Optimization and performance evaluation of double-stranded probe in real-time PCR.
TaqMan probe-based real-time PCR (qPCR/RT-qPCR) has been widely used in various fields because of its high sensitivity and specificity. However, TaqMan probes are associated with a relatively higher background signal, and hence negatively affect the detection results. Double-stranded probes (DSPs) were designed for the high sensitive detection of hepatitis B virus (HBV) DNA and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA using qPCR/RT-qPCR. Old DSPs (ODSPs) consist of different lengths of positive and negative strands with complementary oligonucleotides. We systematically optimized ODSPs length, the free energy of hybridization (ΔG) between complementary oligonucleotides, and the length of sticky ends, and the novel DSPs performances were evaluated in comparison with other types of probes. By using similar length positive and negative strands, controlling ΔG between complementary oligonucleotides to approximately -30 kcal/mol, and maintaining the sticky end length at 4-6 nt, the analytical performances of DSPs were significantly improved. Compared with other types of probes, DSPs are advantageous in fluorescence signal intensity and sensitivity. DSPs can further improve the detection sensitivity and the detection rate of low-concentration samples in molecular diagnosis.


A Simulation Framework for Benchmarking EEG-Based Brain Connectivity Estimation Methodologies.
Due to its high temporal resolution, electroencephalography (EEG) is widely used to study functional and effective brain connectivity. Yet, there is currently a mismatch between the vastness of studies conducted and the degree to which the employed analyses are theoretically understood and empirically validated. We here provide a simulation framework that enables researchers to test their analysis pipelines on realistic pseudo-EEG data. We construct a minimal example of brain interaction, which we propose as a benchmark for assessing a methodology's general eligibility for EEG-based connectivity estimation. We envision that this benchmark be extended in a collaborative effort to validate methods in more complex scenarios. Quantitative metrics are defined to assess a method's performance in terms of source localization, connectivity detection and directionality estimation. All data and code needed for generating pseudo-EEG data, conducting source reconstruction and connectivity estimation using baseline methods from the literature, evaluating performance metrics, as well as plotting results, are made publicly available. While this article covers only EEG modeling, we will also provide a magnetoencephalography version of our framework online.


Effect of glucose on fermentation heat in sheep rumen fluid in vitro.
1. Heat production rate (H) of rumen fluid was measured in a direct calorimeter. Basal H of samples of 15 ml rumen fluid mixed with 45 ml buffer was 0.4 mW/ml rumen fluid. 2. Addition of glucose (0.4-6.4 mg/sample) was followed by a dose-dependent increase in H. Maximal H was 1.1 mW/ml and lasted up to 5 min, returning thereafter to the basal level. 3. Expression of fermentation heat (Hf; kJ/mol substrate added) against glucose dose indicated an asymptotic dose response. 4. Maximal Hf (at infinite dilution) agreed with stoichiometric calculations whereas minimal Hf suggested a partial fermentation of the substrate at a high-glucose dose in the rumen environment.


Biological and clinical effects of anti-TNFalpha treatment.
Tumor necrosis factor alpha (TNFalpha) is implicated in the pathogenesis of many chronic inflammatory diseases such as rheumatoid arthritis (RA), psoriasis and psoriatic arthritis (PsA), ankylosing spondylitis (AS), Crohn's disease, ulcerative colitis and uveitis. The availability of new pharmacological agents (infliximab, etanercept, adalimumab), able to selectively block the TNFalpha, has recently offered new opportunity for the treatment of these diseases. TNFalpha antagonists are different in the mechanism of action and are all effective agents in the treatment of RA and several chronic inflammatory diseases as a large number of controlled clinical trials have shown. Among biological effects of TNFalpha antagonists, the production of autoantibodies has been emphasized. This phenomenon is not correlated with the disease background, since anti-nuclear antibodies (ANA) and anti-double stranded-DNA antibodies (anti-dsDNA) induction is observed in RA as well as in spondyloarthritis (SpA) patients. Nonetheless, recent studies had reported a significant reduction in the serum titre of rheumatoid factor (RF) and/or anti-cyclic citrullinated peptide antibodies (anti-CCP) during anti-TNFalpha therapy. The TNFalpha antagonists represent a significant advance in the therapy of active RA and other chronic inflammatory diseases. However, they have distinct biological, clinical, and pharmacological properties that must be considered when selecting a drug for therapy.


A novel exposure maneuver in laparoscopic right hepatectomy.
Laparoscopic access to the posterosuperior and lateral parts of the right liver is difficult for blocked and deep surgical situations. We invented a novel water bag device (WBD) to improve the exposure of the right liver. Eighteen consecutive patients with lesions isolated to the posterosuperior or lateral right liver were included in our research. They underwent laparoscopic right hepatectomy with the help of the device and were compared with previous similar laparoscopic cases of our operating surgeon. The device was successfully employed without related complications and provided enhanced and stable surgical exposure. All patients were operated on without the need for blood transfusions or laparotomy conversion. The median operation time and estimated blood loss were 227 minutes (range, 114-568) and 88 mL (range, 25-250), respectively. In all cases, tumor-free surgical margins were confirmed and no major complications were observed. The results were better than those in previous similar laparoscopic cases. The WBD is safe and effective for laparoscopic exposure when lesions are located in the posterosuperior and lateral parts of the right liver. With the help of the device, laparoscopic right liver resection is easier to perform instead of undergoing open hepatectomy.


A simple experimental approach to the determination of carrier transport parameters for unlabeled substrate analogs.
A method is described by which affinities and transport rates for unlabeled substrate analogs are readily determined, and which is based on the effect of an unlabeled analog upon the rate of transport of a labeled substrate present at a low concentration on the trans side of the membrane. The procedure is widely applicable since it does not depend on assumptions about rrate-limiting steps and holds for both active and non-active systems. Here it is applied in an experimental study of the facilitated diffusion system for choline in erythrocytes, and it is shown that the transport parameters for a test substrate obtained by this method are the same as those found when the transport of the substrate is followed directly.


Persistent estrus rat models of polycystic ovary disease: an update.
To critically review published articles on polycystic ovary (PCO) disease in rat models, with a focus on delineating its pathophysiology. Review of the English-language literature published from 1966 to March 2005 was performed through PubMed search. Keywords or phrases used were persistent estrus, chronic anovulation, polycystic ovary, polycystic ovary disease, and polycystic ovary syndrome. Articles were also located via bibliographies of published literature. University Health Sciences Center. Articles on persistent estrus and PCO in rats were selected and reviewed regarding the methods for induction of PCO disease. Changes in the reproductive cycle, ovarian morphology, hormonal parameters, and factors associated with the development of PCO disease in rat models were analyzed. Principal methods for inducing PCO in the rat include exposure to constant light, anterior hypothalamic and amygdaloidal lesions, and the use of androgens, estrogens, antiprogestin, and mifepristone. The validated rat PCO models provide useful information on morphologic and hormonal disturbances in the pathogenesis of chronic anovulation in this condition. These studies have aimed to replicate the morphologic and hormonal characteristics observed in the human PCO syndrome. The implications of these studies to human condition are discussed.


Complete nucleotide sequence of a Japanese isolate of Chrysanthemum virus B (genus Carlavirus).
The complete nucleotide sequence of a Chrysanthemum virus B isolate from Japan (CVB-S) has been determined. The genomic RNA of CVB-S is 8,990 nucleotides long, excluding the poly(A) tail and, like that of other carlaviruses, contains six open reading frames (ORFs). Multiple alignment and phylogenetic analyses indicated that the phylogenetic relationship among members of the genus Carlavirus is very diverse, with phlox virus S being the closest relative of CVB. In aphid transmission tests, CVB-S was transmitted at a very low rate by Aphis gossypii, a new vector of the virus.


Herpes simplex keratitis following Smart Pulse Technology assisted transepithelial photorefractive keratectomy: a case report.
Herpes simplex keratitis (HSK) is a rare and sight-threatening complication following refractive surgery. SmartSurfACE surgery is the result of combining transepithelial photorefractive keratectomy (trans-PRK) with Smart Pulse Technology (SPT) to diminish surface irregularities of the residual stromal bed after surgery with less pain, faster re-epithelialization, and better postoperative visual acuity. In this article, we report the first case of HSK following SmartSurf ACE without history of herpetic eye disease. A 21-year-old woman underwent bilateral SmartSurfACE without history of clinical herpetic infection, active eye disease, or systemic disease. Mild superficial punctate keratitis occurred on the tenth postoperative day. The condition was not improved by ophthalmic drugs of anti-inflammation or epithelial healings. Dendritic corneal ulcer appeared within one month, which is the commonly recognized clinical manifestation of herpes simplex keratitis. The patient was managed with topical and systemic antiviral agents. After nine days of antiviral therapy, the lesion healed up, remaining mild stromal scarring in both eyes ultimately. Herpes simplex keratitis is a rare but sight-threatening complication following refractive surgery. For the ocular irritation symptoms of postoperative patients, we should consider the possibility of HSK and give timely treatment.


Genomic determinants of cancer immunotherapy.
Cancer immunotherapies - including therapeutic vaccines, adoptive cell transfer, oncolytic viruses, and immune checkpoint blockade - yield durable responses in many cancer types, but understanding of predictors of response is incomplete. Genomic characterization of human cancers has already contributed to the success of targeted therapies; in cancer immunotherapy, identification of tumor-specific antigens through whole-exome sequencing may be key to designing individualized, highly immunogenic therapeutic vaccines. Additionally, pre-treatment tumor mutational and gene expression signatures can predict which patients are most likely to benefit from cancer immunotherapy. Continued work in harnessing genomic, transcriptomic, and immunological data from clinical cohorts of immunotherapy-treated patients will bring the promises of precision medicine to immuno-oncology.


Apoptosis induction activity of polysaccharide from Lentinus edodes in H22-bearing mice through ROS-mediated mitochondrial pathway and inhibition of tubulin polymerization.
Lentinus edodes is a medicinal mushroom widely used in Asian countries for protecting people against some types of cancer and other diseases. The objective of the present study was to investigate the direct antiproliferation activity and the antitumor mechanisms of water-extracted polysaccharide (WEP1) purified from L. edodes in H22 cells and H22-bearing mice. The extraction, isolation, purification, and structure determination of the water-soluted L. edodes polysaccharide WEP1 were performed. The growth inhibitory effects of WEP1 on H22 cells and H22-bearing mice were determined by 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) method and animal studies. Flow cytometry, scanning electron microscopy, and laser scanning confocal microscopy were used to observe the morphological characteristics of apoptotic cells. The levels of intracellular reactive oxygen species (ROS) were detected by flow cytometry using 2',7'-dichlorofluorescein-3',6'-diacetate (DCFH-DA). Western blot was used to determine the expressions of cell cycle proteins and apoptosis-related proteins. Results showed that WEP1 with a molecular weight of 662.1 kDa exhibited direct antiproliferation activity on H22 cells in a dose-dependent manner. In vivo, WEP1 significantly inhibited the growth of tumor at different doses (50, 100, and 200 mg/kg) and the inhibition rates were 28.27, 35.17, and 51.72%, respectively. Furthermore, morphological changes of apoptosis and ROS overproduction were observed in H22 cells by WEP1 treatment. Cell cycle assay and western blot analyses indicated that the apoptosis induction activity of WEP1 was associated with arresting cell cycle at G2/M phase and activating mitochondrial-apoptotic pathway. Besides, WEP1 disrupted the microtubule network accompanied by alteration of cellular morphology. Results suggested that the antitumor mechanisms of WEP1 might be related to arresting cell cycle at G2/M phase, inhibiting tubulin polymerization and inducing mitochondrial apoptosis. Therefore, WEP1 possibly could be used as a promising functional food for preventing or treating liver cancer.


Trending autoregulatory indices during treatment for traumatic brain injury.
Our goal is to use automatic data monitoring for reliable prediction of episodes of intracranial hypertension in patients with traumatic brain injury. Here we test the validity of our method on retrospective patient data. We developed the Continuous Hemodynamic Autoregulatory Monitor (CHARM), that siphons and stores signals from existing monitors in the surgical intensive care unit (SICU), efficiently compresses them, and standardizes the search for statistical relationships between any proposed index and adverse events. CHARM uses an automated event detector to reliably locate episodes of elevated intracranial pressure (ICP), while eliminating artifacts within retrospective patient data. A graphical user interface allowed data scanning, selection of criteria for events, and calculating indices. The pressure reactivity index (PRx), defined as the least square linear regression slope of intracranial pressure versus arterial BP, was calculated for a single case that spanned 259 h. CHARM collected continuous records of ABP, ICP, ECG, SpO2, and ventilation from 29 patients with TBI over an 18-month period. Analysis of a single patient showed that PRx data distribution in the single hours immediately prior to all 16 intracranial hypertensive events, significantly differed from that in the 243 h that did not precede such events (p < 0.0001). The PRx index, however, lacked sufficient resolution as a real-time predictor of IH in this patient. CHARM streamlines the search for reliable predictors of intracranial hypertension. We report statistical evidence supporting the predictive potential of the pressure reactivity index.


The implant thread as a retention element in cortical bone: the effect of thread size and thread profile: a finite element study.
Assuming that high stress peaks in the bone can trigger bone resorption a screw-shaped bone implant should be given such a design that the peak stresses arising in the bone, as a result of a certain load, are minimized. Using idealized assumptions the aim of the study was to analyse the effect of variations of the size and the profile of the thread of an axially loaded, screw-shaped, bone implant upon the magnitude of the stress peaks in cortical bone. The investigation was performed by means of axisymmetric finite element analysis. It was found that the shape of the thread profile has a profound effect upon the magnitude of the stresses in the bone and that very small threads of a favourable profile can be quite effective.


Metabolic interaction between toluene and ethanol in rabbits.
The metabolic interaction of toluene and ethanol was studied in male rabbits having received ethanol (26.0 mmol/kg PO), toluene (5.4 mmol/kg PO) or both. Compared with ethanol alone, toluene given 2 h after ethanol caused a significantly higher and more prolonged concentration of blood alcohol. A similar trend of blood alcohol was observed at the later stage with toluene given prior to ethanol. On the other hand, with simultaneous doses of the two substances, the blood toluene concentration was higher for the first 15-30 min than the ethanol control and the urinary excretion of hippuric acid, a main metabolite of toluene, was markedly decreased for the first 2 h. The blood ethanol in this group, on the contrary, was reduced until 1 h after administration. These results indicate that toluene and ethanol act reciprocally as competitive inhibitors in their metabolism after single administrations.


Therapeutic drug monitoring of immunosuppressant drugs in clinical practice.
Therapeutic drug monitoring (TDM) is essential to maintain the efficacy of many immunosuppressant drugs while minimizing their toxicity. TDM has become more refined with the development of new monitoring techniques and more specific assays. This article summarizes current data on TDM of the following immunosuppressant drugs used in organ transplantation: cyclosporine, tacrolimus, sirolimus, everolimus, and mycophenolate mofetil. Published data were identified by a MEDLINE search of the English-language literature through March 2001 using the terms therapeutic drug monitoring, cyclosporine, tacrolimus, sirolimus, everolimus, and mycophenolate mofetil. Relevant conference abstracts were also included. TDM of cyclosporine has been well studied, and recent findings indicate that monitoring of drug levels 2 hours after dosing is a more sensitive predictor of outcome than trough (C0) monitoring. C0 levels are being used more widely in TDM of tacrolimus; however, the relationship between C0 and area under the curve has varied widely in clinical trials, with correlations ranging from 0.11 to 0.92. The use of TDM of sirolimus, everolimus, and mycophenolate mofetil is evolving rapidly. TDM of immunosuppressant drugs that have a narrow therapeutic index is an increasingly useful tool for minimizing drug toxicity while maximizing prevention of graft loss and organ rejection.


Pooling of Upper Respiratory Specimens Using a SARS-CoV-2 Real-time RT-PCR Assay Authorized for Emergency Use in Low-Prevalence Populations for High-Throughput Testing.
Nucleic acid amplification testing is a critical tool for addressing the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Specimen pooling can increase throughput and conserve testing resources but requires validation to ensure that reduced sensitivity does not increase the false-negative rate. We evaluated the performance of a real-time reverse transcription polymerase chain reaction (RT-PCR) test authorized by the US Food and Drug Administration (FDA) for emergency use for pooled testing of upper respiratory specimens. Positive specimens were selected from 3 prevalence groups, 1%-3%, >3%-6%, and >6%-10%. Positive percent agreement (PPA) was assessed by pooling single-positive specimens with 3 negative specimens; performance was assessed using Passing-Bablok regression. Additionally, we assessed the distributions of RT-PCR cycle threshold (Ct) values for 3091 positive specimens. PPA was 100% for the 101 pooled specimens. There was a linear relationship between Ct values for pooled and single-tested specimens (r = 0.96-0.99; slope ≈ 1). The mean pooled Ct shifts at 40 cycles were 2.38 and 1.90, respectively, for the N1 and N3 targets. The median Cts for 3091 positive specimens were 25.9 (N1) and 24.7 (N3). The percentage of positive specimens with Cts between 40 and the shifted Ct was 1.42% (N1) and 0.0% (N3). Pooled and individual testing of specimens positive for SARS-CoV-2 demonstrated 100% agreement, which demonstrates the viability of pooled specimens for SARS-COV-2 testing using a dual-target RT-PCR system. Pooled specimen testing can help increase testing capacity for SARS-CoV-2 with a low risk of false-negative results.


Characterization of a novel HLA-B*40 allele, HLA-B*40:186:02, by cloning and sequencing.
A novel HLA-B*40 variant, HLA-B*40:186:02, has been identified by cloning and sequencing in a southern Chinese Han population. Aligned with HLA-B*40:01:01, HLA-B*40:186:02 has a nonsynonymous cytosine mutation at nucleotide position 165 in exon 2, leading to amino acid change from glycine to arginine at codon 56. It differs from HLA-B*40:186:01 by a synonymous change (adenine to cytosine) at position 165 in exon 2.


Status of infection prevention and control capacity in Korean hospitals: implications for disaster response and pandemic preparedness.
This study aims to explore the association of hospital infection prevention and control (IPC) structure (i.e. a dedicated IPC team and/or IPC committee) and IPC capacity in Korean hospitals, as well as its implications in the response and preparedness to COVID-19. This was a cross-sectional study using data collected through a nationwide survey. Participating hospitals completed an online questionnaire. Participation was voluntary. The survey questionnaire was developed by the government in consultation with IPC experts. The questionnaire was distributed to 2108 hospitals, including both acute and long-term care hospitals. The independent variables were the presence of an IPC team and/or IPC committee. The dependent variables were IPC activities and capacity measures, which were based on the World Health Organisation (WHO) recommendations on the core components in IPC. A total of 1442 hospitals completed the survey. Hospitals with IPC structures conducted significantly more IPC activities in all outcome measures compared with hospitals without IPC structures, with the exceptions of monitoring hand hygiene and screening for infectious diseases that showed non-significant differences. Hospitals with IPC structures showed a significant difference in performance in IPC risk assessment, operating outbreak response teams and appraisal of hospital IPC policies compared with hospitals without IPC structures. The presence of a dedicated IPC team and IPC committee was associated with increased IPC activities and IPC capacity. Hospitals with IPC teams and IPC committees showed strong implementation of planning, appraisal, resource management and outbreak response, indicating that strengthening IPC structures within hospitals is the key to more effective IPC and disaster response.


Oxidative stress, evident in antioxidant defences and damage products, in rainbow trout caged outside a sewage treatment plant.
Sewage treatment plants (STPs) are common sources of a wide variety of chemicals entering into the aquatic environment. We have investigated the effects of STP effluent on oxidative stress parameters in rainbow trout via measurements of several enzymatic antioxidants as well as the molecular antioxidant glutathione (GSH). In addition, we have measured levels of oxidative damage, i.e. protein carbonyls and lipid peroxides. Our findings indicate that STP effluent does contain prooxidants that affect fish. Oxidative damage was a more consistent indicator of exposure to prooxidants than antioxidant enzymes. Protein carbonyls and lipid peroxides were both elevated (1.3- and 1.8-fold increases, respectively) in fish caged at the STP site compared with reference site as was the amount of oxidized GSH (GSSG), which showed a 3-fold increase. Antioxidant enzymes showed no clear-cut response. Additionally, older pollutants that have been present in down stream sediment for decades had no effects on the parameters measured here.


Adsorption behaviors of poly(amido amine) dendrimers with an azacrown core and long alkyl chain spacers on solid substrates.
Adsorption behaviors of functional poly(amido amine) dendrimers with an azacrown core and long alkyl chain spacers were investigated on gold and self-assembled monolayer (SAM) by means of time course attenuated total reflection-surface enhanced infrared absorption and surface plasmon resonance spectroscopies. While 1.5th and 2.5th generation (G1.5 and G2.5) ester-terminated dendrimers were slightly adsorbed on all substrates examined, the adsorption of G2 amine-terminated dendrimer increased in the order dodecanethiol SAM<bare gold<3-mercaptopropionic acid (MPA) SAM. The time course results also revealed that the G2 dendrimer displayed the steep increase of adsorption on the bare gold, followed by the slight progress of adsorption. G2 dendrimers adsorbed on MPA SAM involved their subsequent conformational rearrangement, while the adsorption equilibrium was quickly attained for G3 dendrimer on the MPA SAM. It was supported that G3 dendrimer was adsorbed with the perpendicularly extended structure on the MPA SAM substrate, and G2 dendrimer varied its conformation from the globular structure to the extended one perpendicular to the substrate.


Effect of noradrenergic denervation on task-related visual evoked potentials in rats.
The present study examines whether destruction of the noradrenergic innervation of the forebrain interferes with the processing of sensory information in a manner that results in impaired selective attention. Electro-cortical responses to task-relevant and irrelevant stimuli were found to be sensitive indicators of the rat's attention to the stimuli. The amplitude of the response to the task-relevant stimulus increased as the rat's performance improved. The response to irrelevant flashes of light depended on the predictability of the flashes and on the rat's level of arousal. Noradrenergic denervation (with the selective neurotoxin DSP4) did not affect either the behavioural response to a visual stimulus which the rat had been trained to respond to for a food reward, or the late positive potential evoked by this stimulus. Neither did it affect the response to continuous (temporally predictable) flashes of light that were irrelevant to the task. Although the response to unpredictable flashes was also largely unaffected, we did find an additional late component in this response after DSP4 treatment. These results show that the noradrenergic innervation of the occipital cortex does not always regulate the extent to which visual stimuli are processed, but that noradrenergic neurotransmission may be activated in order to diminish excessive processing of unexpected stimuli.


The effect of mechanical forces (vibration or external compression) on the dermal water content of the upper dermis and epidermis, assessed by high frequency ultrasound.
An ultrasound scanner, is used to detect changes in water content of the upper dermis. This has previously been found to vary with age and to show diurnal variation. Furthermore, oedema due to venous disease can be shown, using this technique, to respond to elevation. In this study, the water content of the upper dermis and epidermis of the leg in 16 subjects is increased following vibration for 10 minutes using a passive exercise system. A study of pressure applied to the skin of the heel for 10 minutes in 14 volunteers also showed an increase in water content of epidermis and dermis in young persons, but less so in the elderly. It is postulated that the anatomical structure of the vascular bed of the upper dermis predisposes to transsudation when pressure to the skin is applied, thereby maintaining the resilience of the skin in the young, but less so in the elderly.


Association of Sex With Postoperative Mortality Among Patients With Heart Failure Who Underwent Elective Noncardiac Operations.
Sex differences in postoperative outcomes in patients with heart failure (HF) have not been well characterized. Women generally experience a lower postoperative mortality risk after noncardiac operations. It is unclear if this pattern holds among patients with HF. To determine if the risk of postoperative mortality is associated with sex among patients with HF who underwent noncardiac operations and to determine if sex is associated with the relationship between HF and postoperative mortality. This multisite cohort study used data from the US Department of Veterans Affairs Surgical Quality Improvement Project database for all patients who underwent elective noncardiac operations from October 1, 2009, to September 30, 2016, with a minimum of 1 year follow-up. The data analysis was conducted from May 1, 2018, to August 31, 2018. Heart failure, left ventricular ejection fraction, and sex. Postoperative mortality at 90 days. Among 609 735 patients who underwent elective noncardiac operations from 2009 to 2016, 47 997 patients had HF (7.9%; mean [SD] age, 68.6 [10.1] years; 1391 [2.9%] women) and 561 738 patients did not have HF (92.1%; mean [SD] age, 59.4 [13.4] years; 50 862 [9.1%] women). Among patients with HF, female sex was not independently associated with 90-day postoperative mortality (adjusted odds ratio [aOR], 0.97; 95% CI, 0.71-1.32). Although HF was associated with increased odds of postoperative mortality in both sexes compared with their peers without HF, the odds of postoperative mortality were higher among women with HF (aOR, 2.44; 95% CI, 1.73-3.45) than men with HF (aOR, 1.64; 95% CI, 1.54-1.74), suggesting that HF may negate the general protective association of female sex with postoperative mortality (P for interaction of HF × sex = .03). This pattern was consistent across all levels of left ventricular ejection fraction. Although HF was associated with increased odds of postoperative mortality in both sexes compared with their peers without HF, the odds of postoperative mortality were higher among women with HF than men with HF, suggesting that HF may negate the general protective association of female sex with postoperative mortality risk in noncardiac operations.


Promotion of initial cell adhesion on trisuccinimidyl citrate-modified nickel-free high-nitrogen stainless steel.
The surface of nickel-free high-nitrogen stainless steel (HNS) was modified with a citric acid-based cross-linker, trisuccinimidyl citrate (TSC), to promote initial cell adhesion in external skeletal fixation pins. The remaining active ester groups on TSC-immobilized HNS reacted with the amino groups of serum proteins. The immobilized serum proteins formed cell recognition sites to promote the initial cell adhesion immediately after cell seeding. The amount of fibronectin, which is a typical cell adhesion protein, immobilized on the TSC-immobilized HNS surface was threefold greater than on the original HNS after only 15 min. The fibroblastic cell culture experiments showed that the initial cell adhesion was significantly enhanced on the TSC-immobilized HNS compared with the original HNS at 3 h. Furthermore, the cell adhesion activity of the TSC-immobilized HNS continued to promote cell proliferation even at 7 days. Therefore, TSC-immobilized HNS may enable the rapid integration of soft tissues through its reaction with the patient's serum proteins and extracellular proteins around the surgical site.


Effect of sodium octanoate and sodium perfluorooctanoate on gel-to-liquid-crystalline phase transition of dipalmitoylphosphatidylcholine vesicle membrane.
The gel-to-liquid-crystalline phase transition of dipalmitoylphosphatidylcholine (DPPC) vesicle membrane was measured in the presence of sodium octanoate (SO) (pH 3 and 10) and sodium perfluorooctanoate (SPFO) (pH uncontrolled) by monitoring the scattered light intensity of the vesicle suspension. The phase transition temperature, Tm, decreased linearly with the concentration of added SO within the measured concentration range; the uncharged form of SO (pH 3) was much more effective for the depression of Tm than the charged form (pH 10). On the other hand, with increasing SPFO concentration, levelling off of Tm was observed after depression at an initial stage. From the depression of Tm, the partition coefficients, K, of these surfactants between bulk solution and DPPC vesicle membrane were estimated and compared with those obtained previously for other surfactant systems. The value of K for charged SO fell on the straight line of log K vs. Nc plot for anionic surfactants, where Nc is the carbon number of the hydrocarbon chain of surfactants, whereas K for uncharged SO showed a large positive deviation from the straight line of the plot for non-ionic surfactants. The latter suggested that some specific interaction, presumably hydrogen bond formation, may act between the protonated carboxyl group of SO and the lipid head group. The K value estimated for SPFO was much larger than that for charged SO. This difference in the affinity for the lipid bilayer between fluorocarbon surfactant and hydrocarbon surfactant may be attributed to the difference in their hydrophobicity.


Evoked pressure curves from the external anal sphincter following transcranial magnetic stimulation in healthy volunteers and patients with faecal incontinence.
The hypothesis was tested that evoked pressure curves (EPCs) after transcranial magnetic stimulation (TMS) would provide additional neuropathophysiological information on the descending pathways to the external anal sphincter (EAS) in patients with faecal incontinence (FI). Twenty-five healthy subjects and 69 patients with FI were investigated. TMS was applied to the vertex, and EPCs were recorded with a probe placed through the EAS. TMS was performed with the EAS at rest and during contraction (facilitated responses). At least three responses were recorded for each modality. Clinical data and anorectal manometric, electrophysiological perineal and transanal ultrasound recordings were compared with respect to the EPC results. There was no statistically significant difference between the EPCs of healthy subjects and FI patients. Twenty-three per cent of the FI patients had abnormal EPC latencies, with significantly lower voluntary contraction amplitudes (P = 0.03) and significantly higher rectal sensation (P = 0.04) than the other group. We found no significant difference between FI patients with and without abnormal EPC latencies in terms of clinical characteristics and electrophysiological and endoanal ultrasound parameters. There was no difference in the identified causes of the FI between the two groups. As abnormal EPC latencies were found in 23% of FI patients with no known central neurological disease, abnormal EPC latencies might reveal undetected lesions of descending pathways in patients with FI.


Moderating effect of comorbid anxiety disorders on treatment outcome in a randomized controlled psychotherapy trial in early-onset persistently depressed outpatients.
Persistent depressive disorder (PDD) is associated with high rates of comorbid psychiatric disorders, mostly anxiety disorders (ADs). Comorbid AD was found to be associated with poorer treatment outcome in PDD patients. The effect of comorbid AD on disorder-specific treatment for PDD (Cognitive Behavioral Analysis System of Psychotherapy [CBASP]) has not been studied yet. We analyzed whether the presence of a comorbid AD was moderating the effectiveness of disorder-specific (CBASP) versus nonspecific psychotherapy (supportive therapy [SP]) on depressive symptoms (24-item Hamilton Rating Scale for Depression [HRSD-24]) in a sample of unmedicated early-onset PDD outpatients (N = 268). Secondary outcomes were response and remission of depressive symptoms and the extent of interpersonal problems (Inventory of Interpersonal Problems [IIP-64]). The superiority of CBASP over SP was significantly stronger in PDD patients with comorbid AD compared to patients without AD (in HRSD-24 and IIP-64). There was no significant moderation for remission or response of depressive symptoms. Our hypothesis of a moderating effect of comorbid AD was confirmed. The main limitation might be the exclusion criteria of our sample limiting the generalizability. The major strength is the systematic analysis of the effect of AD in treating early-onset PDD with high quality of psychotherapy in both arms of this trial. Patients suffering from PDD comorbid with AD might experience greater benefit when they are treated with specific as opposed to unspecific therapy. Analyzing subgroups of patients with PDD seems worthwhile to improve treatment effectiveness even within disorder-specific treatment programms.


Biomechanical Study on Three Screw-Based Atlantoaxial Fixation Techniques: A Finite Element Study.
This is a finite element study. This study is aimed to compare the biomechanical behaviors of three screw-based atlantoaxial fixation techniques. Screw-based constructs that are widely used to stabilize the atlantoaxial joint come with their own challenges in surgery. Clinical and in vitro studies have compared the effectiveness of screw-based constructs in joint fixation. Nevertheless, there is limited information regarding the biomechanical behavior of these constructs, such as the stresses and strains they experience. A finite element model of the upper cervical spine was developed. A type II dens fracture was induced in the intact model to produce the injured model. The following three constructs were simulated on the intact and injured models: transarticular screw (C1- C2TA), lateral mass screw in C1 and pedicle screw in C2 (C1LM1-C2PD), and lateral mass screw in C1 and translaminar screw in C2 (C1LM1-C2TL). In the intact model, flexion-extension range of motion (ROM) was reduced by up to 99% with C11-C2TA and 98% with C1LM1-C2PD and C1LM1-C2TL. The lateral bending ROM in the intact model was reduced by 100%, 95%, and 75% with C11-C2TA, C1LM1-C2PD, and C1LM1-C2TL, respectively. The axial rotation ROM in the intact model was reduced by 99%, 98%, and 99% with C11-C2TA, C1LM1-C2PD, and C1LM1-C2TL, respectively. The largest maximum von Mises stress was predicted for C1LM1-C2TL (332 MPa) followed by C1LM1-C2PD (307 MPa) and C11-C2TA (133 MPa). Maximum stress was predicted to be at the lateral mass screw head of the C1LM1-C2TL construct. Our model indicates that the biomechanical stability of the atlantoaxial joint in lateral bending with translaminar screws is not as reliable as that with transarticular and pedicle screws. Translaminar screws experience large stresses that may lead to failure of the construct before the required bony fusion occurs.


Donation of stem cells from blood or bone marrow: results of a randomised study of safety and complaints.
Biological consequences and physical complaints were compared for donors randomly assigned either to blood stem cell (BSC) or bone marrow (BM) donation. In the period 1994-1999, 61 consecutive donors were included. The BSC donors were given G-CSF 10 microg/kg s.c., daily during 5 days before the first leukapheresis. Nineteen donors had one leukapheresis, 10 required two and one donor needed three leukaphereses in order to reach the target cell number of 2 x 10(6) CD34(+) cells/kg bw of the recipient. A median platelet nadir of 102 x 10(9)/l was reached shortly after the last leukapheresis. Three weeks post harvest, 17 of 30 BSC donors had a mild leukopenia. Six had a leukopenia lasting more than a year before returning to normal values. Both groups were monitored prospectively through a standardised questionnaire completed by the donors. BSC donation was significantly less burdensome than BM donation and was preferred by the donors. The short-term risks of BSC mobilisation and harvest seem negligible. The potential long-term effects of G-CSF are unresolved and the donors must be followed closely.


A new family of proteins related to the HEAT-like repeat DNA glycosylases with affinity for branched DNA structures.
The recently discovered HEAT-like repeat (HLR) DNA glycosylase superfamily is widely distributed in all domains of life. The present bioinformatics and phylogenetic analysis shows that HLR DNA glycosylase superfamily members in the genus Bacillus form three subfamilies: AlkC, AlkD and AlkF/AlkG. The crystal structure of AlkF shows structural similarity with the DNA glycosylases AlkC and AlkD, however neither AlkF nor AlkG display any DNA glycosylase activity. Instead, both proteins have affinity to branched DNA structures such as three-way and Holliday junctions. A unique β-hairpin in the AlkF/AlkG subfamily is most likely inserted into the DNA major groove, and could be a structural determinant regulating DNA substrate affinity. We conclude that AlkF and AlkG represent a new family of HLR proteins with affinity for branched DNA structures.


Differential effects of state and trait anxiety on heart rate responses and reaction time.
College students were tested on 10 trials of a reaction time task, using a variable interval warning signal, while heart rate was recorded. Subjects were divided into high and low anxiety groups using a median split based on their scores on a modification of the trait scale of the State-Trait Anxiety Inventory. The low anxiety group exhibited faster reaction times, shorter latency heart rate deceleration following warning signal onset, and a more consistent deceleration immediately prior to the respond signal than the high anxiety group. When the same subjects were divided into groups according to state anxiety, no group differences were found in either heart rate or reaction time. In Part II of the study, subjects listened to six 15 s presentations of a 70 dbA, 250 Hz tone. No group differences in heart rate responses to the tones were found for either state or trait anxiety groups.


The accumulation of three yeast ribosomal proteins under conditions of excess mRNA is determined primarily by fast protein decay.
The suggestion that compensation for overabundant mRNA of the genes for Saccharomyces cerevisiae ribosomal protein (r-protein) L3, L29, or rp59 occurs by translation repression has been reinvestigated. First, analysis of the distribution of these three mRNAs in polysome profiles revealed no differences between normal and mRNA-overproducing strains, indicating that initiation of r-protein translation is not repressed under conditions of mRNA overaccumulation. Second, experiments involving radioactive pulse-labeling of proteins were done by using a modified method of data collection and analysis that allows quantitation and correction for fast decay during the pulse. These measurements revealed that the synthesis rate of the three r-proteins is increased when their mRNA levels are elevated and that their decay rate is also high, with half-lives ranging from a fraction of a minute to more than 10 min. We conclude that accumulation of excess r-protein mRNA has no effect on translation rate; rapid decay of protein during the course of the labeling period can account for the apparent discrepancy between mRNA levels and protein synthesis rates. Yeast r-proteins, when produced in excess, are among the most rapidly degraded proteins so far described.


Discovery of the disubstituted oxazole analogues as a novel class anti-tuberculotic agents against MDR- and XDR-MTB.
A high-throughput screening effort on 45,000 compounds resulted in the discovery of a disubstituted oxazole as a new structural class inhibitor of Mycobacterium tuberculosis (Mtb). In order to improve the activity and investigate the SAR of this scaffold, a series of disubstituted azole analogues have been designed and synthesized. The newly synthesized compounds 1a-y were evaluated for their in vitro anti-TB activity versus replicating, multi- and extensive drug resistant Mtb strains. All the compounds, except 1o, 1p and 1q, showed potent anti-TB activity with MIC of 1-64 mg/L. The test of broad spectrum panel revealed that this series are specific to Mtb. The cytotoxicity assessment indicated that the compounds were not cytotoxic against HEK 293 cells. The compounds could have a novel mechanism to anti-Mtb as they can inhibit drug sensitive and drug resistant Mtb.


Immunohistological detection of human parvovirus B19 in formalin-fixed, paraffin-embedded tissues.
Human parvovirus B19 is a cause of aplastic crises in patients with haemolytic anaemias, prolonged bone marrow failure in the immunosuppressed, and fetal death secondary to non-immune hydrops. The immunohistological detection of parvovirus B19 in formalin-fixed, paraffin-embedded tissues has not previously been reported, and definitive diagnosis of infection in such specimens has relied on the use of specialized DNA hybridization and amplification techniques. A new monoclonal antibody to B19 capsid proteins, R92F6, was found to be capable of labelling infected cells in paraffin-embedded tissues from all 19 cases of parvovirus-related fetal hydrops tested, and in bone marrow from a child with congenital immunodeficiency and chronic parvovirus infection. Viral antigen was detected both in cytoplasmic and in nuclear distributions using the alkaline phosphatase anti-alkaline phosphatase (APAAP) technique without preceding proteolytic digestion. The viral epitope recognized appears to be highly conserved, as specimens were obtained over a 13-year period from widely spaced locations in the U.K. Antibody R92F6 should facilitate rapid diagnosis of parvovirus B19 infection in routinely processed and archival specimens.


Visual acuity after iridectomy or aspiration for congenital cataract: Experimental and clinical studies.
Studies with a camera and a model eye showed that the image produced by peripheral light rays is inferior to the image produced by paraxial rays. In a retrospective clinical study, we determined the best corrected visual acuity in 33 patients (52 eyes) who had had iridectomy (group I) or aspiration (group II) or both for central cataracts. Vision was significantly better (p less than 0.01) in group II than in group I, and postoperative complications of aspiration were minimal. Some patients had unilateral amblyopia or pendular nystagmus. We conclude that if vision is to be improved with a single procedure, aspiration - not optical iridectomy - is indicated. We could not identify any factors in early infancy that would have predicted which patients with congenital central cataracts required early surgery.


Mannose-binding lectin (MBL) therapy in an MBL-deficient patient with severe cystic fibrosis lung disease.
Deficiency of mannose-binding lectin has been shown to be a risk factor for cystic fibrosis (CF) patients. We, therefore, decided to treat a patient with CF, mannose-binding lectin deficiency, severe bronchopulmonary Pseudomonas aeruginosa infection, and rapid deterioration of lung function with purified mannose-binding lectin in an attempt to ameliorate the course of the lung disease. The mannose-binding lectin used originated from pooled human donor plasma and was given as an intravenous infusion twice a week for a period of 3 months. The patients's clinical condition was stabilized during the treatment period, but was not improved. No adverse events were observed. However, the lung function assessed as percent forced expiratory volume in 1 sec (FEV1%) and percent forced vital capacirt (FVC%) correlated significantly with the mannose-binding serum lectin levels (rho=+0.68, P=0.008, and rho=+0.73, P=0.004). Additionally, an inverse correlation with the acute phase-reactant C-reactive protein and the proinflammatory cytokine IL-6 was observed (rho=-0.49, P=0.007 and rho=-0.41, P=0.04, respectively). The results emphasize the importance of mannose-binding lectin as a secondary disease modifier in CF. Moreover, purified mannose-binding lectin can safely be administered to chronically ill patients, and may be a potential treatment in CF and other diseases in which mannose-binding lectin deficiency plays a pathophysiological role.


A Brief Analysis on Clinical Severity of Mandibulofacial Dysostosis Guion-Almeida Type.
Genetic variants in EFTUD2 were proven to influence variable phenotypic expressivity in mandibulofacial dysostosis Guion-Almeida type (MFDGA) or mandibulofacial dysostosis with microcephaly (MFDM). Yet, the association between the severity of clinical findings with variants within the EFTUD2 gene has not been established. Thus, we aim to elucidate a possible genotype-phenotype correlation in MFDM. Forty articles comprising 156 patients were evaluated. The genotype-phenotype correlation was analyzed using a chi-square or Fisher's exact test. The proportion of patients with MFDM was higher in Caucasian relative to Asian populations. Although, in general, there was no apparent genotype-phenotype correlation in patients with MFDM, Asians tended to have more severe clinical manifestations than Caucasians. In addition, cardiac abnormality presented in patients with intronic variants located in canonical splice sites was a predisposing factor in affecting MFDM severity. Altogether, this article provides the pathogenic variants observed in EFTUD2 and possible genotype-phenotype relationships in this disease.


The powerful Solanaceae: Food and nutraceutical applications in a sustainable world.
The Solanaceae family is considered one of the most important families among plant species because, on one hand encompasses many staple food crops of the human diet while, on the other hand, it includes species rich in powerful secondary metabolites that could be valorized in medicine or drug formulation as well as nutraceuticals and food supplements. The main genera are Solanum, Capsicum, Physalis, and Lycium which comprise several important cultivated crops (e.g., tomato, pepper, eggplant, tomatillo, and goji berry), as well as genera notable for species with several pharmaceutical properties (e.g., Datura, Nicotiana, Atropa, Mandragora, etc.). This chapter discusses the nutritional value of the most important Solanaceae species commonly used for their edible fruit, as well as those used in the development of functional foods, food supplements, and nutraceuticals due to their bioactive constituents. The toxic and poisonous effects are also discussed aiming to highlight possible detrimental consequences due to irrational use. Finally, considering the high amount of waste and by-products generated through the value chain of the main crops, the sustainable management practices implemented so far are presented with the aim to increase the added-value of these crops.


Combining the anion gap with the sequential organ failure assessment score to evaluate the short-term prognosis of patients in the cardiac intensive care unit.
We attempted to determine the predictive ability of the first-day Sequential Organ Failure Assessment (SOFA) score in the cardiac intensive care unit, as well as a new score combining the anion gap (AG) with the SOFA score (SOFA-AG). Information was obtained from the Medical Information Mart for Intensive Care III (MIMIC III 1.4) database. We plotted the relationship between the maximum first-day AG and 90-day mortality after admission to the care unit. Patients were divided into five groups based on the hazard ratio (HR) and assigned scores of 0, 1, 2, 3, or 4 points. We compared the area under the curve (AUC) for the receiver-operating characteristic curve of the SOFA and that of the SOFA-AG. A total of 1316 patients were identified and divided into the following five groups: AG 8 to <16 mmol/L; AG 16 to <17 mmol/L; AG 17 to <19 mmol/L; AG 19 to <21 mmol/L; and AG ≥ 21 mmol/L. The SOFA-AG score had a greater AUC than the SOFA score at 7 days (0.770 vs. 0.711; P < 0.001), 14 days (0.751 vs. 0.692; P < 0.001), 28 days (0.741 vs. 0.684; P < 0.001), and 90 days (0.727 vs. 0.667; P < 0.001). The SOFA score showed moderate predictive value only for 7-day mortality after admission to the cardiac intensive care unit, but the SOFA-AG score had improved predictive ability for up to 90 days after admission.


A systematic study of wound contraction in mammalian skin.
The phenomenon of wound contraction has been studied in the skin of rats, rabbits and guinea pigs with the aim of obtaining a standardized wound system to be used for biological assay of in vivo agonists and antagonists of granulation tissue contraction. The sex of the animal, the time of day of wounding, the size and the shape of the wounds all had no apparent influence on the wound contraction curves expressed as per cent of original area against time. The test animal's age and species did have a significant influence on contraction of skin wounds which by analogy with results obtained with rigid splints attached to the skin could be attributed to variations in the thickness, rigidity and fixation of the skin to deep tissues. The shape of the final scar was influenced by the position of the wound on the body surface. The major factor in this instance was considered to be tensile forces acting within the skin. Wound contraction curves in rats and rabbits showed three distinct phases, namely early closure, stationary and logarithmic closure. Guinea pigs lacked the early closure phase. The exact role of the panniculus carnosus in wound healing is considered to be worthy of detailed study.


Comparison of diltiazem and metoprolol for atrial fibrillation with rapid ventricular rate: Systematic review and meta-analysis.
Intravenous calcium channel blockers or beta-blockers are the preferred rate control medications for hemodynamically stable patients with atrial fibrillation with rapid ventricular rate (AF-RVR) in the emergency department. To compare the efficacy of intravenous diltiazem and metoprolol for rate control and safety with respect to development of hypotension and bradycardia in patients with AF-RVR. For this systematic review and meta-analysis, we searched PubMed, Embase, Cochrane databases, and the clinicaltrials.gov registry between database inception and 30th May 2021. Articles were included if they compared efficacy and safety of diltiazem versus metoprolol in critically ill adult patients hospitalized with AF-RVR. Outcome measures were achievement of rate control, development of new hypotension, and bradycardia after drug administration. Of 86 records identified, 14 were eligible, all of which had a low to moderate risk of overall bias. The meta-analysis (Mantel-Haenszel, random-effects model) showed that diltiazem use was associated with increased achievement of rate control target compared to metoprolol [14 studies, n = 1732, Odds Ratio (OR): 1.92; 95% Confidence Intervals (CI):1.26 to 2.90; I2 = 61%]. In the pooled analysis, no differences were seen in hypotension using diltiazem vs metoprolol [12 studies, n = 1477, OR: 0.96; 95% CI:0.61 to 1.52; I2 = 35%] or bradycardia [9 studies, n = 1203, OR: 2.44; 95% CI: 0.82 to 7.31; I2 = 48%]. Intravenous diltiazem is associated with increased achievement of rate control target in patients with AF-RVR compared to metoprolol, while both medications are associated with similar incidence of hypotension and bradycardia.


Feasibility of external cranial cooling during out-of-hospital cardiac arrest.
Hypothermia during brain ischemia can improve neurological outcome. This study tested whether local cranial cooling during the low-flow state of cardiopulmonary resuscitation (CPR) could produce clinically significant cerebral cooling. Ice was applied to the heads and necks of subjects (hypothermia group) with out-of-hospital cardiac arrest (OOHCA) during CPR. Nasopharyngeal and tympanic temperatures were measured as surrogates for cerebral temperature. The rate of cranial cooling in the hypothermia group (-0.06 +/- 0.06 degrees C/min) was not significantly increased compared with a control group without ice (-0.04 +/- 0.07 degrees C/min), although older age was associated with more rapid cranial cooling. Of note, many subjects with OOHCA are already mildly hypothermic (mean cranial temperature= 35.0 +/- 1.2 degrees C) when they are first encountered in the field. This study suggests that brief cranial cooling is ineffective for rapidly lowering brain temperature. However, most cardiac arrest victims are spontaneously mildly hypothermic and preventing rewarming may provide some of the desired benefits of cerebral hypothermia.


The effects of an adopted narrow gait on the external adduction moment at the knee joint during level walking: evidence of asymmetry.
The external knee adduction moment is an accurate estimation of the load distribution of the knee and is a valid predictor for the presence, severity and progression rate of medial compartment knee osteoarthritis. Gait modification strategies have been shown to be an effective means of reducing the external adduction moment. The purpose of this study was to test narrow gait as a mechanism to reduce the external adduction moment and investigate if limb dominance affects this pattern. Fifteen healthy male participants (mean age: 23.8 (SD=3.1) years, mean height: 1.8 (SD=0.1) m, and mean body mass: 82.9 (SD=16.1 kg) took part in this study. Five walking trials were performed for each of the three different gait conditions: normal gait, toe-out gait, and narrow gait. Adoption of the narrow gait strategy significantly reduced the early stance phase external knee adduction moment compared to normal and toe-out gait (p<.002). However, it was observed that this reduction only occurred in the non-dominant limb. Gait modification can reduce the external knee adduction moment. However, asymmetrical patterns between the dominant and non-dominant limbs, specifically during gait modification, may attenuate the effectiveness of this intervention. The mechanism of limb dominance and the specific roles of each limb during gait may account for an asymmetrical pattern in the moment arm and center of mass displacement during stance. This new insight into how limb-dominance effects gait modification strategies will be useful in the clinical setting when identifying appropriate patients, when indicating a gait modification strategy and in future research methodology.


All in the family: times are changing.
General practitioners are familiar with the fact that individual use of care is determined by the family. This paper describes to what extent patterns found in the 1970s can still be replicated in the early 21(st) century, analyzing the impact of the family on frequency of presenting minor illnesses in general practice. We used the data of the families in the CMR database during two five-year periods: 1971-1975 and 2001-2005. All episodes of minor illness were registered using the same classification criteria in both studies. The population under study was divided in low and medium/high social class. Pearson correlation coefficients and a hierarchical linear model were used to analyse the impact of family on the frequency of presenting minor illnesses. Correlations between parents (M/F) were stronger in 1971-1975 than in the 2001-2005 period. The influence of the family on presenting minor illness in general practice has decreased from 27.7 to 20.3%. Though the influence of the family on presenting minor illness in general practice has decreased, nowadays (2001-2005) still we found strong correlations between family members in presenting morbidity to the GP.


Amitriptyline-induced prolonged cholestasis.
We report the case of a patient in whom amitriptyline administration for 5 wk was followed by prolonged cholestasis. Jaundice and pruritus lasted 19 and 20 mo, respectively. Three liver biopsies were performed at different stages of the disease showing the course of liver lesions. Cholestasis initially located in the region of the hepatic venule came to be associated with the progressive development of portal tract lesions consisting of inflammatory infiltration, fibrosis, and disappearance of interlobular bile ducts. Amitriptyline hydroxylation and dextromethorphan O-demethylation are deficient in subjects with the poor metabolizer phenotype of debrisoquine. Drug oxidation phenotyping with dextromethorphan showed that this patient had the extensive metabolizer phenotype. This observation demonstrates that amitriptyline can induce prolonged cholestasis and suggests that the susceptibility to develop liver injury while taking this drug may not be related to a genetic deficiency of its hydroxylation.


The endocrine pharmacology of testosterone therapy in men.
The review starts off by outlining the history of the discovery of the male sex hormone testosterone and the historical background to the various, often dubious, approaches to the treatment of age-related endocrine disorders in older men. A discussion of congenital androgen deficiency in young men is followed by methods of diagnosing hypogonadism in older men. Among therapeutic options, the alternatives to direct testosterone replacement are discussed, although none of them have proved to be particularly successful in clinical practice. For testosterone replacement itself, various routes of administration and pharmaceutical formulations are now available, facilitating good monitoring and individualized therapy.


Variation in optical coherence tomography signal quality as an indicator of retinal nerve fibre layer segmentation error.
Commercial optical coherence tomography (OCT) systems use global signal quality indices to quantify scan quality. Signal quality can vary throughout a scan, contributing to local retinal nerve fibre layer segmentation errors (SegE). The purpose of this study was to develop an automated method, using local scan quality, to predict SegE. Good-quality (global signal strength (SS) ≥ 6; manufacturer specification) peripapillary circular OCT scans (fast retinal nerve fibre layer scan protocol; Stratus OCT; Carl Zeiss Meditec, Dublin, California, USA) were obtained from 6 healthy, 19 glaucoma-suspect and 43 glaucoma subjects. Scans were grouped based on SegE. Quality index (QI) values were computed for each A-scan using software of our own design. Logistic mixed-effects regression modelling was applied to evaluate SS, global mean and SD of QI, and the probability of SegE. The difference between local mean QI in SegE regions and No-SegE regions was -5.06 (95% CI -6.38 to 3.734) (p<0.001). Using global mean QI, QI SD and their interaction term resulted in the model of best fit (Akaike information criterion=191.8) for predicting SegE. Global mean QI ≥ 20 or SS ≥ 8 shows little chance for SegE. Once mean QI<20 or SS<8, the probability of SegE increases as QI SD increases. When combined with a signal quality parameter, the variation of signal quality between A-scans provides significant information about the quality of an OCT scan and can be used as a predictor of segmentation error.


Magnetic resonance imaging-guided conventional catheter ablation of isthmus-dependent atrial flutter using active catheter imaging.
Interventional cardiac magnetic resonance (iCMR) has been established as a radiation-free alternative compared to standard fluoroscopy-guided catheter ablation for cavotricuspid isthmus (CTI)-dependent atrial flutter to image anatomy, structural alterations, and further catheter guidance. The purpose of this study was to explore the safety, feasibility, and efficacy of CTI ablations performed completely in the iCMR suite using active catheter imaging. Consecutive patients underwent iCMR-guided catheter ablation for CTI-dependent atrial flutter. Procedures were performed in a 1.5-T magnetic resonance (MR) imaging unit with MR-conditional ablation catheters. Catheter guidance was achieved using active catheter imaging via integrated MR receive tip coils. Acute success, periprocedural complications, and short-term follow-up were collected for further analysis. All patients (N = 15; 73% male; median age 70 years; interquartile range [67-82]) achieved acute procedural success without any complication. Median procedural time was 43 minutes [33-58] with median radiofrequency delivery time of 18 minutes [12-26]. Postprocedural lesion visualization scanning was completed in a median of 32 minutes [10-42]. None of the patients with 6-month follow-up had atrial flutter recurrence. In the iCMR suite, CTI-dependent atrial flutter ablation could be achieved safely using active catheter imaging without any complication. It further allows detailed anatomic visualization of the CTI, intraprocedural lesion visualization, and exclusion of pericardial effusion.


Evolution of chinook salmon (Oncorhynchus tshawytscha) populations in New Zealand: pattern, rate, and process.
Chinook salmon, Oncorhynchus tshawytscha, from the Sacramento River, California, USA were introduced to New Zealand between 1901 and 1907, and colonized most of their present-day range within about 10 years. The New Zealand populations now vary in phenotypic traits typically used to differentiate salmon populations within their natural range: growth in freshwater and at sea, age at maturity, dates of return to fresh water and reproduction, morphology, and reproductive allocation. This paper reviews a large research program designed to determine the relative contributions of phenotypic plasticity and genetic adaptation to this variation, in an effort to understand the processes underlying the natural evolution of new populations. We found strong evidence of trait divergence between populations within at most 30 generations, particularly in freshwater growth rate, date of return, and reproductive output, with plausible adaptive bases for these differences. Importantly, we also demonstrated not only a genetic basis for post-release survival but higher survival, and hence fitness, of a population released from its established site compared to another population released from the same site. We conclude that divergence of salmon in different rivers probably resulted initially from phenotypic plasticity (e.g., habitat-specific growth rates, and effects of upriver migration on ovarian investment). Philopatry (homing to natal streams) combined with rapid evolution of distinct breeding periods to restrict gene flow, facilitating divergence in other traits. We also suggest that in addition to genetic divergence resulting from random founder effects, divergence may also arise during the very early stages of colonization when the original colonists are a non-random, pre-adapted subset of the source population. This 'favored founders effect' immediately improves the fitness of the new population. Overall, this research reveals the complex interplay of environmental and genetic controls over behavior, physiology and life history that characterize the early stages of population differentiation, a process that has taken place repeatedly during the history of salmon populations.


Clinical evaluation of removable partial dentures with digitally fabricated metal framework after 4 years of clinical service.
This case report describes the clinical outcomes of three patients who received removable partial dentures with a completely digitally designed and manufactured metal framework. The initial intraoral impressions were prepared, and the resulting standard tessellation language files were sent to a dental laboratory, where the alloy framework was designed using inLab software and printed using a 3D printer or milled directly from a Co-Cr disc. The quality of fit of the framework was evaluated intraorally to confirm the laboratory design. The acrylic teeth were set, and the definitive partial dentures were delivered after the acrylic resin bases were processed. The follow-up time was 4 years. No complications or failures related to the components of the partial dentures were observed.


IDF Diabetes Atlas: A review of studies utilising retinal photography on the global prevalence of diabetes related retinopathy between 2015 and 2018.
The purpose of this study is to assess the prevalence of diabetic retinopathy (DR) world-wide from articles published since 2015 where the assessment of the presence and severity of DR was based on retinal images. A total of 4 databases were searched for the MESH terms diabetic retinopathy and prevalence. Of 112 publications 32 studies were included and individual data pooled for analysis. The presence of any DR or diabetic macular edema (DME) was recorded and severity as mild, moderate or severe non-proliferative DR (NPDR), proliferative DR (PDR) and DME and/or clinically significant macular edema (CSME). The level of severity of DR reported refer to persons with diabetes and not individual eyes. The global prevalence of DR and DME, for the period 2015 to 2019 were 27.0% for any DR comprising of 25.2%, NPDR, 1.4% PDR and 4.6% DME. The lowest prevalence was in Europe at 20.6% and South East Asia at 12.5% and highest in Africa at 33.8%, Middle East and North Africa 33.8%, and the Western Pacific region at 36.2%. This study illustrated difficulties in deriving a meaningful global prevalence rate for DR and DME due to the lack of uniformity in defining the study populations, methodological differences, retinal image capture and grading criteria. Therefore, international consensus is required using a minimal data set for future studies.


A study of the prevalence and clinical significance of venom-specific IgE.
The prevalence of unrecognized Hymenoptera-venom sensitization, assessed by venom skin tests (VSTs) in adults with no history of adverse reactions to sting, has been as high as 12% in previous epidemiologic studies. To assess further the clinical importance of positive VSTs in such individuals, we skin tested 33 subjects stung in the field during the preceding 12 months without reaction, and 33 persons who denied being stung in the preceding 3 years. Among the recently stung group, 12/33 had at least one positive VST (greater than or equal to 2+) at 1.0 microgram/ml, whereas 5/33 had positive VST at 0.1 microgram/ml. In contrast, only 2/33 nonstung subjects had positive VST at 1.0 microgram/ml, and none were positive at 0.1 microgram/ml. To estimate, prospectively, the sensitization rate after insect stings, we studied a third group of 11 nonsensitive patients with negative skin tests to Hymenoptera. After a deliberate in-hospital honeybee sting, only 1/11 developed a persistently positive honeybee VST. From among the three groups, we then performed nine sting challenges in eight patients with positive VSTs, and all stings were tolerated without significant reaction. We also measured IgE antibodies to Hymenoptera venoms in random blood bank donors. During April to May, 2/216 sera contained elevated venom-specific IgE antibodies, whereas 14/201 sera collected from October to November contained elevated venom-specific IgE antibodies. We conclude that a small but appreciable portion of the population has venom-specific IgE antibodies and that the prevalence is seasonably variable. Our data indicate that persons recently stung without significant reaction contribute to this group but that only a small portion of this group is at risk for a systemic reaction with a future sting.


First diagnostic criteria for atopic myelitis with special reference to discrimination from myelitis-onset multiple sclerosis.
To establish the first evidence-based diagnostic criteria for atopic myelitis (AM) enabling it to be discriminated from myelitis-onset multiple sclerosis (MS), which is a difficult differential diagnosis. Sixty-nine consecutive AM patients examined from 1996 to 2010 at Kyushu University hospital, who fulfilled the empirical definition of AM (2003), and 51 myelitis-onset MS patients in whom allergen-specific IgE was measured, were enrolled. The first available brain MRI findings were compared between the two. Then, we compared the clinical and laboratory features between the 16 AM cases who did not meet the Barkhof brain MRI criteria for MS after more than 5 years follow-up and 51 myelitis-onset MS cases. Based on the discriminative findings, we established diagnostic criteria for AM and calculated the sensitivity and specificity. AM patients had a significantly lower frequency of Barkhof brain lesions on baseline MRI than myelitis-onset MS patients. AM patients had a significantly higher frequency of present and/or past history of atopic disease and hyperIgEemia, and higher cerebrospinal fluid levels of interleukin 9 and CCL11/eotaxin, but a lower frequency of oligoclonal IgG bands than myelitis-onset MS patients. Our proposed diagnostic criteria for AM demonstrated 93.3% sensitivity and 93.3% specificity for AM against myelitis-onset MS, with 82.4% positive predictive value and 97.7% negative predictive value. Our first evidence-based criteria for AM show high sensitivity and specificity, and would be useful clinically.


Synthesis, characterization of novel injectable drug carriers and the antitumor efficacy in mice bearing Sarcoma-180 tumor.
New unsaturated polyesters of poly(fumaric acid-glycol-dodecanedioic acid) P(FA-GLY-DDDA) copolymers, poly(fumaric acid-glycol-brassylic acid) P(FA-GLY-BA) copolymers, poly(fumaric acid-glycol-tetradecanedioic acid) P(FA-GLY-TA) copolymers and poly(fumaric acid-glycol-pentadecanedioic acid) P(FA-GLY-PA) copolymers were prepared by melt polycondensation of the corresponding mixed monomers: fumaric acid, glycol and one of C(12-15) dibasic acids. The copolymers were characterized by FT-IR, gel permeation chromatography (GPC), and the surface structure of unsaturated polyesters after solidify were studied by atomic force microscopy (AFM). The molecular structure and composition of the unsaturated polyesters were determined by 1H NMR spectroscopy. In vitro studies showed that some of the copolymers are degradable in phosphate buffer at 37 degrees C and have properly drug release rate as drug carriers. The biocompatibility of P(FA-GLY-DDDA) and P(FA-GLY-BA) copolymers under mice skin was also evaluated, macroscopic observation and microscopic analysis demonstrated that the copolymer is biocompatible and well tolerated in vivo. Antitumor efficacy of P(FA-GLY-DDDA) copolymers and P(FA-GLY-BA) copolymers containing 5% adriamycin hydrochloride (ADM) in mice bearing Sarcoma-180 tumor exhibited increased volume doubling time (VDT) (22+/-1.5 days and 24+/-2.5 days) compared to plain subcutaneous injection of ADM (7+/-0.9 days). The antitumor efficacy of injecting P(FA-GLY-DDDA)-ADM inside tumor twice intervened in 22 days exhibited an especially increased cytotoxic effect as revealed by increased VDT (33+/-2.5 days), and the antitumor efficacy of injecting P(FA-GLY-BA)-ADM inside tumor twice intervened in 24 days exhibited an especially increased cytotoxic effect as revealed by increased VDT (35+/-1.5 days). The studies suggested that P(FA-GLY-DDDA) copolymers and P(FA-GLY-BA) copolymers as effective and injectable carriers for antineoplastic drug like adriamycin hydrochloride have a very good foreground in the treatment of noumenon tumor.


Hydrophilic modification of silicone elastomer films: thermal, mechanical and theophylline permeability properties.
The improvement of the hydrophilicity of silicone rubber (SR) is sought in many biomedical applications. In the present work, we have prepared neat films coming from condensation-type SR, modified either through blending or end-linking reaction with low molecular weight poly(ethylene glycol) (PEG). The films were studied with respect to their water uptake capacity, stability of embedment of ethylenoxy moieties, and mechanical and thermal properties. Subsequently, we prepared drug-loaded films with theophylline, a relatively hydrophilic model drug, and studied the release performance of these systems. Results on neat films showed that blending increases significantly the overall water uptake of the films and, at the same time, has a limited detrimental effect on their mechanical properties. On the other hand, end-linked films loaded with theophylline exhibited better rate-controlling properties in vitro, due to better dispersion of the sorbed water.


[Cardiovascular diseases in female workers and methods of their prevention].
The article covers actual incidence of coronary heart disease and arterial hypertension, atherosclerosis risk factors among female workers, its dependence on age and occupation type. The authors studied efficiency and safety of antihypertensive therapy in hypertensive women.


Paracoccidioidomycosis in nude mice: presence of filamentous forms of the fungus.
Congenitally athymic nude mice (nu/nu) and their phenotypically normal littermates (nu/+) were intraperitoneally infected with yeast cells of a strain of Paracoccidioides brasiliensis. The nude mice developed a severe and generalized infection with an intense parasitism of several organs, accompanied by a low-grade of tissue reaction. The lesions were characterized by abundant yeast-like cells of the fungus, and in some animals, numerous hyphal forms could be well visualized. In control animals, infection was moderate, almost exclusively restricted to the area of inoculation, and the lesions presented few parasites surrounded by an inflammatory response. Filamentous forms of the fungus were never encountered in these animals.


[Use of plastic isolators in the prevention of infection in high risk patients with hematologic diseases].
The authors report a microbiological study of the flora of high risk adults with blood diseases isolated in hermetically sealed plastic isolater. This type of isolation is very effective in the prevention of infections due to hospital germs as such contamination occurred only twice in 292 days isolation and was found in only 0.8% on samples. The efficacy of methods of body decontamination and the incidence of infective complications in our series is also reported.


Regulation of the deficiencies along the proximal distal axis of the chick wing-bud: a quantitative analysis.
The wing-bud shows zero regulation following removal of a whole slice of the proximal-distal axis from stage 22 or later. Prior to this it is possible that it may show some regulation. Some of the apparent regulation may be explained by the way in which the limb grows. The bud never shows perfect size regulation or morphallaxis. The proximal-distal axis of the bud between shoulder and wrist expands uniformly from stage 20.


Effect of voluntary activation on age-related muscle fatigue resistance.
Ageing is associated with a higher fatigue resistance during submaximal or maximal fatiguing contractions. The present study aimed to investigate the contribution of the central and peripheral fatigue to the age-related differences in fatigue development of the plantar flexor muscles. Therefore, the voluntary activation, rest twitch moment and voluntary plantar flexor moment were examined before during as well as 2, 5 and 10min after a fatiguing task. This consisted of intermittent isometric submaximal plantar flexor contractions at equal intensity for both young and old adults (considering the age-related differences in muscle inhibition). Consequently, possible differences between young and old adults in voluntary activation during the maximal contraction utilised for determining the intensity of the fatiguing task, which can influence fatigue development, have been taken into account. The plantar flexors moment was calculated using inverse dynamics and the voluntary activation was measured using the twitch interpolation technique. Changes in voluntary activation and rest twitch moment during the fatiguing task were used to assess central and peripheral fatigue, respectively. In both young and old adults, peripheral ( approximately 20%) as well as central fatigue ( approximately 9%) contributed to the time to task failure. Old adults demonstrated greater time to task failure than young ones, but similar voluntary activation behaviour during the fatiguing task. We concluded that, the age-related enhancement in fatigue resistance is not attributable to voluntary activation but is linked to mechanisms located within the working muscle.


Model-based metabolic engineering enables high yield itaconic acid production by Escherichia coli.
Itaconic acid is a high potential platform chemical which is currently industrially produced by Aspergillus terreus. Heterologous production of itaconic acid with Escherichia coli could help to overcome limitations of A. terreus regarding slow growth and high sensitivity to oxygen supply. However, the performance achieved so far with E. coli strains is still low. We introduced a plasmid (pCadCS) carrying genes for itaconic acid production into E. coli and applied a model-based approach to construct a high yield production strain. Based on the concept of minimal cut sets, we identified intervention strategies that guarantee high itaconic acid yield while still allowing growth. One cut set was selected and the corresponding genes were iteratively knocked-out. As a conceptual novelty, we pursued an adaptive approach allowing changes in the model and initially calculated intervention strategy if a genetic modification induces changes in byproduct formation. Using this approach, we iteratively implemented five interventions leading to high yield itaconic acid production in minimal medium with glucose as substrate supplemented with small amounts of glutamic acid. The derived E. coli strain (ita23: MG1655 ∆aceA ∆sucCD ∆pykA ∆pykF ∆pta ∆Picd::cam_BBa_J23115 pCadCS) synthesized 2.27g/l itaconic acid with an excellent yield of 0.77mol/(mol glucose). In a fed-batch cultivation, this strain produced 32g/l itaconic acid with an overall yield of 0.68mol/(mol glucose) and a peak productivity of 0.45g/l/h. These values are by far the highest that have ever been achieved for heterologous itaconic acid production and indicate that realistic applications come into reach.


DSS-induced colitis is exacerbated in STAT-6 knockout mice.
Several transcription factors have been proposed to regulate IBD including the signal transducer and activator of transcription-6 (STAT-6). The role of STAT-6 was examined in the 5% dextran sulfate sodium (DSS)-induced murine model of colitis using STAT-6 and wildtype mice. The disease activity index (DAI) revealed a significant increase in DAI in STAT-6 mice over STAT-6 mice given DSS. Both STAT-6 and wildtype mice displayed severe inflammation and crypt damage. Additionally, STAT-6 mice showed significant injury to the proximal colon compared with their littermate controls. Furthermore, STAT-6 mice receiving DSS had dramatically higher levels of serum nitrite/nitrate than all other groups. STAT-6 animals also displayed higher levels of inteferon-gamma than wildtype mice. Because STAT-6 has been reported to regulate the expression and activity of inducible NO synthase (iNOS), our data suggest that, in DSS colitis, STAT-6 may modulate iNOS, to limit NO formation and control the extent of inflammation in the colon. We conclude that STAT-6 may normally play an important regulatory role in the pathogenesis of inflammatory bowel disease, possibly through modulation of iNOS and interferon-gamma.


Development of integrated jaw-neck motor function in children at 6, 10 and 13 years of age compared to adults: A kinematic longitudinal study.
The functional integration of the jaw and neck motor systems, of great importance to everyday oral activities, is established in early childhood. Detailed characterisation of this developmental progress is largely unknown. To establish developmental changes in jaw-neck motor function in children over the ages 6-13 years compared to adults. Jaw and head movement kinematics during jaw opening-closing and chewing were longitudinally recorded in 20 Swedish children (8 girls) at 6 (6.3 ± 0.4), 10 (10.3 ± 0.3) and 13 (13.5 ± 0.7) years of age and 20 adults (9 women, 28.2 ± 6.7). Movement amplitudes, jaw movement cycle time (CT), coefficient of variation (CV) and head/jaw ratio for amplitudes were analysed. Linear mixed effect analysis and Welch's t-test were used. Children showed pronounced movement variability and longer CT at 6 and 10 years old during opening and chewing (p < .001). Compared to adults, 6-year-olds showed higher head/jaw ratios (p < .02) and longer CT (p < .001) during opening and chewing, and higher CV-head (p < .001) during chewing. Whereas 10-year-olds showed larger jaw and head amplitudes (p < .02) and longer CT (p < .001) during opening, and longer CT (p < .001) and higher CV-head (p < .001) during chewing. For 13-year-olds, longer CT (p < .001) during chewing was found. Children showed pronounced movement variability and longer movement cycle time at 6-10 years and developmental progress in jaw-neck integration from 6 to 13 years, with 13-year-olds displaying adult-like movements. These results add new detailed understanding to the typical development of integrated jaw-neck motor function.


Primary care sector versus treatment centre follow up for post-treatment cervical squamous intraepithelial neoplasia (SIL).
The NHSCSP 2004 guideline recommends that the best practice for the follow up of post-treatment squamous intraepithelial neoplasia (SIL) is by cytological surveillance in the treatment centre or alternatively, in the primary care sector as a minimum standard. We undertook this study to test this recommendation. In a colposcopy clinic of a district general hospital of UK. Retrospective analysis of 418 patients treated for squamous intraepithelial neoplasia whose post treatment cytological surveillance was followed up for 18 months after treatment. A prospective patient preference questionnaire was also undertaken in 50 randomly chosen patients having treatment to elicit their preferences for the place of follow up. At 8 months 66.4% had their first smear in the primary care sector as compared to 88.6% in the treatment centre. The post-treatment follow up smear performance graph seems to be different for the primary care sector compared to treatment centre. In 12 months nearly 90% of the patients had at least one smear and by 18 months 96.9% of the treated patients had a follow up smear in the community which would detect residual disease. There was no statistically significant difference in the detection of residual disease in the community compared to the treatment centre during the follow up period. A total of 76% of patients preferred to have their follow up smears in the primary care. Targets set by the NHSCSP of UK were not met either by the primary care sector or the treatment centre in the 8 months but was satisfactorily achieved by both in 12 months. These preliminary results indicate a need for a larger multi centre randomised trial to establish if the primary sector is truly a satisfactory alternative for patient follow up. Cytological surveillance is best carried out in the place desired by the patients to allow good compliance with the monitoring process.


Mild insomnia induced by environmental perturbations in the rat: a study of this new model and of its possible applications in pharmacological research.
This paper describes an experimental model for testing new products in a situation where sleep is impaired by a nonpharmacological procedure. This model consists in studying the effect of hypnogenic substances on sleep perturbations which take place when chronically implanted rats are moved to a novel individual cage, connected for the first time to their recording cable and dosed intragastrically with test products, vehicle or placebo. Continuous polygraphic recordings were taken throughout the daily treatment period (15 days) and scored for the presence of slow wave sleep (SWS), and rapid eye movement sleep (REM). The hourly vigilance stages were also computed to determine the parameters of their nycthemeral rhythms. In vehicle-treated rats, a decrease in SWS and REM values and a delay of SWS and REM phases of nycthemeral rhythms occurred on the first day as compared with the end of the recording period. Phenobarbital (5 mg/kg p.o.), mecloqualone (5 mg/kg p.o.) and a new benzodiazepine CM 7302 (7 mg/kg p.o.) led to an immediate or fast recovery, whereas no improvement was observed with placebo and with clorazepate dipotassium (9 mg/kg p.o.). This model can be considered as an attractive model for studying mild insomnia because it allows "true" hypnogenic drugs to be selected in a minimum amount of time.


Skin penetration of infective hookworm larvae. II. The path of migration of infective larvae of Ancylostoma braziliense in the metacarpal foot pads of dogs.
The hairless metacarpal foot pads of six hookworm-free puppies were exposed to infective larvae of Ancylostoma braziliense. Serial sections of the biopts stained with Harris' haematoxylin and eosin showed that the infective larvae are able to penetrate the toughest region of canine skin. Pores of eccrine sweat glands did not seem to constitute sites of entry and no larvae were detected in these glands. Larvae were only observed in the epidermis. The histopathology of the infected skin of the foot pads of the puppies was similar to that in human skin with "creeping eruption" as described by Fülleborn (1927). The biopts appeared to consist of hairy skin as well. In the unexposed adjacent hairy skin of the foot pads, larvae were also observed. They were found in the epidermis, hair follicle systems and dermis, suggesting that the migration from the epidermis into deeper tissue depends on the presence of the hair follicle systems.


Delay in the maturation of muscle fibers in infants with congenital hypotonia.
Muscle biopsies of hypotonic children have shown delayed maturation of a fetal type of muscle fibers: subsarcolemmal halo devoid of activity for mitochondrial dehydrogenases, type II predominance and in some cases abnormal dispersion of fiber diameter. Fiber subtypes within group II were also abnormal. One case has definite embryonic characteristics with presence of myoblasts. Not a single clinical pattern was present in these patients and a variety of associated disorders were recognized. Some patients had a clinical picture corresponding to congenital benign hypotonia as described by Walton.


Chairside measurement of the marginal and internal fit of crowns: a new intraoral scan-based approach.
To investigate the marginal and internal fit of crowns, a new intraoral scanner-based digital approach for chairside measurement (D-IOS) was systematically analysed and compared with the conventional silicone replica technique (CV-SR) and digital 3D analysis software (D-GOM). Two models, representing different clinical situations, were constructed, and the first molar was prepared for a full-coverage crown. On the basis of an intraoral scan (Trios 3), copings of three different materials (non-precious alloy, zirconium dioxide, and resin composite) with two different cement spaces (80 μm, 50 μm) were manufactured. The fit of the copings was investigated by all three methods (CV-SR, D-IOS, D-GOM). Therefore, the cement space was visualized with low-viscosity silicone and digitalized with a second intraoral scan. Evaluation of fit by the D-IOS-method was measured in the intraoral scanner software, whereas for analysis by D-GOM, both intraoral scan datasets were transferred to 3D analysis software (GOM Inspect). The CV-SR-method was used as a control group. For all copings, the measurements were repeated five times. The data were analysed with ANOVA. No significant differences between the three evaluation methods and the coping materials were shown. However, in the occlusal area, the internal gap was significantly higher compared to the internal gap in the marginal and axial areas regardless of the cement space setting (p < .05). The target parameter of the cement spaces did not match the actual measured internal gaps. All three evaluation methods and coping materials can be used for the measurement of fit within different clinical situations. The digital chairside measurement implemented in the intraoral scanner software enables an easy, applicable evaluation of fit of crowns without additional laboratory devices or special software applications.


Weak bases increase surface IgM expression in 70Z/3 B lymphoid cell line without increasing kappa gene expression.
A murine B lymphoid cell line, 70Z/3, has proven to be a useful model to study the developmental transition from membrane immunoglobulin (mIgM)-negative to mIgM-positive B cells. 70Z/3 cells have normal intracellular levels of mu heavy chain but kappa mRNA is transcribed at a very low rate. Because both kappa and mu chains are required for IgM to localize on the external membrane the cells remain essentially mIgM negative. A number of biologic agents can stimulate kappa mRNA transcription leading to increased surface IgM expression. Pharmacologic agents which cause cytoplasmic alkalinization have also been shown to increase the level of surface IgM expression when assessed by FITC-labeled anti-IgM antibodies. This increase has been used to argue that cytoplasmic alkalinization is sufficient to stimulate 70Z/3 differentiation but previous studies did not measure kappa mRNA levels or rate of synthesis. In this study we set out to determine whether the increased FITC staining that follows cytoplasmic alkalinization results from increased surface IgM expression and whether that increase results from activation of kappa mRNA transcription. When cells were treated with methylamine, NH4Cl, or monensin there was transient cytoplasmic alkalinization. The levels of surface IgM expression were measured by flow cytometry of cells stained with FITC-labeled anti-mouse kappa and mu. Using this criterion, a 24-hr treatment with either weak bases or monensin increased the level of surface IgM. The increase in fluorescence was not the result of nonspecific binding or uptake by the cell because there was no increase in fluorescence when methylamine-treated cells were stained with FITC-labeled antibody directed against an antigen not found on these cells. Iodination of surface proteins confirmed that the increase in fluorescence was the result of increased levels of IgM protein on the cell surface. However, exposure of the cells to weak bases or monensin caused no increase in either the steady-state level of kappa light chain mRNA, or in the level of kappa protein. We conclude that the transient alkalinization is not sufficient to induce differentiation of the 70Z/3 cells.


Ganglioneurofibroma arising within the extralobar pulmonary sequestration.
Neurogenic tumor arising within the pulmonary sequestration (PS) is rare. A 42-year-old asymptomatic female was referred to our hospital for work-up of extralobar PS. The independent feeding artery from the thoracic aorta was confirmed by three-dimensional computed tomography angiography (3D-CTA). Uniportal thoracoscopic resection of the sequestrated lung with mediastinal lymph node sampling was performed successfully. Ganglioneurofibroma within the PS was diagnosed as the specimen revealed positive expression of SRY-related HMG-box 10 protein, neuron-specific enolase, S-100, chromogranin A and synuclein. Tumor recurrence was not recorded 1 year after the surgery. Preoperative 3D-CTA is useful to identify the aberrant vessels of PS. An elaborate diagnostic work-up after a timely resection is necessary for subsequent management and follow-up plan.


Interaction between cell sodium and the amiloride-sensitive sodium entry step in rabbit colon.
Ouabain abolishes the short-circuit current (Isc) and decreases the transepithelial conductance (Gt) of rabbit colon. In contrast, amphotericin B elicits a maximum Isc and markedly increases Gt. However, in both instances the amiloride-sensitive Na entry step is completely blocked, presumably due to an increase in cell Na. Conversely, when Na-depleted tissues are suddenly exposed to 140 mM Na, the amiloride-sensitive Isc and the amiloride-sensitive component of Gt (alphaGNa) increase abruptly to their maximum values and then decline to steady-state plateaus with a half time of approximately 6 min; throughout the decline (Isc/alphaGNa) = ENa is constant at a value of 95 mV. In the presence of amphotericin B, the Isc abruptly rises to the same maximum but does not decline. These findings indicate that in the presence of 140 mM Na the conductance of the amiloride-sensitive Na entry step can vary from a maximum value of approximately 1.6 mmhos/cm2 when cell Na is depleted, to zero when cell Na is abnormally elevated (e.g., in the presence of ouabain or amphotericin B). Our findings are consistent with a system in which the pathway responsible for transcellular Na transport parallels another cellular compartment with which it communicates. The Na capacity of the active transport pathway appears to be very small so that this compartment fills rapidly after exposure of Na-depleted cells to 140 mM Na, and active transepithelial Na transport is initiated and reaches steady-state levels quickly. The Na capacity of the second compartment is much larger; the Na content of this compartment appears to be responsible for the negative feedback effect on the permeability of the amiloride-sensitive entry step.


Aneuploidy induced by bleomycin in oocytes of Drosophila melanogaster: studies with the aneuploidy pattern method.
The induction of aneuploidy (numerical chromosome aberrations) in oocytes of Drosophila melanogaster by the antitumor drug bleomycin (BLM) was studied with the "aneuploidy pattern method." The aneuploidy pattern obtained after feeding BLM at a concentration of 50 micrograms/ml to Drosophila females resembles the pattern obtained after X-irradiation (3,000 R). This emphasizes the radiomimetic nature of BLM. In addition, new control material is presented. The new control pattern agrees well with that obtained earlier.