changes of IP production. The long-term progesterone treatment paradigm enhanced GnRH-induced IP formation, while it decreased Ca2+ signals and LH secretion. Alpha T3-1 cells were used to perform more detailed analysis of IP formation. The actions of estradiol and progesterone on the production of inositol mono-, bis-, and trisphosphates were similar to those observed in the mixed cell population. It is concluded that estradiol and progesterone modulate both peak and plateau phases of GnRH-stimulated LH secretory responses, effects which are associated with their impact on Ca2+ signals. Our findings argue against a role of IP modulation in the mechanism of progesterone actions on Ca2+ signaling and LH secretion in gonadotrophs. Such a mechanism might be involved in the positive effects of estradiol in these cells.


xbx-4, a homolog of the Joubert syndrome gene FAM149B1, acts via the CCRK and RCK kinase cascade to regulate cilia morphology.
Primary cilia are microtubule (MT)-based organelles that mediate sensory functions in multiple cell types. Disruption of cilia structure or function leads to a diverse collection of diseases termed ciliopathies.1-3 The highly conserved CCRK and RCK kinases (ICK/MOK/MAK) negatively regulate cilia length and structure in Chlamydomonas, C. elegans, and mammalian cells.4-10 How the activity of this kinase cascade is tuned to precisely regulate cilia architecture is unclear. Mutations in the Domain of Unknown Function 3719 (DUF3719)-containing protein FAM149B1 have recently been shown to elongate cilia via unknown mechanisms and result in the ciliopathy Joubert syndrome.11 Here we identify XBX-4, a DUF3719-containing protein related to human FAM149B1, as a regulator of the DYF-18 CCRK and DYF-5 MAK kinase pathway in C. elegans. As in dyf-18 and dyf-5 mutants,10 sensory neuron cilia are elongated in xbx-4 mutants and exhibit stabilized axonemal MTs. XBX-4 promotes DYF-18 CCRK function to regulate localization and function of DYF-5 MAK. We find that Joubert syndrome-associated mutations in the XBX-4 DUF3719 domain also elongate cilia in C. elegans. Our results identify a new metazoan-specific regulator of this highly conserved kinase pathway and suggest that FAM149B1 may similarly act via the CCRK/RCK kinase pathway to regulate ciliary homeostasis in humans.


Diagnosis and interventional radiological treatment of vascular and biliary complications after liver transplantation in children with biliary atresia.
Early diagnosis and appropriate management of vascular and biliary complications after living donor liver transplantation (LDLT) result in longer survival. We report our institutional experience regarding radiological management of these complications among patients with biliary atresia (BA) who underwent LDLT. We analyzed the records of 116 children. All patients underwent Doppler ultrasound (US) at operation, daily for the first 2 postoperative weeks, and when necessary thereafter. After primary evaluation using US, the definite diagnosis of postoperative complication was confirmed using computed tomography, magnetic resonance imaging, and/or operation. There were 61 boys and 55 girls. The overall mean age was 2.69 years. The overall mean preoperative weight and height were 13.06 kg and 83.79 cm, respectively. There were 28 (24.13%) biliary and vascular complications. These were cases of biliary stricture (n = 5), bile leakage (n = 3), hepatic artery stenosis (n = 6), hepatic vein stenosis (n = 4), and portal vein thrombosis (n = 17). The diagnostic accuracy of US in detecting biliary complication, hepatic artery stenosis, hepatic venous stenosis, and portal vein thrombosis was 95.69%, 97.41%, 100%, and 100%, respectively. US in combination with multiple imaging modalities and clinical suspicion resulted in 100% diagnostic accuracy. Percutaneous transhepatic cholangiography, thrombolysis, balloon angioplasty, and stent placement were performed for the complications noted. There was an early mortality due to multiple-organ failure after failed radiological invention and subsequent surgical management. Doppler US is accurate in detecting postoperative complications after pediatric LDLT for BA. Radiological interventions for vascular and biliary complications are effective and safe alternatives to reconstructive surgery.