Percutaneous endoscopic discectomy and thermal annuloplasty for professional athletes.
Percutaneous endoscopic discectomy (PED) for herniated nucleus pulposus has become increasingly popular since first reported by Yeung and Tsou. Additionally, radiofrequency thermal annuloplasty (TA) with PED (PED/TA) has been reported to be effective for discogenic low back pain. We used the PED/TA procedure to successfully treat four professional athletes with discogenic low back pain. All patients were men; their age at operation was 35, 35, 34, and 28 years. Daily low back pain was provoked by discography in all four patients, who had been receiving conservative but ineffective treatment for over 1 year. Surgery was conducted with the patients under local anesthesia. A cannula was placed posterior to the disc and ventral to the posterior annulus fibrosus through a 7-mm skin incision. Degenerated nucleus pulposus at the site was removed percutaneously and the posterior annulus was modulated by bipolar radiofrequency TA. After surgery, low back pain was resolved in all cases. After effective post-surgical rehabilitation with a physical trainer or physical therapist, all patients returned to their original competitive level. PED/TA is a minimally invasive and effective procedure for discogenic low back pain, especially in athletes.


Functional impacts of 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxycytosine at a single hemi-modified CpG dinucleotide in a gene promoter.
Enzymatic oxidation of 5-methylcytosine (5-mC) in the CpG dinucleotides to 5-hydroxymethylcytosine (5-hmC), 5-formylcytosine (5-fC) and 5-carboxycytosine (5-caC) has central role in the process of active DNA demethylation and epigenetic reprogramming in mammals. However, it is not known whether the 5-mC oxidation products have autonomous epigenetic or regulatory functions in the genome. We used an artificial upstream promoter constituted of one cAMP response element (CRE) to measure the impact of 5-mC in a hemi-methylated CpG on the promoter activity and further explored the consequences of 5-hmC, 5-fC, and 5-caC in the same system. All modifications induced mild impairment of the CREB transcription factor binding to the consensus 5'-TGACGTCA-3' CRE sequence. The decrease of the gene expression by 5-mC or 5-hmC was proportional to the impairment of CREB binding and had a steady character over at least 48 h. In contrast, promoters containing single 5-fC or 5-caC underwent further progressive loss of activity, up to an almost complete repression. This decline was dependent on the thymine-DNA glycosylase (TDG). The results thus indicate that 5-fC and 5-caC can provide a signal for perpetuation and enhancement of the repressed transcriptional state by a mechanism that requires base excision repair.


Pseudogenes for human small nuclear RNA U3 appear to arise by integration of self-primed reverse transcripts of the RNA into new chromosomal sites.
We find that both human and rat U3 snRNA can function as self-priming templates for AMV reverse transcriptase in vitro. The 74 base cDNA is primed by the 3' end of intact U3 snRNA, and spans the characteristically truncated 69 or 70 base U3 sequence found in four different human U3 pseudogenes. The ability of human and rat U3 snRNA to self-prime is consistent with a U3 secondary structure model derived by a comparison between rat U3 snRNA and the homologous D2 snRNA from Dictyostelium discoideum. We propose that U3 pseudogenes are generated in vivo by integration of a self-primed cDNA copy of U3 snRNA at new chromosomal sites. We also consider the possibility that the same cDNA mediates gene conversion at the 5' end of bona fide U3 genes where, over the entire region spanned by the U3 cDNA, the two rat U3 sequence variants U3A and U3B are identical.


Prorenin/renin receptor, signals, and therapeutic efficacy of receptor blocker in end-organ damage.
Discovery of prorenin/renin, or (pro)renin, receptor uncovered a novel function of (pro)renin as receptor ligand in addition to enzyme and its precursor; the same receptor was shown to promote reversible activation of prorenin and enhance the enzyme activity of mature renin. Stimulating the receptor activates mitogen-activated protein kinase and hypertrophic, hyperplastic, profibrotic, and cyclooxygenase-2-activating signals. These receptor signals were transmitted independently of angiotensin (Ang) II receptor. A specific blocker of the receptor was discovered-a peptide segment in prorenin that binds to the receptor and blocks ligand binding. Its infusion in animal models of hypertension and diabetes not only prevented nephropathy and cardiac hypertrophy, but also caused regression of nephropathy, whereas Ang II receptor gene deletion and angiotensin-converting enzyme inhibition merely delayed the onset or ameliorated pathologic phenotypes. These results suggest that (pro)renin receptor is responsible for end-organ damage.


Study of secretory component in patients with metastatic breast cancer.
A radioimmunoassay was developed for secretory component, a glandular epithelial cell product secreted as an accessory protein with dimeric immunoglobulins A and M. Forty-four normal women between the ages of 40 and 70 had plasma concentrations of secretory component of 7.195 microgram/ml +/- 3.590 (+/-2 SD). Of the patients with metastic breast cancer of epithelial origin, 34 (91.9%) had plasma concentrations greater than 8.990 microgram/ml; 29 (78.1%) had plasma concentrations greater than 10.785 microgram/ml; 10 (27%) had concentrations greater than 15.000 microgram/ml; and 3 (8.1%) had concentrations greater than 22.710 microgram/ml. Serial plasma concentrations (averaging 3.48 per patient) of 35 (94.6%) of these patients reflected the clinical course of the disease. All patients had some combination of surgery, radiation therapy, and chemotherapy. No direct correlation was noted between the type of therapy and concentration of secretory component. However, plasma secretory component served as a marker in the systemic circulation of a change in the status of the metastatic breast cancer. The impact of this research may lie in its incorporation into clinical management.


CD45 exon 4 point mutation does not confer susceptibility to type 1 diabetes mellitus or Graves' disease.
A point mutation in the leukocyte common antigen (CD45, C-->G77, exon 4) was investigated in patients with type 1 diabetes (IDDM), patients with Graves' disease and controls. The distribution did not differ significantly between patients and controls. This CD45 variant does not therefore confer susceptibility to either IDDM or Graves' disease.


The effects of compulsory health insurance on birth outcomes: evidence from China's UEBMI scheme.
Despite extensive research concerning the impact of health insurance on the advancement of infant health in developed countries, few studies have adjusted their results for potential confounding due to adverse selection in insurance coverage, wherein those who anticipate a need for health services tend to be the ones that acquire insurance. The presence of compulsory health insurance in China, such as the Urban Employee Basic Medical Insurance (UEBMI) scheme may provide an opportunity to estimate the effect of health insurance on infant health, by reducing the endogeneity problem into insurance due to the adverse selection. The objective is to assess the relationship between UEBMI and infant health outcomes in one sizeable municipal-level obstetrics hospital in Shanghai, East China. Medical records data from the Shanghai First Maternity and Infant Hospital from January 1, 2013 to April 30, 2019 were used to form an analysis dataset of 160,429 live births which was comprised of Shanghai residents with UEBMI coverage (n = 101,153) and women without any insurance coverage (n = 59,276). A propensity score matching approach using conjoint quantile regression and probit regression models was used to eliminate latent endogeneity of UEBMI coverage in order to garner robust results. Further analysis stratified by maternal migrant status was conducted to further assess the sensitivity of the findings to distinct patient subgroups. The UEBMI scheme was shown to be associated with improvements in infant birth outcomes. The scheme was associated with: an increase in birth weight of about 30 g (p < 0.001, 95% CI 23.908-35.295). This finding was evident in other five different birth outcomes (premature birth, low birth weight, very low birth weight, low Apgar score, and an abnormal health condition at birth). After stratifying by migrant status, the UEBMI was shown to have a greater effect on migrants compared to local residents of Shanghai. Our findings suggest that health insurance coverage for pregnant women, especially for migrants, has the potential to significantly and directly improve infant health outcomes. Further research is required to determine whether these findings can be replicated for other Chinese jurisdictions.


Evaluation of Modified Sequential Multiplex PCR for Streptococcus pneumoniae Serotyping.
The aims of this study were to develop modified sequential multiplex PCR (SM-PCR) primer sets and to evaluate their ability and efficiency for serotype determination. We selected target serotypes for SM-PCR testing according to serotype prevalence as reported in Asian publications. The modified SM-PCR consisted of 6 groups of PCR reactions, and each reaction was performed using 5 primer pairs. We evaluated the efficiency and performance of this modified multiplex PCR using 378 pneumococcal strains by comparing the findings with the results of the Quellung reaction. A total of 30 primer pairs were used in a consecutive set of 6 reactions. All results were concordant with those of the Quellung reaction and there was no cross-reactivity to unintended serotypes. We could identify the final serotypes of 370 isolates (97.9%). The coverage rates of modified SM-PCR were 42.6%, 65.9%, and 79.4% in reactions1, 2, and 3, respectively. The modified SM-PCR showed acceptable performance for detecting pneumococcal serotypes and can serve as useful alternative to the Quellung reaction.


Working towards a conservation plan for fish parasites: Cyprinid parasites from the south African cape fold freshwater ecoregion as a case study.
The preservation of the world's biodiversity for future generations has been a global objective for many years, with the establishment of the International Union for Conservation of Nature (IUCN) Red List of Threatened Species in 1964. However, the conservation of parasites is a more recent development and, due to the difficulty in obtaining data and studying some of the parasitic species, comes its own series of challenges. Using parasites of cyprinid hosts (one critically endangered, one endangered and three near threatened) collected from South Africa's Cape Fold freshwater ecoregion (CF) as a case study, this paper discusses the challenges and possible solutions for implementing a fish parasite conservation project. Novel data on the fish parasites (1819 metazoan parasite individuals, representing the Acanthocephala, Cestoda, Copepoda, Digenea, Monogenea and Nematoda) of the CF are provided from the five endemic hosts, Cheilobarbus serra (Peters, 1864), Labeobarbus seeberi (Gilchrist et Thompson, 1913), Pseudobarbus phlegethon (Barnard, 1938), Sedercypris calidus (Barnard, 1938), and Sedercypris erubescens (Skelton, 1974). Conservation statuses for selected parasite taxa are also proposed based on the conservation statuses of the fish hosts, according to the Conservation Assessment Methodology for Animal Parasites (CAMAP).


The effects of orthobiologics in the treatment of tendon pathologies: a systematic review of preclinical evidence.
The aim of this systematic review is to explore the current available knowledge about tendon disorders and orthobiologics derived by preclinical experiments to evaluate their role and efficacy in the different stages and conditions related to the tendon healing processes. The systematic review was performed according to the PRISMA guidelines. Different electronic databases (MEDLINE, Web of Science, EMBASE) were searched for studies investigating orthobiologics (PRP and cell-based products from adipose tissue or bone marrow) in animal models or veterinary clinical trials for tendon pathologies (complete/partial tendon ruptures, rotator cuff tears, tendinopathy, enthesis-related injuries). Data regarding the specific product used, the treatment site/pathology, the host and the model were collected. The results were classified into the following categories: histological, biomechanical, molecular and imaging. A large pool of preclinical studies on tendon disorders have been found on platelet-rich plasma (PRP), while data about stromal vascular fraction (SVF) and bone marrow concentrate (BMAC) are still limited and frequently focused on expanded cells, rather than orthobiologics prepared at the point of care. The effect of PRP is related to an acceleration of the healing process, without improvements in the final structure and properties of repaired tendon. Cell-based products have been reported to produce more durable results, but the level of evidence is currently insufficient to draw clear indications. The preclinical results about orthobiologics applications to tendon pathologies would support the rationale of their clinical use and encourage the performance of clinical trials aimed to confirm these data in human subjects.


Pre- and postprandial alterations of portal venous flow: evaluation with single breath-hold three-dimensional half-Fourier fast spin-echo MR imaging and a selective inversion recovery tagging pulse.
To evaluate the influence of food intake on portal flow using unenhanced magnetic resonance imaging (MRI). The study population included 29 healthy subjects. A selective inversion recovery tagging pulse was used on the superior mesenteric vein (SMV) and splenic vein (SpV) to study the correlation of tagged blood in the portal vein (PV). MRI was performed before and 60-90 min after a meal. The flow signal from the SMV increased in 97% of the subjects after the meal. Before the meal the portal flow was dominated by flow from the SpV in 59% of the subjects, while it was dominated by flow from the SMV in 76% of the subjects after the meal. The most common distribution pattern of the flow signal from the SpV before the meal was in the central part of the main PV (55%), while it was in the left side (45%) after the meal. The most common distribution pattern of the flow signal from the SMV was in the bilateral sides of the main PV both before and after the meal (62%). This technique shows potential for evaluating pre- and postprandial alterations of flow from the SpV and SMV in the PV under physiological conditions.


Thyrotropin releasing hormone analogs: a building block approach to the construction of tetracyclic peptidomimetics.
A building block based approach was used to synthesize a pair of tetracyclic peptidomimetics that constrain all but one of the rotational degrees of freedom of the hypothalamic tripeptide hormone thyroliberin. One of the analogs bound to the thyroliberin endocrine receptor (TRH-R) with an affinity greater than that of an analog without constraints. The tetracyclic peptidomimetics were found to be partial agonists for the TRH-R receptor.


Maternal ethanol administration inhibits 5-hydroxytryptamine synthesis and tryptophan hydroxylase expression in the dorsal raphe of rat offspring.
Maternal ethanol consumption during pregnancy has a detrimental effect on the central nervous system (CNS) development of fetus. 5-Hydroxytryptamine (5-HT) is an important neurotransmitter and/or neuromodulator in the mammalian CNS. Tryptophan hydroxylase (TPH) is the rate limiting enzyme of 5-HT synthesis. Ethanol is known to induce neuropsychiatric disorders by alteration of the central serotonergic system. In the present study, the effects of maternal ethanol intake on the 5-HT synthesis and the TPH expression in the dorsal raphe of rat offspring were investigated. The present results show that the synthesis of 5-HT and the expression of TPH in the dorsal raphe of rat offspring were suppressed by maternal ethanol intake and that the suppressive effect of alcohol was more potent in the 5 weeks old rat pups compared to the 3 weeks old rat pups. Based on the present study, it can be suggested that the pathogenesis of ethanol-induced neuropsychological disorders involves ethanol-induced suppression on the 5-HT synthesis and the TPH expression in the dorsal raphe of offspring.


Toxicity mitigation and solidification of municipal solid waste incinerator fly ash using alkaline activated coal ash.
Municipal solid waste (MSW) incineration is a common and effective practice to reduce the volume of solid waste in urban areas. However, the byproduct of this process is a fly ash (IFA), which contains large quantities of toxic contaminants. The purpose of this research study was to analyze the chemical, physical and mechanical behaviors resulting from the gradual introduction of IFA to an alkaline activated coal fly ash (CFA) matrix, as a mean of stabilizing the incinerator ash for use in industrial construction applications, where human exposure potential is limited. IFA and CFA were analyzed via X-ray fluorescence (XRF), X-ray diffraction (XRD) and Inductive coupled plasma (ICP) to obtain a full chemical analysis of the samples, its crystallographic characteristics and a detailed count of the eight heavy metals contemplated in US Title 40 of the Code of Federal Regulations (40 CFR). The particle size distribution of IFA and CFA was also recorded. EPA's Toxicity Characteristic Leaching Procedure (TCLP) was followed to monitor the leachability of the contaminants before and after the activation. Also images obtained via Scanning Electron Microscopy (SEM), before and after the activation, are presented. Concrete made from IFA, CFA and IFA-CFA mixes was subjected to a full mechanical characterization; tests include compressive strength, flexural strength, elastic modulus, Poisson's ratio and setting time. The leachable heavy metal contents (except for Se) were below the maximum allowable limits and in many cases even below the reporting limit. The leachable Chromium was reduced from 0.153 down to 0.0045 mg/L, Arsenic from 0.256 down to 0.132 mg/L, Selenium from 1.05 down to 0.29 mg/L, Silver from 0.011 down to .001 mg/L, Barium from 2.06 down to 0.314 mg/L and Mercury from 0.007 down to 0.001 mg/L. Although the leachable Cd exhibited an increase from 0.49 up to 0.805 mg/L and Pd from 0.002 up to 0.029 mg/L, these were well below the maximum limits of 1.00 and 5.00 mg/L, respectively.


The level of action of 2,4-D on transcription.
In mature cells, 2,4-D retards transcription when it is applied before 3H-uridine incorporation. But transcription is accelerated when it is applied after feeding with 3H-uridine. It has been suggested that the action of 2,4-D on transcription is only at the level of initiation.


Intermittency in mastication and apomorphine-induced gnawing.
Rhythmic behaviors like mastication, gnawing, and locomotion, are characterized by temporal segmentation or intermittency. That is, they frequently occur as a series of short bursts interrupted by pauses rather than as one long uninterrupted burst. The function of intermittency as well as the mechanisms that produce it are unknown. Biogenic amine systems may play a role in producing intermittency; however, experimental work to confirm this is only in its infancy. The current study evaluates the structure of intermittency associated with mastication and apomorphine-induced gnawing in the guinea pig. Thirteen free-roaming animals were videotaped while masticating or gnawing. Eight animals were given 0.5 mg/kg i.m. apomorphine and videotaped while gnawing. The remaining five animals received no apomorphine injections, but were taped while feeding on alfalfa pellets. Custom software was used to score instances of maximum jaw closures in videotaped mastication and gnawing sequences. The time between successive maximum jaw closures, called the interocclude interval (IOI), was calculated for all scored sequences. A cutoff IOI value of 0.26 s differentiated pauses (IOI values equal or greater than 0.26 s) from chews or gnaws (IOI values less than 0.26 s). Two or more successive chews or gnaws, without intervening pauses, defined behavior bursts. Chew, gnaw, and burst durations were quantified and compared. Chew and gnaw durations were similar. However, chewing bursts were significantly longer than gnawing bursts. The significance of these results is presented in light of previous neurophysiological work on rhythmic jaw movements and intermittency.


Association between carpel tunnel syndrome and arteriovenous fistula in hemodialysis patients.
Carpal tunnel syndrome is more common in hemodialysis patients than in the general population. In addition to uremic neuropathy, arteriovenous fistula has been identified as one of the possible causes for the development of carpal tunnel syndrome. Wrists of 279 hemodialysis patients who had arteriovenous fistula for at least 6 months served as the case group and the other intact wrists of the same patients served as the control group. Carpal tunnel syndrome was diagnosed clinically; however, 116 random patients underwent electrodiagnostic studies the day after hemodialysis. Clinically, carpal tunnel syndrome was diagnosed less frequently in the contralateral wrist than in the wrist with arteriovenous fistula (12.2 percent versus 30.5 percent, p < 0.0001). Meanwhile, the authors found a positive correlation between the duration of fistula and the development of carpal tunnel syndrome (p < 0.028). The site of fistula (snuffbox, radiocephalic) used, however, had no effect on the rate of development of carpal tunnel syndrome (p > 0.2). Contrary to the clinical assessment, electrodiagnostic studies did not indicate any significant association between the frequency of carpal tunnel syndrome and arteriovenous fistula or its duration. With the nerve conduction velocity abnormalities observed in patients with uremic polyneuropathy, the authors believe that clinical diagnosis of carpal tunnel syndrome is more indicative of the development of the condition than the electrodiagnostic results. The authors' clinical results have revealed that hemodialysis patients are at considerable risk of developing carpal tunnel syndrome in the wrist with an arteriovenous fistula. These patients should be under close observation and receive routine checkup.


A novel transgenic mouse model reveals deregulation of the ubiquitin-proteasome system in the heart by doxorubicin.
Ubiquitin-proteasome system (UPS) mediated proteolysis is responsible for the degradation of majority of cellular proteins, thereby playing essential roles in maintaining cellular homeostasis and regulating a number of cellular functions. UPS dysfunction was implicated in the pathogenesis of numerous disorders, including neurodegenerative disease, muscular dystrophy, and a subset of cardiomyopathies. However, monitoring in vivo functional changes of the UPS remains a challenge, which hinders the elucidation of UPS pathophysiology. We have recently created a novel transgenic mouse model that ubiquitously expresses a surrogate protein substrate for the UPS. The present study validates its suitability to monitor in vivo changes of UPS proteolytic function in virtually all major organs. Primary culture of cells derived from the adult transgenic mice was also developed and tested for their applications in probing UPS involvement in pathogenesis. Applying these newly established in vivo and in vitro approaches, we have proven in the present study that doxorubicin enhances UPS function in the heart and in cultured cardiomyocytes, suggesting that UPS hyper-function may play an important role in the acute cardiotoxicity of doxorubicin therapy.


The purification and characterization of ADP-glucose pyrophosphorylase A from developing maize seeds.
ADPglucose pyrophosphorylase A (ATP:alpha-D-glucose-1-phosphate adenylyltransferase, EC 126.96.36.199) from developing maize (Zea mays) endosperm was purified 129 fold to apparent homogeneity. The molecular weight estimated by gel filtration and by polyacrylamide gel electrophoresis was 375 000 and 400 000, respectively. The preparation gave a single protein band after SDS-polyacrylamide gel electrophoresis suggesting a monomer mol. wt. of 96 000. It was concluded that ADPglucose pyrophosphorylase A in maize endosperm is a tetramer of four similar molecular weight subunits. Values for the Km for glucose 1-phosphate and ATP were 3.8 . 10(-5) and 1.8 . 10(-4) M, respectively (using the homogeneous preparation).


Long-term nutrition support in gastrointestinal disease--a systematic review of the evidence.
Patients with gastrointestinal disease or those having undergone gastrointestinal surgery are often at nutritional risk for extended periods after they are discharged from the acute care setting. This systematic review aims to assess the evidence for the use of oral nutrition support, in the form of oral nutrition supplementation or individualized dietary advice in people with gastrointestinal illness, for a prolonged period of time, deemed greater than 2 mo. A systematic review of relevant databases was conducted from January 1980 until December 2009. Suitable articles were appraised looking at oral nutrition support, for greater than 2 mo, and the association with nutritional outcomes in people with gastrointestinal illness. Six studies were identified for this systematic review. The range of patient groups, interventions, and outcomes differed between studies. Overall evidence supports the use of oral nutrition support for improving outcomes such as weight, function, caloric intake, and quality of life. There was a trend toward improved outcomes with individualized dietary advice as the study period increased. There is strong evidence to support the use of oral nutrition support in a long-term setting for gastrointestinal patients. It appears that as the timeframe increases, there may be a greater role for the use of individualized dietary advice over oral nutrition supplementation alone. However, more research is needed in this area.


[Large bowel preparation in colon surgery: elemental diet (author's transl)].
None of the usual methods of bowel preparation guarantee an aseptic large bowel. The intention is to reduce intestinal flora quantitatively, which can be achieved by mechanical cleansing alone. During the preparation period caloric and amino acid intake is insufficient. Therefore alimentation with elemental diet is necessary. The patient gets an adequate caloric and protein supply. The diet is totally absorbed in the small intestine, the large bowel remains clean, and the bacterial flora is reduced.


Chromatic discrimination with variation in chromaticity and luminance: data and theory.
Boynton and Kambe developed a model of chromatic discrimination in which thresholds are mediated by two independent mechanisms: the short-wavelength sensitive (S-) cones (S-cone axis), and the middle-wavelength sensitive (M-) and long-wavelength sensitive (L-) cones (M/L-cone axis). In this study, we used a Maxwellian view optical system to investigate fundamental properties of the model as a function of chromaticity and luminance. We confirmed that discriminations along the S-cone axis were dependent on S-cone excitation level. However, changes in chromaticity and changes in mean luminance were not described by a single threshold-vs-radiance (TVR) template. We developed a model to account for the different effects of changing S-cone excitation by varying mean chromaticity and by varying mean luminance. M/L-cone discriminations showed a minimum at the L-cone excitation to white, indicating strong opponency. The thresholds increased with luminance approaching a Weber region and showing parallel functions for differing chromaticities. These data are fit by a model allowing retinal gain controls and spectral opponency.


Personal medical electronic devices and walk-through metal detector security systems: assessing electromagnetic interference effects.
There have been concerns that Electromagnetic security systems such as walk-through metal detectors (WTMDs) can potentially cause electromagnetic interference (EMI) in certain active medical devices including implantable cardiac pacemakers and implantable neurostimulators. Incidents of EMI between WTMDs and active medical devices also known as personal medical electronic devices (PMED) continue to be reported. This paper reports on emission measurements of sample WTMDs and testing of 20 PMEDs in a WTMD simulation system. Magnetic fields from sample WTMD systems were characterized for emissions and exposure of certain PMEDs. A WTMD simulator system designed and evaluated by FDA in previous studies was used to mimic the PMED exposures to the waveform from sample WTMDs. The simulation system allows for controlled PMED exposure enabling careful study with adjustable magnetic field strengths and exposure duration, and provides flexibility for PMED exposure at elevated levels in order to study EMI effects on the PMED. The PMED samples consisted of six implantable cardiac pacemakers, six implantable cardioverter defibrillators (ICD), five implantable neurostimulators, and three insulin pumps. Each PMED was exposed in the simulator to the sample WTMD waveforms using methods based on appropriate consensus test standards for each of the device type. Testing the sample PMEDs using the WTMD simulator revealed EMI effects on two implantable pacemakers and one implantable neurostimulator for exposure field strength comparable to actual WTMD field strength. The observed effects were transient and the PMEDs returned to pre-exposure operation within a few seconds after removal from the simulated WTMD exposure fields. No EMI was observed for the sample ICDs or insulin pumps. The findings are consistent with earlier studies where certain sample PMEDs exhibited EMI effects. Clinical implications were not addressed in this study. Additional studies are needed to evaluate potential PMED EMI susceptibilities over a broader range of security systems.


Functional outcomes and complications in patients with bladder cancer undergoing robotic-assisted radical cystectomy with extracorporeal Indiana pouch continent cutaneous urinary diversion.
To evaluate the functional outcomes and complications for patients with bladder cancer undergoing robotic-assisted laparoscopic radical cystectomy with Indiana pouch continent cutaneous urinary diversion. From February 2004 to March 2010, 34 patients underwent robotic-assisted laparoscopic radical cystectomy with Indiana pouch continent cutaneous urinary diversion reconstruction. After surgery, the complications were identified, categorized, and graded using an established 5-grade modification of the original Clavien grading system, and continence was assessed. Descriptive statistics were used in evaluating the outcomes. Fischer's exact test was used in the comparison of early and late Clavien grade III complications. Overall, 175 (123 early and 52 late) complications after surgery were reported in 32 (94%) of 34 patients. Within 90 days of surgery, 31 (91%) of 34 patients experienced ≥ 1 early complication. Of 34 patients, 15 (44%) reported ≥ 1 late complications (>90 days). Most (85% and 69%, respectively) early and late complications were graded as minor (grade II or less). Fewer patients with early complications required an additional intervention (grade III) compared with patients with late complications (14% vs 31%; P = .116). The most common complication in both intervals was infection, reported in 22% and 37% of patients with early and late complications, respectively. The continence data for 31 patients at a mean follow-up of 20.1 months (median 12.0) showed that all but 1 patient (97%) had daytime and nighttime continence. Patients undergoing robotic-assisted laparoscopic radical cystectomy with Indiana pouch continent cutaneous urinary diversion reconstruction have comparable complication rates and functional outcomes compared with patients in the open series.


Cell dynamics in the pulpal healing process following cavity preparation in rat molars.
Odontoblast-lineage cells acquire heat-shock protein (HSP)-25-immunoreactivity (IR) after they complete their cell division, suggesting that this protein acts as a switch between cell proliferation and differentiation during tooth development. However, there are few available data concerning the relationship between cell proliferation and differentiation following cavity preparation. The present study aims to clarify the expression of HSP-25 in the odontoblast-lineage cells with their proliferative activity after cavity preparation by immunocytochemistry for HSP-25 and cell proliferation assay using 5-bromo-2'-deoxyuridine (BrdU) labeling. In untreated control teeth, intense HSP-25-IR was found in odontoblasts and some subodontoblastic mesenchymal cells. Cavity preparation caused the destruction of odontoblasts and the disappearance of HSP-25-IR was conspicuous at the affected site, although some cells retained HSP-25-IR and subsequently most of them disappeared from the pulp-dentin border by postoperative day 1. Contrary, some subodontoblastic mesenchymal cells with weak HSP-25-IR began to take the place of degenerated cells, although no proliferative activity was recognizable in the dental pulp. Interestingly, proliferative cells in the dental pulp significantly increased in number on day 2 when the newly differentiating cells already arranged along the pulp-dentin border, and continued their proliferative activity in the wide range of the pulp tissue until day 5. These findings indicate that progenitor cells equipped in the subodontoblastic layer firstly migrate and differentiate into new odontoblast-like cells to compensate for the loss of the odontoblast layer, and subsequently the reorganization of dental pulp was completed by active proliferation of the mesenchymal cells occurring in a wide range of pulp tissue.


Optimized Mn-doped iron oxide nanoparticles entrapped in dendrimer for dual contrasting role in MRI.
Magnetic resonance imaging has acquired importance as a major tool for diagnosis and staging of cancers in humans. Injection of certain imaging agents have proved to improve contrast between normal and cancer cells on magnetic resonance imaging (MRI). Using the principles of MR contrast imaging, we have designed a dual mode (T1 and T2) contrast agent based on folic acid functionalized manganese ferrite nanoparticles (MNP) entrapped in 3G polyamidoamide (PAMAM) dendrimers. The ratio of Mn:Fe was tuned to achieve optimal performance. This multifunctional nanocarrier system was developed for targeting cancer cells to produce both T1 and T2 contrast which in turn helps in better diagnosis and staging of cancer. FTIR spectroscopy, X-Ray diffraction, atomic absorption spectroscopy, UV-Visible spectroscopy, and dynamic light scattering measurements were employed to characterize the multifunctional system at different stages of engineering. The ratio of relaxivities r2/r1 is 4.6 at 1.5 T for the MNP prepared with 0.5 molar ratio of Mn/Fe based on MR images obtained from phantom and tumor bearing mouse. The value of r2/r1 shows that the 0.5 molar ratio of Mn/Fe can be used to prepare MNP for the production of dual mode contrast in MR imaging.


High-throughput and trace analysis of diazepam in plasma using DART-MS/MS and its pharmacokinetic application.
A high-throughput quantitative analytical method based on Direct Analysis in Real Time tandem mass spectrometry (DART-MS/MS) has been developed and validated for the determination of diazepam in rat plasma, whereby analyzing of each sample needs merely 25 μL plasma, simple solid phase extraction sample preparation and 15 s acquisition time. The multiple reaction monitoring (MRM) transitions at m/z 285.2 → 193.1 and 316.0 → 270.0 were selected for the monitoring of diazepam and its internal standard clonazepam respectively. A good linearity within the range of 10-2000 ng/mL, an intra- and inter-day precisions within <7.78% as to an accuracy ranging from 1.04% to 7.92% have been achieved. The method has been successfully applied to the pharmacokinetic study of diazepam in rats' plasma after a single intragastric administration at a dose of 10 mg/kg. The results indicate that this method fulfills the requirements of the bioanalysis in sensitivity and accuracy. It shows considerable promise for application of DART-MS to the quantitative investigation of other drugs.


A comparison of smoking behaviour characteristics between Caucasian smokers in the United Kingdom and Malay smokers in Malaysia.
There is evidence that smoking behaviour differs by ethnicity. This study aims to compare smoking behaviour characteristics between Caucasian and Malay smokers. A cross sectional survey, involving 175 smokers attending smoking cessation clinics at the Institute of Psychiatry, London, United Kingdom and University Malaya, Kuala Lumpur, Malaysia between May 2005 and February 2007. Data on demographics, smoking history, nicotine dependence and smoking behaviour were collected. All participants were males, mean age 30.7 ± 10.3 years. Caucasians initiated smoking significantly earlier (mean age 14.8 ± 2.8 years) (p = 0.001) and smoked regularly significantly earlier (mean age 17.3 ± 3.5) (p = 0.003) than Malays (mean starting age 16.9 ± 4.4 years and mean age regular use 19.5 ± 4.5 years), respectively. Caucasians smoked less for social integration than Malays (p = 0.03) but smoked more for regulation of negative affect than Malays (p = 0.008) and smoked more for hedonism than Malays (p < 0.001). Malays smoke as a means of socially integrating. This has important public health implications. Social reasons and the social environment play a role in smoking uptake, smoking maintenance and smoking cessation and this should be borne in mind for strategies planning to promote smoking cessation.


Toxic effects of dietary polybrominated biphenyls on mink.
Serial levels of fireMaster FF-1, a commercial mixture of polybrominated biphenyls (PBB), and tissues from chickens and a cow that had previously consumed PBB were fed to mink to ascertain the chronic effects of the commercial and "metabolized" form of this compound on mink. Diets that contained 6.25 ppm (or more) PBB were lethal to adult mink within 10 months. One to 2.5 ppm dietary PBB fed for 9 months had an adverse effect on litter size, kit weight at birth, and kit survival. The data suggest that the PBB derived from contaminated beef and poultry was more toxic than the original PBB. The clinical signs of PBB poisoning in mink were food rejection, weight loss, and unthrifty appearance, and fatty infiltration of the liver. Based on these findings, mink must be considered highly susceptible to PBB toxicity. PBB residue levels 60 times the amount in the diet were found in the adipose tissue of the PBB-treated animals.


Healthy people 2010 leading health indicators: how children with special health care needs fared.
We compared estimates for children with and without special health care needs (SHCN) at 2 time periods for national health objectives related to the Healthy People 2010 leading health indicators (LHIs). Data were from the 2003 and 2007 National Surveys of Children's Health. Seven survey items were relevant to the LHIs and available in both survey years: physical activity, obesity, household tobacco use, current insurance, personal health care provider, past-month depressive symptoms, and past-year emergency department visits. In 2003 and 2007, children with SHCN fared worse than those without SHCN with respect to physical activity, obesity, household tobacco exposure, depressive symptomology, and emergency department visits, but fared better on current insurance and having a personal health care provider. Physical activity and access to a personal health care provider increased for all children, whereas the absolute disparity in personal provider access decreased 4.9%. Significant disparities exist for key population health indicators between children with and without SHCN. Analyses illustrated how population-based initiatives could be used to frame health challenges among vulnerable populations.


Rapid down modulation of P-selectin glycoprotein ligand-1 (PSGL-1, CD162) by G-CSF in humans.
In vitro and animal studies suggest a critical role for P-selectin glycoprotein ligand-1 (PSGL-1) in the regulation of WBC adhesion and neutrophil counts. As WBC activation decreases PSGL-1 expression on WBCs in vitro, the effects of G-CSF on PSGL-1 expression were examined. Two different G-CSF doses (1 and 5 microg/kg IV) were compared with high-dose dexamethasone (1 mg/kg twice daily) and placebo in a randomized, double-blind, four-way cross-over trial in eight healthy volunteers. Surface expression of WBC adhesion molecules was quantified by flow cytometry. Both G-CSF and dexamethasone led to a delayed down regulation of L-selectin. In contrast, G-CSF rapidly down regulated PSGL-1 expression on neutrophils within 90 minutes, whereas neither dexamethasone nor placebo had an effect. Similarly, incubation of WBCs with clinically relevant G-CSF concentrations (60 microg/L) for 90 minutes down modulated PSGL-1 expression on neutrophils and enhanced CD11b expression, compatible with a direct PSGL-1 down regulation by G-CSF-induced neutrophil activation. Similar to G-CSF, GM-CSF down regulated PSGL-1 in vitro. Both drugs induced shedding of soluble PSGL-1, supporting the concept that proteolytic cleavage is a potential mechanism of PSGL-1 down regulation on neutrophils. G-CSF, but not dexamethasone, down regulates PSGL-1 expression on the surface of neutrophils in humans. This could also partly explain the synergistic effects when both drugs are combined for optimal mobilization of neutrophils for clinical granulocyte transfusion programs.


Polymorphisms in lncRNA CCAT1 on the susceptibility of lung cancer in a Chinese northeast population: A case-control study.
To explore the association of rs1948915, rs7013433 in long noncoding RNA (lncRNA) CCAT1 and rs6983267 in MYC enhancer region with the risk of lung cancer in a Chinese northeast population, a case-control study was conducted. The hospital-based case-control study contained 669 lung cancer patients and 697 healthy controls. Taqman® Probe allele resolution was used for genotyping. The differences between the case-control groups were analyzed using Student t-test and chi-square test. Logistic regression analysis was used to assess the relationship between the genotypes and the risk of lung cancer. Cross-generation analysis was used to explore the relationship between gene-environment interaction and lung cancer. There was no association between the three selected single-nucleotide polymorphisms (SNPs) and the susceptibility of lung cancer. Rs1948915 CT was correlated with lung adenocarcinoma. In female stratification, rs1948915 CT/CC was associated with a decreased susceptibility of lung cancer significantly. Additionally, the additive and multiplicative interaction models showed that there was no interaction between the three selected SNPs and smoking status in lung cancer. There may be an association between lung adenocarcinoma and rs1948915 polymorphism in the Chinese northeast population, while rs7013433 and rs6983267 might have no association. There was no interaction between the three selected SNPs and smoking status.


Rod pathways in mammalian retinae.
A variety of recent experiments has resolved the way in which signals are transmitted from rod photoreceptors to ganglion cells in the mammalian retina. Rods connect to a single class of rod bipolar cell, which depolarize in response to light. Rod bipolar cells are not connected directly to ganglion cells: they synapse onto rod amacrine cells, which excite ON-centre ganglion cells via gap junctions, and inhibit OFF-centre ganglion cells via inhibitory glycine synapses. Monoamines have particular influences on the rod system, through synapses with rod amacrine and rod bipolar cells, and a function for dopamine and indoleamines within this system can be hypothesized from recent experiments. There is evidence to suggest that dopaminergic amacrine cells bring the surround response into the rod system through synapses with the rod amacrine cell, and that an indoleamine, probably serotonin, increases the signal in the ON pathway through a feedback synapse onto the rod bipolar terminal.


A Phase 3, Multicenter, Randomized, Two-Arm, Open-Label Study of Intermittent Oral Dosing of Roxadustat for the Treatment of Anemia in Japanese Erythropoiesis-Stimulating Agent-Naïve Chronic Kidney Disease Patients Not on Dialysis.
Roxadustat is an oral hypoxia-inducible factor prolyl hydroxylase inhibitor approved for the treatment of anemia in Japan for patients with dialysis-dependent (DD) chronic kidney disease (CKD). Multicenter, randomized, open-label, noncomparative, phase 3 study to evaluate roxadustat for anemia of non-dialysis-dependent (NDD) CKD in Japan. Erythropoiesis stimulating agent (ESA)-naïve NDD-CKD patients were randomized to roxadustat (initial dose, 50 or 70 mg 3 times weekly), titrated to maintain hemoglobin (Hb) within 10.0-12.0 g/dL, for ≤24 weeks. Patients with either transferrin saturation of ≥5% or serum ferritin of ≥30 ng/mL during the screening period were eligible. Endpoints included response rate (proportion of patients achieving Hb ≥10.0 or ≥10.5 g/dL and Hb increase ≥1.0 g/dL from baseline) at end of treatment; average Hb (weeks 18-24); change of average Hb from baseline to weeks 18-24; maintenance rate (proportion of patients achieving Hb 10.0-12.0 g/dL at weeks 18-24); rate of rise (RoR) of Hb from weeks 0-4, discontinuation, or dose adjustment. Adverse events were monitored throughout the study. Of 135 patients who provided informed consent, 100 were randomized and 99 received roxadustat (50 mg, n = 49; 70 mg, n = 50). The mean (SD) dose of roxadustat per intake at week 22 was 36.3 (22.7) mg in the roxadustat 50 mg group and 36.8 (16.0) mg in the roxadustat 70 mg group. Prior medications included oral iron therapy (20.2%) and intravenous iron therapy (1.0%). Overall response rate (95% CI) was 97.0% (91.4, 99.4; Hb ≥10.0 g/dL) and 94.9% (88.6, 98.3; Hb ≥10.5 g/dL). Mean (SD) Hb (weeks 18-24) was 11.17 (0.62) g/dL. Mean (SD) change of Hb from baseline (weeks 18-24) was 1.34 (0.86) g/dL. Maintenance rate (95% CI) was 88.8% (80.3, 94.5) among patients with ≥1 Hb measurement during weeks 18-24. Mean (SD) RoR of Hb was 0.291 (0.197) g/dL/week (50 mg) and 0.373 (0.235) g/dL/week (70 mg). Nasopharyngitis and hypertension were the most common adverse events. Roxadustat increased and maintained Hb in ESA-naïve, partially iron-depleted NDD-CKD patients with anemia.


[Ecological data on house dust mites in the city of Semipalatinsk].
Distribution of mites of the genus Dermatophagoides in flats of allergic patients depending on the type of the building structure, relative air humidity and season of the year was studied. The mites are most frequently encountered in houses of wood or adobe with high air humidity. Autumn is an optimum season for reproduction of mites.


Clinical aspects of sperm bank.
The ability to preserve human sperm by cryogenic methods has been developed and successfully applied to reproductive biology. Some of the advantages include 1) availability of sperm for heterologous (donor) insemination, 2) accumulation of multiple oligospermic specimens for homologous (husband) insemination, 3) preservation of sperm prior to vasectomy, 4) collection and preservation of sperm from men who face permanent injury to spermatogenesis through an operation, chemotherapy or radiation, 5) to obviate the potential genetic danger of man's exposure to environmental pollutants such as radiation and 6) availability of husband's sperm at ovulation time.


Visualization and quantification of myocardial mass at risk using three-dimensional contrast echocardiography.
Three-dimensional echocardiographic assessment of myocardial ischemia using contrast echocardiography has been hampered by limitations of available contrast agents and analytic software. In the study presented, a three-dimensional perfusion imaging method was evaluated in the porcine model of myocardial ischemia using a novel contrast agent. Three-dimensional echocardiography was performed in eight open-chested pigs before, during and after left anterior descending (six animals) or circumflex (two animals) coronary artery occlusion. The intramyocardial contrast effect was obtained by left atrial injection of Myomap, a deposit contrast agent. Myocardial opacification was visible in all studies and retained in all three-dimensional datasets. Three-dimensional intensity analysis demonstrated a significant difference, exceeding 20 intensity units in every animal (in 127-level scale), between perfused and non-perfused myocardium. Reperfusion followed by contrast reinjection resulted in homogenous myocardial enhancement. Myocardial mass at risk was clearly delineated in all studies and measured with a mean error of -0.1 +/- 2.0 g against real mass (p = non-significant). Spatial extent of ischemia could be displayed in volume-rendered reconstruction of separate perfusion territories. Quantitative analysis of myocardial contrast enhancement from three-dimensional datasets is feasible and allows accurate measurement of myocardial mass at risk.


Long non-coding RNA ANRIL is up-regulated in bladder cancer and regulates bladder cancer cell proliferation and apoptosis through the intrinsic pathway.
Antisense non-coding RNA in the INK4 locus (ANRIL) is a member of long non-coding RNAs and has been reported to be dysregulated in several human cancers. However, the role of ANRIL in bladder cancer remains unclear. This present study aimed to investigate whether and how ANRIL involved in bladder cancer. Our results showed up-regulation of ANRIL in bladder cancer tissues versus the corresponding adjacent non-tumor tissues. To explore the specific mechanisms, ANRIL was silenced by small interfering RNA or short hairpin RNA transfection in human bladder cancer T24 and EJ cells. Knockdown of ANRIL repressed cell proliferation and increased cell apoptosis, along with decreased expression of Bcl-2 and increased expressions of Bax, cytoplasmic cytochrome c and Smac and cleaved caspase-9, caspase-3 and PARP. However, no change of cleaved caspase-8 level was observed. Furthermore, in vivo experiment confirmed that knockdown of ANRIL inhibited tumorigenic ability of EJ cells in nude mice. Meanwhile, in accordance with in vitro study, knockdown of ANRIL inhibited expression of Bcl-2 and up-regulated expressions of Bax and cleaved caspase-9, but did not affect cleaved caspase-8 level. In conclusion, we first report that ANRIL possibly serves as an oncogene in bladder cancer and regulates bladder cancer cell proliferation and apoptosis through the intrinsic apoptosis pathway.


The psychotropic self/imaginary: subjectivity and psychopharmaceutical use among heroin users with co-occurring mental illness.
Many people diagnosed with mental illnesses struggle with illicit drug addiction. These individuals are often treated with psychiatric medications, yet little is known about how they experience this treatment. Research on the subjective experience of psychiatric medication use highlights the complex, contradictory, and ambiguous feelings often associated with this treatment. However, for those with mental illness and addiction, this experience is complicated by the need to manage both psychiatric medication and illicit drug use. Using ethnographic data from a study of heroin use in Northeast Ohio, we explore this experience by expanding the pharmaceutical self/imaginary (Jenkins, Pharmaceutical Self: The Global Shaping of Experience in an Age of Psychopharmacology, School for Advanced Research Press, Santa Fe, NM, 2010b) to include psychopharmaceuticals and illicit drugs, what we call the psychotropic self/imaginary. Through this lens we explore the ways participants interpret and manage their psychotropic drug use in relation to sociocultural, institutional, and political-economic contexts. This analysis reveals how participants seek desired effects of legally prescribed and illicit drugs to treat mental illness, manage heroin addiction, and maintain a perceived "normal" self. Participants manage their drug use using active strategies, such as selective use of psychiatric medications, in the context of structural constraints, such as restricted access to mental health care, and cultural contexts that blur distinctions between "good" medicines and "bad" drugs.


Utilization of in vivo ultrafiltration in biomedical research and clinical applications.
Ultrafiltration (UF) is a filtrate selection method with a wide range of biomedical and clinical applications, including detoxification of blood in hemodialysis and peritoneal dialysis. New is, however, the use of UF as a convenient in vivo sampling method that, for example, has been used in diabetics. Ultrafiltration avoids complicated and time-consuming recovery calculations that are necessary when using in vivo microdialysis, as recoveries of low molecular weight molecules are near 100%. The subcutaneously or intravenously placed UF probes have been studied for off-line sample analysis and for continuous on-line monitoring, in a wide variety of species, including dogs, rats, pigs and humans. This review discusses the potential of in vivo UF as a continuous tissue sampling technique in clinical research areas, and in several major biomedical applications including glucose and lactate monitoring and drug kinetic studies.


Renal Oxygen Saturation as an Early Indicator of Shock in Children.
Shock is a life-threatening syndrome in which tissue perfusion and oxygen delivery are inadequate. Near-infrared spectroscopy (NIRS) has been suggested as a noninvasive tool for monitoring and detecting the state of inadequate tissue perfusion. Renal and mesenteric oximetry show decreased cardiac output earlier than systemic or global parameters of tissue oxygenation or cerebral oximetry. However, until now there has been no study on the validity of regional renal oxygen saturation (rRSO2) by NIRS for diagnosing shock in children. To analyze the validity of rRSO2 by NIRS to diagnose shock in children. This cross-sectional study was conducted in critically ill children (aged 1 month-18 years) who were admitted to the pediatric intensive care unit (PICU), from September to November 2020, consecutively. Patients were classified into two groups: shock and non-shock. The diagnosis of shock is based on clinical criteria (tachycardia, sign of hypoperfusion and decrease systolic blood pressure <P5 according to age). Measurement of rRSO2 by NIRS was performed by the doctor in charge when the patient came to PICU. The baseline rRSO2 value (%) made a receiver operating characteristic (ROC) curve and was used to find the optimal cut-off value and calculated sensitivity and specificity. We enrolled 20 critically ill patients. The baseline rRSO2 in the shock (n=10) and non-shock (n=10) groups were, 44.00±4.95 vs 78.70±4.52 (p 0.003). The optimal cutoff value of the baseline rRSO2 to predict shock is less than 58.5% with area under the curve (AUC) value is 94.4% (95% CI of 84.4-100%), p 0.001, sensitivity 90% and specificity 90% in critically ill children. The rRSO2 value by NIRS can differentiate between shock and non-shock in critically ill patients accurately.


Dehydroevodiamine attenuates calyculin A-induced tau hyperphosphorylation in rat brain slices.
This study was to investigate the effect of dehydroevodiamine (DHED) on Alzheimer's disease (AD)-like tau hyperphosphorylation induced by calyculin A (CA), an inhibitor of protein phosphatase (PP)-2A and PP-1, and the involvement of PP-2A in metabolically competent rat brain slices. Rat brain slices were pre-incubated at 33 degree centigrade in the presence (10, 100, and 200 micromol/L, respectively) or absence of DHED for 1 h. Then, CA 0.1 micromol/L was added and the slices were treated for another 2 h. Western blotting and/or immunohistochemistry were used to measure the phosphorylation level of tau and PP-2A. CA treatment could remarkably increase the immunoreactivity of pS262 and decrease the staining of Tau-1, representing tau hyperphosphorylation at Ser262 (pS262) and Ser198/ 199/202 (Tau-1, as the antibody reacts with unphosphorylated tau, therefore, decreased staining represents increased phosphorylation). Pre-incubation of the brain slices with DHED could efficiently attenuate the CA-induced tau hyperphosphorylation at the above AD-related sites. Additionally, DHED also decreased the basal phosphorylation level of tau at Ser396, although CA failed to induce tau hyperphosphorylation at this site. Furthermore, CA treatment induced an increased level of Tyr307-phosphorylated PP-2A, which represents inactivation of the phosphatase, whereas DHED arrested the elevation of the inhibitory modification of PP-2A. DHED can attenuate CA-induced tau hyperphosphorylation at multiple AD-related sites in metabolically active rat brain slices. The underlying mechanism may involve a decreased inhibitory phosphorylation of PP-2A at Tyr307.


Necroptosis is dispensable for motor neuron degeneration in a mouse model of ALS.
Motor neuron degeneration in amyotrophic lateral sclerosis (ALS) is proposed to occur by necroptosis, an inflammatory form of regulated cell death. Prior studies implicated necroptosis in ALS based on accumulation of necroptotic markers in affected tissues of patients and mouse models, and amelioration of disease in mutant superoxide dismutase 1 (SOD1G93A) mice with inhibition of the upstream necroptotic mediators, receptor interacting protein kinase 1 (RIPK1), and RIPK3. To definitively address the pathogenic role of necroptosis in ALS, we genetically ablated the critical terminal executioner of necroptosis, mixed lineage kinase domain-like protein (MLKL), in SOD1G93A mice. Disease onset, progression, and survival were not affected in SOD1G93A mice lacking MLKL. Motor neuron degeneration and activation of neuroinflammatory cells, astrocytes, and microglia, were independent of MLKL expression in SOD1G93A mice. While RIPK1 accumulation occurred in spinal cords of SOD1G93A mice in late stage disease, RIPK3 and MLKL expression levels were not detected in central nervous system tissues from normal or SOD1G93A mice at any disease stage. These findings demonstrate that necroptosis does not play an important role in motor neuron death in ALS, which may limit the potential of therapeutic targeting of necroptosis in the treatment of neurological disorders.


The substrates for self-stimulation of the lateral hypothalamus and medial prefrontal cortex: a comparison of strength-duration characteristics.
The directly activated substrates for self-stimulation of the lateral hypothalamus (LH) and medial prefrontal cortex (MPFC) were described by comparing their strength-duration characteristics. The current required to maintain a half-maximal rate of lever pressing was traded off against the pulse duration while all other stimulation parameters were kept constant. In this manner, cathodal strength-duration curves were obtained at four LH and eight MPFC sites; anodal curves were obtained at two of the LH and six of the MPFC sites. In general, the cathodal LH curves had lower rheobases than the cathodal MPFC curves and continued to descend after the MPFC curves had levelled off. At short pulse durations, the anodal curves lay above the cathodal curves, a finding more pronounced in the LH data. The two sets of curves converged at the longer pulse durations. The differences in the strength-duration curves are consistent with the notion that different directly stimulated neurons are responsible for the rewarding effects of LH and MPFC stimulation. Anatomical and physiological properties that could account for these differences are discussed.


Do autistic patients change healthcare services utilisation through the transition age? An Italian longitudinal retrospective study.
This paper aims to provide an estimate of the prevalence rate of autism spectrum disorder (ASD) in 8-year-olds in 2017 based on administrative databases and to investigate the change in healthcare service use during the healthcare transition age of 18. This research is based on a longitudinal retrospective cohort study. The data is drawn from the Italian Administrative Healthcare Database (2010-2017). We identified 5607 ASD patients; 331 ASD patients from 2012 to 2015 in the calendar year of their 18th birthday were selected and their health service utilisation during a 5-year period-ranging from 2 years preceding and succeeding their 18th year-were investigated. None. Prevalence, incidence and proportion of ASD patients receiving specific healthcare services were included in the outcome measures. Prevalence of ASD at age 8 was 5.4/1000. Global access to health and social services was lower both before and after age 18 (46.5% at 16; 68.0% at 18; 54.1% at 20). The percentage of patients receiving a neuropsychiatric consultation decreased after age 18 (30.8% at 18; 5.4% at 20). Community mental health services (CMHS) utilisation rate increased above 18 years of age. Regarding psychiatric visits, for both outpatient and CMHS, an increase was observed from 17.8% at age 18 to 25.4% at age 20. The utilisation of rehabilitation services decreased with age, dropping from 17.8% at age 16 to 1.8% at age 20. Psychiatric outpatient services remained stable across ages at about 14%. Our findings suggest that ASD patients changed clinical reference services with age from neuropsychiatric and rehabilitative services towards psychiatric and community-based services as they transitioned from paediatric to adult healthcare services.


Plant sex chromosomes: a non-degenerated Y?
Animal Y chromosomes have undergone chromosome-wide degeneration in response to a lack of recombination, and ancient Ys contain few functional genes. Recent research suggests that plant Y chromosomes may evolve differently and retain most of their ancestral genes.


Anthropometric and craniofacial patterns in mentally retarded males with emphasis on the fragile X syndrome.
Anthropometric and craniofacial profile patterns indicating the percent difference from the overall mean were developed on 34 physical parameters with 31 white, mentally retarded males (23 adults and 8 children) with the fra(X) syndrome matched for age with 31 white, mentally retarded males without a known cause of their retardation. The fra(X) syndrome males consistently showed larger dimensions for all anthropometric variables, with significant differences for height, sitting height, arm span, hand length, middle finger length, hand breadth, foot length, foot breadth, and testicular volume. A craniofacial pattern did emerge between the two groups of mentally retarded males, but with overlap of several variables. Significant differences were noted for head circumference, head breadth, lower face height, bizygomatic diameter, inner canthal distance, ear length and ear width, with the fra(X) syndrome males having larger head dimensions (head circumference, head breadth, head length, face height and lower face height), but smaller measurements for minimal frontal diameter, bizygomatic diameter, bigonial diameter, and inner canthal distance. Several significant correlations were found with the variables for both mentally retarded males with and without the fra(X) syndrome. In a combined anthropometric and craniofacial profile of 19 variables comparing 26 white fra(X) syndrome males (13 with high expression (> 30%) and 13 with low expression (< 30%), but matched for age), a relatively flat profile was observed with no significant differences for any of the variables. Generally, fra(X) syndrome males with increased fragile X chromosome expression have larger amplifications of the CGG trinucleotide repeat of the FMR-1 gene. No physical differences were detectable in our study between fra(X) males with high expression and apparently larger amplifications of the CGG trinucleotide repeats compared with those patients with low expression. Our research illustrates the use of anthropometry in identifying differences between mentally retarded males with or without the fra(X) syndrome and offers a comprehensive approach for screening males for the fra(X) syndrome and selecting those individuals for cytogenetic and/or molecular genetic testing.


Otolaryngologic Manifestations of Trisomy 13 and Trisomy 18 in Pediatric Patients.
The survival rate of patients with trisomy 13 and trisomy 18 has increased dramatically over the past two decades. We sought to comprehensively describe the otolaryngologic clinical characteristics and procedures required for these patients at our institution. We performed algorithmic identification of patients with a diagnosis of trisomy 13 and trisomy 18 for whom the otolaryngology service provided inpatient or outpatient care at our institution between the dates of February 1997 and March 2021. Of the 47 patients studied, 18 patients had a diagnosis of trisomy 13, and 29 had a diagnosis of trisomy 18. Complete trisomy was present in 44% (8/18) of trisomy 13 patients and 55% (16/29) of trisomy 18 patients. 81% of patients were living at the time of the study. About 94% (44/47) of patients required consultation with another specialty in addition to Otolaryngology. Overall, the most common diagnoses among this cohort were gastroesophageal reflux disease (47%), dysphagia (40%), otitis media (38%), and obstructive sleep apnea (34%). Nearly three-quarters (74%) of patients studied required an otolaryngologic procedure. The most common surgical procedure was tonsillectomy and/or adenoidectomy. Patients with trisomy 18 were significantly more likely to have external auditory canal stenosis and obstructive sleep apnea whereas patients with trisomy 13 were more likely to have cleft lip and palate. Patients with a diagnosis of trisomy 13 or 18 often require multidisciplinary management and the range of required care spans the breadth of otolaryngology. 4 Laryngoscope, 133:1501-1506, 2023.


Shared and distinct anatomical correlates of semantic and phonemic fluency revealed by lesion-symptom mapping in patients with ischemic stroke.
Semantic and phonemic fluency tasks are frequently used to test executive functioning, speed and attention, and access to the mental lexicon. In semantic fluency tasks, subjects are required to generate words belonging to a category (e.g., animals) within a limited time window, whereas in phonemic fluency tasks subjects have to generate words starting with a given letter. Anatomical correlates of semantic and phonemic fluency are currently assumed to overlap in left frontal structures, reflecting shared executive processes, and to be distinct in left temporal and right frontal structures, reflecting involvement of distinct memory processes and search strategies. Definite evidence for this assumption is lacking. To further establish the anatomical correlates of semantic and phonemic fluency, we applied assumption-free voxel-based and region-of-interest-based lesion-symptom mapping in 93 patients with ischemic stroke. Fluency was assessed by asking patients to name animals (semantic), and words starting with the letter N and A (phonemic). Our findings indicate that anatomical correlates of semantic and phonemic fluency overlap in the left inferior frontal gyrus and insula, reflecting shared underlying cognitive processes. Phonemic fluency additionally draws on the left rolandic operculum, which might reflect a search through phonological memory, and the middle frontal gyrus. Semantic fluency additionally draws on left medial temporal regions, probably reflecting a search through semantic memory, and the right inferior frontal gyrus, which might reflect the application of a visuospatial mental imagery strategy in semantic fluency. These findings establish shared and distinct anatomical correlates of semantic and phonemic fluency.


Caloric restriction prevents age-associated accrual of oxidative damage to mouse skeletal muscle mitochondria.
The purpose of this study was to understand the nature of the causes underlying the senescence-related decline in skeletal muscle mass and performance. Protein and lipid oxidative damage to upper hindlimb skeletal muscle mitochondria was compared between mice fed ad libitum and those restricted to 40% fewer calories--a regimen that increases life span by approximately 30-40% and attenuates the senescence-associated decrement in skeletal muscle mass and function. Oxidative damage to mitochondrial proteins, measured as amounts of protein carbonyls and loss of protein sulfhydryl content, and to mitochondrial lipids, determined as concentration of thiobarbituric acid reactive substances, significantly increased with age in the ad libitum-fed (AL) C57BL/6 mice. The rate of superoxide anion radical generation by submitochondrial particles increased whereas the activities of antioxidative enzymes superoxide dismutase, catalase, and glutathione peroxidase in muscle homogenates remained unaltered with age in the AL group. In calorically-restricted (CR) mice there was no age-associated increase in mitochondrial protein or lipid oxidative damage, or in superoxide anion radical generation. Crossover studies, involving the transfer of 18- to 22-month-old mice fed on the AL regimen to the CR regimen, and vice versa, indicated that the mitochondrial oxidative damage could not be reversed by CR or induced by AL feeding within a time frame of 6 weeks. Results of this study indicate that mitochondria in skeletal muscles accumulate significant amounts of oxidative damage during aging. Although such damage is largely irreversible, it can be prevented by restriction of caloric intake.


Laboratory practices and incidence of non-O157 shiga toxin-producing Escherichia coli infections.
We surveyed laboratories in Washington State, USA, and found that increased use of Shiga toxin assays correlated with increased reported incidence of non-O157 Shiga toxin-producing Escherichia coli (STEC) infections during 2005-2010. Despite increased assay use, only half of processed stool specimens underwent Shiga toxin testing during 2010, suggesting substantial underdetection of non-O157 STEC infections.


Comparison of perceptional nasal obstruction with rhinomanometric and acoustic rhinometric assessment.
Perception of nasal stuffiness, together with rhinomanometric and acoustic rhinometric assessments were evaluated before and after nasal and/or sinus surgery in 50 Japanese adult patients. Nasal resistance on both expiration and inspiration correlated well with the patients' perceptions (P<0.05). Nasal volumes (0-4 cm and 0-7 cm) obtained from acoustic rhinometry measurements also correlated well with perception (P<0.05). No significant relationships were found between rhinomanometric and acoustic rhinometric results. Perceptional nasal obstruction, rhinomanometric and acoustic rhinometric results (except for the sectional areas at the I-notch) improved significantly after the operations. Rhinomanometry and acoustic rhinometry are valuable methods for assessing nasal stuffiness, but we must take account of the differences between the two methods.


Effect of low-frequency stimulation on kindling induced changes in rat dentate gyrus: an ultrastructural study.
It has been shown that low-frequency stimulation (LFS) can induce anticonvulsant effects. In this study, the effect of different LFS frequencies on kindling induced behavioral and ultrastructural changes was investigated. For induction of kindled seizures in rats, stimulating and recording electrodes were implanted in perforant path and dentate gyrus, respectively. Animals were stimulated in a rapid kindling manner. Different groups of animals received LFS at different frequencies (0.5, 1 and 5 Hz) following kindling stimulations and their effects on kindling rate were determined using behavioral and ultrastructural studies. Kindling stimulations were applied for 7 days. Then, the animals were sacrificed and their dentate gyrus was sampled for ultrastructural studies under electron microscopy. All three used LFS frequencies (0.5, 1 and 5 Hz) had a significant inhibitory effect on kindling rate and decreased afterdischarge duration and the number of stimulations to achieve stage 4 and 5 seizures significantly. In addition, application of LFS prevented the increase in the post-synaptic density and induction of concave synaptic vesicles following kindling. There was no significant change between anticonvulsant effects of LFS at different frequencies. Obtained results show that LFS application can prevent the neuronal hyperexcitability by preventing the ultrastructural changes during kindling and this may be one of the mechanisms of LFS anticonvulsant effects.


Rectus femoris transfer to improve knee function of children with cerebral palsy.
Stance phase stability and swing phase clearance, prerequisites for normal ambulation, often are lost in the gait of children with cerebral palsy. Lengthening of the hamstrings usually will improve stance-phase knee extension but will not greatly alter swing-phase knee flexion. This paper presents the outcome of transfer of the distal end of the rectus femoris in conjunction with hamstrings lengthening in 37 knees, and compares it with a control group of 24 knees in which only hamstrings lengthening was done. In the first group swing-phase knee flexion was improved by 16.0 +/- 14.4 degrees, compared to 9.5 +/- 7.5 degrees in the control group, and residual knee flexion in stance was reduced to 8.9 +/- 8.1 degrees, compared to 15.1 +/- 13.8 degrees in the controls. Poor outcome in the transfer-plus-lengthening group was associated mainly with foot rotation in excess of 8 degrees internally or externally, or postoperative knee flexion in stance. Criteria for selection of cases and methods of improving surgical outcome are discussed.


Impact of the MIC of piperacillin-tazobactam on the outcome of patients with bacteremia due to extended-spectrum-β-lactamase-producing Escherichia coli.
We investigated the impact of the piperacillin-tazobactam MIC in the outcome of 39 bloodstream infections due to extended-spectrum-β-lactamase-producing Escherichia coli. All 11 patients with urinary tract infections survived, irrespective of the MIC. For other sources, 30-day mortality was lower for isolates with a MIC of ≤ 2 mg/liter than for isolates with a higher MIC (0% versus 41.1%; P = 0.02).


FAD-mediated enzymatic conversion of NAD+ to NADH: application to chiral synthesis of L-lactate.
Electroenzymatic reduction of NAD+ to NADH for subsequent use in enzymatic synthesis has been carried out at carbon electrodes bearing lipoamide dehydrogenase (LiDH) immobilized under a Nafion film. The self-mediated electron transfer was made possible by an excess of flavin adenine dinucleotide (FAD) entrapped together with LiDH. Results were compared to those obtained with a similar electrode containing both LiDH and a polymeric form of FAD (pFAD) prepared by anodic polymerization of FAD.


Sexually transmitted diseases in children and adolescents.
Sexually transmitted diseases have an impact on children's health at three times in their development: during pregnancy or delivery (through maternal-neonatal transmission), during prepubertal childhood (through sexual transmission), and during adolescence (through sexual transmission). These phases must be considered distinctly by the physician, since not only the type of infection but also the management and long-term implications of the disease vary with the time period in which it was acquired.


Understanding group leader affect as a factor for career education programming.
The purpose of this study was to explore how group leader affect impacted facilitation of a career education program. Through a case study design, data were gathered via focus groups and blog posts from 16 program staff members. Five major themes were derived, highlighting group leader affect and experience: Emotions During Intervention, Flexibility, Student Engagement and Connections, Support from Program Staff, and School Culture. The findings encouraged career educators to remain flexible when delivering programming, to consider regular evaluations of affect throughout career programming, and to acknowledge the reciprocity of engagement, affect, and programmatic buy-in between facilitators and participants.


Novel substituted isoxazole FXR agonists with cyclopropyl, hydroxycyclobutyl and hydroxyazetidinyl linkers: Understanding and improving key determinants of pharmacological properties.
Several isoxazole-containing series of FXR agonists have been published over the last 15years, subsequent to the prototypical amphiphilic 'hammerhead'-type structure that was originally laid out by GW4064, the first potent synthetic FXR agonist. A set of novel compounds where the hammerhead is connected to the terminal carboxylic acid-bearing aryl or heteroaryl moiety by either a cyclopropyl, a hydroxycyclobutyl or a hydroxyazetidinyl linker was synthesized in order to improve upon the ADME properties of such isoxazoles. The resulting compounds all demonstrated high potencies at the target receptor FXR but with considerable differences in their physicochemical and in vivo profiles. The structure-activity relationships for key chemical features that have a major impact on the in vivo pharmacology of this series are discussed.


Determination of atomoxetine metabolites in human plasma by liquid chromatography/tandem mass spectrometry and its application to a pharmacokinetic study.
4-Hydroxyatomoxetine (4-HAT) and N-desmethylatomoxetine (N-DAT) are major metabolites of atomoxetine, a potent and selective inhibitor of the presynaptic norepinephrine transporter that is used for the treatment of attention deficit/hyperactivity disorder. The pharmacological activity of 4-HAT is similar to that of atomoxetine. We have developed and validated a simple, rapid and sensitive liquid chromatography analytical method with tandem mass spectrometry (LC-MS/MS) for the determination of 4-HAT and N-DAT in human plasma. After liquid-liquid extraction with methyl t-butyl ether, chromatographic separation of analytes was performed using a reversed-phase Luna C(18) column (2.0mm×100mm, 3μm particles) with a mobile phase of 10mM ammonium formate buffer (pH 3.5)-methanol (10:90, v/v) and quantified by MS/MS detection in ESI positive ion mode. The flow rate of the mobile phase was 250μL/min and the retention times of 4-HAT, N-DAT and internal standard (IS, metoprolol) were 0.9, 1.0 and 1.0min, respectively. The calibration curves were linear over the range of 0.05-20ng/mL for 4-HAT and 0.1-20ng/mL for N-DAT. The lower limits of quantification, using 200μL human plasma, were 0.05 and 0.1ng/mL for 4-HAT and N-DAT, respectively. The mean accuracy and precision for intra- and inter-day validation of 4-HAT and N-DAT were both within the acceptable limits. This LC-MS/MS method showed improved sensitivity for quantification of the two main metabolites of atomoxetine in human plasma compared with previously described analytical methods. The validated method was successfully applied to a pharmacokinetic study in humans.


The stem region of group A transferase is crucial for its specificity, and its alteration promotes heterologous Forssman synthase activity.
Some stem region mutants of human blood group A transferase (hAT) possess Forssman synthase (FS) activity, but very little is known about the mechanisms responsible for this enzymatic crosstalk. We performed confocal microscopy and image analysis to determine whether different intra-Golgi localization was accountable for this acquired activity. We also performed structural modeling and mutational and normal mode analyses. We introduced new mutations in the stem region and tested its FS and AT activities. No differences in subcellular localization were found between hAT and FS-positive mutants. AlphaFold models of hAT and mFS (mouse Forssman synthase) showed that the hAT stem region has a tether-like stem region, while in mFS, it encircles its catalytic domain. In silico analysis of FS-positive mutants indicated that stem region mutations induced structural changes, decreasing interatomic interactions and mobility of hAT that correlated with FS activity. Several additional mutations introduced in that region also bestowed FS activity without altering the AT activity: hAT 37-55 aa substitution by mFS 34-52, 37-55 aa deletion, and missense mutations: S46P, Q278Y, and Q286M. Stem region structure, mobility, and interactions are crucial for hAT specificity. Moreover, stem region mutations can lead to heterologous Forssman activity without changes in the catalytic machinery.


Donor lymphocyte infusion-mediated graft-versus-leukemia effects in mixed chimeras established with a nonmyeloablative conditioning regimen: extinction of graft-versus-leukemia effects after conversion to full donor chimerism.
We investigated an approach to separating graft-versus-lymphoma (GVL) effects from graft-versus-host disease (GVHD) in mice receiving a nonmyeloablative conditioning regimen allowing establishment of mixed hematopoietic chimerism. We evaluated the ability of donor lymphocyte infusions (DLI) to mediate GVL effects without GVHD in mixed chimeras prepared with cyclophosphamide, anti-T-cell antibodies, and thymic irradiation. To examine the fate of GVH-reactive donor CD8+ T cells, we used the 2C T-cell receptor (TCR) transgenic mouse strain, which carries an Ld-specific transgenic TCR on the B6 background. Administration of DLI on day 35 post-BMT led to conversion from mixed to full donor chimerism and mediated a powerful GVL effect with complete protection (100% survival) against mortality induced by a host-type lymphoma (EL4) administered 7 days later (100% mortality in non-DLI controls; P<0.001). No GVHD occurred in DLI recipients. Rechallenging the surviving DLI recipients, which had converted to full chimerism, with the same tumor dose 17 weeks later led to rapid tumor mortality. Long-term DLI recipients had anti-host proliferative responses, but not CTL responses in vitro. When given as DLI together with wild-type spleen cells, marked expansion of GVH-reactive 2C CD8+ T cells was observed on day 10, followed by a marked decline in their numbers by week 10 post-DLI. Nonmyeloablative induction of mixed chimerism followed by administration of DLI can mediate powerful GVL effects. The late loss of DLI-mediated GVL effects may reflect the eventual loss of donor-derived GVH-reactive CTL, which occurs in association with conversion to full donor chimerism.


Vitamin A transport between retina and pigment epithelium--an interstitial protein carrying endogenous retinol (interstitial retinol-binding protein).
We have demonstrated and partially characterized an interstitial retinol-binding protein (IRBP) confined to bovine interphotoreceptor matrix (IPM). The native protein is a concanavalin A-binding glycoprotein with a mol. wt of 260 k as measured by gel-filtration and size-exclusion high-performance liquid chromatography. On SDS-gels, its mol. wt is 140-145 k. Since the protein is glycosylated, this value is probably too high. Hence, the native protein may be a dimer consisting of two identical subunits. The endogenous ligand has been analyzed by high-performance liquid chromatography--it consists mainly of all-trans retinol. Occasionally, retinal and 11-cis retinol are also associated with it. The amount of retinol bound to IRBP increases when the eyes are illuminated. The total binding capacity was estimated to represent 4-5% of the retinol released from a total rhodopsin bleach. We have established that, like serum retinol-binding protein, IRBP can be also bind retinoic acid, although it has not been established that retinoic acid is an endogenous ligand. The fluorescence emission lambda max for IRBP with its native ligand is at 470 nm and the excitation lambda max for this fluorescence is at 333 nm. Other retinoid carriers in the interphotoreceptor matrix have molecular weights of about 15 and 33 k. These probably correspond to cellular retinol- and retinal-binding proteins, respectively. Since both proteins have been identified in the pigment epithelium and retina cytosols, their presence in the IPM could be a result of cell damage. We conclude that interstitial retinol-binding protein is the best candidate for a transport protein carrying retinol between the rod outer segments and the pigment epithelium.


Cold Hands or Feet: Is It Raynaud's or Not?
Raynaud's phenomenon is an exaggerated response to cold stimuli that may be primary or secondary. The diagnosis relies on patient history and physical examination to distinguish RP from other vasomotor dysfunction (e.g. acrocyanosis, pernio, small fiber neuropathy with vasomotor symptoms, and complex regional pain syndrome). Achenbach syndrome, or spontaneous venous hemorrhage, may also be mistaken for RP but is a self-limiting phenomenon. Laboratory evaluation and vascular diagnostic testing may identify SRP causes. Regardless of etiology, treatment includes warming with trigger avoidance, and consideration of vasodilators (eg. calcium channel, alpha-1 blockers). SRP with digital ulceration may require PDE5i, endothelin-1 receptor blockers, and prostanoids. Refractory cases may require pneumatic arterial pumps, botulinum toxin administration, or surgical sympathectomy.


Two intertwined methylation activities of the MmeI restriction-modification class-IIS system from Methylophilus methylotrophus.
The class-IIS restriction endonuclease, R.MmeI, was isolated from Methylophilus methylotrophus. It was originally described as a monomeric enzyme, with the native Mr 105000+/-7000, which did not cleave DNA efficiently [Boyd et al. (1986) Nucleic Acids Res. 14, 5255-5274; Tucholski et al. (1995) Gene 157, 87-92]. However, it was discovered that R.MmeI endonucleolytic activity is enhanced by S-adenosyl-l-methionine (AdoMet) and sinefungin, an analogue of AdoMet. Surprisingly, the purified R.MmeI endonuclease was found to have a second enzymatic activity, namely methylation of the adenine residue to N6-methyladenine in the top strand of the MmeI-recognition sequence, 5'-TCCR*AC-3' (*A=meA. The R.MmeI methylating activity requires AdoMet and is increased in the presence of several divalent cations, 20-fold by Mg2+ or Ca2+, and less by Mn2+, Zn2+ and Co2+; however, methylation is inhibited entirely by sinefungin, at concentrations above 9microM. The latter observation shows that the enhancing effect of AdoMet or sinefungin on the DNA cleavage was not related to the process of DNA methylation. Furthermore, a second component of the MmeI restriction-modification system, a M.MmeI methyltransferase, was isolated and purified. The M.MmeI protein was found to have an Mr of 48000+/-2000 (under denaturing conditions) and to methylate both adenine residues (*A) in the MmeI-recognition sequence 5'-TCCR*AC-3'/3'-*AGGYTG-5'. Methylation of the top strand does not inhibit the DNA cleavage by R.MmeI, whereas methylation of both DNA strands blocks the cleavage process.


Stress analysis of the human temporomandibular joint.
Stress analysis of the human temporomandibular joint (TMJ) consisting of mandibular disc, condyle and fossa-eminence complex during normal sagittal jaw closure was performed using non-linear finite element analysis (FEA). The geometry of the TMJ was obtained from magnetic resonance imaging (MRI). The tissue proportion was measured from a cadaver TMJ. Contact surfaces were defined to represent the interaction between the mandibular disc and the condyle, and between the mandibular disc and the fossa-eminence complex so that finite sliding was allowed between contact bodies. Stresses in the TMJ components (disc, condyle and fossa-eminence complex), and forces in capsular ligaments were obtained. The results demonstrated that, with the given condylar displacement, the stress in the condyle was dominantly compressive and in the fossa-eminence complex was dominantly tensile. The cancellous bone was shielded by the shell shaped cortical bone from the external loading. The results illustrate the stress distributions in the TMJ during a normal jaw closure.


Membrane properties of rat medial vestibular nucleus neurons in vivo.
In our previous study using the whole-cell patch clamp technique combined with reverse transcription-polymerase chain reaction analysis in rat brainstem slices, we demonstrated that the classification of neurons in the medial vestibular nucleus (MVN) based on three membrane properties detected as voltage response properties to depolarizing and hyperpolarizing current pulses, namely afterhyperpolarization (AHP) profiles, firing patterns, and response patterns to hyperpolarizing current pulses, is useful for clarifying the relationship between membrane properties and cellular markers for excitatory and inhibitory neurons. These membrane properties characterized in vitro, however, have not been ascertained in vivo. To address this issue, we applied the whole-cell patch clamp recording method to in vivo preparations of young adult rats and investigated voltage responses to depolarizing and hyperpolarizing current pulses. We found three AHP profiles, three firing patterns, and three response patterns to hyperpolarizing current pulses in MVN neurons in vivo that were characterized in our previous in vitro study. The MVN neuronal populations classified on the basis of the three membrane properties in vivo were comparable to those obtained in vitro. This finding indicates that the classification of MVN neurons based on the three membrane properties is applicable to in vivo preparations.


P/Q-type voltage-gated calcium channel antibodies in paraneoplastic disorders of the central nervous system.
Whether P/Q-type voltage-gated calcium channel (VGCC) antibodies are present in the serum of patients with paraneoplastic syndromes other than the Lambert-Eaton myasthenic syndrome (LEMS) and tumors other than small-cell lung cancer (SCLC) is controversial. Using a commercially available radioimmunoprecipitation assay kit, we examined the sera of 93 patients with paraneoplastic syndromes of the central nervous system (CNS), including 27 patients with paraneoplastic cerebellar degeneration (PCD) associated with tumors other than SCLC and 66 SCLC patients with paraneoplastic encephalomyelitis and sensory neuronopathy (PEM/SN). All PCD sera from patients with tumors other than SCLC were negative for P/Q-type VGCC antibodies. Eight of 66 (12%) SCLC patients with PEM/SN had P/Q-type VGCC antibodies; 4 had LEMS and the other 4 had no symptoms of LEMS or they were overlooked and, therefore, not examined electrophysiologically. In patients with paraneoplastic syndromes of the CNS, the detection of P/Q-type VGCC antibodies supports the diagnosis of LEMS; in our series, only 6% of patients with SCLC and PEM/SN may have had a false positive antibody result, or undiagnosed LEMS.


Carbamazepine promotes specific stimuli-induced NLRP3 inflammasome activation and causes idiosyncratic liver injury in mice.
The occurrence of idiosyncratic drug-induced liver injury (IDILI) is a leading cause of post-marketing safety warnings and withdrawals of drugs. Carbamazepine (CBZ), widely used as an antiepileptic agent, could cause rare but severe idiosyncratic liver injury in humans. Although recent studies have shown that inflammasome is implicated in CBZ-induced hepatocellular injury in vitro, the precise pathogenesis of hepatotoxicity remains largely unexplored. Here we report that CBZ causes idiosyncratic liver injury through promoting specific stimuli-induced NLRP3 inflammasome activation. CBZ (40 μM) enhances NLRP3 inflammasome activation triggered by adenosine triphosphate (ATP) or nigericin, rather than SiO2, monosodium urate crystal or intracellular lipopolysaccharide (LPS). In addition, CBZ has no effect on NLRC4 or AIM2 inflammasome activation. Mechanistically, synergistic induction of mitochondrial reactive oxygen species (mtROS) is a crucial event in the enhancement effect of CBZ on ATP- or nigericin-induced NLRP3 inflammasome activation. Moreover, the "C=C" on the seven-membered ring and "C=O" on the nitrogen of CBZ may be contribute to NLRP3 inflammasome hyperactivation and hepatotoxicity. Notably, in vivo data indicate that CBZ (50 mg/kg) causes liver injury in an LPS (2 mg/kg)-mediated susceptibility mouse model of IDILI, accompanied by an increase in caspase-1 activity and IL-1β production, whereas the combination of CBZ and LPS does not exhibit the effect in NLRP3-knockout mice. In conclusion, CBZ specifically promotes ATP- or nigericin-induced NLRP3 inflammasome activation and causes idiosyncratic liver injury. Our findings also suggest that CBZ may be avoided in patients with NLRP3 inflammasome activation-related diseases that are triggered by ATP or nigericin, which may be risk factors for IDILI.


An update on imaging of tarsal tunnel syndrome.
Tarsal tunnel syndrome (TTS) is an entrapment neuropathy of the tibial nerve (TN) within the tarsal tunnel (TT) at the level of the tibio-talar and/or talo-calcaneal joints. Making a diagnosis of TTS can be challenging, especially when symptoms overlap with other conditions and electrophysiological studies lack specificity. Imaging, in particular MRI, can help identify causative factors in individuals with suspected TTS and help aid surgical management. In this article, we review the anatomy of the TT, the diagnosis of TTS, aetiological factors implicated in TTS and imaging findings, with an emphasis on MRI.


What are LGBTQI+ parental experiences of healthcare support and decision-making regarding infant feeding options? A grounded theory study.
To explore LGBTQI+ parental experiences regarding their interactions with healthcare professionals as a resource for feeding options during the prenatal-to-neonatal period. This single-center, interview-based qualitative study of LGBTQI+ parents utilized grounded theory to identify and verify emergent themes and subthemes. We developed a conceptual framework to illustrate relationships among themes and subthemes, as well as opportunities for healthcare professionals and families to improve LGBTQI+ parental support. Thematic saturation was attained after interviewing 21 parents from 12 families. Analyses revealed four main themes representing opportunities for improvement: education, continuity of care, parental engagement and open communication. Personal and interpersonal factors influenced parental experiences and decisions, which shaped ultimate feeding outcomes. LGBTQI+ parents revealed challenges of establishing feeding practices that best aligned with their values and goals. Recognizing these factors can help healthcare professionals improve their counseling, engagement and support of LGBTQI+ parents.


Reexamination ofFusarium graminearum NRRL-13820and NRRL-13852 reported as type A trichothecene producers.
NRRL-13820 and NRRL-13852 are reported to be two atypicalFusarium graminearum strains type A trichothecene producers [T-2 toxin (T-2) and diacetoxy-scirpenol (DAS)]. These two strains were reexamined by morphological, genetical (DNA / DNA relatedness) and toxicological techniques and compared with 28 wildF graminearum isolates obtained from corn in Italy and the USA. The isolate NRRL-13820 was morphologically confirmed as a typical isolate ofF graminearum, while the isolate NRRL-13852 showed some peculiar characteristics. Nuclear DNA comparison between NRRL-13820 and NRRL-13852 displayed 49% similarity and showed 94 % and 44 % relatedness, respectively, when compared withF graminearum NRRL-13833, which is a well assessed type B trichothecene producer [deoxynivalenol (DON) and 15-acetyldeoxynivalenol]. NRRL-13820, NRRL-13852, and NRRL-13833, as well as the 28 wild isolates, were not able to synthesize T-2, HT-2 nor DAS. Finally, NRRL-13820 and NRRL-13833, but not NRRL-13852, were able to produce DON (120 and 40/μg/g, respectively). The data support the concept that the production of examined type A trichothecenes is very rare inF graminearum.


A Saccharomyces cerevisiae cell-based quantitative beta-galactosidase assay compatible with robotic handling and high-throughput screening.
Reporter-gene assays that employ the Escherichia coli lacZ gene are ubiquitously employed in biological research. However, we were not able to readily identify a quantitative method that worked reliably with yeast (Saccharomyces cerevisiae) cells and that was compatible with high-throughput screening and robotic liquid handling tools. We have therefore adapted a commercially available assay employing a 6-O-beta-galactopyranosyl-luciferin substrate to provide the required sensitivity with minimal sample handling times. Our assay uses only one-tenth of the reagents suggested by the reagent manufacturer (Promega) for equivalent assays with mammalian cell cultures and produces rapid, sensitive and reproducible analysis with as little as 1 microl yeast cell culture and with < 100 cells. We demonstrate that the assay is compatible with yeast strains generated by the systematic yeast deletion project and functions equally well with genomically integrated or plasmid-encoded lacZ reporters and with cells grown in complex or defined media. The high-sensitivity, miniaturized format reduced sample handling required will make this assay useful for a wide range of applications.