Singapore Undiagnosed Disease Program: Genomic Analysis aids Diagnosis and Clinical Management.
Use next-generation sequencing (NGS) technology to improve our diagnostic yield in patients with suspected genetic disorders in the Asian setting. A diagnostic study conducted between 2014 and 2019 (and ongoing) under the Singapore Undiagnosed Disease Program. Date of last analysis was 1 July 2019. Inpatient and outpatient genetics service at two large academic centres in Singapore. Inclusion criteria: patients suspected of genetic disorders, based on abnormal antenatal ultrasound, multiple congenital anomalies and developmental delay. patients with known genetic disorders, either after clinical assessment or investigations (such as karyotype or chromosomal microarray). Use of NGS technology-whole exome sequencing (WES) or whole genome sequencing (WGS). (1) Diagnostic yield by sequencing type, (2) diagnostic yield by phenotypical categories, (3) reduction in time to diagnosis and (4) change in clinical outcomes and management. We demonstrate a 37.8% diagnostic yield for WES (n=172) and a 33.3% yield for WGS (n=24). The yield was higher when sequencing was conducted on trios (40.2%), as well as for certain phenotypes (neuromuscular, 54%, and skeletal dysplasia, 50%). In addition to aiding genetic counselling in 100% of the families, a positive result led to a change in treatment in 27% of patients. Genomic sequencing is an effective method for diagnosing rare disease or previous 'undiagnosed' disease. The clinical utility of WES/WGS is seen in the shortened time to diagnosis and the discovery of novel variants. Additionally, reaching a diagnosis significantly impacts families and leads to alteration in management of these patients.


Alteration of blood oxygenation level-dependent signaling by local circulatory condition.
To determine regional differences in the relationship between neuronal activation and blood oxygenation level-dependent (BOLD) signal changes. We performed BOLD and perfusion-based studies on healthy adult volunteers (40 for BOLD and 20 for perfusion-based studies) with visual stimulation of varied extent in the visual field of subjects. Regions with a significant negative correlation between the extent of visual stimuli and BOLD and perfusion responses were found bilaterally on the lateral surface of the occipital lobe just anterior to the occipital pole. These regions were located in the border zone between the territories of the middle and posterior cerebral arteries. Insufficient perfusion pressure in these regions may have led to attenuation of BOLD and perfusion responses to stimulation of a large portion of the visual field, or a condition in which a large extent of the visual areas was activated in the proximal regions of the territories perfused by the middle and posterior cerebral arteries. This finding supports the hypothesis that regional differences in circulatory condition can result in alterations of the relationship between neuronal activation and BOLD signal changes in the normal human brain.


Silver nanoparticles coated with dodecanethiol used as fillers in non-cytotoxic and antifungal PBAT surface based on nanocomposites.
In the present study, we report the preparation of antifungal and non-cytotoxic polymer nanocomposites with potential application in biomedical materials. Dodecanethiol-protected silver nanoparticles (AgNPs-DDT) were synthesized by a reduction/precipitation method and dispersed in chloroform to obtain stable colloidal dispersions. PBAT-based nanocomposites containing 0.25, 0.5 and 2 wt% AgNPs-DDT were prepared by casting method. The incorporation of AgNPs-DDT in PBAT matrix resulted in nanocomposites which combine improved mechanical performance and antifungal properties with a non-cytotoxic characteristic.


Bone grafting to the jaws with special reference to reconstructive preprosthetic surgery. A historical review.
The first report on the transfer of fresh bone was in 1668 when van Meekeren described how a Russian surgeon had repaired a soldier's skull defect by implanting dog bone with good results. Discovery of the anesthetics, nitrous oxide and ether, in addition to the introduction of antisepsis opened up new surgical possibilities and made bone grafting a possible clinical procedure. Around the turn of the last century, papers began to appear on the restoration of continuity defects of the mandibula. In the future it will be possible to acquire a living graft in the exact size desired based on a 3D reconstruction of a CT scan of the defect.


Effects of sodium bicarbonate correction of metabolic acidosis on regional tissue oxygenation in very low birth weight neonates.
To determine the effects of sodium bicarbonate (NaHCO3) correction of metabolic acidosis on cardiopulmonary, laboratory, and cerebral, renal and splanchnic regional oxygen saturation (rSO2) and fractional tissue oxygen extraction (FTOE) in extremely premature neonates during the first postnatal week. Observational cohort data were collected from 500 to 1250 g neonates who received NaHCO3 'half' corrections (0.3 * Weight (kg) * Base Deficit (mmol l(-1))) for presumed renal losses. Twelve subjects with normal blood pressure and heart rate received 17 NaHCO3 corrections. Mean (±s.d.) gestational age was 27±2 week and birth weight was 912±157 g. NaHCO3 corrections provided a mean (±s.d.) 4.5±1.0 ml kg(-1) fluid bolus, shifted mean (±s.d.) base deficit from 7.6±1.8 to 3.4±2.1 mmol l(-1) (P<0.05), and increased median (±s.d.) pH from 7.23±0.06 to 7.31±0.05 (P<0.05). No significant changes in blood pressure, pulse oximetry, PCO2, lactate, sodium, blood urea nitrogen, creatinine or hematocrit were observed. Cerebral, renal and splanchnic rSO2 (74%, 66% and 44%, respectively, at baseline) and FTOE (0.21, 0.29 and 0.52, respectively, at baseline) were unchanged following NaHCO3 correction. NaHCO3 infusions decreased base deficits and increased pH though produced no discernible effects or benefits on cardiopulmonary parameters including rSO2 and FTOE. These findings warrant further prospective evaluation in larger populations with more significant metabolic acidosis to determine the utility of tissue oxygenation monitoring in differentiating metabolic acidosis due to oxygen delivery/consumption imbalance versus renal bicarbonate losses.


Rerefinement of poly[di-aqua-bis-(μ3-2-methyl-pro-pano-ato-κ4 O:O,O':O')bis-(μ3-2-methyl-propano-ato-κ3 O:O:O)(μ2-2-methyl-propano-ato-κ3 O:O,O')(2-methyl-propano-ato-κ2 O,O')trilead(II)].
The crystal structure of the title complex, [Pb3(C4H7O2)6(H2O)2] n , was redetermined on basis of modern CCD-based single-crystal X-ray data at 120 K. The current study basically confirms the previous report [Fallon et al. (1997 ▸). Polyhedron, 16, 19-23] at 190 K, but with higher accuracy and precision. In particular, positional disorder of one of the 2-methyl-propano-ate anions over two sets of sites was resolved, showing a refined ratio of the disorder components of 0.535 (9):0.465 (9). The three independent cations in the structure have coordination numbers of [7 + 1], [6 + 1], and [5 + 3], with O atoms belonging either to carboxyl-ate groups or water mol-ecules. This arrangement leads to the formation of sheets parallel to (01), whereby the hydro-phobic 2-methyl-propanyl groups of the anions are oriented above and below the hydro-philic sheets to form a layered structure. Within a sheet, hydrogen bonds of the type Owater-H⋯O are formed, whereas the hydro-phobic groups between adjacent layers inter-act through van der Waals forces.


Mucositis in head and neck cancer patients treated with radiotherapy and systemic therapies: Literature review and consensus statements.
Oral mucositis (OM) due to radiotherapy and systemic therapies in head and neck cancer treatment represents a major problem causing a wide spectrum of clinical signs and symptoms. This adverse event may reduce quality of life, resulting from debilitating oral pain, bleeding, dysphagia, infections, impairment of food intake, high rate of hospitalization and may interfere with the delivery of programmed treatment plans, ultimately jeopardizing patient outcome. Globally, there is a lack of evidence on effective measures for the prevention and treatment of OM, and only scant uniform conclusions and recommendations can be derived from the existing literature and guidelines. A multidisciplinary team of Italian head and neck cancer experts met in Milan 17-18 February 2013 with the aim of reaching consensus on prophylaxis and management of mucositis. The results of the literature review and the statements that achieved consensus are reported and discussed in this paper. The Delphi Appropriateness Method was used as a structured communication method for achieving consensus. Subsequently, external expert reviewers evaluated the conclusions carefully according to their area of expertise. This paper presents 13 clusters of statements on prophylaxis and treatment of mucositis that achieved consensus. OM represents a very stressful situation for head and neck cancer patients submitted to chemo-radiation or exclusive radiation treatment. A multidisciplinary approach is mandatory, but there is still no gold-standard protocol that is prominently better than others.


Genotoxic effects of starvation and dimethoate in haemocytes and midgut gland cells of wolf spider Xerolycosa nemoralis (Lycosidae).
The aim of this study was to assess the genotoxic effects of starvation and dimethoate (organophosphate insecticide) in female and male wolf spiders Xerolycosa nemoralis (Lycosidae) exposed to the stressors under laboratory conditions. DNA damage was measured in haemocytes and midgut gland cells using the comet assay. In response to the two stressing factors, both cell types showed %TDNA, tail length (TL) and OTM values higher in males than in females. Level of DNA damage in haemocytes was greater than in midgut gland cells. In both sexes, the strongest genotoxicity was recorded at single application of dimethoate. After five-time exposure to the pesticide, genotoxic effects of a single dose were sustained in males and reduced to the control level in females. Starvation stress was well tolerated by the females, in which neither cell type was affected by DNA damage. However, in male haemocytes food deprivation induced severe DNA damage, what suggests suppression of the defence potential at prolonged starvation periods.


Torsadogenic potential of a novel remyelinating drug clemastine for multiple sclerosis assessed in the rabbit proarrhythmia model.
We assessed the torsadogenic effects of a novel remyelinating drug clemastine for multiple sclerosis using an in vivo proarrhythmia model of acute atrioventricular block rabbit, since the drug has been demonstrated to suppress the human ether-á-go-go related gene (hERG) K+ channels. Bradycardia was induced by atrioventricular node ablation in isoflurane-anesthetized New Zealand White rabbits (n = 5), and the ventricle was electrically driven at 60 beats/min throughout the experiment, except when extrasystoles appeared. Intravenous administration of clinically relevant dose of 0.03 mg/kg of clemastine and 10-times higher dose of 0.3 mg/kg hardly affected the QT interval or duration of the monophasic action potential (MAP) of the ventricle. Additional administration of clemastine at 3 mg/kg significantly increased the QT interval, MAP duration and the short-term variability of repolarization. Meanwhile, the premature ventricular contractions with R on T phenomenon were observed in 3 out of 5 animals, and torsades de pointes arrhythmias were detected in 1 out of 5 animals. These results suggest that the torsadogenic potential of clemastine is obviously observed in the acute atrioventricular block rabbit, which will not appear within the prescribed dose for multiple sclerosis.


Alternatives to prokinetics to move the pylorus and colon.
Gastrointestinal motility disorders (GMDs) are common in the ICU. When encountering these problems, one typically thinks of prokinetics. This review summarizes current evidence of treatments. Prokinetics are not the first-line therapy for GMDs. In fact, the clinical implications of using prokinetic agents are rather controversial. Current evidence on alternative treatment modalities such as fluid and electrolyte management, laxatives, opioid antagonists, purgative enemas, acupuncture, physical therapies and probiotics is growing. Current state of the art to treat GMDs is primarily focused at the elimination of underlying trigger factors. Fluid and electrolyte management as well as laxatives and peripherally acting μ-opioid receptor antagonists are the recommended first-line therapies that can be complemented with prokinetics. Acupuncture as well as physical modalities, such as massage or warming of the abdomen, is promising with few side-effects and should be considered as well.


Causation and prevention of solely estrogen-induced oncogenesis: similarities to human ductal breast cancer.
Estrogens are intimately involved in the causation of some of the most prevalent cancers afflicting women, particularly, breast, endometrial, cervico-vaginal, and possibly ovarian. Therefore, it has become particularly pertinent to elucidate the molecular changes and mechanisms whereby estrogens elicit their oncogenic actions so that better prevention strategies can be developed. The estrogen-induced Syrian hamster tumors of the kidney have emerged as one of the most intensively studied in-vivo models in solely estrogen-induced oncogenesis. An advantage of this model is that the tumors occur in the absence of any intervening morphologic changes, but rather they are the result of the continuous progression of a subset of interstitial stem cells in the kidney leading to tumor formation. Evidence is presented that the origin of these tumors is derived from ectopic "uterine" stem cells, which are responsive to estrogenic hormones. The other animal tumor model studied is the highly sensitive estrogen-induced mammary tumors of female ACI rats. Their steroid receptor and other gene alterations have been delineated. Importantly, a crucial early event in this solely estrogen-induced oncogenic process, common to both animal tumor models, is the overexpression and amplification of c-myc and its protein product. Chromosomal instability, in both early and large well-established frank tumors, is another important characteristic found during early E-induced oncogenesis. These features have been shown to be characteristic of human ductal carcinomas in-situ and in primary invasive ductal breast carcinomas. The molecular alterations seen are considered crucial in eliciting estrogen-induced oncogenesis and have established for the first time a direct causal link between estrogen and the induction of chromosomal instability and aneuploidy in these estrogen-associated neoplasms.


Elucidating the roles of Cr(VI)-Cu(II) Co-pollution in the stress of aniline degradation stress: Insights into metabolic pathways and functional genes.
In order to examine the impact of Cu(II)-Cr(VI) co-pollution in printing and dyeing wastewater on the aniline biodegradation system (ABS), loading experiments were conducted on ABS at varying concentrations of Cu(II)-Cr(VI). The synergistic stress imposed by Cu(II)-Cr(VI) accelerated the deterioration of the systems, with only the C2-3 (2 mg/L Cr(VI)-3 mg/L Cu(II)) sustaining stable operation for 42 days. However, its nitrogen removal performance remained significantly impaired, resulting in a total nitrogen (TN) removal rate below 40%. High-throughput sequencing analysis revealed a stronger correlation between Cr(VI) and microbial diversity compared to Cu(II). Metagenomic sequencing results demonstrated that Cu(II) emerged as the dominant factor influencing the distribution of dominant bacteria in C2-3, as well as its contribution to contaminant degradation. The complex co-pollution systems hindered aniline degradation and nitrogen metabolism through the combined bio-toxicity of heavy metals and aniline, thereby disrupting the transport chain within the systems matrix.


Affinity labelling of the tick-borne encephalitis virus RNA replicase proteins by 4-N-exo-base-substituted photoreactive CTP analogs.
4-N-exo-base-substituted photoreactive analogs of CTP were designed and synthesized. Two flavivirus proteins NS5 and NS3 are shown to be labelled after RNA synthesis in the presence of the analogs, irradiation by UV-light (313 nm) and subsequent [alpha-32P]NTP incorporation.


Stochastic model of the formation of cancer metastases via cancer stem cells.
The author presents Monte Carlo simulations of the temporal kinetics of the formation of cancer metastases with emphasis on cancer stem cells. The model used implies the existence of premetastatic niches. The population of cancer stem cells located outside tumors and inducing the formation of new tumors in niches is considered to be heterogeneous. If the niches are equivalent with respect to the formation of metastases, the kinetics are predicted to exhibit an induction period and then rapid growth of the number of metastases. If the niches are heterogeneous, the kinetics are found to be more gradual.


Patterns of heroin and cocaine injection and plasma HIV-1 RNA suppression among a long-term cohort of injection drug users.
Previous studies suggest that active drug use may compromise HIV treatment among HIV-positive injection drug users (IDU). However, little is known about the differential impacts of cocaine injection, heroin injection, and combined cocaine and heroin injection on plasma HIV-1 RNA suppression. Data were derived from a longstanding open prospective cohort of HIV-positive IDU in Vancouver, Canada. Kaplan-Meier methods and Cox proportional hazards regression were used to examine the impacts of different drug use patterns on rates of plasma HIV-1 RNA suppression. Between May 1996 and April 2008, 267 antiretroviral (ART) naïve participants were seen for a median follow-up duration of 50.6 months after initiating ART. The incidence density of HIV-1 RNA suppression was 65.2 (95%CI: 57.0-74.2) per 100 person-years. In Kaplan-Meier analyses, compared to those who abstained from injecting, individuals injecting heroin, cocaine, or combined heroin/cocaine at baseline were significantly less likely to achieve viral suppression (all p<0.01). However, none of the drug use categories remained associated with a reduced rate of viral suppression when considered as time-updated variables (all p>0.05). Active injecting at the time of ART initiation was associated with lower plasma HIV-1 RNA suppression rates; however, there was no difference in suppression rates when drug use patterns were examined over time. These findings imply that adherence interventions for active injectors should optimally be applied at the time of ART initiation.


Vascular cell adhesion molecule-1 gene expression during human smooth muscle cell differentiation is independent of NF-kappaB activation.
Vascular cell adhesion molecule-1 (VCAM-1) gene expression in cytokine-activated cells depends on two kappaB elements. Since VCAM-1 expression appears developmentally regulated and cytokine-inducible in smooth muscle cells (SMCs), we have studied the role of NF-kappaB in differentiated SMC VCAM-1 expression. Confluent SMCs were cultured either in a serum-free medium in order to induce differentiation, or in medium with serum, stimulated or not by tumor necrosis factor alpha (TNF-alpha). The expression of smooth muscle myosin heavy chain, a SMC marker, and VCAM-1 was induced concomitantly in serum-free medium, whereas only VCAM-1 expression was induced by cytokine-treatment. We showed that the p50 and p65 subunits of NF-kappaB were localized in the cytoplasm of differentiating SMCs, whereas they were translocated into the nucleus of TNF-alpha-activated SMCs. Electrophoretic mobility shift assays with VCAM-1 gene kappaB elements failed to detect any induction of DNA-protein complex with nuclear extracts of differentiating SMCs in contrast to the cytokine-activated SMC nuclear extracts. Furthermore, VCAM-1 mRNA induction was inhibited in TNF-alpha-stimulated SMCs, but not in differentiating SMCs, by pyrrolidine dithiocarbamate, an inhibitor of NF-kappaB protein activation. Taken together, these findings suggest that in contrast to TNF-alpha activation, NF-kappaB is not involved in VCAM-1 gene expression during SMC differentiation.


pH and NIR responsive polydopamine-doped dendritic silica carriers for pesticide delivery.
The stimuli-responsive pesticide delivery system provides a powerful strategy for enhancing the effective utilization of pesticides and reducing environmental pollution. Here, we prepared a new polydopamine doped dendritic silica (SiO2/PDA) nanocarriers for pesticide delivery. The SiO2/PDA nanocarriers present uniform spheres with an average diameter of 250 nm and carry center-radial inner pores. After loading dinotefuran (DNF) insecticide, polyethyleneimine (PEI) was used to cap the loaded-pesticide pores. The resulting SiO2/PDA@PEI displays high photothermal conversion effect (η = 35.1 %) and pH and near infrared (NIR) light-responsive release behavior. Meanwhile, the SiO2/PDA and SiO2/PDA@PEI carriers displayed high adhesion and wettability to leaves, and the photostability of DNF encapsulated in SiO2/PDA@PEI was improved by nearly 10 times greater than for free DNF. Importantly, the SiO2/PDA carriers possessed a benign biocompatibility on Escherichia coli (E. coli), seed and cells. Therefore, this work provides a promising approach to improve the utilization of pesticide.


Megasphaera elsdenii and Saccharomyces Cerevisiae as direct fed microbials during an in vitro acute ruminal acidosis challenge.
This study aimed to evaluate the effects of Saccharomyces cerevisiae and Megasphaera elsdenii as direct fed microbials (DFM) in beef cattle finishing diets to alleviate acute ruminal lactic acidosis in vitro. A dual-flow continuous culture system was used. Treatments were a Control, no DFM; YM1, S. cerevisiae and M. elsdenii strain 1; YM2, S. cerevisiae and M. elsdenii strain 2; and YMM, S. cerevisiae and half of the doses of M. elsdenii strain 1 and strain 2. Each DFM dose had a concentration of 1 × 108 CFU/mL. Four experimental periods lasted 11 days each. For the non-acidotic days (day 1-8), diet contained 50:50 forage to concentrate ratio. For the challenge days (day 9-11), diet contained 10:90 forage to concentrate ratio. Acute ruminal acidosis was successfully established. No differences in pH, D-, L-, or total lactate were observed among treatments. Propionic acid increased in treatments containing DFM. For N metabolism, the YMM treatment decreased protein degradation and microbial protein synthesis. No treatment effects were observed on NH3-N concentration; however, efficiency of N utilization by ruminal bacteria was greater than 80% during the challenge period and NH3-N concentration was reduced to approximately 2 mg/dL as the challenge progressed.


Neural correlates of hemianopic completion across the vertical meridian.
Hemianopic completion refers to the perceptual completion of figures located across the vertical meridian in the context of hemianopia, such that one half of the figure falls within the blind hemifield. It can occur whether the figure is itself complete (veridical completion) or incomplete (paracompletion). Psychophysical evidence suggests that this phenomenon may be a constructive one, and may share features with completion phenomena in normal vision. The neural structures mediating hemianopic completion are unknown. Here we studied the neural activity evoked by hemianopic completion using event-related fMRI in an individual (POV) with a large right visual field homonymous hemianopic scotoma due to left occipital damage. Either a large achromatic circular contour straddling the vertical meridian or a semicircular contour within the left hemifield just crossing the vertical meridian was presented to POV on each trial. POV indicated by button press whether he perceived a semicircular contour, a patchy circular contour or a complete circular contour. On trials where he reported perceiving a complete circular contour despite being presented with a semicircular contour (paracompletion), activity was increased in a region of ipsilateral extrastriate cortex (contralateral to the lesion, ipsilateral to the illusory edge of the circle). These results are discussed in the context of illusory contour completion in healthy subjects and more generally in the recovery of function after brain damage.


Effects of Intradermal Administration Volume Using a Hollow Microneedle on the Pharmacokinetics of Fluorescein Isothiocyanate Dextran (M.W. 4,000).
Hollow microneedles (hMNs) have been gaining attention as a tool to enable the intradermal (i.d.) administration of pharmaceutical products. However, few reports have examined the effect of administration volume on distribution in the skin and pharmacokinetics parameters after i.d. injection. In the present study, a model middle molecular weight compound, fluorescein isothiocyanate dextran (M.W. 4,000, FD-4), was selected, and blood concentration-time profiles after i.d. and subcutaneous (s.c.) injections with different administration volumes were compared. FD-4 solution was injected i.d. using a hMN or injected s.c. with a 27 G needle. Pharmacokinetics and dermatokinetics of FD-4 were analyzed using a compartment model. The skin distribution of iodine, as an X ray tracer, was used to evaluate drug disposition. With the administered drug assumed to be absorbed from the broad injection site into blood vessels in the upper and lower dermis by rapid (krapid) and slow (kslow) first-order absorption rate constants, respectively, better agreement of observed and theoretical values was obtained. Furthermore, the fraction, F, of the administered dose absorbed with krapid decreased with the increase in injection volume after i.d. injection, although the pharmacokinetics parameters were almost the same regardless of administration volume after s.c. injection. The drug distribution in the skin may be related to the obtained pharmacokinetics parameters suggested that the number of needles in the MN system and the total administration volume should be considered in designing hMN systems. The present results provide useful information that may support effective drug delivery with hMNs.


Atraumatic restorative treatment and Carisolv use for root caries in the elderly: 2-year follow-up randomized clinical trial.
New preventive and treatment strategies are required to address the high prevalence of caries among the elderly. The main objective of this study was to analyze the effectiveness of Carisolv® gel to improve the clinical behavior of restorations obtained by atraumatic restorative treatment (ART) in root caries of elderly patients. A secondary objective was to determine the factors associated with the failure of ART restorations after a 2-year follow up. A randomized controlled trial with 2-year follow-up was designed for this purpose. Candidate caries lesions were randomly assigned to an ART group for root caries treatment with the conventional ART technique, filling with glass ionomer, or an ART + Carisolv® gel for the same ART plus the application of a caries solvent (Carisolv®). Evaluations were conducted at 6, 12, and 24 months. A total of 81 restorations were performed, 37 in the ART group and 44 in the ART + Carisolv® gel group, with 22 and 26 restorations, respectively, surviving at the end of the 24-month follow-up. Survival rates at 24 months did not significantly differ between ART (63 %) and ART + Carisolv® gel (62 %) restorations. The best model for predicting the failure of the restorations included the number of tooth-brushings/day, the presence or not of prosthesis, the anterior or posterior location of the tooth, and the baseline plaque level. The application of Carisolv does not modify the survival rate of ART restorations in elderly patients. The use of Carisolv gel does not improve the outcomes of atraumatic restorative treatment.


Effect of naloxone, haloperidol and propranolol on cyclic 3',5'-adenosine monophosphate content of rat amygdala.
Naloxone (0.4 mg/kg, i.p.) causes an increase of cyclic adenosine monophosphate levels in the amygdala, but not in the hippocampus, caudate, or hypothalamus, of rats. The effect is antagonized by haloperidol (0.5 mg/kg, i.p.) and by propranolol (0.5 mg/kg, i.p.). This is consistent with the hypothesis of a tonic inhibitory influence of endogenous opiates on central dopaminergic and beta-noradrenergic systems. Haloperidol had an effect of its own on amygdala cyclic adenosine monophosphate levels which was blocked by propranolol. This suggests the possibility of an antagonistic interaction between dopaminergic and beta-noradrenergic innervation on this structure.


Retained foreign body in a child.
We present the case of a child who ingested or inhaled a foreign body that lodged between the esophagus and the trachea for more than 23 months. The only presenting complaint was of recurrent upper respiratory tract infections. The patient, a 3-year-old girl, required a tracheostomy to alleviate respiratory distress. She made a complete recovery after surgical removal of the foreign body through a neck incision.


Patients' and Health Care Professionals' Perceptions of the Potential of Using the Digital Diabetes Questionnaire to Prepare for Diabetes Care Meetings: Qualitative Focus Group Interview Study.
In effective diabetes management, it is important that providers and health care systems prioritize the delivery of patient-centered care and that they are respectful of and responsive to individual patient preferences and barriers. The objective of the study was to conduct focus group interviews to capture patients' and health care professionals' perceptions and attitudes regarding digital technology and to explore how the digital Diabetes Questionnaire can be used to support patient participation in diabetes care, as a basis for an implementation study. A qualitative study was conducted with six focus group discussions with diabetes specialist nurses and medical doctors (n=29) and four focus group discussions with individuals with diabetes (n=23). A semistructured focus group interview guide was developed, including probing questions. The data were transcribed verbatim, and qualitative content analysis was performed using an inductive approach. Two main categories were revealed by the qualitative analysis: perceptions of digital technology and the digital questionnaire in diabetes management and care and perceptions of participation in diabetes care. An overarching theme that emerged from the focus group interviews was patients' and professionals' involvement in diabetes care using digital tools. The analysis identified important factors to consider when introducing the digital Diabetes Questionnaire in clinical use. Both professionals and patients need support and training in the practical implementation of the digital questionnaire, as well as the opportunity to provide feedback on the questionnaire answers.


Intermittent-hypoxia induced autophagy attenuates contractile dysfunction and myocardial injury in rat heart.
Sleep apnea syndrome (SAS) is considered to be associated with heart failure (HF). It is known that autophagy is induced in various heart diseases thereby promotes survival, but its excess may be maladaptive. Intermittent hypoxia (IH) plays pivotal role in the pathogenesis of SAS. We aimed to clarify the relationships among IH, autophagy, and HF. Rats underwent IH at a rate of 20cycles/h (nadir of 4% O2 to peak of 21% O2 with 0% CO2) or normal air breathing (control) for 8h/d for 3weeks. IH increased the cardiac LC3II/LC3I ratio. The IH induced upregulation of LC3II was attenuated by the administration of an inhibitor of autophagosome formation 3-methyladenine (3-MA), but enhanced by an inhibitor of autophagosome-lysosome fusion chloroquine (CQ), showing enhanced autophagic flux in IH hearts. Electron microscopy confirmed an increase in autophagosomes and lysosomes in IH. With 3-MA or CQ, IH induced progressive deterioration of fractional shortening (FS) on echocardiography over 3weeks, although IH, 3-MA, or CQ alone had no effects. With CQ, IH for 4weeks increased serum troponin T levels, reflecting necrosis. Western blotting analyses showed dephosphorylation of Akt and mammalian target of rapamycin (mTOR) at Akt (Ser2448, 2481) sites, suggesting the activation of autophagy via Akt inactivation. Conclusions. IH-mediated autophagy maintains contractile function, whereas when autophagy is inhibited, IH induces systolic dysfunction due to myocyte necrosis. General significance. This study highlighted the potential implications of autophagy in cardio-protection in early SAS patients without comorbidity, reproduced in normal rats by 3~4weeks of IH.


Identifying and addressing gaps in the implementation of a community care team for care of Patients with multiple chronic conditions.
Patients with multiple chronic conditions represent a growing segment for healthcare. The Chronic Care Model (CCM) supports leveraging community programs to support patients and their caregivers overwhelmed by their treatment plans, but this component has lagged behind the adoption of other model elements. Community Care Teams (CCTs) leverage partnerships between healthcare delivery systems and existing community programs to address this deficiency. There remains a gap in moving CCTs from pilot phase to sustainable full-scale programs. Therefore, the purpose of this study was to identify the cognitive and structural needs of clinicians, social workers, and nurse care coordinators to effectively refer appropriate patients to the CCT and the value these stakeholders derived from referring to and receiving feedback from the CCT. We then sought to translate this knowledge into an implementation toolkit to bridge implementation gaps. Our research process was guided by the Assess, Innovate, Develop, Engage, and Devolve (AIDED) implementation science framework. During the Assess process we conducted chart reviews, interviews, and observations and in Innovate and Develop phases, we worked with stakeholders to develop an implementation toolkit. The Engage and Devolve phases disseminate the toolkit through social networks of clinical champions and are ongoing. We completed 14 chart reviews, 11 interviews, and 2 observations. From these, facilitators and barriers to CCT referrals and patient re-integration into primary care were identified. These insights informed the development of a toolkit with seven components to address implementation gaps identified by the researchers and stakeholders. We identified implementation gaps to sustaining the CCT program, a community-healthcare partnership, and used this information to build an implementation toolkit. We established liaisons with clinical champions to diffuse this information. The AIDED Model, not previously used in high-income countries' primary care settings, proved adaptable and useful.


Overview of the main methods used for estimating catastrophic health expenditure.
Out-of-pocket payments are expenditures borne directly by an individual/household for health services that are not reimbursed by any third-party. Households can experience financial hardship when the burden of such out-of-pocket payments is significant. This financial hardship is commonly measured using the "catastrophic health expenditure" (CHE) metric. CHE has been applied as an indicator in several health sectors and health policies. However, despite its importance, the methods used to measure the incidence of CHE vary across different studies and the terminology used can be inconsistent. In this paper, we introduce and raise awareness of the main approaches used to calculate CHE and discuss critical areas of methodological variation in a global health context. We outline the key features, foundation and differences between the two main methods used for estimating CHE: the budget share and the capacity-to-pay approach. We discuss key sources of variation within CHE calculation and using data from Ethiopia as a case study, illustrate how different approaches can lead to notably different CHE estimates. This variation could lead to challenges when decisionmakers and policymakers need to compare different studies' CHE estimates. This overview is intended to better understand how to interpret and compare CHE estimates and the potential variation across different studies.


Diabetes, hyperglycemia, and central corneal thickness: the Singapore Malay Eye Study.
To examine the relationship of diabetes and hyperglycemia with central corneal thickness (CCT) in Malay adults in Singapore. Population-based cross-sectional study. Three thousand two hundred eighty Malay adults ages 40-80 years living in Singapore. The study population was selected using an age-stratified random sampling procedure of Malay 40- to 80-year-olds living in the southwestern part of Singapore. Participants had a standardized interview, examination, and ocular imaging at a centralized study clinic. Central corneal thickness was measured with an ultrasound pachymeter, and nonfasting serum glucose and glycosylated hemoglobin (Hb A(1C)) was obtained from all participants. Diabetes was defined as having nonfasting glucose levels of > or =200 mg/dl (11.1 mmol/l), a self-report of diabetic medication use, or physician diagnosis of diabetes. Central corneal thickness. Of the 3280 (78.7% response) participants, data on CCT were available on 3239 right eyes. Central corneal thickness was normally distributed, with a mean of 541.2 microm. There were 748 persons with diabetes (23.0%). After controlling for age and gender, central corneas were significantly thicker in persons with diabetes than in those without diabetes (547.2 microm vs. 539.3 microm, P<0.001) and, in the total population, with higher serum glucose (539.6, 540.2, 541.3, and 544.4, comparing increasing glucose quartiles; P = 0.023) and higher Hb A(1C) (537.8, 541.0, 541.4, and 545.5, comparing increasing Hb A(1C) quartiles; P<0.001) levels. In multiple linear regression models adjusting for age, intraocular pressure (IOP), body mass index, and axial length, persons with diabetes had, on average, central corneas 6.50 microm thicker than those of persons without diabetes. This population-based study among Malays showed that diabetes and hyperglycemia are associated with thicker central corneas, independent of age and IOP levels. These findings may have implications for understanding the relationship between diabetes and glaucoma.


Unfolding of iron and copper complexes of human lactoferrin and transferrin.
1. Human lactoferrin and transferrin are capable of binding two iron or copper ions into specific binding sites in the presence of bicarbonate. 2. Urea and several alkyl ureas have been effective in unfolding these metal-protein complexes. 3. Biphasic transitions are observed for the unfolding of each of the metal complexes of these proteins as determined by direct visible spectroscopy suggesting the release of iron(III) and Cu(II) ions from both of these metal-binding proteins during the unfolding process. 4. Greater stabilization and increased resistance to protein unfolding is observed for all iron(III) complexes compared to Cu(II) complexes of lactoferrin and transferrin as determined by isothermal unfolding and thermal denaturation. 5. Relative stabilization of the different metal-protein complexes investigated within this study were determined to be as follows: Lf-Fe(III) greater than Lf-Cu(II); Tf-Fe(III) greater than Tf-Cu(II), and Lf-Fe(III) greater than Tf-Fe(III); Lf-Cu(II) greater than Tf-Cu(II).


Expression of the GluA2 subunit of glutamate receptors is required for the normal dendritic differentiation of cerebellar Purkinje cells.
Cerebellar Purkinje cells differentiate the most elaborate dendritic trees among neurons in the brain and constitute the principal part of cerebellar neuronal circuitry. In the present study, we examined the role of the GluA2 subunit of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors in the dendritic differentiation of Purkinje cells. Since mature Purkinje cells express the GluA2 subunit, AMPA receptors on them exhibit a low Ca2+ permeability. Does this expression of GluA2, leading to the loss of Ca2+ permeability of AMPA receptors, have a positive significance in the dendritic differentiation of Purkinje cells? To answer this question, we introduced GluA2 siRNA into immature Purkinje cells in cerebellar cell cultures using a single-cell electroporation technique. The dendritic elongation and branching, as well as spine formation, were inhibited by GluA2 knockdown in Purkinje cells. GluA2 knockdown augmented the elevation of intracellular Ca2+ concentrations and a higher incidence of oscillation of intracellular Ca2+ concentrations in response to glutamate. These findings suggest that excessive elevation of intracellular Ca2+ concentrations has a negative effect on the dendritic differentiation of Purkinje cells and that the expression of GluA2 inhibits this negative effect in the development of Purkinje cells.


Characterization of a rhabdomyosarcoma reveals a critical role for SMG7 in cancer cell viability and tumor growth.
Soft-tissue sarcomas (STSs) are a rare and diverse group of mesenchymal cancers plagued with aggression, poor response to systemic therapy, and high rates of recurrence. Although STSs generally have low mutational burdens, the most commonly mutated genes are tumor suppressors, which frequently acquire mutations inducing nonsense-mediated mRNA decay (NMD). This suggests that STS cells may exploit NMD to suppress these anti-cancer genes. To examine the role that the NMD factor SMG7 plays in STS, we developed an inducible knockout mouse model in the Trp53-/- background. Here, we isolated a subcutaneous STS and identified it as a rhabdomyosarcoma (RMS). We report that knockout of SMG7 significantly inhibited NMD in our RMS cells, which led to the induction of NMD targets GADD45b and the tumor suppressor GAS5. The loss of NMD and upregulation of these anti-cancer genes were concomitant with the loss of RMS cell viability and inhibited tumor growth. Importantly, SMG7 was dispensable for homeostasis in our mouse embryonic fibroblasts and adult mice. Overall, our data show that the loss of SMG7 induces a strong anti-cancer effect both in vitro and in vivo. We present here the first evidence that disrupting SMG7 function may be tolerable and provide a therapeutic benefit for STS treatment.


Attitudes toward sex in the aged.
Eighty university students and 40 staff members of a nursing home for the aged rated their reactions to three stories: a neutral (decision-making) story, a coital story, and a masturbatory story. For each story, the age and gender of the main character were varied. Results indicated an absence of negative attitudes toward sex in the aged. However, sex in the aged was less credible than sex in young people. Nursing home staff members had more negative attitudes toward sexual stories than did the students. Gender of rater had little effect. Coitus was rated more favorably for the male character than for the female character, and the reverse was true for masturbation.


Applying single-image super-resolution to enhancment of deep-water bathymetry.
We present research using single-image super-resolution (SISR) algorithms to enhance knowledge of the seafloor using the 1-minute GEBCO 2014 grid when 100m grids from high-resolution sonar systems are available for training. We performed numerical experiments of x15 upscaling along three midocean ridge areas in the Eastern Pacific Ocean. We show that four SISR algorithms can enhance this low-resolution knowledge of bathymetry versus bicubic or Splines-In-Tension algorithms through upscaling under these conditions: 1) rough topography is present in both training and testing areas and 2) the range of depths and features in the training area contains the range of depths in the enhancement area. We quantitatively judged successful SISR enhancement versus bicubic interpolation when Student's hypothesis testing show significant improvement of the root-mean squared error (RMSE) between upscaled bathymetry and 100m gridded ground-truth bathymetry at p < 0.05. In addition, we found evidence that random forest based SISR methods may provide more robust enhancements versus non-forest based SISR algorithms.


High accuracy of single-molecule real-time sequencing in detecting a rare α-globin fusion gene in carrier screening population.
The α-globin fusion gene between the HBA2 and HBAP1 genes becomes clinically important in thalassemia screening because this fusion gene can cause severe hemoglobin (Hb) H disease when combining with α0 -thalassemia (α0 -thal). Due to its uncommon rearrangement in the α gene cluster without dosage changes, this fusion gene is undetectable by common molecular testing approaches used for α-thal diagnosis. In this study, we used the single-molecule real-time (SMRT) sequencing technique to detect this fusion gene in 23 carriers identified by next-generation sequencing (NGS) among 16,504 screened individuals. Five primers for α and β thalassemia were utilized. According to the NGS results, the 23 carriers include 14 pure heterozygotes, eight compound heterozygotes with common α-thal alleles, and one homozygote. By using SMRT, the fusion mutant was successfully detected in all 23 carriers. Furthermore, SMRT corrected the diagnosis in two "pure" heterozygotes: one was compound heterozygote with anti-3.7 triplication, and the other was homozygote. Our results indicate that SMRT is a superior method compared to NGS in detecting the α fusion gene, attributing to its efficient, accurate, and one-step properties.


Modular Nucleic Acid Scaffolds for Synthesizing Monodisperse and Sequence-Encoded Antibody Oligomers.
Synthesizing protein oligomers that contain exact numbers of multiple different proteins in defined architectures is challenging. DNA-DNA interactions can be used to program protein assembly into oligomers; however, existing methods require changes to DNA design to achieve different numbers and oligomeric sequences of proteins. Herein, we develop a modular DNA scaffold that uses only six synthetic oligonucleotides to organize proteins into defined oligomers. As a proof-of-concept, model proteins (antibodies) are oligomerized into dimers and trimers, where antibody function is retained. Illustrating the modularity of this technique, dimer and trimer building blocks are then assembled into pentamers containing three different antibodies in an exact stoichiometry and oligomeric sequence. In sum, this report describes a generalizable method for organizing proteins into monodisperse, sequence-encoded oligomers using DNA. This advance will enable studies into how oligomeric protein sequences affect material properties in areas spanning pharmaceutical development, cascade catalysis, synthetic photosynthesis, and membrane transport.


Risk factors for complications of laparoscopic Nissen fundoplication.
Although the rate of complications from laparoscopic Nissen fundoplication is low and the adverse postoperative sequelae are well known, both are disturbing for the patient. Identifying risk factors could be helpful in the better selection of patients for this procedure. A retrospective review of 126 patients with a mean follow-up period of 3.5 years was conducted. The patients' demographics, pre- and postoperative symptoms, and outcomes were analyzed and compared. Three groups of patients were distinguished: (group 1) 9 patients with intraoperative complications (5 with perforation, 3 with hemorrhage, 1 with pneumothorax); (group 2) 16 patients with postoperative complications (5 with severe dysphagia, 4 with failure, 2 with pneumonia, 2 with incisional hernia, 1 with intestine perforation, 1 with fundoplication herniation, 1 with infection, 1 with gastric ulcer); and (group 3) 101 patients without any complications. The patients'demographics, preoperative symptoms, and preoperative studies were comparable in all three groups. The body mass index (BMI) was significantly higher (p < 0.05) statistically in group 1 (32.4 kg/m2) and group 2 (33.6 kg/m2) than in group 3 (28.7 kg/m2). However, the Visick grade and the subjective outcome were similarly good in all three groups. Although preoperative studies and symptoms do not seem to predict complications of laparoscopic Nissen fundoplication, patients with an increased BMI were at increased risk for complications in this study. Therefore, such patients should be counseled appropriately regarding the greater likelihood of intraoperative and postoperative complications.


Assortment, but not knowledge of assortment, affects cooperation and individual success in human groups.
The success or failure of human collective action often depends on the cooperation tendencies of individuals in groups, and on the information that individuals have about each other's cooperativeness. However, it is unclear whether these two factors have an interactive effect on cooperation dynamics. Using a decision-making experiment, we confirm that groups comprising individuals with higher cooperation tendencies cooperate at a higher level than groups comprising individuals with low cooperation tendencies. Moreover, assorting individuals with similar cooperation tendency together affected behaviour so that the most cooperative individuals tended to cooperate more and the least cooperative individuals cooperated less, compared to their behaviour in randomly formed groups. In line with predictions of evolutionary models of cooperation, there was a strong positive association between individuals' cooperation tendency and success when groups were formed assortatively, whereas such association did not exist when groups were formed randomly. Surprisingly, information about group members' cooperativeness in assorted groups had no effect on cooperation levels. We discuss potential explanations for why information about cooperativeness of others may be disregarded in certain circumstances.


Anatomic and clinical correlates of uterine perforation.
We reviewed cases of uterine perforations which occurred at or were referred to the Boston Hospital for Women, Lying-In Division, over the 2 year period from mid-1975 to mid-1977. There were 25 uterine perforations; twenty patients were pregnant and five were not. In the pregnant patients, 16 perforations involved the cervix or the lower uterine segment, whereas only four were located in the uterine fundus; in the nonpregnant patients, all five were fundal perforations. In the pregnant patients, 12 required laparotomy, eight had serious lacerations of the uterine artery, and three had hysterectomy while none of the nonpregnant patients had lacerations or required subsequent procedures. Thus, there is a significant anatomic difference between those perforations which occurred in the pregnant patients and in the nonpregnant patients, the manner in which they presented clinically, and the need for intervention via laparotomy, subsequent morbidity, and outcome as reflected in future reproductive capability. Perforation at or near the cervix may be more common than previously assumed. Furthermore, two distinct clinical entities of cervical perforation exist in pregnant patients as based on the anatomic location of the defect. The anatomy of cervical perforations, their recognition, and their management are discussed.


Dynamic CT myocardial perfusion imaging identifies early perfusion abnormalities in diabetes and hypertension: Insights from a multicenter registry.
To identify patients with early signs of myocardial perfusion reduction, a reference base for perfusion measures is needed. To analyze perfusion parameters derived from dynamic computed tomography perfusion imaging (CTPI) in patients with suspected coronary artery disease (CAD), and relationship with risk factors. In this multicenter study, coronary CT angiography (cCTA) and dynamic CTPI were performed by second-generation dual-source CT in patients suspected of CAD. Risk factors were collected from hospital records. Patients with visual perfusion defects on CTPI, previous coronary intervention, or missing risk factor details were excluded. This analysis included 98 patients (mean age ± standard deviation [SD], 59.0 ± 8.6yrs; 73 male). Global measures of left ventricular myocardial blood flow (MBF), myocardial blood volume (MBV) and volume transfer constant (K(trans)) were calculated. Mean MBF was 139.3 ± 31.4 mL/100 mL/min, MBV 19.1 ± 2.7 mL/100 mL, and Ktrans 85.0 ± 17.5 mL/100 mL/min. No significant differences in perfusion parameters were found by gender or age category. Hypertension and diabetes mellitus resulted in lower perfusion parameters (hypertension vs normotension: MBV 18.5 ± 3.0 vs 19.7 ± 2.3 mL/100 mL and K(trans) 82.0 ± 18.0 vs 89.0 ± 16.0, p < 0.05; diabetes vs no diabetes: MBF 128.5 ± 31.5 vs 144.0 ± 30.5 mL/100 mL/min and MBV 17.9 ± 2.4 vs 19.4 ± 2.8 mL/100 mL, p < 0.05). In patients with hyperlipidemia, MBF was higher (146.8 ± 34.4 vs 130.7 ± 24.3 mL/100 mL/min, p < 0.05). Smoking and family history did not show perfusion parameter differences. Dynamic CTPI identifies early perfusion disturbances in conditions like diabetes and hypertension. With further standardization, absolute perfusion measures may improve CAD risk stratification in patients without visual perfusion defects.


The methylene blue colorimetric microassay for determining cell line response to growth factors.
The validity of the methylene blue colorimetric microassay for determining the response of monolayers of human ovarian tumour cell lines to different growth factors was investigated. Linearity of the relationship between cell density and optical density was confirmed for each cell line (r=0.989-0.999,p<0.001), and when initial cell density was optimised to give exponential growth over the assay period, differences in response to medium supplements were obvious. The response of target cells to growth factors, obtained using the methylene blue assay, were compared with, and found to parallel, previously documented responses obtained non-colorimetrically. Thus Mink lung epithelial cells (MLEC) were inhibited by TGβ (Holleyet al., 1983), EGF had an inhibitory effect on A431 cells (Gill & Lazar, 1981; Barnes, 1982), and the mesothelial cell line showed a proliferative response to EGF and hydrocortisone (Connell and Rheinwald, 1983).The methylene blue colorimetric microssay was found to be a simple, reliable, sensitive method with low variability, for determining the response of cultured cells to growth factors.


A handheld multifunctional smartphone platform integrated with 3D printing portable device: On-site evaluation for glutathione and azodicarbonamide with machine learning.
Azodicarbonamide (ADA) in flour can be easily decomposed to semi-carbazide and biuret, exhibiting strong genotoxicity in vitro and carcinogenicity. Glutathione (GSH) can be conjugated with some ketone-containing compounds and unsaturated aldehydes to form toxic metabolites. Here, a novel ratio fluorescence probe based on blue emitting biomass-derived carbon dots (BCDs) and yellow emitting 2,3-diaminophenazine (OxOPD) was prepared for the bifunctional determination of glutathione (GSH) and ADA. This strategy includes three processes: (1) Ag+ oxidizes o-phenylenediamine (OPD) to produce OxOPD. The peak at 562 nm was enhanced, and the peak at 442 nm was reduced due to fluorescence resonance energy transfer (FRET), (2) glutathione binds Ag+ and inhibits the production of OxOPD, (3) ADA oxidizes GSH to form GSSG, resulting in the release of Ag+ by GSH. Therefore, the newly designed ratio fluorescence probe can be based on the intensity ratio (I442/I562) changes and significant fluorescent color changes to detect GSH and ADA. Moreover, a smartphone WeChat applet and a yolov3-assisted deep learning classification model have been developed to quickly detect GSH and ADA on-site based on an image processing algorithm. These results indicate that smartphone ratiometric fluorescence sensing combined with machine learning has broad prospects for biomedical analysis.


Inhibitory effects of desflurane and sevoflurane on oxytocin-induced contractions of isolated pregnant human myometrium.
In this study, we investigated the inhibitory effects of desflurane and sevoflurane on oxytocin-induced contractions of isolated human myometrium. Following delivery of the infant and placenta, a small segment of myometrium was excised from the upper incisional surface of the lower uterine segment and 20 strips, randomly assigned into two groups (n = 10), were obtained from 20 non-laboring term parturients. The study protocol consisted of a 60-min period of spontaneous contractions, control recording with oxytocin 2 x 10(9) m (10-min period), washout interval of 10 min, volatile administration (three times per 15-min period) of 0.5, 1 and 2 minimum alveolar concentration (MAC), response to oxytocin (10-min period), a further washout interval (10-min period) and subsequent control recording with oxytocin without anesthetics. After oxytocin administration, the frequency and amplitude of contractions increased (P < 0.05) and the duration decreased (P < 0.05). The frequency and amplitude of contractions induced with oxytocin decreased significantly at 0.5, 1 and 2 MAC of desflurane and sevoflurane (P < 0.05). The amplitude of contractions was significantly different at 1 MAC between the two groups (P < 0.05). The duration of contractions at 2 MAC decreased in both groups (P < 0.05). Desflurane and sevoflurane at 0.5, 1 and 2 MAC inhibit the frequency and amplitude of myometrial contractions induced with oxytocin in a dose-dependent manner. However, desflurane inhibits the amplitude less than sevoflurane at 1 MAC. We suggest that 0.5 MAC of both agents and 1 MAC of desflurane may be safely used in the presence of oxytocin following delivery of the infant and placenta during Cesarean section without fear of uterine atony and hemorrhage.


Selective reactions in the analytical characterisation of steroids by gas chromatography-mass spectrometry.
The analytical characterization, by GC-MS, of individual compounds in mixtures of steroids, such as occur frequently in biological extracts, is difficult because of the close similarities in structure and properties of many components. The improved separating power of capillary (open-tubular) columns alleviates the problem, but does not solve it fully: for example, the coincidence of retention times of two different compounds may still be virtually complete. Comparative analyses on two distinctively different phases afford one valuable application of selectivity, but may not always be feasible when costly columns are required. Comparative analyses of the sample, before and after effecting its modification by well-defined reactions, are inexpensive and are particularly when selective transformations are used. The use of the microbial enzyme cholesterol oxidase as a selective oxidant for 3 beta-hydroxysteroids (chiefly limited to 4-ene, 5-ene and 5 alpha-types) is illustrated for a model mixture of androstanols related to the boar pheromone (5 alpha, 16-androsten-3 alpha-ol). Retention regularities and changes in mass spectra enhance the reliability of identifications. An exploratory application of cholesterol oxidase in the analysis of minor "polar" sterols in human serum is reported. Most of the known minor sterols are good substrates for the enzyme, and their transformation products yield distinctive GC-MS data, as exemplified for the 7 alpha- and 7 beta-hydroxycholesterols. Another convenient and versatile selective reagent is methaneboronic acid, which yields cyclic esters of suitably constituted diols. These derivatives have shorter retention times (on "non-polar" phases) than the di-TMS ethers, chiefly by virtue of their much lower molecular weights. The mass spectra of cyclic boronates generally show clear molecular ions, also fragmentations that complement the information obtainable from the di-TMS ethers. These features are illustrated for a group of diols and triols of the 5 alpha-pregnane series.


Associations Between Musculoskeletal Pain Experience and Pressure and Cold Pain Sensitivity: A Community-based Cross-sectional Study of Young Adults in the Raine Study.
To investigate the cross-sectional associations between musculoskeletal pain experience and measures of pressure and cold pain sensitivity in young adults from the Western Australian Pregnancy Cohort (Raine) Study. In total, 917 participants were eligible for analysis if they provided data pertaining to musculoskeletal pain status at the 22-year follow-up and had data for at least 1 valid pain sensitivity test. Standardized protocols were used to assess pressure pain threshold (4 sites: lumbar spine, tibialis anterior, upper trapezius, and wrist) and cold pain threshold (wrist). Four pain experience groups ("No pain" [n=562, 61.3%], "Low" [n=84, 9.2%], "Medium" [n=147, 16.0%], "High" [n=124, 13.5%]) were determined by latent class analysis using parameters of pain chronicity, frequency, intensity, and number of pain areas. Variables considered as confounders included sex, age, ethnicity, waist-hip ratio, psychological symptoms, sleep quality, physical activity, sedentary behavior, smoking, and income. There were no associations between pain experience and pressure pain sensitivity after adjusting for confounders. The "Medium" and "High" pain experience groups demonstrated heightened cold pain sensitivity compared with the "No pain" group (P=0.023), adjusted for sex and smoking. This study provides the most extensive investigation of the relationship between musculoskeletal pain experience and pressure and cold pain sensitivity in young adults. Heightened cold pain sensitivity in those classified as "Medium" and "High" pain experience may suggest altered nociceptive processing and has implications for clinical management.


Influence of water content and drying on the physical structure of native hyaluronan.
Hydration properties of semi-diluted hyaluronan were studied by means of time domain nuclear magnetic resonance. Based on the transverse proton relaxation times T2, the plasticization of hyaluronan which was precipitated by isopropylalcohol and dried in the oven have been determined at water content 0.4 g of water per g of hyaluronan. Above this water content, the relaxation times increased and levelled off around 0.8 g of water per g of hyaluronan which agrees well with values determined earlier by differential scanning calorimetry and dielectric relaxometry. The freeze dried and oven dried samples showed differences in their physical structure such as glass transition, plasticization concentration and sample topography which influenced their kinetics and mechanisms of hydration. Results confirmed earlier hypothesis that some native biopolymer structures can be easily modified by manipulation of preparation conditions, e.g. drying, giving fractions with specific physicochemical properties without necessity of their chemical modification.


Factors that adolescent males take into account in decisions about an unplanned pregnancy.
Little is known about what factors adolescent males consider important when making decisions concerning the resolution of an unplanned pregnancy with a teenage partner. Young men's influence on pregnancy outcome decisions can play an important part in the subsequent psychological adjustment of the female. The present report draws on data from a larger study with teenage males [Condon, J. T., Corkindale, C. J., Russell, A., & Quinlivan, J. A. (2006). Processes and factors underlying adolescent males' attitudes and decision-making in relation to an unplanned pregnancy. Journal of Youth and Adolescence, 35, 447-458], and extends the findings of that study. Using a 25-item scale embedded in an electronic role-play, data were obtained from 330 male Australian school students on their priorities and concerns in relation to possible outcomes of a partner's pregnancy. Common factors taken into account by almost the entire adolescent sample related to the negative effects of becoming a teenage father. The differences between participants were identified using cluster analysis, which produced three groupings. The majority group was characterised as 'well-balanced' (80.6%), and the two minority groups as 'unwilling/unready' (10.9%) and 'family-centred' (8.5%). Group membership was strongly predictive of the males' final decision regarding the hypothetical pregnancy outcome. Understanding adolescent attitudes and beliefs when faced with this decision may assist practitioners in their guidance of the young couple and help prevent negative psychological sequelae.


Fluorescent g-C3N4 nanosheets enhanced photosynthetic efficiency in maize.
Nano-enabled agriculture becomes a new and rapidly evolving area of research, particularly, nanomaterials (NMs) with light-harvesting capacities for enhancing photosynthesis. However, mechanisms for the interactions between these NMs and plants are not fully understood. Herein, fluorescent and water-soluble graphitic carbon nitride (g-C3N4) nanosheets were prepared and used as artificial antenna to amplify light harvesting ability and enhance photosynthesis in maize. Upon root exposure to 10 mg·L-1 g-C3N4 nanosheets, the g-C3N4 can be taken up and distributed in leaves. Also, the nutrients (Mg, P, Fe, and Mn), chlorophyll content, electron transfer rate, net photosynthetic rate, and carbohydrates content in maize were increased significantly by 1.1%, 51.8%, 44.6%, 121.8%, 12.1%, 44.5%, 30.0% and 32.3%, respectively. In addition, the gene expressions of psbA (photosystem II reaction center protein A) and psaA (photosystem I P700 chlorophyll A apoprotein A1) were up-regulated by 56.3% and 26.8%, respectively. Moreover, the activities of phosphoenolpyruvate carboxylase (PEPC) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) were significantly increased by 242.3% and 156.3%, respectively. This study provides a new perspective on the use of g-C3N4 nanosheets to promote plant growth and develop nano-enabled agricultural technology.


Enzyme-inducing and cytotoxic effects of wood-based materials used as bedding for laboratory animals. Comparison by a cell culture study.
Enzyme-inducing and cytotoxic effects of wood-based materials used as bedding for laboratory animals were studied in a cell culture system. Mouse hepatoma cell line, Hepa-1, was exposed to acetone extracts of hardwoods (alder and aspen), softwoods (pine and a mixture of pine and spruce) and cellulose materials. Cytotoxicity and induction of cytochrome P450IA1 (aryl hydrocarbon hydroxylase) and aldehyde dehydrogenase were measured. Both softwood and hardwood extracts were shown to contain inducers of these enzymes. Pine appeared to be the most potent inducer and softwoods more potent than hardwoods. The softwoods and alder were clearly more cytotoxic than aspen. The two bleached cellulose materials were found to contain inducers of aryl hydrocarbon hydroxylase. Unlike the wood beddings, the extracts of the cellulose materials were not found to be toxic to the cells. Hepa-1 cell culture system was found to be a rapid and sensitive method for screening and comparative purposes.


Return-to-work intervention versus usual care for sick-listed employees: health-economic investment appraisal alongside a cluster randomised trial.
To evaluate the health-economic costs and benefits of a guided eHealth intervention (E-health module embedded in Collaborative Occupational healthcare (ECO)) encouraging sick-listed employees to a faster return to work. A two-armed cluster randomised trial with occupational physicians (OPs) (n=62), clustered and randomised by region into an experimental and a control group, to conduct a health-economic investment appraisal. Online self-reported data were collected from employees at baseline, after 3, 6, 9 and 12 months. Occupational health care in the Netherlands. Employees from small-sized and medium-sized companies (≥18 years), sick-listed between 4 and 26 weeks with (symptoms of) common mental disorders visiting their OP. In the intervention group, employees (N=131) received an eHealth module aimed at changing cognitions regarding return to work, while OPs were supported by a decision aid for treatment and referral options. Employees in the control condition (N=89) received usual sickness guidance. Net benefits and return on investment based on absenteeism, presenteeism, health care use and quality-adjusted life years (QALYs) gained. From the employer's perspective, the incremental net benefits were €3187 per employee over a single year, representing a return of investment of €11 per invested Euro, with a break-even point at 6 months. The economic case was also favourable from the employee's perspective, partly because of QALY health gains. The intervention was costing €234 per employee from a health service financier's perspective. The incremental net benefits from a social perspective were €4210. This amount dropped to €3559 in the sensitivity analysis trimming the 5% highest costs. The data suggest that the ECO intervention offers good value for money for virtually all stakeholders involved, because initial investments were more than recouped within a single year. The sometimes wide 95% CIs suggest that the costs and benefits are not always very precise estimates and real benefits could vary considerably. NTR2108; Results.


Factors predictive of papillary thyroid micro-carcinoma with bilateral involvement and central lymph node metastasis: a retrospective study.
The optimal resection extent for papillary thyroid microcarcinoma (PTMC) remains controversial. The objective of the study was to investigate risk factors of bilateral PTMC and central lymph node metastasis (CLNM) to guide surgical strategies for PTMC patients. We retrospectively reviewed 211 PTMC patients who underwent total thyroidectomy (TT) and 122 clinical lymph node-negative (cN0) cases that underwent prophylactic central lymph node dissection (CLND) between 2010 and 2011. The frequency, pattern, and predictive factors for bilateral PTMC and CLNM in these patients were studied using univariate and multivariate analysis with respect to the following variables: age, gender, extrathyroidal extension (ETE), T stage, with Hashimoto thyroiditis (HT), tumor size and multifocality based on final pathology, and preoperative evaluation using ultrasonography (US). Fifty-four of 211 (25.6%) patients had bilateral PTMC. In multivariate analysis, multifocality (P < 0.001, OR = 23.900) and tumor size ≥7 mm (P = 0.014, OR = 2.398) based on US were independent predictive factors for bilateral PTMC which was also independently associated with multifocality (P < 0.001, OR = 29.657) and tumor size ≥7 mm (P = 0.005, OR = 2.863) based on final pathology. Among 122 cN0 patients who underwent prophylactic CLND, we found 49.2% of patients had CLNM. CLNM was independently associated with men, age <50 years and tumor size ≥7 mm based on final pathology or preoperative US. TT should be considered for PTMC patients who are found multifocality and tumor size ≥7 mm based on preoperative US. CLND need be considered in cN0 patients who are men, aged <50 years or tumor size ≥7 mm based on preoperative US.


Protein Kinase C at the Crossroad of Mutations, Cancer, Targeted Therapy and Immune Response.
The frequent PKC dysregulations observed in many tumors have made these enzymes natural targets for anticancer applications. Nevertheless, this considerable interest in the development of PKC modulators has not led to the expected therapeutic benefits, likely due to the complex biological activities regulated by PKC isoenzymes, often playing ambiguous and protective functions, further driven by the occurrence of mutations. The structure, regulation and functions of PKCs have been extensively covered in other publications. Herein, we focused on PKC alterations mostly associated with complete functional loss. We also addressed the modest yet encouraging results obtained targeting PKC in selected malignancies and the more frequent negative clinical outcomes. The reported observations advocate the need for more selective molecules and a better understanding of the involved pathways. Furthermore, we underlined the most relevant immune mechanisms controlled by PKC isoforms potentially impacting the immune checkpoint inhibitor blockade-mediated immune recovery. We believe that a comprehensive examination of the molecular features of the tumor microenvironment might improve clinical outcomes by tailoring PKC modulation. This approach can be further supported by the identification of potential response biomarkers, which may indicate patients who may benefit from the manipulation of distinctive PKC isoforms.


Management Strategies for Patients Presenting With Symptomatic Lymphadenopathy and Breast Edema After Recent COVID-19 Vaccination.
Ipsilateral axillary lymphadenopathy is a well-documented finding associated with COVID-19 vaccination. Varying guidelines have been published for the management of asymptomatic patients who have a history of recent vaccination and present with incidental lymphadenopathy at screening mammography. Some experts recommend follow-up imaging, and others suggest that clinical management, rather than repeat imaging or biopsy, is appropriate. Symptomatic patients with lymphadenopathy and/or additional abnormal imaging findings should be treated differently depending on risk factors and clinical scenarios. Although ipsilateral lymphadenopathy is well documented, ipsilateral breast edema after COVID-19 vaccination has been rarely reported. The combination of ipsilateral lymphadenopathy and diffuse breast edema after COVID-19 vaccination presents a clinical management challenge because edema can obscure underlying abnormalities at imaging. For symptomatic patients with lymphadenopathy and associated breast parenchymal abnormality, prompt action is appropriate, including diagnostic evaluation and consideration of tissue sampling. This approach may prevent delays in diagnosis and treatment of patients with malignancy masked by symptoms from the vaccination.


Low level of glucocorticoid receptor messenger ribonucleic acid in pituitary adenomas manifesting Cushing's disease with resistance to a high dose-dexamethasone suppression test.
The overnight 8-mg dexamethasone suppression test is often used to differentiate Cushing's disease, due to an oversecretion of ACTH from the pituitary gland, from other kinds of Cushing's syndrome. However, a few patients with ACTH-producing pituitary adenoma show no suppression of plasma cortisol after the administration of 8 mg of dexamethasone. To clarify the relationship between the level of glucocorticoid receptor (GR) in the pituitary adenoma and the sensitivity to dexamethasone in Cushing's disease, we thus examined the levels of GR alpha and GR beta mRNAs in the pituitary adenomas in six patients who were proven at surgery to have pituitary ACTH-producing adenomas. Total RNA was extracted from six pituitary adenomas and pituitary tissue adjacent to one of the adenomas, and the mRNA levels of GR alpha, GR beta, pro-opiomelanocortin (POMC) and beta-actin in these samples were sampled by quantitative RT-PCR. The GR alpha mRNA levels in the adenomas from the two patients who showed no response to the 8-mg dexamethasone suppression test were significantly lower than those in the adenomas of four patients who showed suppression. The GR beta mRNA level was much lower than that of GR alpha mRNA but not significantly different among the six adenomas. These results suggest strongly that decreased expression of GR alpha in pituitary adenomas may be the major reason for the marked insensitivity to the 8-mg dexamethasone suppression test observed in two patients with Cushing's disease.


Experience of commencing Carvedilol in elderly patients with heart failure in a routine outpatient clinic.
Carvedilol has been shown to be beneficial for patients with heart failure, but it is not clear how it should be initiated in routine clinical practice, particularly in the elderly. This study is a retrospective review of 19 patients of age 80+/-4 years with heart failure, who had Carvedilol treatment initiated in a routine outpatient clinic. No patient experienced a worsening of their heart failure or required admission to hospital. There was a mean reduction in heart rate of 11 beats/min. Carvedilol was tolerated by 68% of the patients. Thus, Carvedilol can be safely initiated in a routine outpatient clinic in appropriately selected elderly patients.


Tissue estimation of protein gene product 9.5 (PGP 9.5) expression and apoptosis in vitiligo.
Vitiligo is probably the end result of different interacting processes. To determine the possible roles of neural and apoptotic mechanisms in the pathogenesis of vitiligo. Fifty-six biopsies from 28 patients with generalized vitiligo (28 from depigmented lesional areas and 28 from clinically nondepigmented skin at the periphery of the same areas) were examined; the panaxonal marker neuropeptide protein gene product 9.5 (PGP 9.5) and apoptosis were investigated using immunohistochemistry. Statistically significant differences were detected in the numbers of PGP 9.5-positive nerve fibers/axons in the papillary dermis between the center and periphery of the lesions (i.e. increased at the center in comparison with the periphery). A statistically significant inverse association was found between PGP 9.5 immunostaining in the dermis at the lesion center and the duration of the disease. When apoptosis and PGP 9.5 expression were compared, there was an identical distribution of PGP 9.5-positive nerve fibers/axons and apoptotic cells in the epidermis (i.e. basal in the lesion center; diffuse at the lesion periphery). There is a possible connection between the neural and apoptotic pathogenetic theories of vitiligo.


Stereoscopic acuity measurement in aphakia.
We performed a variety of tests to assess binocular function on monocular aphakic patients wearing contact lenses, monocular pseudophakic patients, bilateral spectacle-wearing aphakic patients, bilateral aphakic contact lens wearers, and bilateral pseudophakic patients. Stereoscopic acuity levels at distance and near when visual acuity was at least 6/12 (20/40) provided the most useful data. Pseudophakic patients and contact lens wearers performed equally well and outperformed spectacle wearers. Implant patients did not have superior stereoscopic acuity.


Incidence of Neonatal Hypothermia in the Newborn Nursery and Associated Factors.
Thermoregulation is a key component of well-newborn care. There is limited epidemiologic data on hypothermia in late preterm and term infants admitted to the nursery. Expanding on these data is essential for advancing evidence-based care in a population that represents more than 3.5 million births per year in the US. To examine the incidence and factors associated with hypothermia in otherwise healthy infants admitted to the newborn nursery following delivery. A retrospective cohort study using electronic health record data from May 1, 2015, to August 31, 2021, was conducted at a newborn nursery at a university-affiliated children's hospital. Participants included 23 549 infants admitted to the newborn nursery, from which 321 060 axillary and rectal temperature values were analyzed. Infant and maternal clinical and demographic factors. Neonatal hypothermia was defined according to the World Health Organization threshold of temperature less than 36.5 °C. Hypothermia was further classified by severity (mild: single episode, temperature 36.0-36.4 °C; moderate/severe: persistent or recurrent hypothermia and/or temperature <36.0 °C) and timing (early: all hypothermic episodes occurred within the first 24 hours after birth; late: any episode extended beyond the first 24 hours). Of 23 549 included infants (male, 12 220 [51.9%]), 5.6% were late preterm (35-36 weeks' gestation) and 4.3% were low birth weight (≤2500 g). The incidence of mild hypothermia was 17.1% and the incidence of moderate/severe hypothermia was 4.6%. Late hypothermia occurred in 1.8% of infants. Lower birth weight and gestational age and Black and Asian maternal race and ethnicity had the highest adjusted odds across all classifications of hypothermia. The adjusted odds ratios of moderate/severe hypothermia were 5.97 (95% CI 4.45-8.00) in infants with a birth weight less than or equal to 2500 vs 3001 to 3500 g, 3.17 (95% CI 2.24-4.49) in 35 week' vs 39 weeks' gestation, and 2.65 (95% CI 1.78-3.96) in infants born to Black mothers and 1.94 (95% CI 1.61-2.34) in infants born to Asian mothers vs non-Hispanic White mothers. In this cohort study of infants in the inpatient nursery, hypothermia was common, and the incidence varied by hypothermia definition applied. Infants of lower gestational age and birth weight and those born to Black and Asian mothers carried the highest odds of hypothermia. These findings suggest that identifying biological, structural, and social determinants of hypothermia is essential for advancing evidence-based equitable thermoregulatory care.


Aurora kinase B/C inhibition impairs malignant glioma growth in vivo.
Inhibition of Aurora kinase B has been evaluated as a therapy to block solid tumor growth in breast cancer, hepatocellular carcinoma, lung adenocarcinoma, and colorectal cancer models. Aurora kinase inhibitors are in early clinical trials for the treatment of leukemia. We hypothesized that Aurora B inhibition would reduce malignant glioma cell viability and result in impaired tumor growth in vivo. Aurora B expression is greater in cultured malignant glioma U251 cells compared to proliferating normal human astrocytes, and expression is maintained in U251 flank xenografts. Aurora B inhibition with AZD1152-HQPA blocked cell division in four different p53-mutant glioma cell lines (U251, T98G, U373, and U118). AZD1152-HQPA also inhibited Aurora C activation loop threonine autophosphorylation at the effective antiproliferative concentrations in vitro. Reduction in cell viability of U251 (p53(R273H)) cells was secondary to cytokinesis blockade and apoptosis induction following endoreplication. AZD1152-HQPA inhibited the growth of U251 tumor xenografts and resulted in an increase in tumor cell apoptosis both in vitro and in vivo. Subcutaneous administration of AZD1152-HQPA (25 mg/kg/day × 4 days; 2 cycles spaced 7 days apart) resulted in a prolongation in median survival after intracranial inoculation of U251 cells in mice (P = 0.025). This is the first demonstration that an Aurora kinase inhibitor can inhibit malignant glioma growth in vivo at drug doses that are clinically relevant.


EEG analysis with simulated neuronal cell models helps to detect pre-seizure changes.
To test if a method for real-time detection of epileptic seizures based on electroencephalographic (EEG) analysis with simulated neuronal cell models can be modified to identify pre-seizure changes. Our EEG analysis method consists of two simulated leaky integrate and fire units (LIFU) connected to a signal preprocessing stage that marks parts of the EEG signals with slopes larger than a preset threshold Hth with unit pulses. The LIFUs change their spiking frequency depending on the rate and the synchrony of the impinging pulse trains. Here, we use our method in a high-sensitivity mode by setting Hth to low values, which causes the LIFUs to continuously spike during the interictal state. We test if the LIFUs spiking rates change before seizure onset. We used 9 long-term EEGs (16+/-7 h) of 7 patients with drug resistant epilepsy. Fifteen seizures were analyzed and all were preceded by an increase of the time-averaged spiking rates SR(av) of the LIFUs. We defined a function F(Sz), which quantifies the changes of SR(av). F(Sz) increased and stayed above an individually set and fixed threshold 83+/-91 min (range: 4-330 min) before EEG seizure onset. Only two false alarms occurred. We conclude that EEG analysis with simulated neuronal cell models may be used to detect pre-seizure changes with high sensitivity and specificity.


Adaptive flood risk management planning based on a comprehensive flood risk conceptualisation.
Densely populated deltas are so vulnerable to sea level rise and climate change that they cannot wait for global mitigation to become effective. The Netherlands therefore puts huge efforts in adaptation research and planning for the future, for example through the national research programme Knowledge for Climate and the Delta Programme for the Twenty-first century. Flood risk is one of the key issues addressed in both programmes. Adaptive management planning should rely on a sound ex-ante policy analysis which encompasses a future outlook, establishing whether a policy transition is required, an assessment of alternative flood risk management strategies, and their planning in anticipation without running the risk of regret of doing too little too late or too much too early. This endeavour, addressed as adaptive delta management, calls for new approaches, especially because of uncertainties about long-term future developments. For flood risk management, it also entails reconsideration of the underlying principles and of the application of portfolios of technical measures versus spatial planning and other policy instruments. To this end, we first developed a conceptualisation of flood risk which reconciles the different approaches of flood defence management practice and spatial planning practice in order to bridge the gap between these previously detached fields. Secondly, we looked abroad in order to be better able to reflect critically on a possible Dutch bias which could have resulted from many centuries of experience of successful adaptation to increasing flood risk, but which may no longer be sustainable into the future. In this paper, we explain the multiple conceptualisation of flood risk and argue that explicitly distinguishing exposure determinants as a new concept may help to bridge the gap between engineers and spatial planners, wherefore we show how their different conceptualisations influence the framing of the adaptation challenge. Also, we identify what the Netherlands may learn from neighbouring countries with a different framing of the future flood risk challenge.


Isolation of histone proteins from rat normal and tumour blood plasma.
Histone proteins were isolated from both normal and tumour rat blood plasma: H1, (H2b + H2a) for normal plasma and H1, H3, (H2b + H2a) and H4 for tumour one.


Low-Dose Naltrexone (LDN) for Chronic Pain at a Single Institution: A Case Series.
Low-dose naltrexone (LDN) has increased in popularity as a non-opioid medication that may decrease chronic pain symptoms. LDN is most commonly used to treat fibromyalgia, complex regional pain syndrome (CRPS), and painful diabetic neuropathy. Other studies suggest that LDN provides general symptom reduction in inflammatory conditions such as Crohn's disease and multiple sclerosis. We reviewed our experience with patients to whom we have prescribed LDN to see what types of painful conditions were most responsive to LDN in our patient population. Charts from patients who came to the Pain Center between 2014 and 2021 were reviewed. Of the n = 137 patients who were prescribed LDN, 44% had no evidence of ever filling the prescription, and 4.4% of the responses were not charted. Of the remaining who took LDN (n = 70), 64% had some relief and were designated as 'Responders'. The most common pain diagnosis was neuropathic pain which, when added to the diagnosis of complex regional pain syndrome, accounted for 51% of responders to LDN. Patients who experienced greater than 50% pain relief from LDN were more likely to have the diagnosis of neuropathic pain or complex regional pain syndrome (p = 0.038, Fisher's Exact Test). There was a significant difference in the diagnosis of patients who responded to LDN. Patients with spondylosis were much less likely to respond to LDN when compared with other diagnoses (p = 0.00435, Chi-Square Test). Patients with all types of neuropathic pain, including CRPS, were significantly more likely to have pain relief from LDN than patients with spondylosis (p=0.018). The diagnosis of spondylosis was more often associated with a lack of response to LDN than any other diagnosis. Patients may need to have a trial of several weeks before analgesic effects are seen with LDN.


Distribution of growth factors in subfoveal neovascular membranes in age-related macular degeneration and presumed ocular histoplasmosis syndrome.
We performed a histopathologic and immunohistologic study to determine the macromolecular and cellular components of subfoveal neovascular membranes removed at the time of submacular surgery. Subfoveal neovascular membranes were surgically removed from ten patients (seven with age-related macular degeneration and three with presumed ocular histoplasmosis syndrome). Tissues obtained were examined by light and electron microscopy to identify structural components. Immunohistochemical staining was then performed with monoclonal antibodies to various growth factors, including transforming growth factor-beta 1, basic fibroblast growth factor, platelet-derived growth factor, and epidermal growth factor, as well as antibodies against procollagen 1 and phosphotyrosine residues. Most cells in subfoveal neovascular membranes are retinal pigment epithelial cells and cells resembling fibroblasts, with some vascular endothelial cells, lymphocytes, and macrophages. Basic fibroblasts growth factor was found in the extracellular matrix and in endothelial cells. Transforming growth factor-beta 1 was found in endothelial cells, fibroblasts, and retinal pigment epithelial cells. Procollagen 1 was found in protein-synthesizing fibroblasts, and phosphotyrosine residues were detected within fibroblasts, endothelial cells, and retinal pigment epithelial cells. Subfoveal neovascular membranes are neovascular complexes composed of retinal pigment epithelial cells, fibroblasts, vascular endothelial cells, and chronic inflammatory cells. Furthermore, transforming growth factor-beta 1 and basic fibroblast growth factor are present within the major cell types, which suggests a possible pathogenic role in the development of the neovascular complex.


The role of magnetic resonance imaging in the preoperative assessment of anorectal anomalies.
The role of magnetic resonance imaging (MRI) compared to other investigations in providing information relevant to and affecting surgical management in patients with anorectal anomalies (ARA) was studied in two groups of patients. In group 1 a pilot study was done in 9 patients with colostomy for high ARA awaiting posterior sagittal anorectoplasty. These patients had MRI, plain radiographs, a distal loopogram through the mucous fistula, ultrasound scans of the spinal cord and urinary tract, and a micturating cystourethrogram. The level of bowel and spinal-cord termination was correctly determined in all patients by MRI and conventional investigations. The presence or absence of a fistula, urological abnormalities, and bony abnormalities of the pelvis and spine were better visualized by conventional investigations than with MRI. The striated-muscle complex (SMC) was well-visualized by MRI, the thickness accurately graded, and the findings confirmed at operation. The information on muscle thickness and the presence or absence of a fistula was interesting, but played no part in decisions made about surgical management, and the operative technique was not altered. In group 2 the patients were much older and had MRI for persistent problems, mainly soiling; 6 had low lesions and 6 had high lesions. Three patients with low lesions were found to have intraspinal abnormalities on MRI; 4 were found to have malpositioned bowel in the SMC, which was again accurately visualized and graded. Therefore, in 7/12 patients of this group a surgically correctable abnormality was found. We conclude that MRI has no role as a primary investigation in patients with high ARA, but it is useful if there are suspected spinal-cord problems or persistent problems after definitive surgery.


Pharmacokinetics and pharmacodynamics of meropenem in critically ill patients.
The pharmacokinetics and pharmacodynamics of meropenem were investigated in 14 critically ill patients with sepsis. Patients with creatinine clearance (CrCl) higher than 50 ml/min received 1 g meropenem three times daily (Group I) and patients with CrCl lower than 50 ml/min received 1 g meropenem twice daily (Group II). Meropenum concentrations in plasma were determined by high performance liquid chromatography with UV detection. The pharmacokinetic parameters differed between the two groups as follows, Group I, maximal concentration 56.3 +/- 19.1 microg/ml; trough concentration 3.3 +/- 2.5 microg/ml; elimination half life 2.5 +/- 1.2 h; clearance (Cl) 155.8 +/- 40.6 ml/min; MRT 2.2 +/- 0.4 h; steady state volume of distribution (V(ss)) 21.7 +/- 5.7 l, and AUC(-8) 119.4 +/- 32.6 microg/ml h. Group II, maximal concentration 71.1 +/- 5.1 microg/ml; trough concentration 3.4 +/- 1.8 microg/ml; elimination half life 3.9 +/- 1.6 h; Cl 77.7 +/- 15.8 ml/min; MRT 3.5 +/- 0.6 h; V(ss), 17.1 +/- 2.1 l, and AUC(0-12) 230.2 +/- 43.3 microg/ml h. The most frequently isolated bacteria from blood and wound infections were Acinetobacter baumanii, Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli; their meropenem minimal inhibitory concentrations (MICs) ranged from 0.064 to 3.0 mg/l. In most cases the pharmacodynamic parameters, measured as T>MIC index, were higher than 75%. In both groups, patients with susceptible pathogens (MIC<1 mg/l) had meropenem plasma levels which exceeded the MIC for the whole dosing interval. When pathogens were highly resistant (A. baumanii or P. aeruginosa) the T>MIC indices were lower.


Evaluation of a new nitrosoureido water-soluble sugar, ecomustine or CY 233 (NSC 609224), on human xenografts.
Because of its wide spectrum of activity against murine models regardless of the route of administration, CY 233 (NSC 609624) was investigated in human tumours xenografted in nude mice, namely one amelanotic melanoma, MeXF 274, and three colon adenocarcinomas, i.e. CXF 233, CXF 243 and CXF 609. CY 233 was highly active against the MeXF 274 melanoma and produced a complete response on day 14 at 20 mg/kg given i.v. on days 0 and 7. After three intermittent i.v. injections of 10 mg/kg, it produced a partial response of long duration against the colon adenocarcinoma CXF 243. The interesting fact that resistant lines responded to CY 233 warrants further investigation of other schedules of treatment and of the sensitivity profile of CY 233 in a wider panel of tumour xenografts.


Pomegranate polyphenols down-regulate expression of androgen-synthesizing genes in human prostate cancer cells overexpressing the androgen receptor.
Prostate cancer is dependent on circulating testosterone in its early stages and is treatable with radiation and surgery. However, recurrent prostate tumors advance to an androgen-independent state in which they progress in the absence of circulating testosterone, leading to metastasis and death. During the development of androgen independence, prostate cancer cells are known to increase intracellular testosterone synthesis, which maintains cancer cell growth in the absence of significant amounts of circulating testosterone. Overexpression of the androgen receptor (AR) occurs in androgen-independent prostate cancer and has been proposed as another mechanism promoting the development of androgen independence. The LNCaP-AR cell line is engineered to overexpress AR but is otherwise similar to the widely studied LNCaP cell line. We have previously shown that pomegranate extracts inhibit both androgen-dependent and androgen-independent prostate cancer cell growth. In this study, we examined the effects of pomegranate polyphenols, ellagitannin-rich extract and whole juice extract on the expression of genes for key androgen-synthesizing enzymes and the AR. We measured expression of the HSD3B2 (3beta-hydroxysteroid dehydrogenase type 2), AKR1C3 (aldo-keto reductase family 1 member C3) and SRD5A1 (steroid 5alpha reductase type 1) genes for the respective androgen-synthesizing enzymes in LNCaP, LNCaP-AR and DU-145 human prostate cancer cells. A twofold suppression of gene expression was considered statistically significant. Pomegranate polyphenols inhibited gene expression and AR most consistently in the LNCaP-AR cell line (P=.05). Therefore, inhibition by pomegranate polyphenols of gene expression involved in androgen-synthesizing enzymes and the AR may be of particular importance in androgen-independent prostate cancer cells and the subset of human prostate cancers where AR is up-regulated.


Encrusted Ureteral Stent Retrieval Using Flexible Ureteroscopy with a Ho: YAG Laser.
A 23-year-old female had bilateral ureteral stents placed due to bilateral renal stones and hydronephrosis. The bilateral ureteral stents were changed every 3 months. A kidney ureter bladder (KUB) film showed left encrustation along the ureteral stent thus necessitating removal; however, the ureteral stent could not be removed cystoscopically. The ureteral stent was, therefore, extracted using flexible ureteroscopy (URS) with a holmium (Ho): yttrium aluminum garnet (YAG) laser.