[Effect of infra-low-frequency transcranial magnetic stimulation on motor function in children with spastic cerebral palsy].
To study the therapeutic effects of infra-low-frequency transcranial magnetic stimulation in children with spastic cerebral palsy. Seventy-five children with spastic cerebral palsy were randomly divided into two groups: control (n=33) and treatment groups (n=42). The treatment group accepted infra-low-frequency transcranial magnetic stimulation besides conventional comprehensive rehabilitation therapy. The control group only accepted conventional comprehensive rehabilitation therapy. Motor functions were assessed by gross motor function measure (GMFM) and fine motor function measure (FMFM) at one and three months after treatment. Improvement in the ability to sit in the treatment was better than in the control group at one month after treatment (P<0.05). Improvement in the ability to sit, crawl and kneel, total score of GMFM, and improvement of joint active ability of limbs, grasping ability and operating ability in the treatment group were better than the control group at three months after treatment (P<0.05). Infra-low-frequency transcranial magnetic stimulation can effectively improve motor function in children with spastic cerebral palsy.


Systems-Wide High-Dimensional Data Acquisition and Informatics Using Structural Mass Spectrometry Strategies.
Untargeted multiomics data sets are obtained for samples in systems, synthetic, and chemical biology by integrating chromatographic separations with ion mobility-mass spectrometry (IM-MS) analysis. The data sets are interrogated using bioinformatics strategies to organize the data for identification prioritization. The use of big data approaches for data mining of massive data sets in systems-wide analyses is presented. Untargeted biological data across multiomics dimensions are obtained using a variety of chromatography strategies with structural MS. Separation timescales for different techniques and the resulting data deluge when combined with IM-MS are presented. Data mining self-organizing map strategies are used to rapidly filter the data, highlighting those features describing uniqueness to the query. Examples are provided in longitudinal analyses in synthetic biology and human liver exposure to acetaminophen, and in chemical biology for natural product discovery from bacterial biomes. Matching the separation timescales of different forms of chromatography with IM-MS provides sufficient multiomics selectivity to perform untargeted systems-wide analyses. New data mining strategies provide a means for rapidly interrogating these data sets for feature prioritization and discovery in a range of applications in systems, synthetic, and chemical biology.


Managing pain in feline patients.
This article reviews the current knowledge of pain assessment in cats and the most effective methods for its alleviation. Excellent acute pain management is achievable in cats by using opioids, nonsteroidal anti-inflammatory drugs (NSAIDs), alpha(2)-agonists, and local anesthetics. A multimodal approach using agents that work at different places in the pain pathway is encouraged because this can have added benefits. Management of chronic pain in cats can be challenging, but there is now an approved NSAID for long-term use. As we gain experience with less traditional analgesics, such as gabapentin, and critically evaluate complimentary therapies, our ability to provide comfort to this population of cats should improve.


T7 RNA polymerase bypass of large gaps on the template strand reveals a critical role of the nontemplate strand in elongation.
We show that T7 RNA polymerase can efficiently transcribe DNA containing gaps from one to five bases in the template strand. Surprisingly, broken template strands missing up to 24 bases can still be transcribed, although at reduced efficiency. The resulting transcripts contain the full template sequence with the RNA deleted for the gapped region missing on the template strand. These findings indicate that the end of a downstream template strand can be brought into the polymerase and transcribed as if it were a part of an intact polynucleotide chain by utilizing the unpaired nontemplate strand. This, as well as transcription of an intact template strand, relies heavily upon the non-template strand, suggesting that a duplex DNA-binding site on the leading edge of RNA polymerase is required for RNA chain elongation on DNA templates. This work contributes substantially to the emerging picture that the nontemplate strand is an important element of the transcription elongation complex.


Evidence that the Gh protein is a signal mediator from alpha 1-adrenoceptor to a phospholipase C. I. Identification of alpha 1-adrenoceptor-coupled Gh family and purification of Gh7 from bovine heart.
Our previous studies on alpha 1-adrenoceptor-mediated signaling suggested that Gh is a signal mediator. Gh consists of a 74-kDa GTP-binding alpha-subunit and a 50-kDa beta-subunit. Studies using the alpha 1-agonist-receptor-G-protein ternary complexes from various tissues and species revealed that the intensity (GTP-binding) of the [alpha-32P]GTP-labeled proteins resulting from activating the alpha 1-receptor was significantly attenuated by phentolamine. The molecular masses of GTP-binding proteins were 74 kDa in rat heart and liver, 77 kDa in dog heart, 78 kDa (Gh7 alpha) in bovine heart and liver, and 80 kDa in human heart. Supporting these observations, a specific antibody to Gh7 alpha not only recognized these GTP-binding proteins in the ternary complex preparations, but also co-immunoprecipitated alpha 1-adrenoceptors, indicating a tight association of these GTP-binding proteins with the alpha 1-adrenoceptor. These results also demonstrate that functional and structural similarities exist among these GTP-binding proteins. Additionally, one of the identified G-proteins (termed Gh7) was purified from bovine heart. Gh7 consisted of the 78-kDa GTP-binding protein and a 50-kDa protein.


Spongiosolysis: a new surgical treatment of impotence caused by distal venous leakage.
Until recently ligation of the dorsal veins of the penis had been the only effective surgical treatment in cases of erectile dysfunction caused by venous insufficiency of the corpora cavernosa. Failure of this operation can be owing to persistent distal venous leakage consisting of venous shunts between the distal corpora cavernosa and corpus spongiosum, which can be demonstrated by cavernosography. These shunts can be closed successfully by spongiosolysis, that is by dissecting the distal half of the corpus spongiosum and by isolating the tips of the corpora cavernosa. Of 5 patients who underwent spongiosolysis after previous ligation of the dorsal vein of the penis 4 regained erectile ability with the help of intracavernous injection of a vasoactive drug mixture (15 mg. per ml. papaverine hydrochloride plus 0.5 mg. per ml. phentolamine mesylate, 0.5 to 2 ml. per injection), which was necessary because of concomitant arterial lesions demonstrated by arteriography. The only failure proved to be persistent venous insufficiency of the deep dorsal vein of the penis. Since none of the patients had any serious complication spongiosolysis seems to be a safe procedure in the treatment of distal venous leakage.


Effect of disk maturation on diffusion of low-molecular-weight gadolinium complexes: an experimental study in rabbits.
To measure the rate of enhancement in immature intervertebral disks in comparison with that of mature disks after intravenous administration of gadolinium-containing contrast media. Four rabbits were imaged with MR for 120 minutes after the injection of gadopentetate dimeglumine, gadoteridol, or gadodiamide. Contrast enhancement was measured in the lumbar intervertebral disks on each image. The rate and magnitude of enhancement in the immature and mature disks were compared. Contrast enhancement was detected in intervertebral disks with all three contrast media. Enhancement was significantly greater in immature than in mature animals. Greater enhancement in immature disks is consistent with a lower concentration of fixed negative charges in the glycosaminoglycans of the disk.


Transcatheter arterial embolization for traumatic injury to the pharyngeal branch of the ascending pharyngeal artery: Two case reports.
The ascending pharyngeal artery (APhA) comprises the pharyngeal trunk (PT) and neuromeningeal trunk. The PT feeds the nasopharynx and adjacent tissue, which potentially connects with the sphenopalatine artery (SPA), branched from the internal maxillary artery (IMA). Due to its location deep inside the body, the PT is rarely injured by trauma. Here, we present two cases that underwent transcatheter arterial embolization (TAE) of the PT of the APhA due to trauma and iatrogenic procedure. Case 1 is a 49-year-old Japanese woman who underwent transoral endoscopy under sedation for a medical check-up. The nasal airway was inserted as glossoptosis occurred during sedation. Bleeding from the nasopharynx was observed during the endoscopic procedure. As the bleeding continued, the patient was referred to our hospital for further treatment. Contrast-enhanced computed tomography (CT) demonstrated extravasation in the nasopharynx originating from the right Rosenmuller fossa. TAE was performed and the extravasation disappeared after embolization. Case 2 is a 28-year-old Japanese woman who fell from the sixth floor of a building and was transported to our hospital. Contrast-enhanced CT demonstrated a complex facial fracture accompanying extravasation in the left pterygopalatine fossa to the nasopharynx. Angiography demonstrated an irregular third portion of the IMA. As angiography after TAE of the IMA demonstrated extravasation from the PT of the APhA, additional TAE to the artery was performed. The bleeding stopped after the procedure. Radiologists should be aware that the PT of the APhA can be a bleeding source, which has a potential connection with the SPA.


Effect of very old age on anconeus motor unit loss and compensatory remodelling.
It is not known how the process of compensatory remodeling through collateral reinnervation continues into very old age (>80 years) or whether there is a limit to effective motor unit (MU) reinnervation. Therefore, we explore electrophysiological properties related to motor unit number estimates (MUNEs) in very old participants (79-90 years of age) compared with young controls (25-29 years of age). Decomposition-enhanced spike-triggered averaging was used to collect surface and intramuscular electromyography information from the anconeus to derive a MUNE. Young participants had a MUNE of ∼38 and ∼25 at 30% and 50% root mean squared maximum voluntary contraction (RMSMVC ) with surface motor unit potentials (S-MUPs) of ∼145 μV and 236 μV, respectively. Older participants had a MUNE of ∼23 and ∼16 at 30% and 50% RMSMVC with S-MUPs of 168 μV and 232 μV, respectively. In this muscle, an age limit to successful remodeling through collateral reinnervation, to compensate for the presumed ongoing losses of MUs, may have been surpassed. Muscle Nerve 57: 659-663, 2018.


Refinement of an ovine-based immunoglobulin therapy against SARS-CoV-2, with comparison of whole IgG versus F(ab')2 fragments.
The development of new therapies against SARS-CoV-2 is required to extend the toolkit of intervention strategies to combat the global pandemic. In this study, hyperimmune plasma from sheep immunised with whole spike SARS-CoV-2 recombinant protein has been used to generate candidate products. In addition to purified IgG, we have refined candidate therapies by removing non-specific IgG via affinity binding along with fragmentation to eliminate the Fc region to create F(ab')2 fragments. These preparations were evaluated for in vitro activity and demonstrated to be strongly neutralising against a range of SARS-CoV-2 strains, including Omicron B2.2. In addition, their protection against disease manifestations and viral loads were assessed using a hamster SARS-CoV-2 infection model. Results demonstrated protective effects of both IgG and F(ab')2, with the latter requiring sequential dosing to maintain in vivo activity due to rapid clearance from the circulation.


Methodology for fitting and updating predictive accident models with trend.
Reliable predictive accident models (PAMs) (also referred to as Safety Performance Functions (SPFs)) have a variety of important uses in traffic safety research and practice. They are used to help identify sites in need of remedial treatment, in the design of transport schemes to assess safety implications, and to estimate the effectiveness of remedial treatments. The PAMs currently in use in the UK are now quite old; the data used in their development was gathered up to 30 years ago. Many changes have occurred over that period in road and vehicle design, in road safety campaigns and legislation, and the national accident rate has fallen substantially. It seems unlikely that these ageing models can be relied upon to provide accurate and reliable predictions of accident frequencies on the roads today. This paper addresses a number of methodological issues that arise in seeking practical and efficient ways to update PAMs, whether by re-calibration or by re-fitting. Models for accidents on rural single carriageway roads have been chosen to illustrate these issues, including the choice of distributional assumption for overdispersion, the choice of goodness of fit measures, questions of independence between observations in different years, and between links on the same scheme, the estimation of trends in the models, the uncertainty of predictions, as well as considerations about the most efficient and convenient ways to fit the required models.


[Pustular psoriasis].
The pustular forms o f psoriasis make up a heterogeneous entity from a clinical point of view. However, the existence of clinically detectable aseptic pustular lesions is common to all these forms. How they are related to plaque psoriasis, also called psoriasis vulgaris, the most frequent form of psoriasis, and the genetic and molecular mechanisms recently updated in some forms, have renewed interest in these pustular forms, resulting in reconsideration of their place within cutaneous and systemic inflammatory diseases.


Toxoplasma invasion delayed by TgERK7 eradication.
Toxoplasma gondii causes serious clinical toxoplasmosis in humans mostly due to its asexual life cycles, which can be artificially divided into five tightly coterminous stages. Any radical or delay for the stage will result in tremendous changes immediately behind. We previously demonstrated that TgERK7 is associated with the intracellular proliferation of T. gondii, but during the process, other stages before were not meanwhile determined. To further clarify the function of ERK7 gene in T. gondii, the complemental strain of ΔTgERK7 tachyzoites created previously was engineered via electric transfection with the recombinant pUC/Tgerk7 plasmid, named pUC/TgERK7 strain in this study, and was used together with ΔTgERK7 and wild-type GT1 strains to retrospect the phenotypic changes including invasion and attachment. The results showed that TgERK7 protein can be re-expressed in the ΔTgERK7 tachyzoites and eradication of this protein leads to significantly lower invasion of T. gondii at 1 h and 2 h post-infection (P < 0.05), which is the key factor causing the following slow intracellular proliferation, in comparison with wild-type GT1 and pUC/TgERK7 parasites; noteworthily, at other early time points including 15 min for attachment assay was no statistical difference (P > 0.05). The data suggested that ERK7 protein in T. gondii is an important virulence factor that participates in the invasion of this parasite.


Structural determinants of the stability of thermolysin-like proteinases.
Thermolysin is a member of a family of homologous proteinases which differ in their resistance to thermally induced unfolding and subsequent autolytic degradation. Site-directed mutagenesis studies of the thermolysin-like proteinase (TLP) from Bacillus stearothermophilus (TLP-ste) show that its reduced resistance to thermally induced autolysis, as compared to thermolysin, is due to only some of the 44 naturally occurring amino-acid differences between them. In fact TLP-ste becomes more resistant than thermolysin by mutation of just a few of these amino-acids. The crucial differences are all localized to a solvent-exposed region in the N-terminal domain of TLP-ste.


Determination of organic contaminants in L'Albufera Natural Park using microporous polyethylene tube passive samplers: An environmental risk assessment.
L'Albufera Natural Park (Valencia, Spain) is a protected wetland of international significance that provides critical habitats to endemic and threatened bird and plant species. This study aims to use multiple cross-validation techniques to generate an accurate estimation of the environmental risk of organic contaminants (OCs) in an internationally important coastal wetland, to identify compounds of concern and their potential sources and risk factors. Microporous polyethylene tube (MPT) passive samplers were deployed at 12 locations across L'Albufera Natural Park with concurrent grab samples collected. A subset of MPT samplers were also analysed by an additional laboratory in Australia to widen the range of contaminants and assess interlaboratory reproducibility of results. Forty-three pesticides, 20 pharmaceuticals and personal care products (PPCPs), 20 per-and polyfluoroalkyl substances (PFAS) and 4 organophosphorus flame retardants (OPFRs) were detected in the MPT samplers. The fungicides tebuconazole and difenoconazole were detected at the highest concentrations in passive samplers (maximum concentrations, 153 ng sampler-1 and 106 ng sampler-1, respectively). Several other pesticides were detected in all locations (mean concentrations >1 ng sampler-1). The compounds fenamiphos, propyzamide, difenoconazole, propiconazole, metsulfuron methyl, sodium bis (perfluorohexyl) phosphinate (6:6 PFPiA), 6:2 fluorotelomer sulfonamide alkylbetaine (6:2 FTAB), 6:2 fluorotelomersulfonate (6:2 FTS), citalopram desmethyl and citalopram were reported in the wetland for the first time. Spatial distribution analysis revealed higher pesticide concentrations in the North of L'Albufera. A risk quotient (RQ) analysis showed that ibuprofen is of concern in the area. Overall, the MPT sampling approach is promising as a risk assessment tool for better understanding the transport and fate of OCs in protected areas.


Why and How European Farmers Are Dedicated to Breeding the Dwarf Dahomey Cattle.
This study investigates the motivations and breeding practices of farmers keeping Dahomey cattle in European countries. Data were collected using a web-based open-closed questionnaire survey targeting 55 farmers from Germany, Switzerland and Austria. Descriptive analyses revealed that the earliest European Dahomey herds were established in 2005. Moreover, interest in the breed recently increased as 63.7% of the investigated farmers established their herds between 2016 and 2020. The average herd size comprises seven Dahomey cattle, kept for managing grassland (59.3%), for production of meat or as breeding stock (32.1%) and for a hobby (8.6%). The animals are mostly kept in grazing systems throughout the year, partly fattened with supplement feeds. The low disease incidence and no need for extra health care in the herds indicate the robustness of the breed. Furthermore, meat quality, calving ease, small size, calm character and low feed requirements of Dahomey cattle were valued by the farmers. For the preservation of these features, farmers confirmed their enthusiasm to support any breeding and conservation program of this smallholder breed in Europe and Benin. This study highlights the importance of small-sized cattle for sustainable breeding systems and with regard to ecosystem management practices.


[Morphological and mechanical peculiarities of erythrocytes and neutrophils from patients of different age groups with chronic obstructive pulmonary disease and asthma].
Modern scanning probe microscopy methods make possible to study the peculiarities of the cell response in chronic obstructive pulmonary disease and asthma, as well as to diagnose the severity of the diseases which is important in senile age, particularly.


A novel electron paramagnetic resonance spin-probe technique demonstrates the relation between the production of hydroxyl radicals and ischemia-reperfusion injury.
Many previous studies have suggested an increase in hydroxyl radical (OH) production after myocardial ischemia-reperfusion; however, traditional techniques have not been able to conclusively prove this phenomenon. We investigated whether the production of OH was increased during myocardial reperfusion using a novel electron paramagnetic resonance (EPR) technique using an OH-specific spin probe. An OH scavenger, 3-methyl-1-phenyl-2-pyrazolin-5-one (MCI-186), was used to examine the relationship between OH production and post-ischemic functional recovery or the degree of myocardial injury. We used an isolated rabbit-heart preparation perfused with support-rabbit blood, and the heart was reperfused after normothermic global ischemia. Heart samples were reacted with the OH-specific spin probe, 4-hydroxy-2,2,6,6-tetramethyl-piperidine-N-oxyl (hydroxyl-TEMPO). The rate of decay of the EPR signal showed OH production. We investigated the rate of EPR signal decay and cardiac function. The rate of signal decay was significantly increased just after reperfusion compared with that of pre-ischemia (2.00×10(-2)±0.77×10(-2)min(-1) vs 0.11×10(-2)±0.02×10(-2)min(-1), p<0.01). Administration of MCI-186 reduced the rate of decay to 0.86×10(-2)±0.14×10(-2)min(-1) just after reperfusion (p<0.01). Cardiac function was significantly improved 60 min after reperfusion using MCI-186 compared without MCI-186 (left ventricular developed pressure was 95±9 mm Hg vs 60±6 mm Hg and the first derivative of the left ventricular pressure (dP/dt) was 1843±200 mm Hg s(-1) vs 1182±127 mm Hg s(-1)). A novel EPR spin-probe technique demonstrated the relation between the production of OH and ischemia-reperfusion injury. We confirmed that OH production influenced cardiac function and myocardial ischemia-reperfusion injury.


Purification and partial characterization of camel (Camelus Dromedarius) ceruloplasmin.
Adult and young camel ceruloplasmin (Cp) were isolated and purified using the single-step chromatography on amino ethyl-activated sepharose. There are no differences between the adult and the young camel protein. The molecular mass of the protein, as estimated by SDS-PAGE (denaturant conditions), was approximately 130000 Da. The electrophoretic mobility of camel Cp is slightly higher as compared to human and sheep protein suggesting that the camel Cp is homogeneous, compact and more acid. The copper content was estimated to be 5.8+/-0.3 atoms per molecule. The spectroscopic feature includes an absorption maximum at 610 nm, which could be attributed to type 1 copper. The EPR spectrum was completely devoid of any typical signal of the type 2 copper. The kinetic parameters of the adult camel Cp for the specific activity as p-phenylendiamine oxidase were determined as K(m)=0.42 mM and V(max)=0.93 microM NADH/mn/mg Cp. The optimum pH for the activity was 5.7.


Clock gene defect disrupts light-dependency of autonomic nerve activity.
The discovery of clock genes revealed the major molecular components responsible for circadian time-keeping in mammals, but the mechanism by which autonomic nervous system may control circadian rhythm and its relationship to metabolism is unclear. As the Cry1 and Cry2 genes are indispensable for molecular core oscillator function, we investigated autonomic nervous system activity and metabolism in Cry1-/-Cry2-/- mice. The mice were kept in a light-dark cycle, and showed normal circadian locomotor activities including feeding. However, the circadian rhythmicity of oxygen consumption, heart rate, and body temperature were abolished, suggesting hypermetabolism in these mice. Cry1-/-Cry2-/- mice also showed impaired glucose tolerance due to decreased insulin secretion. These results indicate that sympathetic neural activity in Cry1-/-Cry2-/- mice is elevated, reducing adiposity and impairing insulin secretion and suggest that dysregulation of the autonomic nervous system may induce metabolic disorders.


Venous Thromboembolism After Blood Transfusions in Women Undergoing Hysterectomy for Non-Malignant Indications: A Retrospective Cohort Study.
To quantify the effect of blood transfusion on the risk of venous thromboembolism (VTE) among women undergoing hysterectomy for non-malignant indications. A retrospective cohort study using the American College of Surgeons National Surgical Quality Improvement Program (ACS NSQIP) was conducted. Women who underwent hysterectomy for non-malignant indications between 2011 and 2016 were identified using the Current Procedural Terminology and Internationally Classification of Diseases codes. The primary outcome was development of VTE. Data on patient demographics and perioperative variables were obtained. Pair-wise comparison using χ2 tests were performed to compare women with and without VTE. Multivariable logistic regression was performed to adjust for potential confounders and identify independent predictors of VTE. Between 2011 and 2016, 169 593 women underwent hysterectomy for non-malignant indications. The overall incidence of VTE was 0.32%. Patient characteristics associated with VTE included obesity and higher American Society of Anesthesiologists (ASA) status. Associated operative factors included abdominal surgery, blood transfusion, and prolonged operative time (P < 0.05 for all). Following adjustment for potential confounders, abdominal hysterectomy was associated with greater odds of VTE than laparoscopic or vaginal approaches (adjusted odds ratio [aOR] 1.81; 95% CI 1.48-2.21 and aOR 2.31; 95% CI 1.62-3.28, respectively). Greater odds of VTE were also observed with OR time >150 minutes (aOR 1.88; 95% CI 1.46-2.42), ASA class ≥III (aOR 1.53; 95% CI 1.05-2.26), and intra- and postoperative transfusion (aOR 2.65; 95% CI 1.78-3.95 and aOR 2.98; 95% CI 1.95-4.55, respectively). The risk of VTE is low in women undergoing hysterectomy for non-malignant indications. Blood transfusion was associated with the highest risk of VTE.


Evaluation of univariate ranges with a multivariate standard.
In medical decision making generally many test results are produced. Several authors have discussed the problems involved and concluded that a multivariate approach often has to be preferred. Because until now such an approach has not often been used in practice and qualitative arguments for a multivariate approach are seldom supported by quantitative results, in this paper the use of univariate reference ranges is evaluated with a multivariate range as the standard, taking into account correlation coefficients. Probabilities of false positive and false negative results and predictive values are computed and displayed for decision rules based on two separate test results with an elliptic region as the gold standard. The conclusion is that the quality of the univariate approach is dependent on the correlation coefficient and on the choice of the decision rule. In the most important cases, however, it falls short compared with a multivariate approach. For example, the probability that a univariate positive result is false is around 50% for all values of the correlation coefficient, which is unacceptably high.


Penetration, permeation, and absorption of triamcinolone acetonide in normal and psoriatic skin.
Penetration studies of radiolabelled Triamcinolone acetonide from ointment or cream preparations revealed that in cases of normal as well as psoriatic skin 70-90% of the applied substance remains on the surface. Normal horny layer stores up to 30% of the steroid. Nevertheless, a rapid penetration into the living layers of the skin is observed, whereby the epidermal concentrations reach levels between 5-10(-6) and 3-10(-5) M (mol per liter of tissue). The excretion in the urine took more than 72 h after removal of the excess of substance from the skin. In psoriatic skin, the epidermal and dermal concentrations were 3-10 times higher than in normal skin. This increase lies in the same range as the one resulting from removal of the horny layer by stripping prior to the application, as reported earlier.


HIF prolyl hydroxylase 2/3 deletion disrupts astrocytic integrity and exacerbates neuroinflammation.
Astrocytes constitute the parenchymal border of the blood-brain barrier (BBB), modulate the exchange of soluble and cellular elements, and are essential for neuronal metabolic support. Thus, astrocytes critically influence neuronal network integrity. In hypoxia, astrocytes upregulate a transcriptional program that has been shown to boost neuroprotection in several models of neurological diseases. We investigated transgenic mice with astrocyte-specific activation of the hypoxia-response program by deleting the oxygen sensors, HIF prolyl-hydroxylase domains 2 and 3 (Phd2/3). We induced astrocytic Phd2/3 deletion after onset of clinical signs in experimental autoimmune encephalomyelitis (EAE) that led to an exacerbation of the disease mediated by massive immune cell infiltration. We found that Phd2/3-ko astrocytes, though expressing a neuroprotective signature, exhibited a gradual loss of gap-junctional Connexin-43 (Cx43), which was induced by vascular endothelial growth factor-alpha (Vegf-a) expression. These results provide mechanistic insights into astrocyte biology, their critical role in hypoxic states, and in chronic inflammatory CNS diseases.


Effect of porcine placental extract on menopausal symptoms in postmenopausal women: A prospective, randomized, double-blind, placebo-controlled trial.
To determine the effect of oral administration of porcine placental extract (PPE) on the menopausal symptoms of Korean women. This study was a multicenter, randomized double-blind placebo-controlled trial. A hundred menopausal Korean women were randomly allocated either to the PPE 400 mg per day or the control group during 12 weeks of the study period. Menopausal symptoms were evaluated using the Kupperman Index (KMI). The KMI decreased after 12 weeks in both groups; however, the difference in the changes was not statistically significant. Among overweight or obese women whose body mass index (BMI) was 23 kg/m2 or higher, the KMI decreased more significantly in the PPE group than placebo group after 12 weeks (-18.52 ± 9.09 vs -11.40 ± 10.68, P < 0.05). Among 49 early menopausal women whose duration of menopause was less than 3 years, the KMI decreased more significantly in the PPE group after 12 weeks (-17.29 ± 9.07 vs -11.29 ± 10.68, P < 0.05). The baseline estradiol level was not statistically different between the two groups before trial, but the estradiol level of the PPE group was significantly increased at 12 weeks (11.13 ± 35.13 pg/m, P < 0.05). Oral administration of 400 mg PPE per day decreases menopausal symptoms in women with BMI 23 kg/m2 or higher or in early menopausal women. PPE may be considered as a short-term complementary treatment to reduce menopausal symptoms, especially in overweight or early menopausal women.


Subthalamic DBS replaces levodopa in Parkinson's disease: two-year follow-up.
Subthalamic nucleus (STN) deep brain stimulation (DBS) of patients with PD allows reduction of antiparkinsonian medication but has only a mild direct effect on dyskinesia. Since antiparkinsonian medication has short- and long-term effects that may prevent an estimate of the maximal possible impact of STN DBS, such medication was used at the lowest possible dosage after DBS implantation. To study the maximal and long-term effects of STN DBS using the lowest dose of medication. Twenty consecutive patients with PD with motor fluctuations and dyskinesia underwent bilateral implantation under stereotactic guidance, microrecording, and clinical control. All medications were stopped before implantation and reintroduced, at the lowest dosage needed, only if the postoperative motor score did not reach the baseline level. Unified PD Rating Scale (UPDRS) motor (subscale III) scores were measured at baseline and after 3, 6, 12, and 24 months. After 21 plus minus 8 months, the UPDRS III "off-medication" score was decreased by 45% and was similar to the preoperative UPDRS III "on" score. Overall, medication was reduced by 79%, being completely withdrawn in 10 patients. Fluctuations and dyskinesia showed an overall reduction of >90%, disappearing completely in patients without medication. These improvements were maintained for 2 years. These results show that STN DBS could replace levodopa and allowed all antiparkinsonian medication to be discontinued in 50% of patients with PD. Fluctuations and dyskinesia disappeared completely in these patients but persisted in those still on medication. These improvements were maintained for 2 years.


[Complications of corneal lamellar refractive surgery].
Techniques available for corneal lamellar refractive surgery are laser-assisted in situ keratomileusis (LASIK) using a microkeratome or femtosecond laser incision followed by excimer laser corneal ablation, and femtosecond laser-assisted refractive lenticule extraction (ReLEx). These treatments are nowadays considered to be safe and effective standard procedures for surgical correction of mild to moderate ametropia. Possible complications include too small or decentered optical zones, intraoperative flap cutting errors and postoperative inflammation (e.g. diffuse lamellar keratitis, DLK), epithelial or flap folds, epithelial ingrowths or iatrogenic ectasia. The occurrence of complications may be significantly reduced by compliance to corresponding standards of indication and treatment that are based on current scientific knowledge.


Combination of antidepressants in the treatment of major depressive disorder: a systematic review and meta-analysis.
The objective was to perform a systematic review and meta-analysis of studies that assessed the effect of the combination of antidepressants from the beginning of the treatment of major depressive disorder. Studies were retrieved from PubMed (1966 to August 2010), Cochrane Library (August 2010), Embase (1980 to August 2010), PsycINFO (1980 to August 2010), Lilacs (1982 to August 2010), clinical trials registry, thesis database (www.capes.gov.br), and secondary references. All randomized controlled trials that compared a combination of antidepressants with a single antidepressant from the beginning of the treatment of major depressive disorder in adults were included. Data analysis was performed using the Review Manager 5.0. Of 3492 studies retrieved, five satisfied the inclusion criteria. In one study, only data about dropouts were included. Antidepressant combination was shown to be better than a single antidepressant considering remission (relative risk [RR], 2.71; 95% confidence interval [CI], 1.69-4.35) and response (RR, 1.55; 95% CI, 1.21-1.97). Mirtazapine plus selective serotonin reuptake inhibitor (SSRI) was superior to an isolated SSRI for remission (RR, 1.88; 95% CI, 1.06-3.33). Tricyclic antidepressant plus SSRI was superior to SSRI for remission and response (RR, 8.58; 95% CI, 1.70-43.32 and RR, 1.78; 95% CI, 1.07-2.93, respectively). There was no difference between combined and monotherapy groups in dropouts owing to adverse effects. The results suggest that antidepressant combination is more efficient than a single antidepressant without a significant decrease in tolerability. However, the small number of clinical trials and methodological problems precludes definitive conclusions.


School scoliosis screening and the United States Preventive Services Task Force. An examination of long-term results.
Analysis of scoliosis screening in Dublin (1979-1990) of patients who passed significant levels (25 degrees and 40 degrees) and received orthotic treatment or surgery. To reexamine the basis of school scoliosis screening. The United States Preventive Services Task Force questioned the validity of school screening for adolescent idiopathic scoliosis because there is insufficient scientific evidence to justify it. New entrants (1986-87) to the screening program were used to test the validity of the screening methods. The total screened population, proportions reaching significant Cobb angles or undergoing treatment, were compared across the time period. The sensitivity of the forward bend test in detecting or predicting 40 degrees scoliosis was 0.83, the specificity was 0.99, and the predictive value was 0.08. The proportion prescribed braces declined significantly (P < 0.01), but there was no significant change in those with scoliosis greater than 24 degrees (0.18%) or 39 degrees (.08%) nor was there a significant change in the numbers needing surgery (.045%). The incidence of significant scoliosis and of surgery is low and independent of changes in bracing policy. The validity of screening to cause significant change in prevalence must be challenged.


A reliable method of high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry for determining selenoamino acids in selenoproteins from Lactococcus lactis.
A reliable method for simultaneous determination of four organic selenium species by HPLC-ICP-MS was developed and implemented in determining organic selenoamino acids (Se-AAs) in selenoproteins from Lactococcus lactis (L. lactis) NZ9000. The method consisted of liberating Se-AAs from selenoproteins using ultrasound-assisted protease hydrolysis, and quantitatively detecting Se-AA speciations by HPLC-ICP-MS. After optimizations of proteolysis conditions, chromatographic conditions and determination conditions, the established method could efficiently separate the four Se-AAs, including SeCys, SeCys2, SeMeCys and SeMet within 10 min. It presented high sensitivity with the limits of detection and quantitation in the range of 0.197∼0.240 μg∙L-1 and 0.788∼0.960 μg∙L-1, respectively, good repeatability with a relative standard deviation (RSD) of less than 5%, and good recovery in the desired floating range of 90%∼105%, verifying the good accuracy. The method successfully detected four selenium species in the purified glutathione peroxidase (LlGPx) overexpressed in L. lactis NZ9000, SeCys (0.9716∼1.6784 μg∙g-1), SeCys2 (1.0695∼1.2124 μg∙g-1), SeMeCys (0.7288∼0.7984 μg∙g-1) and SeMet (1.0058∼1.9571 μg∙g-1), accounting for up to 80.14% of total selenium. There was no difference of order of magnitude in the four Se-AAs, indirectly indicating the random incorporation of selenium into selenoprotein LlGPx in L. lactis NZ9000. This work throws new light on the identification and biosynthesis of organic selenium species in selenoproteins and selenium-riched organisms like L. lactis.


Six putative IQ motifs of the recombinant chicken intestinal brush border myosin I are involved in calmodulin binding.
Chicken brush border myosin I has up to six IQ sequence motifs at which it may bind calmodulin. To determine the relative contributions of these motifs to calmodulin binding, fusion deletion fragments were expressed in Escherichia coli and their ability to bind calmodulin was assessed by the gel overlay technique. The first three N-terminal IQ sites showed strong binding with calmodulin. Surprisingly, the last three incomplete IQ motifs also contributed substantial calmodulin binding. The first and fourth IQ sites bound calmodulin but tended to reduce binding in combination with other sites. The data indicate that interactions among all six IQ motifs contribute to the ability of myosin I to bind calmodulin.


Alternative reproductive tactics in male Cape ground squirrels Xerus inauris.
In some animal societies, males vary in the strategies and tactics that they use for reproduction. Explanations for the evolution of alternative tactics have usually focussed on extrinsic factors such as social status, the environment or population density and have rarely examined proximate differences between individuals. Anecdotal evidence suggests that two alternative reproductive tactics occur in cooperatively breeding male Cape ground squirrels. Here we show that there is strong empirical support for physiological and behavioural differences to uphold this claim. 'Dispersed' males have higher resting metabolic rates and a heightened pituitary activity, compared with philopatric 'natal' males that have higher circulating cortisol levels. Dispersed males also spend more time moving and less time feeding than natal males. Additionally, lone males spend a greater proportion of their time vigilant and less of their time foraging than those that were in groups. The choice of whether to stay natal or become a disperser may depend on a number of factors such as age, natal group kin structure and reproductive suppression, and the likelihood of successful reproduction whilst remaining natal. Measuring proximate factors, such as behavioural and endocrine function, may provide valuable insights into mechanisms that underlie the evolution of alternative reproductive tactics.


[Handling of COVID-19 Outbreak on a Non-invasive Ventilation Ward].
COPD patients have a higher risk of experiencing severe COVID-19 illness. The outbreak of COVID-19 on an in-patient ward for non-invasive ventilation (NIV) furthermore demonstrated high mortality (32 %) for COPD patients with ongoing NIV and indicated enhanced contagiousness by used equipment.Prophylactic and therapeutic measures taken against COVID-19 are hereby displayed.


Actinic folliculitis.
We report a case of actinic folliculitis in a 29-year-old woman who presented with a 10-year history of a recurrent pustular eruption affecting her face, typically appearing 4-6 h after exposure to sunlight. Actinic follicuitis is a rare photodermatosis, which falls into the same spectrum as acne aestivalis and actinic superficial folliculitis.


Targeting gastrin-releasing peptide receptors of prostate cancer cells for photodynamic therapy with a phthalocyanine-bombesin conjugate.
Sulfonated aluminum phthalocyanines (AlPcS) are potent photosensitizers for the photodynamic therapy (PDT) of cancer. In this study we evaluate the possibility to improve the efficacy of AlPcS-PDT for prostate cancer by targeting tetrasulfonated aluminum phthalocyanines (AlPcS(4)) to the gastrin-releasing peptide receptor (GRPR) through coupling to bombesin. A mono-carbohexyl derivative of AlPcS(4) is attached to 8-Aoc-bombesin(7-14)NH(2) via an amide bridge to yield a bombesin-AlPcS(4) conjugate linked by a C-14 spacer chain. The conjugate is characterized by mass spectroscopy and shown to bind to the GRPR with a relative binding affinity (RBA) of 2.3, taking bombesin (RBA=100) as unity. The in vitro photodynamic efficacy of the conjugate against PC-3 human prostate cancer cells is improved by a factor 2.5 over the non-conjugated mono-carbohexyl derivative of AlPcS(4).


Long-term outcomes of drug-induced autoimmune-like hepatitis after pulse steroid therapy.
Pulse steroid therapy occasionally causes drug-induced autoimmune-like hepatitis (DI-ALH), but the long-term outcome of treated patients is not well known. In this study, we investigated the long-term outcomes of DI-ALH due to pulse steroid therapy. We retrospectively reviewed the medical records of 405 patients treated with pulse high-dose methylprednisolone in Kurashiki Central Hospital. The frequency and clinicopathological characteristics of acute liver injury that occurred within 3 months after the therapy were analyzed. The diagnosis of DI-ALH was made according to the revised international autoimmune hepatitis group criteria. Among the 405 patients treated with methylprednisolone, 61 (15.1%) had acute liver injury after the pulse steroid therapy, and DI-ALH was diagnosed in five patients (1.2%). Absence of oral prednisolone tapering after the pulse steroid therapy was a significant risk factor for the subsequent development of DI-ALH (odds ratio 11.9; p = 0.017). One patient was treated with 3 days of intravenous methylprednisolone followed by oral prednisolone. Two patients were treated with glycyrrhizin followed by oral prednisolone due to ineffectiveness of glycyrrhizin. Remission was achieved with glycyrrhizin alone, and spontaneous remission without drug therapy occurred in one patient each. During the median follow-up period of 34 months, no relapse was evident in all the patients without maintenance therapy. Pulse steroid therapy can cause DI-ALH, especially when subsequent prednisolone is not tapered. Prednisolone is effective for DI-ALH due to pulse steroid therapy, and can be safely withdrawn once remission is achieved.


Femtosecond cataract surgery: A review of current literature and the experience from an initial installation.
Cataract surgery remains the most widely performed intraocular procedure throughout the world. Safety and accuracy of the procedure are paramount and techniques should remain under constant review. Recently, the introduction of the femtosecond laser to assist cataract surgery has provided ophthalmologists with an exciting tool that may further improve outcomes. We review the existing literature and discuss the installation and initial experience of a femtosecond laser into our practice.


Endoscopic laser therapy for duodenal villous adenoma.
A 72-year-old woman had a 20-mm supraampullary villoglandular adenoma with a focal carcinoma in situ. The neurological status of the patient did not allow any major surgical procedure. Therefore laser therapy was performed using the Nd:YAG laser. Six laser sessions were necessary to destroy the whole adenoma. After 30 months of follow-up, there was neither endoscopic nor histologic recurrence. The laser therapy should be therefore considered in treating large sessile duodenal villous adenoma occurring in inoperable patients. However, a repeated endoscopic surveillance must be performed.


Comparative assessment of anti-cancer drugs against NUDT15 variants to prevent leucopenia side effect in leukemia patients.
Human nucleotide triphosphate diphosphatase (NUDT15) is one of the essential proteins involved in the hydrolysis of anti-cancer drugs against leukemia. Polymorphisms in NUDT15 significantly affect the hydrolysis activity that leads to side effects, including leucopenia. Drugs having a better affinity with NUDT15 protein and contributing stable conformation may benefit patients from leucopenia. Most frequent NUDT15 polymorphisms causing structure variability and their association with leukemia were screened. The selected protein variants and anti-cancer drug structures were collected. Further, molecular docking was performed between drugs and NUDT15 variants along with the wild-type. Finally, molecular dynamics were executed for 100 ns to understand the stability of the protein with the anti-cancer drug based on molecular trajectories. Three-dimensional structures of NUDT15 wild, the most frequent variants (Val18Ile, Arg139Cys, and Arg139), and the anti-cancer drugs (azathioprine, mercaptopurine, and thioguanine) were selected and retrieved from structure databases. On molecular docking the binding energies of anti-cancer drugs against NUDT15 structures ranged from - 5.0 to - 5.9 kcal/mol. Among them, azathioprine showed the highest affinities (- 7.3 kcal/mol) for the wild and variant structures. Additionally, the molecular dynamics suggest all analyzed NUDT15 were stable with azathioprine based on the dynamic trajectories. Our results suggest azathioprine could be the preferable anti-cancer drug for the population with NUDT15 variants that could effectively be hydrolyzed as evidenced by molecular docking and dynamic simulation.


Gender and age affect the levels of exhaled nitric oxide in healthy children.
Asthma is a chronic inflammatory disorder of the lung and diagnosis is difficult in children. The measurement of fractional exhaled nitric oxide (FeNO) may be useful in the diagnosis and monitoring of treatments. A number of factors affect FeNO levels and their influence varies across countries and regions. This study included 300 healthy students, aged from 6 to 14 years, who participated voluntarily. A comprehensive medical survey was used and measurements of FeNO levels and spirometric parameters were recorded in Shenyang, China. We observed that the median FeNO was 11 ppb (range, 8-16 ppb) in children from the northern areas of China. For males, the median level was 13 ppb (range, 9-18 ppb) and the median level was 10 ppb (range, 8-14 ppb) for females. There was a significant difference between males and females (P= 0.007) and age was correlated with FeNO (R2= 0.6554), while weight, height, body mass index (BMI), forced vital capacity (FVC), forced expiratory volume (FEV1), FEV1/FVC and peak expiratory flow (PEF) had no correlation with FeNO. In conclusion, the median FeNO is 11 ppb (range, 8-16 ppb) in male and female healthy children from northern areas of China and is affected by gender and age.


Evaluation of factors influencing annual occurrence, bioaccumulation, and biomagnification of antibiotics in planktonic food webs of a large subtropical river in South China.
Biological pump is important to control the fate and distribution of organic contaminants, particularly in temperate and cold oligotrophic waters. However, it remains largely unknown how factors affect the long-term occurrence and fate of ionogenic organic compounds in subtropical eutrophic waters. The present study aimed to assess biogeochemical and physical factors affecting the annual occurrence, bioaccumulation, and trophic transfer of 14 antibiotics through planktonic food webs in the Pearl River, a large subtropical eutrophic river in China. This was done by carrying out 1-year simultaneous field observations of antibiotic concentrations in five water column compartments and assessing the variability of bioconcentration (BCF), bioaccumulation (BAF), and biomagnification (BMF) factors, which were influenced by plankton biomass, pH and temperature of water columns. The annual mean antibiotic concentration per site ranged from 1014.66 ± 535.66 ng L-1 to 1464.63 ± 1075.91 ng L-1, and was positively correlated with phytoplankton biomass, but independent of the proximity of the sites to urban areas. Antibiotic occurrences in both phytoplankton and zooplankton were greatly influenced by a biodilution effect. The annual occurrence of antibiotics in the water column was modulated by biological pumps as well as their equilibrium partitioning, and indirectly influenced of eutrophication with pH increased with phytoplankton biomass and phytoplankton life cycling. BAF of antibiotics by plankton had biphasic correlations with temperature (n = 150, R2 = 0.17-0.60, p < 0.001) and decreased with plankton biomass (n = 105-147, R2 = 0.10-0.22, p < 0.001). The trophic transfer of antibiotics from phytoplankton to zooplankton (BMFs) were positively correlated with both phytoplankton biomass (n = 30, R2 = 0.58, p < 0.001) and temperature (n = 132-150, R2 = 0.12-0.43, p < 0.001). Mean BMFs of ciprofloxacin, lomefloxacin, ofloxacin, oxytetracycline, and tetracycline ranged between 0.18 and 2.25, implying these chemicals can undergo biomagnification along planktonic food webs. The present research demonstrates the important role of biogeochemical and physical factors in the environmental fate of antibiotics at large spatiotemporal scales.


Prenatal diagnosis as a test for genodermatoses: its past, present and future.
The prenatal diagnosis (PND) of severe hereditary skin diseases started in the early 1980s using fetal skin biopsy techniques based on ultrastructural and immunohistochemical abnormalities of the fetal skin. Recent success in identifying responsible genes and demonstrating mutations in such genes has set the stage for DNA-based PND in the 1990s. Common examples of skin conditions which can be prenatally diagnosed include epidermolysis bullosa, oculocutaneous albinism and Harlequin ichthyosis in which the severity of the clinical phenotype appears to justify PND in families at risk. More recently, preimplantation diagnoses of inherited diseases have become possible using in vitro fertilization techniques. The diagnosis consists of a blastomere biopsy of the six to ten-cell embryo and a DNA analysis of single blastomeres. Disease-free embryos are selected for transfer to the uterus, thereby avoiding the need for termination of a fetus found to be affected by conventional PND. Furthermore, carrying out a PND using a single fetal cell from the maternal blood, such as nucleated erythrocytes, has become technically feasible. Although there are many questions that remain unanswered, the outlook for further development of noninvasive PND in the future appears optimistic.


Prevalence of American Heart Association type VI carotid atherosclerotic lesions identified by magnetic resonance imaging for different levels of stenosis as measured by duplex ultrasound.
Via magnetic resonance imaging (MRI), we sought to determine the prevalence of atherosclerotic American Heart Association type VI lesions (AHA-LT6) (lesions with luminal surface defect, hemorrhage/thrombus, or calcified nodule) in carotid arteries that represented all categories of stenosis as measured by duplex ultrasound. Arterial stenosis alone has been shown to be a poor predictor of cardiovascular events. Autopsy studies suggest that features associated with AHA-LT6 lesions, rather than the degree of luminal narrowing, characterize the high-risk plaque. A total of 192 subjects underwent bilateral carotid artery magnetic resonance imaging (MRI) scans at 1.5T after evaluation with ultrasound to determine stenosis. After excluding arteries with a previous endarterectomy, poor image quality, or missing ultrasound data, there were 175 patients with 260 arteries available for analysis. The AHA lesion type was determined by the consensus opinion of 2 experienced carotid MRI reviewers. In total, 96 of 260 (37.0%) arteries had >or=1 location with AHA-LT6. Of the arteries with AHA-LT6, 84.4% had hemorrhage, 45.8% had a ruptured fibrous cap, and 14.6% showed other type of complications. Prevalence of AHA-LT6 was an increasing sequence of 8.1% in the 37 arteries with 1% to 15% stenosis, 21.7% in the 60 arteries with 16% to 49% stenosis, 36.8% in the 114 arteries with 50% to 79% stenosis, and 77.6% in the 49 arteries with 80% to 99% stenosis. Complicated AHA-LT6 are frequently found in arteries with <or=50% stenosis. These findings indicate that complex lesions develop in a substantial number of arteries in the absence of high-grade stenosis. Ongoing prospective studies will determine the predictive value of vulnerable plaque features, as visualized by MRI, for risk of subsequent ischemic events.


Gender differences in subtypes of depression by first incidence and age of onset: a follow-up of the Lundby population.
The Lundby Study is a prospective mental health survey in a community population (N = 3563), in which data were collected in 4 waves of field-work between 1947 and 1997. We investigated gender differences during the follow-up in overall first incidence rates, ages of onset, and incidence by age of onset patterns, in different subtypes of depression. The overall incidence rate in females was higher than males for most subtypes of depression. However, for depression with melancholic and/or psychotic features, the overall first incidence rate did not differ significantly between the genders. The mean age of onset did not differ significantly between females and males in any of the depressive subtypes. Nevertheless, females and males had different first incidence rates by age of onset patterns for unipolar non-melancholic DSM-IV mood disorder and major depressive disorder (MDD), with a consistent gender incidence gap across all ages, but with the most conspicuous gender gap in middle age. The first incidence rates by age of onset patterns for DSM-IV MDD with melancholic and/or psychotic features did not differ significantly between the genders. The findings support that females are more prone than males to develop depression with medium severity, but no gender differences were found in melancholic and/or psychotic depression. The findings may support that unipolar non-melancholic depression and melancholic and/or psychotic depression represents different disorders. Tentative explanations for this are discussed.


Cardiovascular magnetic resonance of acute myocardial infarction at a very early stage.
Very early changes in myocardial tissue composition during acute myocardial infarction (AMI) are difficult to assess in vivo. Cardiovascular magnetic resonance (CMR) imaging provides techniques for visualizing tissue pathology. The diagnostic role of CMR in very acute stages of myocardial infarction is uncertain. We investigated signal intensity changes beginning within 60 min after acute coronary occlusion in patients undergoing therapeutic septal artery embolization. We investigated eight patients with hypertrophic obstructive cardiomyopathy undergoing interventional septal artery embolization by applying microparticles to reduce left ventricular outflow tract obstruction. In a clinical 1.5-tesla (T) CMR system, we visualized infarct-related myocardial signal by T(1)-weighted sequences before and 20 min after administration of contrast media (delayed enhancement) and edema-related signal by T(2)-weighted spin-echo sequences before and 58 +/- 14 min after the intervention as well as on days 1, 3, 7, 14, 28, 90, and 180 during follow-up. Infarct-related changes as defined by contrast enhancement were observed as early as 1 h after the intervention and during six months of follow-up. In contrast, infarct-related myocardial edema, as visualized by high signal intensity in T(2)-weighted spin-echo sequences, was not consistently detectable 1 h after acute arterial occlusion; this was possible in all subsequent studies until day 28. Contrast-enhanced magnetic resonance imaging detected infarct-related signal changes as early as 1 h after AMI in humans, whereas the sensitivity of edema-related signal changes was not sufficient during this very early stage.


Clinical characteristics associated with isolation of small-colony variants of Staphylococcus aureus and Pseudomonas aeruginosa from respiratory secretions of patients with cystic fibrosis.
During a 3-month period, small-colony variant phenotypes of both Staphylococcus aureus and Pseudomonas aeruginosa were isolated from respiratory secretions of 8.2% and 9.2%, respectively, of 98 patients with cystic fibrosis, particularly those with advanced pulmonary disease and prolonged antibiotic exposure.


Pediatric nurses' knowledge and practices of nasogastric tube placement and verification: A cross-sectional study.
The study assesses pediatric nurses' knowledge and routine practices in relation to nasogastric tube (NGT) placement and verification procedure. By investigating these aspects, the study seeks to contribute to the enhancement of patient safety and the improvement of pediatric nursing care in the context of NGT procedures by ensuring accurate NGT placement and minimizing complications in pediatric patients. The study assesses pediatric nurses' knowledge and practices of inserting and confirming the placement of NGTs among pediatric patients in the Asir Province, Saudi Arabia. In this multihospital cross-sectional study, a total of 130 pediatric nurses participated. The study used a validated questionnaire comprising two parts: one focusing on the demographic data of the pediatric nurses and the other assessing their knowledge and practices regarding NGT placement and verification. The collected data were analyzed using descriptive statistics and linear regression tests in the SPSS software. The research question addressed in this study was to determine the knowledge and practice levels of pediatric nurses in relation to NGT placement and verification. This study included 130 registered pediatric nurses, of whom more than half had unsatisfactory knowledge (53.1%) and incompetent practices (58.5%) related to NGT insertion. Linear regression associated with perceived nurses' practices scores explained that knowledge score, years of experience, educational qualification, and training courses had a high-frequency positive effect on the practice score, with P < 0.01. Linear regression associated with perceived nurses' knowledge scores explained that educational qualification, practice score, and training courses had a high-frequency positive effect on the knowledge score, with P < 0.01. The study found that pediatric nurses' knowledge and practices regarding NGT were inadequate. These results highlight the importance of implementing continuous educational programs to improve care practices pertaining to nasogastric tubes and enhance patient safety and quality of care.


Ruggedness testing of chromatographic methods: selection of factors and levels.
The first step in a ruggedness test is the selection of factors to be examined and their levels. In this paper, both topics are discussed, thereby completing a strategy described earlier. It is demonstrated, by means of some examples, that depending on the formulation (definition) of a factor, information that is physically more or less meaningful is extracted from the experimental design results. Among others, the inclusion of the compounds of a buffer and of the components of a mixture in a screening design were examined. A general guideline to select the levels of the factors in a ruggedness test was proposed. Some special cases, i.e. asymmetric intervals around the nominal level, were also discussed.


Correlation Between the Timing of Diagnostic Ureteroscopy and Intravesical Recurrence in Upper Tract Urothelial Cancer.
To identify the effects of diagnostic ureteroscopy on intravesical recurrence, 104 patients who had undergone radical nephroureterectomy were analyzed. The number of patients with intravesical recurrence was 34 (32.6%) at a mean of 37 months. Multivariate Cox model analysis revealed that delayed nephroureterectomy after diagnostic ureteroscopy were independent predictive factors for intravesical recurrence. The purpose of this study was to evaluate the effects of the timing of diagnostic ureteroscopy (URS) relative to radical nephroureterectomy (RNU) on intravesical recurrence (IVR). We retrospectively evaluated 104 patients who had undergone RNU for upper tract urothelial cancer (UTUC) at a single tertiary referral center between March 2003 and December 2012. All patients were divided depending on the timing of diagnostic URS: the no URS group (30 patients, no URS), 1-session group (33 patients, diagnostic URS immediately followed by RNU), and 2-session group (41 patients, RNU after diagnostic URS at a median time of 5 days). We analyzed for IVR-free survival using the Kaplan-Meier and Cox proportional regression methods. Of the 104 patients, 34 (32.6%) developed subsequent IVR at a mean interval of 7.2 months. The Kaplan-Meier curve showed that the IVR rate was significantly greater in the 2-session group than in the other groups (P = .004). Univariate analysis indicated no effect on IVR in the 1-session group compared with the no URS group (hazard ratio [HR], 95% confidence interval [CI] 1.58; 0.517-4.833). However, the 2-session group had a significantly greater number of patients with IVR than the no URS group (HR, 3.82; 95% CI 1.438-10.131). Multivariate Cox proportional analysis revealed that the 2-session group was an independent predictor of IVR in UTUC patients (HR, 3.61; 95% CI 1.039-12.557). Delay of RNU after diagnostic URS significantly increased the risk of IVR in UTUC patients.


Herpes simplex virus type 1 induces nuclear accumulation of hyperphosphorylated tau in neuronal cells.
Herpes simplex virus type 1 (HSV-1) is a neurotropic virus that remains latent in host neurons. Viral DNA replication is a highly structured process in which the redistribution of nuclear proteins plays an important role. Although tau is most widely known as a microtubule-associated protein found in a hyperphosphorylated state in the brains of patients with Alzheimer's disease (AD), this protein has also been detected at other sites such as the nucleolus. Here, we establish that HSV-1 infection gives rise to an increase in tau phosphorylation and that hyperphosphorylated tau accumulates in the nucleus, forming defined structures in HSV-1-infected neuronal cells reminiscent of the common sites of viral DNA replication. When tau expression in human neuroblastoma cells was specifically inhibited using an adenoviral vector expressing a short hairpin RNA to tau, viral DNA replication was not affected, indicating that tau is not required for HSV-1 growth in neuronal cells. Given that HSV-1 is considered a risk factor for AD, our results suggest a new way in which to understand the relationships between HSV-1 infection and the pathogenic mechanisms leading to AD.


Differential ubiquitination of Smad1 mediated by CHIP: implications in the regulation of the bone morphogenetic protein signaling pathway.
Smad1, a downstream regulator of the bone morphogenetic protein (BMP) receptors, is tightly regulated by the ubiquitin-proteasomal degradation system. To dissect the mechanisms that underlie the regulation of Smad1, it is important to investigate the specific ubiquitination site(s) in Smad1. Here we report that the alpha-NH(2) group of the N terminus and the epsilon-NH(2) groups of internal lysine residues 116, 118 and 269 (K116, K118 and K269) of Smad1 are ubiquitin acceptor sites mediated by the carboxyl terminus of Hsc70-interacting protein (CHIP). The in vitro degradation assay indicates that ubiquitination at the N terminus partially contributes to the degradation of Smad1. Furthermore, we demonstrate that the ubiquitination level of pseudo-phosphorylated Smad1 by CHIP is stronger than that of wild-type Smad1 and can be strongly inhibited by a phosphorylated tail of Smad1, PIS(pS)V(pS). Third, our results indicate that Hsp70 facilitates CHIP-mediated poly-ubiquitination of Smad1 whereas it attenuates CHIP-meditated mono-ubiquitination of Smad1. Finally, consistent with the in vitro observation, we show that CHIP preferentially mediates the degradation of phospho-Smad1/5 in vivo. Taken together, these results provide us a hint that CHIP might preferentially regulate phosphorylated Smad1 and thus the BMP signaling.


Using clinically derived human tissue to 3-dimensionally bioprint personalized testicular tubules for in vitro culturing: first report.
To study the feasibility and spermatogenic potential of 3-dimensional (3D) bioprinting personalized human testicular cells derived from a patient with nonobstructive azoospermia (NOA). A human testicular biopsy from a single donor with NOA was dissociated into single cells, expanded in vitro, and 3D bioprinted into tubular structures akin to the seminiferous tubule using AGC-10 bioink and an RX1 bioprinter with a CENTRA coaxial microfluidic printhead from Aspect Biosystems. Three-dimensional organoid cultures were used as a nonbioprinted in vitro control. Academic medical center. A 31-year-old man with NOA with testis biopsy demonstrating Sertoli cell-only syndrome. Three-dimensional bioprinting and in vitro culturing of patient-derived testis cells. Cellular viability after printing was determined, along with the expression of phenotypic and spermatogenic functional genetic markers after 12 days of in vitro culture. Testicular cultures were expandable in vitro and generated sufficiently large numbers for 3D bioprinting at 35 million cells per mL of bioink. Viability 24 hours after printing was determined to be 93.4% ± 2.4%. Immunofluorescence staining for the phenotype markers SRY-Box transcription factor 9, insulin-like 3, actin alpha 2 smooth muscle, and synaptonemal complex protein 3 after 12 days was positive, confirming the presence of Sertoli, Leydig, peritubular myoid, and meiotic germ cells. Reverse transcription qualitative polymerase chain reaction analysis showed that after 12 days in spermatogenic media, the bioprints substantially up-regulated spermatogenic gene expression on par with nonbioprinted controls and showed a particularly significant improvement in genes involved in spermatogonial stem cell maintenance: inhibitor of deoxyribonucleic acid binding 4 by 365-fold; fibroblast growth factor 3 by 94,152-fold; stem cell growth factor receptor KIT by twofold; stimulated by retinoic acid 8 by 125-fold; deleted in azoospermia-like by 114-fold; synaptonemal complex protein 3 by sevenfold; zona pellucida binding protein by twofold; transition protein 1 by 2,908-fold; and protamine 2 by 11-fold. This study demonstrates for the first time the feasibility of 3D bioprinting adult human testicular cells. We show that the bioprinting process is compatible with high testicular cell viability and without loss of the main somatic phenotypes within the testis tissue. We demonstrate an increase in germ cell markers in the 3D bioprinted tubules after 12 days of in vitro culture. This platform may carry future potential for disease modeling and regenerative opportunities in a personalized medicine framework.


Single-stranded DNA loops as fiducial markers for exploring DNA-protein interactions in single molecule imaging.
A polymerase chain reaction (PCR) based method of adding a single-stranded DNA (ssDNA) hairpin loop to one end of linear double-stranded (ds) DNA templates was developed. The loop structure serves as a fiducial marker in single molecule imaging by atomic force microscopy (AFM) and can be applied to study DNA-protein interactions. The nucleic acid end-labels allow discrimination of the polarity of the DNA template in the AFM while limiting non-specific interactions which might occur from non-nucleic acid labels. Homo-polynucleotide ssDNA loops made up of 20 base-pairs (bp) for each of the four bases (A, T, G, C) were investigated to determine the effects of sequence on template labelling. The products were produced with high efficiency and high yield with the loop readily distinguished from the dsDNA template by height and diameter in the AFM. The application of the method to study DNA transcription was investigated by firing Escherichia Coli RNA polymerase (RNAP) from a λPR promoter in the direction of the loop-labelled end. The ssDNA loops captured elongating complexes of RNAP, arresting transcription and preventing dissociation. The dual role of the loop as a polarity marker and retainer of previously active RNAP will allow mechanisms of gene expression to be studied with single molecule sensitivity by AFM. This will enable insight into molecular interactions of RNAP on single DNA templates in convergent or tandem transcription configurations.


[Endoscopic treatment of a gastric phytobezoar using electrohydraulic lithotripsy: a case report].
The Authors report a case of spontaneous large gastric phytobezoar in an unoperated stomach and describe this pathology especially with regards to pathogenesis, diagnosis and endoscopic treatment.


Inhibition of IKK-β by epidioxysterols from the flowers of Calotropis gigantea (Niu jiao gua).
Calotropis gigantea (Asclepiadaceae) (Niu jiao gua) has been used as a poultice in Chinese medicine for treating inflammatory skin diseases, e.g., neurodermatitis. This study aims to isolate the epidioxysterols from the flowers of C. gigantea, elucidate their structures and evaluate their possible inhibitory effects on the NF-κB pathway. The two epidioxysterols 9,11-dehydroergosterol peroxide (1) and ergosterol peroxide (2) were isolated from the powdered flowers of C. gigantea by ultrasonic-assisted extraction, followed by the purification of the crude extract by column chromatography (i.e., silica gel and MCI-gel CHP 20P open columns). The chemical structures of these compounds were identified through a comparison of their HRMS, (1)H and (13)C NMR data with those in the literature. The in vitro IKK-β inhibitory activities of compounds 1 and 2 (1-100 µM) were evaluated using an IKK α and β Assay/Inhibitor Screening Kit, which is a single-site, semi-quantitative immunoassay. Berberine was used as a positive control. The IKK-β inhibitory activities between compounds 1 and 2 were compared by a two-tailed Student's t test to summarize the structure activity relationship. Compounds 1 and 2 exhibited a dose-dependent inhibitory activity towards IKK-β in a similar manner to that of berberine. The IKK-β inhibitory activities of these two epidioxysterols were significantly stronger (P = 0.001 for compound 1 and P = 0.028 for compound 2) than that of berberine at the concentration of 100 µM. Furthermore, at the same concentration the suppressive effect of compound 1 towards IKK-β was greater than that of compound 2 (P = 0.041), while their activities at 10 and 50 µM were comparable. The difference in the results at 100 µM therefore suggested that the double bond between C-9 and C-11 in compound 1 could be responsible for its higher inhibitory activity towards IKK-β at this concentration. 9,11-dehydroergosterol peroxide (1) and ergosterol peroxide (2) were isolated from the flowers of C. gigantea and exhibited in vitro inhibitory activities towards IKK-β.


Nonclassical secretion of human catalase on the surface of CHO cells is more efficient than classical secretion.
There is a great demand for improved production of therapeutic proteins using mammalian cell expression systems and transgenic animals. There have been intensive endeavors to optimize production at the transcriptional and translational levels, but comparatively little attention has been paid to the secretory level, especially to nonclassical secretion. To compare the efficiencies of classical and nonclassical secretion, we expressed GFP-tagged human catalase conjugated with a classical signal peptide and with several short peptides derived from mouse Engrailed 2 (mEN2) homeoprotein for nonclassical secretion and internalization in CHO cells. Surprisingly, the results showed that the secretory efficiency was significantly greater (up to 2.3 fold) than classical secretion when the fusion protein was driven by the secretory sequence (SS) of mEN2, and up to 1.9 fold when the classical secretion process was modified by incorporating the internalization sequence (IS) of mEN2. The effect of these short peptides on nonclassical secretion and internalization may indicate potential applications in the improved production of complex therapeutic proteins in mammalian cell expression systems and transgenic animals.


Prey selection by the copepod Diacyclops thomasi.
1. Adult females of the predaceous copepod, Diacyclops thomasi, consistently selected for the soft-bodied rotifers Synchaeta pectinata, Polyarthra major and P. remata when presented various combinations of 8 rotifer species and 2 crustacean species as prey. Diacyclops did not select for other small, soft-bodied rotifers such as P. vulgaris and Ascomorpha ecaudis and, for loricate species such as Keratella cochlearis, K. crassa and for large soft-bodied adult Asplanchna priodonta. The small cladocerans, Bosmina longirostris and Chydorus sphaericus also were resistant to predation by this copepod. 2. Increased hunger in Diacyclops increased the clearance rates on both vulnerable and Diacyclops-resistant prey but did not greatly increase mortality of resistant prey relative to vulnerable prey. Sated Diacyclops preferred small, vulnerable prey like P. major over larger-bodied Synchaeta. This effect may be attributed to limited gut space when food is abundant. 3. When Diacyclops was presented different relative proportions of Keratella and Synchaeta at a constant total prey density (500 prey/L), it selected Synchaeta over Keratella in all trial proportions. However, Diacyclops selected more strongly for Keratella (but at a much lower clearance rate than for Synchaeta) when the relative abundance of this predator-resistant species was greatest. These results support optimal foraging in this predator. 4. Predator-prey interactions of the kind reported in this study can help identify important food web pathways and can be used to interpret predator-mediated changes in zooplankton communities in mature.


An Emotional Stress as a Trigger for Reverse Takotsubo Cardiomyopathy: A Case Report and Literature Review.
Reverse Takotsubo cardiomyopathy is one of the rarest types of stress-induced cardiomyopathy, which despite sharing similar pathogenic mechanisms with its more common counterpart, has different clinical manifestations, demographics, and laboratory values. We present the case of a 61-year-old woman who came to the hospital with a chief complaint of chest pain and dyspnea. She was found to have elevated troponin and severely depressed left ventricular function. Echocardiography showed normal contracting apex, with the rest of the left ventricle being hypokinetic. Cardiac catheterization revealed mild coronary artery disease and confirmed echocardiographic findings showing hyperkinetic apex and dilated base. She was discharged home on ACE inhibitor and B-blocker. A repeat echocardiogram 2 weeks after the initial presentation showed complete resolution of cardiac dysfunction. Reverse Takotsubo cardiomyopathy is a rare disease mimicking acute coronary syndrome. It is essential to rule out organic coronary disease prior to attributing the presentation to Takotsubo cardiomyopathy. With supportive care, the long-term prognosis is good in the vast majority of patients.


Between- and Within-Individual Sociodemographic and Psychological Determinants of PrEP Adherence Among Men Who have Sex with Men Prescribed a Daily PrEP Regimen in Wales.
We investigated the determinants of daily PrEP use and coverage of condomless anal sex (CAS) by PrEP among men who have sex with men in Wales, UK. We measured PrEP use by electronic monitors and CAS by secure online surveys. We defined PrEP use based on daily medication cap openings and coverage as CAS episodes preceded by ≥ 3 days of PrEP use and followed by ≥ 2 days of PrEP use. We included 57 participants (5463 observations). An STI diagnosis was associated with lower PrEP use but also lower PrEP coverage. Older adults had higher PrEP use. A belief that other PrEP users took PrEP as prescribed was associated with lower PrEP coverage. An STI diagnosis is an important cue for an intervention, reflecting episodes of high-risk sexual behaviour and low PrEP coverage. Other results provide a basis for the development of an evidence-informed intervention for promoting coverage of PrEP.


Extrauterine endometrial stromal sarcoma: A systematic review and outcome analysis.
Endometrial stromal sarcoma (ESS) is the second most common uterine mesenchymal neoplasm. ESS can arise from extrauterine locations without any uterine involvement and is called extrauterine ESS (EESS). The epidemiological features of EESS are not well-known. Moreover, the factors affecting its outcome have not been systemically studied. The treatment of EESS closely follows that of uterine ESS, comprised of different combinations of surgical management, hormone therapy, chemotherapy, and radiation therapy. However, the effectiveness of different treatment protocols for EESS has not been studied. Here, we have performed a systematic review of all reported cases of EESS in the English literature. We further performed a meta-analysis of the outcome data and investigated how the patients' age, tumor site, tumor size, and management affect the overall and progression-free survival of the patients. We found that tumor site and mode of treatment significantly affected the overall survival and progression-free survival of the patients. Tumor size significantly affected overall survival but not progression-free survival, while the age at diagnosis did not affect patient outcome. As far as we know, ours is the first systematic study of this rare malignancy with an emphasis on outcome analysis.


Personal view. Is it reality or an illusion that liquid-based cytology is better than conventional cervical smears?
Liquid-based cytology (LBC) has been heralded as the way forward for cervical screening, and as the answer to many of its problems. It is already used as a sole method of cell preparation in many private clinics in the UK. It is being used for colposcopy smears in many NHS clinics and is now being piloted for primary screening in three screening centres in England, as well as one in Scotland and one in Wales. LBC has been welcomed as a new technology because it deals with the problem of specimen adequacy at source, removing responsibility for slide preparation and fixation from the clinician or nurse. It provides uniformly well-fixed preparations that are free of inflammatory exudate and blood, and seem easier to screen than conventional smears. There are many articles in the world literature suggesting that LBC is more accurate than conventional screening, and it is thought likely to reduce the number of false negative tests. The main reasons for piloting LBC in the NHS Cervical Screening Programme (NHSCSP) lie in its potential for reducing screening times and for reducing the numbers of repeats for inadequate tests. LBC is expensive in terms of equipment, capital costs, maintenance, consumables, training, technical preparation time, transportation and disposal of liquid media. Its costs could be justified if they were offset by the money saved from reduced screening time and repeat tests, but only if its accuracy in terms of sensitivity and specificity were proven to be equal to or better than conventional cytology. Although that is generally held to be true by the public and medical profession alike, there is very little hard evidence to support it.


Incorporation of antifreeze proteins into zebrafish embryos by a non-invasive method.
The cryopreservation of fish embryos is a challenge because of their structure, with multiple compartments and permeability barriers, and their high chilling sensitivity. Vitrification at advanced developmental stages is considered to be the more promising option. Nevertheless, all reported attempts have failed. Previous studies demonstrated a better ability for freezing in species that naturally express antifreeze proteins (AFPs). These proteins have been delivered into other fish embryos using time-consuming techniques like microinjection. In the present study, the introduction of FITC labelled AFPs was assayed in zebrafish embryos at early developmental stages (from 2-cell to high blastula stage), before the formation of the yolk syncytial layer, by an easy and non-invasive method and evaluated by fluorescence and confocal microscopy. Incubation with AFPs at 128-cell or high blastula stage provides incorporation of the protein in 50-90% of embryos without affecting hatching. Incubation in media containing protein is a simple, harmless and effective method which makes it possible to treat several embryos at the same time. AFPs remain located in derivatives from marginal blastomeres: the yolk syncytial layer, the most cryosensitive and impermeable barrier, and different digestive organs. Our findings demonstrate that delivery of AFP type I and AFP type III into zebrafish embryos by incubation in media containing protein is a simple and harmless method that may improve cryoprotection of the cellular compartment.


Effects of succinylacetone on amino acid uptake in the rat kidney.
Infants with hereditary tyrosinemia also have a renal Fanconi syndrome and excrete succinylacetone (SA). We have studied the effects of SA on rat renal tubular amino acid transport in vivo and in vitro using isolated renal tubules. Injection of SA produces increased clearance of several amino acids in the intact animal. In vitro SA causes a reversible inhibition of alpha-aminoisobutyric acid uptake, resulting from depressed low- and high-affinity transport systems. Addition of glutamate, succinate, or glucose, alone or in combination, did not restore transport. These observations suggest the usefulness of SA in the production of a physiologic animal model for the study of the human Fanconi syndrome.


Identification of potentially relevant metals for the etiology of autism by using a Bayesian multivariate approach for partially censored values.
Heavy metals are known to be able to cross the placental and blood brain barriers to affect critical neurodevelopmental processes in the fetus. We measured metal levels (Al, Cd, Hg, Li, Pb and Zn) in the cord blood of newborns and in the serum of the same children at 5 years of age, and compared between individuals with or without (controls) autism spectrum disorder (ASD) diagnosis. The samples were from a biobank associated with the All Babies in Southeast Sweden (ABIS) registry. We proposed a Bayesian multivariate log-normal model for partially censored values to identify potentially relevant metals for the etiology of ASD. Our results in cord blood suggest prenatal Al levels could be indicative of later ASD incidence, which could also be related to an increased possibility of a high, potentially toxic, exposure to Al and Li during pregnancy. In addition, a larger possibility of a high, potentially beneficial, exposure to Zn could occur during pregnancy in controls. Finally, we found decisive evidence for an average increase of Hg in 5-year-old ASD children compared to only weak evidence for controls. This is concordant with previous research showing an impaired ability for eliminating Hg in the ASD group.


Determining and mapping the spatial mismatch between soil and rice cadmium (Cd) pollution based on a decision tree model.
Environmental complexity leads to differences in the spatial distribution of heavy metal pollution in soil and rice. Such spatial differences will seriously affect the safety of planted rice and can impact regional management and control. How to scientifically reveal these spatial differences is an urgent problem. In this study, the spatial mismatch relationship between Cd pollution in soil and rice grains (brown rice) was first explored by the interpolation method. To further reveal the causes of these, the specific recognition rules of the spatial relationship of Cd pollution were extracted based on a decision tree model, and the results were mapped. The results revealed a spatial mismatch in Cd pollution between the soil and rice grains in the study area, and the main results are as follows: (i) slight soil pollution and safe rice accounted for 68.88% of the area; (ii) slight soil pollution and serious rice pollution accounted for 13.39% of the area and (iii) safe soil and serious rice pollution accounted for 11.63% of the area. In addition, 11 recognition rules of Cd spatial pollution relationship between soil and rice were proposed, and the main environmental factors were determined: SOM (soil organic matter), Dis-residence (distance from residential area), soil pH and LAI (leaf area index). The average accuracy of rule recognition was 75.90%. The study reveals the spatial mismatch of heavy metal pollution in soil and crops, providing decision-making references for the spatial accurate identification and targeted prevention of heavy metal pollution spaces.


Characterization of a cDNA encoding a human kidney, cytochrome P-450 4A fatty acid omega-hydroxylase and the cognate enzyme expressed in Escherichia coli.
A cDNA encoding a cytochrome P-450 4A (CYP4AII) was cloned from a human kidney cDNA library. Northern blot analysis and RNase protection assays indicate that related mRNAs occur in kidney and liver with the highest abundance found in kidney. The enzyme was expressed from its cDNA in Escherichia coli. A solubilized preparation of the enzyme reconstituted with cytochrome P-450 reductase catalyzed the omega-hydroxylation of lauric acid, palmitic acid, and arachidonic acid with turnover numbers of 9.8, 2.2 and 0.55 min-1, respectively. Little or no activity was detected toward prostaglandins A1 and E1.


High-affinity and intermediate-affinity forms of the human interleukin 2 receptor, expressed in an interleukin 9-dependent murine T cell line, deliver proliferative signals via differences in their transduction pathways.
Although several reports have claimed that the p70 IL-2R per se transduces a growth signal in lymphoid and non-lymphoid systems, there is no convincing evidence for this in lymphoid T cell lines. In order to investigate the mechanism of IL-2R-dependent signal transduction pathways via the p70 IL-2R in lymphoid T cells, we have established two IL-9-dependent murine (TS1) cell lines stably expressing the human p70 IL-2R subunit or the human p55-p70 IL-2R complex. Whereas the parental T cell line, TS1, proliferated in response to IL-9 or IL-4 but not IL-2, cell lines stably expressing human p70 IL-2R or p55-p70 IL-2R complex proliferated in response to IL-2. This implies that the murine T cell, TS1, contains all the intracellular components necessary for directing IL-2 signalling. In human p55-p70 IL-2R-transfected cells, the expression of a functional murine p55 IL-2R seemed to be induced and regulated by IL-2. In human p70 IL-2R-transfected cells, in the presence of IL-2 an interaction requiring only intermediate-affinity was sufficient for transduction of a proliferative signal. In addition human p70 IL-2R per se, is biologically functional via a transduction pathway not requiring induction of murine p55 IL-2R and consecutive high-affinity complex reconstitution. Thus, although both transfected cell lines can transduce a proliferative signal in the presence of IL-2, a difference in their transduction pathway is probably involved for the induction of p55 IL-2R.


The egg came first, of course! Anterior-posterior pattern formation in Drosophila embryogenesis and oogenesis.
The anterior-posterior body pattern of the Drosophila embryo is initiated through the action of maternal gene products. In particular, three groups of maternally acting genes (the anterior, posterior and terminal groups) have been shown to direct the synthesis and spatial restriction of the three major organizing activities in the egg. The initial spatial localizations of the maternal organizing activities are established during oogenesis. After fertilization these activities regulate zygotic gene activity along the anterior-posterior axis of the egg.


Racial/ethnic distribution of graduates from doctorate and masters epidemiology degree programs in the United States, 2008 to 2018.
To identify trends in racial and ethnic diversity of epidemiology graduate degree recipients in the United States between academic years 2008 and 2018. National-level data from the National Center for Education Statistics was analyzed to assess the change in proportions of epidemiology degrees conferred to each racial/ethnic group - American Indian or Alaska Native; Asian, Native Hawaiian or Other Pacific Islander; Black or African American; Hispanic or Latino; White; and two or more races- over two time periods, Fall 2007- Spring 2012 (Period 1) and Fall 2012 - Spring 2018 (Period 2). During Period 13,837 epidemiology graduate degrees were conferred, and 6960 in Period 2. Within race/ethnicity groups, there was a statistically significant increase in graduate epidemiology degrees awarded over the two time periods to students of Hispanic or Latino ethnicity, and to students reporting two or more races. The proportion of degrees awarded to non-White students in aggregate increased by 4.7 percentage points, from 33.5% to 38.2%, while awards to White students decreased by the same amount. Overall, the racial/ethnic diversity of epidemiology graduates in the United States increased between 2008 and 2018, however, further efforts are needed to increase awards within some racial minority subgroups.


MEDICAL AND SOCIAL ANALYSIS OF PATIENTS WHO DIED OF SARS-COV-2 IN THE SPECIALIZED INFECTIOUS DEPARTMENT OF THE CITY OF SUMY DURING THE FIRST WAVE OF THE EPIDEMIC.
The aim of the article is to analyze demographic indicators, clinical symptoms, concomitant pathology, and the course of acute respiratory viral infection (ARVI) caused by SARS-CoV-2 in patients with a fatal outcome of the disease. Materials and methods: To achieve the goal, a statistical method, an analytical method, and a method of retrospective analysis of the medical histories of patients with fatal cases who were hospitalized with a diagnosis of ARVI caused by SARS-CoV-2 were used. Results: Mortality among patients who were hospitalized with a diagnosis of ARVI caused by SARS-CoV-2 was 8.18 ± 2.17%. Among them, 62% were male and 38% were female. Cardiovascular pathology took first place in the structure of concomitant pathology of all age groups and accounted for 76%. Oncological diseases accounted for 62%, gastrointestinal diseases - 54%, endocrine diseases - 38%, and respiratory system diseases 23% of the total number of patients with fatal cases. Conclusions: Mortality from coronavirus infection in the period March - July 2020 among the male population was 62%, of which 13% - from the age group 18-45 years, 38% - from the age group 46-64 years, and 50% - patients 65 years old and older. Among the female population, the mortality rate was 38%, of which 20% were women in the age group 46-64% and 80% were 65 years and older. The presence of no-hospital polysegmental pneumonia as a complication of ARVI caused by SARS-CoV-2 was 62% among all age groups of the studied patients with fatal cases.


Hydrazino-s-triazine based labelling reagents for highly sensitive glycan analysis via liquid chromatography-electrospray mass spectrometry.
Labelling strategy plays an important role in mass spectrometry (MS) based glycan analysis due to the high hydrophilicity and low ionization efficiency of glycans. Ten hydrazino-s-triazine based labelling reagents were synthesized under facile and controllable conditions for highly sensitive liquid chromatography-electrospray mass spectrometry glycan analysis in this work. Attached to N-glycans through non-reductive reactions, these new labelling reagents were evaluated in aspect of the differently enhanced glycan response to mass spectrometry. Three of the ten labelling reagents demonstrated to be reliable and remarkable for glycan analysis with satisfactory linearity and lowered limits of detection using maltoheptaose (DP7) as model. Furthermore, the most optimal labelling reagent was taken as an example for highly sensitive profiling of N-linked glycans both cleaved from chicken avidin and glycoproteins in human serum, indicating prospective availability for these labelling reagents in frontier of glycomics researches.


Lumbar spinal cord and early cortical evoked potentials after tibial nerve stimulation: effects of stature on normative data.
Short-latency somatosensory evoked responses to tibial nerve stimulation were recorded simultaneously over the spinous processes of the lumbar vertebrae and scalp in normal young subjects. This non-invasive test revealed values which are highly reproducible and could be closely correlated with subject-related variables such as height and leg length. A nomogram describing the close relationship between absolute latency of various response components and stature was derived and shown to be an important variable in the establishment of normative data nad interpretation of SSEP in relation to disorders affecting the peripheral and central nervous system. In contrast, central conduction times of interpeak intervals (N22-P37 and N22-N45) were unrelated to variations in the subjects, stature providing a reliable clinical measure in the assessment of the central somatosensory pathway.


Immunoprint analysis of Calvatia cyathiformis allergens. I. Reactivity with individual sera.
Basidiomycete allergens have received scant attention to date, compared to allergens from deuteromycetes. This is true even though in previous studies, 32% of atopic patients with respiratory allergies were skin test reactive to basidiospore extracts. Since sufficient quantities of Calvatia cyathiformis spores were available, their allergens were sequentially fractionated by gel filtration (GF) and hydrophobic interaction chromatography (HIC). Unfractionated extract (crude), GF, and HIC fractions were electrofocused in polyacrylamide gel (pH 3.5 to 9.5) and immunoprinted onto CNBr-activated nitrocellulose (0.2 microns) filters. Sera from 19 skin prick test positive and 10 negative subjects, with comparable total IgE levels, were used to screen individual strips of each blot. Blots were reacted with 125I-labeled anti-IgE and analyzed by autoradiography. IgE from 79%, 89%, and 89% of the positive sera bound to crude extract, GF, and HIC, respectively; IgE from none of the negative sera bound to crude extract. A series of bands (pH 3.6 to 4.6) reacted with 63%, one band (pH 6.6) reacted with 68%, and one band (pH 9.3) reacted with 63% of the sera tested. These studies demonstrate at least three important groups of allergens in C. cyathiformis spores. This characterization allows assignment of initial C. cyathiformis allergen designations and development of a purification protocol.