Controlled Inhibition of the Mesenchymal Stromal Cell Pro-inflammatory Secretome via Microparticle Engineering.
Mesenchymal stromal cells (MSCs) are promising therapeutic candidates given their potent immunomodulatory and anti-inflammatory secretome. However, controlling the MSC secretome post-transplantation is considered a major challenge that hinders their clinical efficacy. To address this, we used a microparticle-based engineering approach to non-genetically modulate pro-inflammatory pathways in human MSCs (hMSCs) under simulated inflammatory conditions. Here we show that microparticles loaded with TPCA-1, a small-molecule NF-κB inhibitor, when delivered to hMSCs can attenuate secretion of pro-inflammatory factors for at least 6 days in vitro. Conditioned medium (CM) derived from TPCA-1-loaded hMSCs also showed reduced ability to attract human monocytes and prevented differentiation of human cardiac fibroblasts to myofibroblasts, compared with CM from untreated or TPCA-1-preconditioned hMSCs. Thus, we provide a broadly applicable bioengineering solution to facilitate intracellular sustained release of agents that modulate signaling. We propose that this approach could be harnessed to improve control over MSC secretome post-transplantation, especially to prevent adverse remodeling post-myocardial infarction.


Augenblickdiagnose in neurology: Revisiting 'diagnosis in the blink of an eye'.
The art of diagnosis in clinical neurology requires attentive listening and careful observation. In certain situations, a fragment of the history or a physical sign may be so distinctive that it allows clinicians to evoke a specific diagnosis. This quick mental process was previously referred to as 'Augenblickdiagnose' ('diagnosis in the blink of an eye') in a seminal paper by Dr. William Campbell in 1998. We aimed to revisit this concept by providing additional clinical vignettes. The authors wrote clinical vignettes using examples from their own clinical practice and performed a non-systematic review of influential neurology textbooks using the words 'pathognomonic' and 'highly suggestive'. Twenty examples from various fields of neurology are presented in a table, stratified by major fields of neurology. A short educational reflection is provided for each diagnosis considered. 'Augenblickdiagnose' is an engaging teaching resource that also contributes to 'neurophilia', that is, a fascination for neurology, perhaps increasingly in today's modern neurology practice. However, multiple cognitive biases underlying mental shortcuts may lead to an incorrect diagnosis. It is important to stress that good clinical practice in neurology requires taking a thorough history and performing a careful neurological examination.


Transmission potential of the human head louse, Pediculus capitis (Anoplura: Pediculidae).
Millions of people are infested by head lice every year. However, louse transfer between hosts is not well-understood. Our goals were to determine: (1) which stages were most likely to disperse and why, (2) the likelihood of fomites transmission, and (3) if host blood gender affects louse development. Various life stages of lice at differing densities were permitted to cross over a 15-cm hair bridge placed between two artificial blood-feeding arenas. Louse transfer caused by hot air movements, combing, toweling, and passive transfer to fabric was investigated. The ability of lice to oviposit on different foreign substrates and the hatching potential of eggs intermittently incubated for 8 h/night on a host were likewise investigated. Louse in vitro development following feeding on human female or male donor blood was compared. Adult lice were the most likely to disperse. Neither population density nor hunger significantly affected dispersal tendencies. Lice were dislodged by air movement, combs and towels, and passively transferred to fabric within 5 min. Females oviposited on a variety of substrates and 59% of eggs incubated for 8 h/night hatched after 14-16 days. There was no survivorship difference between lice artificially fed on female vs. male blood. Adult lice are the most mobile, indicating that they are most likely to initiate new infestations. Although head-to-head contact may be the primary route of transmission, less direct routes involving fomites may play a role and need further evaluation. Blood-borne factors do not appear to cause any gender-biased host preference.


Retraction: Sensing risk, fearing uncertainty: systems science approach to change.
[This retracts the article on p. 30 in vol. 8, PMID: 24744723.].


Synergistic interaction between 4-nonylphenol and high but not low temperatures in Dendrobaena octaedra.
Studies on joint effects of toxic compounds and temperature have clearly shown that single stressor tests often underestimate the critical limits of the stressor evaluated. In the present study, the joint effects of nonylphenol (NP) and high (25-35 degrees C) or low (1.4 to -6.4 degrees C) temperatures in Dendrobaena octaedra were elucidated. The effects of these combined stresses were analyzed in a multiplicative model using an independent action model as a reference, and there was a strongly synergistic interaction between the effects of NP and high but not low temperatures. This interaction may be caused by NP disrupting membrane stability during thermal stress and/or by a depletion of protective heat shock proteins. While the survival of D. octaedra at high temperatures significantly decreased with increasing NP concentration up to 400 mg kg(-1) dry soil, reproduction was significantly reduced when the earthworms were exposed to a concentration as low as 30 mg NP kg(-1) dry soil.


Early mechanical complications following fixation of proximal femur fractures: From prevention to treatment.
Proximal femur fractures have significant functional repercussions in both older adults (sometimes life-threatening) and younger adults (socioeconomic). This study will review the early mechanical complications (EMC) associated with the fixation of femoral neck (FNF) and trochanteric (TF) fractures. What is the nature of these complications? They consist of incorrect fracture fixation, secondary displacement and/or fixation failure (FNF: 3-10%; TF: 5-7%), and more rarely, secondary fractures distal to the implant. What are the risk factors and how can they be controlled? They are related to the patient (osteoporosis), to the type of fracture (FNF: posterior comminution, magnitude of displacement, vertical fracture line, fracture of calcar/TF: comminution of calcar, lesser trochanter fracture, metaphyseal comminution, lateral cortex fracture, greater trochanter comminution), to the chosen treatment (FNF: triple screw fixation or screw-plate are preferable/TF: all implants suitable for simple fractures, cervicomedullary implants for complex fractures), and to the surgical technique (quality of reduction, implant positioning). How should they be treated? Early revision of internal fixation can be difficult and increases the infection risk. Inter- or subtrochanteric valgus osteotomy leads to satisfactory results but is difficult to carry out. Secondary arthroplasty, which is the preferred option in older adults, leads to good outcomes but is technically more difficult, and has a higher complication rate in EMC of trochanteric fractures. LEVEL OF EVIDENCE: V.


[Expression of long non-coding RNA HOTAIR mRNA in ovarian cancer].
To investigate the long non-coding RNA HOTAIR (long non-coding HOX antisense intergenic RNA) by examining HOTAIR expression levels in ovarian cancer tissue and normal ovarian tissue. The mRNA of long non-coding RNA HOTAIR expressions were detected by real-time fluorescence quantitative PCR in ovarian cancer (44) and normal ovary tissues (14). The expression of HOTAIR in ovarian cancer tissue was higher than that in normal ovarian tissue (1.26 +/- 0.27 vs. 0.14 +/- 0.09, P < 0.05). The expression was statistically higher in poorly differentiated ovarian cancer than poorly-moderately, moderately-well, and well-differentiated ones (1.65 +/- 0.41 vs. 0.39 +/- 0.14, P < 0.05). RNA HOTAIR expressions were higher in ovarian cancer; it may play a role in ovarian cancer, and become a biomarker for malignant degree of ovarian cancer and may provide a novel therapeutic target for ovarian cancer.


Bioassay of kinship in northwestern Europe.
Kinship of Barra (Outer Hebrides) is bioassayed as 0.0096 relative to northwestern Europe, in reasonable agreement with prediction of 0.008 relative to Britain. The exponential decline of kinship with distance is similar to Scandinavia. Kinship of larger populations is consistent with predictions from isolates. Kinship is largely due to local drift rather than Norse admixture, the estimate of which is obscured by drift and appears highly unreliable. Other populations in northwest Europe, including Iceland, Lewis and Orkney, give estimates of kinship which reflect drift and geography, but also do not provide reliable estimates of admixture. Bioassay of kinship from gene frequencies gives observations to be explained in terms of migration and drift, but contributes nothing to the question of whether polymorphism in the species is maintained by neutral mutation or selection. Although the latter is on various grounds more likely, no calculations based on gene frequency distribution can provide critical evidence.


Synchronous occurrence of hepatic mucinous cystic neoplasm and serous ovarian cystadenoma.
The coexistence of hepatic mucinous cystic neoplasm and ovarian serous cystadenoma is rare and can only be confirmed by histological examination of the surgical specimen. Complete surgical resection is the optimal treatment. Mucinous cystic neoplasms (MCN) of the liver are rare premalignant lesions that account for less than 5% of all hepatic cysts. The synchronous occurrence of hepatic MCN with ovarian cystadenomas is rare. Herein, we report a case of a hepatic MCN coexistent with a serous ovarian cystadenoma.


Early studies on the safety and efficacy of thalidomide for symptomatic inflammatory bowel disease.
Thalidomide is clinically effective in the treatment of graft versus host disease in bone marrow transplantation and aphthous ulceration in HIV infection. It appears to exert a selective effect on tumor necrosis factor-alpha (TNF-alpha) production. Tumor necrosis factor-alpha is implicated in the pathogenesis of inflammatory bowel disease (IBD). The aim of this study was to assess the efficacy and safety of thalidomide in symptomatic IBD. Eleven patients (nine males, mean age 33 years, range 20-77 years) with chronic inflammatory bowel disease (six Crohn's disease (CD), four ulcerative colitis (UC), one indeterminate colitis (IC)) who were symptomatic despite standard medical therapy were administered a daily dose of thalidomide for 12 weeks in an open-labeled protocol. Their response was assessed by using clinical, colonoscopic, histological, and immunological methods. Two patients withdrew at 3 weeks because of mood disturbances. Of the remaining nine patients, eight (five CD, two UC and one IC) had a marked clinical response, while one patient with CD had no response. The mean stool frequency decreased from 4.3 to 2.3 per day (P = 0.0012), and the stool consistency increased from 2.1 to 1.2 (P = 0.02). The mean Crohn's Disease Activity Index decreased from 117 to 48 (P = 0.0008). Endoscopic inflammatory and histological grade, C-reactive protein and erythrocyte sedimentation rate (ESR) all decreased significantly (P = 0.011, P = 0.03, P = 0.023 and P = 0.044, respectively). However, the serum TNF-alpha levels did not change. Side-effects included mild sedation, xerostomia and skin dryness in all, constipation in three, and minor abnormalities in nerve conduction in one patient. These data strongly suggest that thalidomide is an effective short-term treatment for symptomatic IBD.


The Impact of Trimetazidine on Disease Severity and Quality of Life in Parkinson's Disease.
Trimetazidine is contraindicated in movement disorders, however, a not negligible part of trimetazidine users is still patients with Parkinson's disease (PD). The present study aimed to objectively determine the impact of trimetazidine on the severity of symptoms and the health-related quality of life of patients with PD by measuring changes after its withdrawal. A consecutive series of 42 patients with PD using trimetazidine underwent detailed neurological and neuropsychological assessments at baseline and three months after the discontinuation of trimetazidine. Clinically relevant improvements were achieved with discontinuation of trimetazidine according to changes in scores of each part of the Movement Disorder Society-sponsored Unified Parkinson's Disease Rating Scale (Part I: -25.7%, p < 0.001; Part II: -23.8%, p < 0.001; Part III: -28.5%, p < 0.001; Part IV: -30.1%, p = 0.004) and total scores of the Non-Motor Symptoms Scale (-25.6%, p = 0.004) and the Montgomery-Asberg-Depression Rating Scale (-20.1%, p = 0.001). Benefits resulting from the withdrawal of the drug also manifested in the improvement of the health-related quality of life based on changes in the summary index of the 39-item Parkinson's Disease Questionnaire (-18.2%, p = 0.031). Our results provide clinical rationale for strictly avoiding the use of trimetazidine in PD. Discontinuation of trimetazidin results in clinically relevant improvements in Parkinsonian symptoms.


Gender differences exist in neuromuscular control patterns during the pre-contact and early stance phase of an unanticipated side-cut and cross-cut maneuver in 15-18 years old adolescent soccer players.
Non-contact ACL injuries generally occur as the foot contacts the ground during cutting or landing maneuvers and the non-contact ACL injury rate is 2-8 times greater in females compared to males. To provide insight into the gender bias of this injury, this study set out to identify gender differences in the neuromuscular response of the quadriceps, hamstrings and gastrocnemii muscles in elite adolescent soccer players during the pre-contact and early stance phases of an unanticipated side-cut and cross-cut. For the early stance phase of the two maneuvers, females demonstrated greater rectus femoris activity compared to males. Throughout the pre-contact phase of the maneuvers, a rectus femoris activation difference was identified with females having an earlier and more rapid rise in muscle activity as initial ground contact approached. Females demonstrated greater lateral and medial gastrocnemii activity for the pre-contact and early stance phases of the side-cut and greater lateral gastrocnemii activity during early stance of the cross-cut. Timing of hamstring activity also differed between genders prior to foot contact. The differences suggest that the activation patterns observed in females might not be providing adequate joint protection and stability, thereby possibly having a contributing role towards increased non-contact ACL injuries in females.


Combined conversion of lignocellulosic biomass into high-value products with ultrasonic cavitation and photocatalytic produced reactive oxygen species - A review.
The production of high-value products from lignocellulosic biomass is carried out through the selective scission of crosslinked CC/CO bonds. Nowadays, several techniques are applied to optimize biomass conversion into desired products with high yields. Photocatalytic technology has been proven to be a valuable tool for valorizing biomass at mild conditions. The photoproduced reactive oxygen species (ROSs) can initiate the scission of crosslinked bonds and form radical intermediates. However, the low mass transfer of the photocatalytic process could limit the production of a high yield of products. The incorporation of ultrasonic cavitation in the photocatalytic system provides an exceptional condition to boost the fragmentation and transformation of biomass into the desired products within a lesser reaction time. This review critically discusses the main factors governing the application of photocatalysis for biomass valorization and tricks to boost the selectivity for enhancing the yield of desired products. Synergistic effects obtained through the combination of sonolysis and photocatalysis were discussed in depth. Under ultrasonic vibration, hot spots could be produced on the surface of the photocatalysts, improving the mass transfer through the jet phenomenon. In addition, shock waves can assist the dissolution and mixing of biomass particles.


Plasma Torquetenovirus (TTV) microRNAs and severity of COVID-19.
Torquetenovirus (TTV), a widespread anellovirus recognized as the main component of the healthy human virome, displays viremia that is highly susceptible to variations in immune competence. TTV possesses microRNA (miRNA)-coding sequences that might be involved in viral immune evasion. Among TTV-encoded miRNAs, miRNA t1a, t3b, and tth8 have been found in biological fluids. Here, the presence of TTV DNA and TTV miRNAs in the plasma of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected subjects was investigated to monitor the possible association with coronavirus disease 2019 (COVID-19) severity. Detection of TTV DNA and miRNA t1a, t3b, and tth8 was investigated in plasma samples of 56 SARS-CoV-2-infected subjects with a spectrum of different COVID-19 outcomes. TTV DNA and TTV miRNAs were assessed with a universal single step real-time TaqMan PCR assay and miRNA quantitative RT-PCR miRNA assay, respectively. The TTV DNA prevalence was 59%, whereas at least one TTV miRNA was found in 94% of the patients tested. miRNA tth8 was detected in 91% of subjects, followed by miRNAs t3b (64%) and miRNAt1a (30%). Remarkably, although TTV DNA was unrelated to COVID-19 severity, miRNA tth8 was significantly associated with the degree of disease (adjusted incidence rate ratio (IRR) 2.04, 95% CI 1.14-3.63, for the subjects in the high severity group compared to those in the low severity group). Our findings encourage further investigation to understand the potential role of TTV miRNAs in the different outcomes of COVID-19 at early and late stages.


Endodontic-periodontal interrelationships: an online study guide.
The Editorial Board of the Journal of Endodontics has developed a literature-based study guide of topical areas related to endodontics. This study guide is intended to give the reader a focused review of the essential endodontic literature and does not cite all possible articles related to each topic. Although citing all articles would be comprehensive, it would defeat the idea of a study guide. This section will cover classification of endodontic-periodontal lesions, root amputations and hemisections, osseous grafting procedures, guided tissue regeneration, and vertical root fractures.


Cell density-enhanced expression of inducible nitric oxide synthase in murine macrophages mediated by interferon-beta.
Nitric oxide (NO) has an important cytotoxic role in host defense processes against invading microorganisms and neoplastic cells. Here we demonstrate the effect of culture density on the expression of NO synthase and NO production by lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. At high cell densities, the LPS-induced expression of iNOS message, protein, and activity is markedly enhanced. We demonstrate the effects to be mediated by a diffusible macrophage product. Increasing cell density correlates with activation of IFN-dependent signaling pathways. We observe enhanced phosphorylation of STAT-1 on tyrosine 701 and serine 727, and an increase in STAT-1 DNA binding. Expression of the IFN-stimulated transcription factor IRF-1 is also enhanced. The data are consistent with the reported involvement of IFN-beta as an autocrine co-activator of iNOS expression. Considering the importance of NO as a cytotoxic mediator of host immunity, the data suggest that macrophage density is important in regulating the magnitude of NO production, and thus, the host response to infection.


AMP-activated protein kinase: ancient energy gauge provides clues to modern understanding of metabolism.
The AMP-activated protein kinase (AMPK) is an evolutionarily conserved sensor of cellular energy status, and recent data demonstrate that it also plays a critical role in systemic energy balance. AMPK integrates nutritional and hormonal signals in peripheral tissues and the hypothalamus. It mediates effects of adipokines (leptin, adiponectin, and possibly resistin) in regulating food intake, body weight, and glucose and lipid homeostasis. AMPK is regulated by upstream kinases of which the tumor suppressor, LKB1, is the first to be identified. Complex signaling networks suggest that AMPK may prevent insulin resistance, in part by inhibiting pathways that antagonize insulin signaling. Through signaling, metabolic, and gene expression effects, AMPK enhances insulin sensitivity and fosters a metabolic milieu that may reduce the risk for obesity and type 2 diabetes.


Identifying UiO-67 Metal-Organic Framework Defects and Binding Sites through Ammonia Adsorption.
Ammonia is a widely used toxic industrial chemical that can cause severe respiratory ailments. Therefore, understanding and developing materials for its efficient capture and controlled release is necessary. One such class of materials is 3D porous metal-organic frameworks (MOFs) with exceptional surface areas and robust structures, ideal for gas storage/transport applications. Herein, interactions between ammonia and UiO-67-X (X: H, NH2 , CH3 ) zirconium MOFs were studied under cryogenic, ultrahigh vacuum (UHV) conditions using temperature-programmed desorption mass spectrometry (TPD-MS) and in-situ temperature-programmed infrared (TP-IR) spectroscopy. Ammonia was observed to interact with μ3 -OH groups present on the secondary building unit of UiO-67-X MOFs via hydrogen bonding. TP-IR studies revealed that under cryogenic UHV conditions, UiO-67-X MOFs are stable towards ammonia sorption. Interestingly, an increase in the intensity of the C-H stretching mode of the MOF linkers was detected upon ammonia exposure, attributed to NH-π interactions with linkers. These same binding interactions were observed in grand canonical Monte Carlo simulations. Based on TPD-MS, binding strength of ammonia to three MOFs was determined to be approximately 60 kJ mol-1 , suggesting physisorption of ammonia to UiO-67-X. In addition, missing linker defect sites, consisting of H2 O coordinated to Zr4+ sites, were detected through the formation of nNH3 ⋅H2 O clusters, characterized through in-situ IR spectroscopy. Structures consistent with these assignments were identified through density functional theory calculations. Tracking these bands through adsorption on thermally activated MOFs gave insight into the dehydroxylation process of UiO-67 MOFs. This highlights an advantage of using NH3 for the structural analysis of MOFs and developing an understanding of interactions between ammonia and UiO-67-X zirconium MOFs, while also providing directions for the development of stable materials for efficient toxic gas sorption.


Single-molecule, quantitative detection of low-abundance somatic mutations by high-throughput sequencing.
Postzygotic somatic mutations have been found associated with human disease, including diseases other than cancer. Most information on somatic mutations has come from studying clonally amplified mutant cells, based on a growth advantage or genetic drift. However, almost all somatic mutations are unique for each cell, and the quantitative analysis of these low-abundance mutations in normal tissues remains a major challenge in biology. Here, we introduce single-molecule mutation sequencing (SMM-seq), a novel approach for quantitative identification of point mutations in normal cells and tissues.


Large artery stiffness according to different assessment methods in adult population of St.Petersburg.
The aim of the present study was to assess the prevalence of increased arterial stiffness by different diagnostic methods and its association with cardiovascular risk in Russian population-based cohort. In terms of Russian epidemiological study ESSE-RF a random selection of 452 apparently healthy Saint-Petersburg inhabitants aged 25-65 years was performed. Fasting lipids, glucose and blood pressure measurements were performed. We used 3 diagnostic methods of arterial stiffness assessment: pulse wave velocity by applanation tonometry (SphygmoCor - PWV-S) and pulse wave velocity by volumetric sphygmography (VaSera - PWV-V), and cardio-ankle vascular index (CAVI) by VaSera. 341 (75,4%) had normal parameters of arterial stiffness assessed by all methods. Spearmen's coefficient of correlation and "kappa" coefficient for PWV-S and CAVI were 0,74 and 0,04, for PWV-S and PWV-V - 0,10 and 0,06, for CAVI and PWV-V - 0,28 and 0,03, respectively. There was a significant correlation between cardiovascular risk (defined by SCORE) and PWV-S (r = 0,38, p < 0,001) and a non-significant trend of increasing CAVI along with cardiovascular risk (r = 0,35, p = 0,14). Different methods of arterial stiffness assessment showed a weak correlation with each other. Carotid-femoral pulse wave velocity detected by applanation tonometry is associated with high cardiovascular risk score and might be considered as better additional risk marker for cardiovascular risk stratification.


Evaluation of long-term stability of SARS-CoV-2 nucleic acid extracted from human nasopharyngeal samples.
The maintenance of SARS-Cov-2 RNA samples poses a new challenge for laboratories and researchers. In addition, it is a requirement in order to identify what strain of the new coronavirus is predominant in a region, for instance. Therefore, it is a must to keep the quality and viability of stored RNA to respond to this and other valid questions. In other to test the quality of our samples and storing protocols, we randomly checked RNA samples four different times over one year using a second RT-PCR assay after the first test. The virus genes, N1 and N2, showed no significant increase in the media of the CT value between the first assay and subsequent times with p > 0.05. However, the human RP gene showed differences in the first three times analyzed, but within the acceptable sample cut-off, according to the test manufacturer. After one year, the RNA extracted from human nasopharyngeal specimens are viable to detect the virus SARS-CoV-2 genes with minor changes.


Targeted isolation of diverse human protective broadly neutralizing antibodies against SARS-like viruses.
The emergence of current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) and potential future spillovers of SARS-like coronaviruses into humans pose a major threat to human health and the global economy. Development of broadly effective coronavirus vaccines that can mitigate these threats is needed. Here, we utilized a targeted donor selection strategy to isolate a large panel of human broadly neutralizing antibodies (bnAbs) to sarbecoviruses. Many of these bnAbs are remarkably effective in neutralizing a diversity of sarbecoviruses and against most SARS-CoV-2 VOCs, including the Omicron variant. Neutralization breadth is achieved by bnAb binding to epitopes on a relatively conserved face of the receptor-binding domain (RBD). Consistent with targeting of conserved sites, select RBD bnAbs exhibited protective efficacy against diverse SARS-like coronaviruses in a prophylaxis challenge model in vivo. These bnAbs provide new opportunities and choices for next-generation antibody prophylactic and therapeutic applications and provide a molecular basis for effective design of pan-sarbecovirus vaccines.


Differential expression of two related organ-specific genes in pea.
We have screened a pea genomic library using a cDNA probe derived from pea shoot RNA. From this screen, we isolated two closely related genes, designated as S2 and P4. An intriguing property of these two genes is the presence in their coding region of a repeated sequence that is conserved between them in sequence but not in the number of the repeating units. The predicted amino acid sequence suggests that these proteins could be exported and glycosylated. 3' S1 analysis reveals that one of the genes, S2, is expressed highly in stem, as expected from previous work. However, mRNA derived from the other gene, P4, is not detectable in stem tissue, but is present in tissue derived from pea pods. The 5' upstream sequence of S2 and P4 are 94% identical up to position -121, suggesting that sequences upstream of -121 are responsible for organ-specific expression of the two genes.


Ascidiacyclamides containing oxazoline and thiazole motifs assume square conformations and show high cytotoxicity.
Four cyclic octapeptides were designed from ascidiacyclamide [cyclo(-Ile-Oxz-D-Val- Thz-)2 ] (ASC, 1) to investigate the effects of oxazoline (Oxz) and thiazole (Thz) rings on the structures and cytotoxicities of the peptides. cyclo(-Ile-Thz-D-Val-Oxz-)2 (2) had the same number of Oxz and Thz rings as ASC, but the ring positions were switched. cyclo(-Ile-Oxz-D-Val-Thz-Ile-Thz-D-Val-Thz-) (3) and cyclo(-Ile-Thz-D-Val-Oxz-Ile-Thz-D-Val-Thz-) (4) contained one Oxz and three Thz rings within the molecule. All Oxz rings were substituted with Thz in cyclo(-Ile-Thz-D-Val-Thz-)2 (5). These analogues had new Oxz and Thz blocks forming the 24-membered ring. Based on CD spectra and X-ray diffraction analyses, the structures of all four analogues were classified as square ASC forms. But the structures of 2 and 5 differed from the original square form of 1, and they showed no cytotoxicity. The structure of 3 was very similar to that of 1, and 3 showed 10 times greater cytotoxicity than 1. Although no definite structure of 4 was obtained, it showed three times greater cytotoxicity than 1. It appears that the position and number of Oxz residues are essential determinants in the structure-cytotoxicity relationship of ASC analogues.


What aspects of vision facilitate haptic processing?
We investigate how vision affects haptic performance when task-relevant visual cues are reduced or excluded. The task was to remember the spatial location of six landmarks that were explored by touch in a tactile map. Here, we use specially designed spectacles that simulate residual peripheral vision, tunnel vision, diffuse light perception, and total blindness. Results for target locations differed, suggesting additional effects from adjacent touch cues. These are discussed. Touch with full vision was most accurate, as expected. Peripheral and tunnel vision, which reduce visuo-spatial cues, differed in error pattern. Both were less accurate than full vision, and significantly more accurate than touch with diffuse light perception, and touch alone. The important finding was that touch with diffuse light perception, which excludes spatial cues, did not differ from touch without vision in performance accuracy, nor in location error pattern. The contrast between spatially relevant versus spatially irrelevant vision provides new, rather decisive, evidence against the hypothesis that vision affects haptic processing even if it does not add task-relevant information. The results support optimal integration theories, and suggest that spatial and non-spatial aspects of vision need explicit distinction in bimodal studies and theories of spatial integration.


Enzyme-controlled reactions giving alkanols of use in the synthesis of biologically active molecules.
(+/-)-Bicyclo[3.2.0]hept-2-en-6-one was reduced using a variety of fungi including yeasts. Baker's yeast gave 6-exo-(1R,5S,6S)-bicyclo[3.2.0]hept-2-en-6-ol and 6-endo-(1S,5R,6S)-bicyclo[3.2.0]hept-2-en-6-ol while Curvularia lunata and Mortierella ramanniana gave only the 6-endo-alcohol and optically active bicycloheptenone. The optically active bicycloheptenols were used to prepare prostaglandin F2 alpha in the natural configuration.


Description of monoclonal antibody defining an HLA allotypic determinant that includes specificities within the B5 cross-reacting group.
The selection and characterization of a cloned murine monoclonal antibody (4D12) that defines an HLA allotypic determinant is described. Antibody 4D12 immunoprecipitates HLA heavy and light chains. From studies on 4D12 reactivity with a large panel of well-characterized target cells, antibody 4D12 defines a public HLA determinant that includes specificities within but not identical to the B5 cross-reacting group. Antibody 4D12 should be useful in the study of HLA-B5--related supratypic antigens.


[Delineation for oral cavity and oropharyngeal cancers].
The aim of this article is to present the determination and the delineation of target volumes for oral cavity and oropharyngeal carcinomas treated with intensity-modulated irradiation. The delineation on the computerized tomography scanner (CT scan) requires a precise method because of the complexity of the head-and-neck anatomy. Different elements are necessary: clinical examination, diagram of the initial tumor, surgical and pathological reports and medical imaging (CT scan, magnetic resonance imaging and fluorodeoxyglucose positron emission tomography). The clinical target volumes, the planning target volumes, the organs at risk and the planning organs at risk volumes are discussed. The concept of selectivity of the potential subclinical disease near the primary tumour and the selection of neck nodal targets are specified according to the literature.


Transcription factor positive regulatory domain 4 (PRDM4) recruits protein arginine methyltransferase 5 (PRMT5) to mediate histone arginine methylation and control neural stem cell proliferation and differentiation.
During development of the cerebral cortex, neural stem cells (NSCs) undergo a temporal switch from proliferative (symmetric) to neuron-generating (asymmetric) divisions. We investigated the role of Schwann cell factor 1 (SC1/PRDM4), a member of the PRDM family of transcription factors, in this critical transition. We discovered that SC1 recruits the chromatin modifier PRMT5, an arginine methyltransferase that catalyzes symmetric dimethylation of histone H4 arginine 3 (H4R3me2s) and that this modification is preferentially associated with undifferentiated cortical NSCs. Overexpressing SC1 in embryonic NSCs led to an increase in the number of Nestin-expressing precursors; mutational analysis of SC1 showed that this was dependent on recruitment of PRMT5. We found that SC1 protein levels are down-regulated at the onset of neurogenesis and that experimental knockdown of SC1 in primary NSCs triggers precocious neuronal differentiation. We propose that SC1 and PRMT5 are components of an epigenetic regulatory complex that maintains the "stem-like" cellular state of the NSC by preserving their proliferative capacity and modulating their cell cycle progression. Our findings provide evidence that histone arginine methylation regulates NSC differentiation.


[Squamous cell carcinoma arising in oral lichen planus : Report of two cases].
Lichen planus is a chronic inflammatory, immunologically mediated mucocutaneous dermatosis. Lichen planus mucosae predominantly affects the oral cavity. Various trigger factors such as bacterial or viral infections, drugs or physical stimuli are discussed in the development of the disease. An association with human papillomavirus infections has also been described, but is not sufficiently proven. Lichen planus mucosae is considered as a premalignant condition, but the malignant transformation rate is low. The risk of malignant transformation is significantly increased in patients with oral lichen planus who smoke, drink alcohol or have hepatitis C. We describe two patients with locally advanced squamous cell carcinoma that developed on a longstanding oral lichen planus. Both cases were successfully treated with radical tumor resection, subsequent tissue reconstruction, and adjuvant radiation/radiochemotherapy.


Feasibility of Including Behavioral Feeding Training Within a Parent Intervention for Young Children With Type 1 Diabetes.
Young children with T1D frequently display challenging eating behaviors interfering with diabetes management. The current study explored the feasibility and acceptability of a behavioral parent feeding training session with young children with type 1 diabetes. As part of a larger intervention pilot focused on healthy eating and physical activity, 9 young children (Mage = 4.22) with type 1 diabetes (T1D) and their mothers participated in a novel/non-preferred food training session. Parents were taught strategies and then were given an opportunity to use the strategies with their child. The paradigm was video recorded and content was coded for parent and child behavior. Feasibility was high, and all parents rated the feeding session as acceptable. All parents demonstrated using at least one behavioral feeding skill (M = 3.38, SD = 1.60). All 9 (100%) children touched at least one of their non-preferred foods (M = 2.05, SD = 0.75), and 5 (56%) ate at least one novel/non-preferred food (M = 1.65, SD = 0.87). Parents of young children with T1D demonstrated use of parenting skills after receiving brief instructions, with more children than not trying at least one novel/non-preferred food.


The roles of colour change and behaviour in the hygrothermal balance of a littoral isopod, Ligia oceanica.
Ligia oceanica can change its colour using melanophores, the animal's reflectance varying between about 2 and 10%. Darker individuals heat up more quickly, and to higher body temperatures, than do pale ones. Colour change shows an underlying circadian rhythm, though the pattern of this rhythm varies with temperature, humidity, background and time of year. In general the rhythm is such as to ensure maximum camouflage at the critical dusk period, but in some conditions hygrothermal needs are overriding and the animals are paler (to stay cool) or darker (to warm up). In addition, animals show short term colour modification; when transferred to differing backgrounds and temperatures their colours initially reflect background matching, but after 30-45 min are modified into thermally appropriate shades, dark at 5° C and pale at 20° C. Field-caught specimens showed body temperatures that varied with colouration, and modification of colour in relation to thermal needs, particularly by being paler than expected when forced into the open by daytime high tides, and darker than expected when active prior to dusk. Animals invariably selected dark backgrounds in choice chambers. However, choice of humidity depended on previous experience; saturated air was normally preferred, but warm animals chose drier air (to allow evaporative cooling) unless also water-stressed. They also tended to disperse to facilitate cooling, whereas aggregation increased with increasing RH and with decreasing temperature. The interactions of colour changes, behavioural choices, and activity patterns in controlling the hygrothermal belance of Ligia in the intertidal environment are discussed in the light of these results.


Sanitary status of oocytes and embryos collected from heifers experimentally exposed to Leptospira borgpetersenii serovar hardjobovis.
In a preliminary trial and three experiments, a total of 30 Holstein heifers were experimentally infected with a culture of Leptospira borgpetersenii serovar hardjobovis via one or more routes (uterine, cervical supraconjunctival, intranasal) and oviductal and uterine fluids recovered post-mortem or in vivo following superovulation with FSH. All routes of administration were effective in establishing Leptospira infection in the reproductive tract and Leptospira were identified in the oviductal and uterine fluids of all 30 heifers by microscopy. The incidence of infection was confirmed by positive identification of serum antibodies by the microscopic agglutination test (MAT). Twenty-one samples of the embryos (n = 59) recovered were cultured using bacteriological procedures and all tested negative for the infectious microorganism. Using polymerase chain reaction (PCR) assay, however, showed that 29% (7/24) of morula and blastocyst stage embryos, and one out of 29 oocytes tested positive for the presence of leptospiral DNA. A single oocyte or embryo collected from the infected heifers was inoculated intravenously to 26 test heifers. None of the test heifers developed antibody titers to Leptospira. It was concluded that, despite the presence of leptospires in the reproductive tract of donor animals and the association of leptospiral DNA with uterine stage embryos, the transmission of this disease is unlikely to occur by transfer of in vivo produced embryos in the bovine.


Multiple LEDT wavelengths modulate the Akt signaling pathways and attenuate pathological events in mdx dystrophic muscle cells.
This study is aimed at investigating the effects of LEDT, at multiple wavelengths, on intracellular calcium concentration; on transient receptor potential canonical channels; on calcium-binding protein; on myogenic factors; on myosin heavy chains; on Akt signaling pathway; on inflammatory markers; and on the angiogenic-inducing factor in dystrophic muscle cell culture experimental model. Dystrophic primary muscle cells were submitted to LEDT, at multiple wavelengths (420 nm, 470 nm, 660 nm, and 850 nm), and evaluated after 48 h for cytotoxic effects and intracellular calcium content. TRPC-1, TRPC-6, Calsequestrin, MyoD, Myogenin, MHC-slow, MHC-fast, p-AKT, p-mTOR, p-FoxO1, Myostatin, NF-κB, TNF-α, and VEGF levels were evaluated in dystrophic primary muscle cells by western blotting. The LEDT, at multiple wavelengths, treated-mdx muscle cells showed no cytotoxic effect and significant lower levels in [Ca2 +]i. The mdx muscle cells treated with LEDT showed a significant reduction of TRPC-1, NF-κB, TNF-α and MyoD levels and a significant increase of Myogenin, MHC-slow, p-AKT, p-mTOR, p-FoxO1 levels, and VEGF levels. Our findings suggest that different LEDT wavelengths modulate the Akt-signaling pathways and attenuate pathological events in dystrophic muscle cells, and a combined multiwavelength irradiation protocol may even provide a potentially therapeutic strategy for muscular dystrophies.


Dynamic intraligamentary stabilization versus conventional ACL reconstruction: A matched study on return to work.
The dynamic intraligamentary stabilization (DIS) technique is based on a different treatment approach than ACL reconstruction in that it intends to promote self-healing of the ligament. It is only recommended for acute injuries (<21days). The purpose of the present study was to compare DIS and ACLR with respect to the extent of work incapacity, revision rates, secondary arthroscopies, and treatment costs during recovery. The study was a post-hoc analysis of prospectively collected data in the Swiss National Accident Insurance Fund (SUVA) database. All registered DIS cases treated until 31 December 2012 were included in the study. ACLR cases were matched to DIS cases using a propensity score approach and analysed in a follow-up period of 2 years after injury. Paired Student's T-test and the Chi-square test were used to compare the outcome measures. All 53 DIS patients were matched to an ACLR pair. The mean time period from injury to surgery was 14days for DIS and 50days for ACLR (p<0.001). Overall work incapacity was 13% for DIS and 17% for ACLR resulting in a difference of nearly 1 month of absence from work (p=0.03). The course of postoperative work incapacity was very similar between the groups, while the work incapacity prior to surgery lower in the DIS group. We found no difference in treatment costs, secondary arthroscopies and revision rates. DIS patients benefited from nearly one month shorter absence from work than ACLR patients. This difference is likely related to the early surgical timing that is recommended for DIS. Since no differences were found between DIS and ACLR in terms of treatment costs, secondary arthroscopies and revision rates, the study supports the choice of DIS as an additional treatment option for acute ACL injuries. Further comparative studies are proposed to improve the evidence about optimal timing and best practice in ACL treatment.


Family caregiver knowledge of treatment intent in a longitudinal study of patients with advanced cancer.
Caregivers have become part of a triad of care and frequently attend patient consultations in the ambulatory cancer setting. Effective caregiving and decision making require that they understand the course of the disease and the changing treatment goals. This study sought to evaluate caregiver perception of treatment intent. A cohort of 317 subjects (181 patients and 136 caregivers) from The Canberra Hospital's Cancer Services were followed for 6 months. Caregiver understanding of patient treatment intent was measured over time together with sources of information. Most caregivers understood that the illness was life-threatening (92% at week 12) and that treatment goals were to control illness and improve quality of life. Only half understood that treatment was noncurative (48% at week 12); 27% were unsure and 25% believed that treatment would cure. A high proportion of caregivers identified the specialist as the source of information (77%) and almost half also included the general practitioner (47%). These figures remained fairly constant over time. There were significant gender and age differences in understanding. At baseline, more women than men had an accurate perception of treatment intent and these numbers increased over time. Men's perception did not change. Caregivers' ability to fully engage in the task of caring for those with a terminal illness may be hampered by their lack of understanding of the treatment patients receive.


Gastric achlorhydria in H/K-ATPase-deficient (Atp4a(-/-)) mice causes severe hyperplasia, mucocystic metaplasia and upregulation of growth factors.
Gastric neoplasia is common in humans, yet controversy remains over contributions of chronic achlorhydria, gastrinemia and hyperplasia, to cancer risk. To study this, mice lacking the gastric H/K-ATPase (Atp4a(-/-) mice) were used to determine whether chronic loss of acid secretion, with attendant hypergastrinemia, predisposes to cancer phenotype. Atp4a(-/-) and Atp4a(+/+) mice, paired for age and gender, were examined at 3, 8, 12 and 20 months for histopathology, and for expression of the trefoil factor family (TFF)1-3, Reg IIIbeta, gamma and delta, osteopontin, CD44, chromogranin A, Crp-ductin, and galectin, all of which are important in cell growth. By 8 months, the glandular stomach of the Atp4a(-/-) mice doubled in weight and thickness, and several modulators of growth were increased. Female Atp4a(-/-) mice were more hyperplastic than Atp4a(-/-) males at 12 and 20 months. By 1 year, severe mucocystic hyperplasia, incomplete intestinal metaplasia, ciliated metaplasia, a shift in mucins from neutral to acidic, and inflammation were widespread. Cells in the mucus pit zone developed a pyloric-type appearance, containing large hyaline-like, periodic acid-Schiff (PAS)-negative/alcian blue-negative inclusions. But critical characteristics of gastric neoplasia, such as nuclear atypia, invasion into the muscularis mucosa, and metastases were absent. In Atp4a(-/-) mice, chromogranin A and histidine decarboxylase, RegIIIgamma and delta, TFF3, osteopontin and CD44 were upregulated while Reg IIIbeta, and TFF1 were reduced. Chronic achlorhydria and hypergastrinemia in aged Atp4a(-/-) mice produced progressive hyperplasia, mucocystic and incomplete intestinal metaplasia, and the upregulation of growth factors without histological evidence of neoplasia.


Application of the Severity Factor and HMF Removal of Red Macroalgae Gracilaria verrucosa to Production of Bioethanol by Pichia stipitis and Kluyveromyces marxianus with Adaptive Evolution.
Gracilaria verrucosa, red seaweed, is a promising biomass for bioethanol production due to its high carbohydrate content. The optimal hyper thermal (HT) acid hydrolysis conditions are 12% (w/v) G. verrucosa with 0.2 M H2SO4 at 130 °C for 15 min, with a severity factor of 1.66. This HT acid hydrolysis produces 50.7 g/L monosaccharides. The maximum monosaccharide concentration of 58.0 g/L was achieved with 96.6% of the theoretical monosaccharide production from 120 g dry weight/L G. verrucosa slurry after HT acid hydrolysis and enzymatic saccharification. Fermentation was carried out by removing an inhibitory compound and via yeast adaptation to galactose. Both Pichia stipitis and Kluyveromyces marxianus adapted to galactose were excellent producers, with the ethanol yield (YEtOH) of 0.50 and 29.0 g/L ethanol production. However, the bioethanol productivity with Pichia stipitis adapted to galactose is higher than that with Kluyveromyces marxianus adapted to galactose, being 0.81 and 0.35 g/L/h, respectively. The results from this study can be applied to industrial scale bioethanol production from seaweed.


SARS-CoV-2 and Dengue virus Co-infection. A Case Report.
Coinfection of SARS-CoV-2 and dengue virus has not been previously reported. We report a confirmed case with favourable outcome, but whether the occurrence of simultaneous infections may alter the usual clinical course of each infection is still unknown.


[Development of a knee joint simulator (author's transl)].
This Knee Joint Simulator serves to investigate the tribological behaviour of total knee prostheses. In order to register the most important influences upon it, it is necessary to derive first of all the requirements upon the simulation of the various movements and loads from the physiological knee joint. These and some other requirements which are of a common sort, serve as the basis of the construction, the function of which is to be described in the following passage.


Graphene wrapped Cu2O nanocubes: non-enzymatic electrochemical sensors for the detection of glucose and hydrogen peroxide with enhanced stability.
In this study, a sensor for the detection of glucose and hydrogen peroxide was developed on the basis of Cu2O nanocubes wrapped by graphene nanosheets (Cu2O/GNs) as electrocatalysts. Cubic Cu2O nanocrystals/graphene hybrid has been successfully fabricated by a chemical reduction method at low temperature. The morphologies of the synthesized materials were characterized by scanning electron microscopy (SEM) and powder X-ray diffraction measurements (XRD). As a non-enzymatic amperometric sensor, the resulting Cu2O/graphene composite exhibited high sensitivity for the detection of glucose and H2O2. Moreover, the graphene coating was found to be able to effectively improve the electrochemical cycling stability of the fabricated sensor. With the Cu2O/GNs modified electrode, amperometric sensing of glucose was realized with a linear response over the concentration range from 0.3 to 3.3mM, a detection limit of 3.3 μM (S/N=3), high selectivity and short response time (<9s). Compared to unsupported Cu2O nanocubes, the graphene-wrapped Cu2O nanocubes exhibited higher catalytic activity for glucose oxidation with higher sensitivity and lower detection limit. The enzymeless sensor also exhibited good response toward H2O2, with the linear response ranging from 0.3 to 7.8mM at -0.4V and the detection limit of 20.8 μM. Moreover, because the surface is covered by graphene nanosheets, the as-synthesized Cu2O/GNs exhibited improved electrochemical stability. Such novel graphene nanosheets wrapped Cu2O nanocubes represent promising enzyme-free glucose and hydrogen peroxide sensors with high sensitivity and selectivity, improved stability and fast amperometric response.


Rapidly progressive outer retinal necrosis caused by varicella zoster virus in a patient infected with human immunodeficiency virus.
We describe a 21-year-old immunocompromised patient with a unique type of necrotizing herpetic retinitis. The retinitis progressed with loss of vision when acyclovir or ganciclovir alone was used for treatment. Examination of a chorioretinal biopsy specimen revealed that varicella zoster virus was the causative agent. Foscarnet monotherapy also failed to prevent progression of the retinal infection. Combination antiviral therapy with ganciclovir and foscarnet appeared to delay progression of disease and helped maintain a visual acuity of 20/200 for at least 6 months after the onset of infection.


Influence of pretreatment with various insecticides on the N-demethylation of dimethylnitrosamine.
The effects of various classes of insecticides were studied on the N-demethylation of dimethylnitrosamine (DMN) by mouse liver enzymes. Organochlorine insecticides, represented by lindane, DDT, and endrin, increased the activities of DMN demethylase I and II. The latter enzyme was more susceptible to the inducive action of the tested chlorinated insecticides. On the other hand, the synthetic pyrethroids, fenvalerate and flucythrinate, did not alter the activity of either enzyme. While pretreatment with carbaryl, a carbamate derivative, was without effect, moderate elevation in the activity of both demethylases was observed following administration of carbofuran. Dimethoate, representing organophosphorus compounds, was the only insecticide tested to inhibit the N-demethylation of DMN, with more pronounced effect on DMN demethylase I. Since DMN requires metabolic activation for its hepatotoxic and carcinogenic actions, alterations in the activities of its metabolizing enzymes as a sequela of exposure to certain insecticides may change susceptibility to its toxicity and/or carcinogenicity.


Local delivery of the Neuregulin1 receptor ecto-domain (ecto-ErbB4) has a positive effect on regenerated nerve fiber maturation.
The Neuregulin/ErbB system plays an important role in the peripheral nervous system, under both normal and pathological conditions. We previously demonstrated that expression of soluble ecto-ErbB4, the released extracellular fragment of the ErbB4 receptor, stimulated glial cell migration in vitro. In this study we examined the possibility of manipulating this system in vivo in order to improve injured peripheral nerve regeneration. Transected rat median nerves of adult female Wistar rats were repaired with a 10-mm-long graft made by muscle-in-vein combined nerve guide previously transduced with either the adeno-associated viral (AAV) vector AAV2-LacZ or AAV2-ecto-ErbB4. Autologous nerve grafts were used as control. Both stereological and functional analyses were performed to assess nerve regeneration. Data show that delivery of soluble ecto-ErbB4 by gene transfer in the muscle-in-vein combined nerve guide has a positive effect on fiber maturation, suggesting that it could represent a potential tool for improving peripheral nerve regeneration.


Effect of genetic polymorphisms of CYP3A5 and MDR1 on cyclosporine concentration during the early stage after renal transplantation in Chinese patients co-treated with diltiazem.
The aim of this study was to assess the influence of the cytochrome (CYP450)3A5 and multidrug resistance (MDR1) gene polymorphisms on cyclosporine A (CsA) trough concentration during the early stage after renal transplantation in Chinese patients co-treated with diltiazem. CYP3A5*3 (A6986G) and MDR1 C1236T, G2677T/A and C3435T polymorphisms were determined by PCR followed by restriction fragment length polymorphism (RFLP) analysis. A total of 112 Chinese renal transplant patients were enrolled in the study. The whole blood trough concentration was measured at 7 days after transplantation, and the dose-adjusted trough levels were compared among the different genotypes. The dose-adjusted trough levels of CsA were significantly higher in MDR1 2677TT carriers than in GG plus GT carriers (59.5 +/- 15.9 vs. 34.5 +/- 9.4 vs. 43.2 +/- 13.6 ng/mL per mg per kg; P < 0.0001). In patients who were co-treated with diltiazem, compared with carriers of haplotype T-T-C, the carriers of haplotype C-G-C and haplotype T-G-T had significantly lower dose-adjusted trough blood concentrations of CsA than the non-carrier group (P = 0.002, P = 0.000 and P = 0.000, respectively). However, no evidence was found that there was a relationship between the CYP3A5*3, MDR1 C1236T and MDR1 C3435T polymorphisms and CsA dose-adjusted trough concentrations. This study demonstrated that the G2677T/A single nucleotide polymorphisms in MDR1 and MDR1 haplotypes C-G-C, T-G-T and T-T-C are associated with the CsA concentration during the very early post-transplant period in Chinese renal transplant patients co-treated with diltiazem. These polymorphisms may be useful for determining the appropriate initial dose of CsA after renal transplantation.


Motor-cortical oscillations in early stages of Parkinson's disease.
Pathophysiological changes in basal ganglia-thalamo-cortical circuits are well established in idiopathic Parkinson's disease (PD). However, it remains open whether such alterations already occur at early stages representing a characteristic neurophysiological marker of PD. Therefore, the present study aims at elucidating changes of synchronised oscillatory activity in early PD patients. In this study, we performed whole-head magnetoencephalography (MEG) in a resting condition and during steady state contraction of the more severely affected forearm in 10 drug–naive, de novo patients, in 10 early-stage patients with chronic medication and in 10 age-matched control subjects. While cortico-muscular coherence (CMC) did not differ between groups, patients showed increased sensori-motor cortical power at beta frequency (13–30 Hz) during rest as well as during isometric contraction compared to controls. In healthy control subjects the power of the contralateral hemisphere was significantly suppressed during isometric contraction. By contrast, both hemispheres were activated equally strongly in de novo patients. In medicated patients, the pattern was found to be reversed. Contralateral beta power was significantly correlated with motor impairment during isometric contraction but not during rest. The present results suggest that the reduced ability of the primary motor cortex to disengage from increased beta band oscillations during the execution of movements is an early marker of PD.


Association between serum sodium and in-hospital mortality among critically ill patients with spontaneous subarachnoid hemorrhage.
The aim of this study was to retrospectively explore the relationship between serum sodium and in-hospital mortality and related factors in critically ill patients with spontaneous subarachnoid hemorrhage (SAH). Data were collected from the Medical Information Mart for Intensive Care IV database. Restricted cubic splines were used to explore the relationship between serum sodium and in-hospital mortality. Receiver operating characteristic analysis was used to calculate the optimal cutoff value of sodium fluctuation, and decision curve analysis was plotted to show the net benefit of different models containing serum sodium. A total of 295 patients with spontaneous SAH were included in the retrospective analysis. The level of sodium on ICU admission and minimum sodium in the ICU had a statistically significant non-linear relationship with in-hospital mortality (non-linear P-value < 0.05, total P-value < 0.001). Serum sodium on ICU admission, minimum serum sodium during ICU, and sodium fluctuation were independently associated with in-hospital mortality with odds ratios being 1.23 (95% confidence interval (CI): 1.04-1.45, P = 0.013), 1.35 (95% CI: 1.18-1.55, P < 0.001), and 1.07 (95% CI: 1.00-1.14, P = 0.047), respectively. The optimal cutoff point was 8.5 mmol/L to identify in-hospital death of patients with spontaneous SAH with sodium fluctuation, with an AUC of 0.659 (95% CI 0.573-0.744). Among patients with spontaneous SAH, we found a J-shaped association between serum sodium on ICU admission and minimum sodium values during ICU with in-hospital mortality. Sodium fluctuation above 8.5 mmol/L was independently associated with in-hospital mortality. These results require being tested in prospective trials.


P53, cyclin-dependent kinase and abnormal amplification of centrosomes.
Centrosomes play a critical role in formation of bipolar mitotic spindles, an essential event for accurate chromosome segregation into daughter cells. Numeral abnormalities of centrosomes (centrosome amplification) occur frequently in cancers, and are considered to be the major cause of chromosome instability, which accelerates acquisition of malignant phenotypes during tumor progression. Loss or mutational inactivation of p53 tumor suppressor protein, one of the most common mutations found in cancers, results in a high frequency of centrosome amplification in part via allowing the activation of the cyclin-dependent kinase (CDK) 2-cyclin E (as well as CDK2-cyclin A) which is a key factor for the initiation of centrosome duplication. In this review, the role of centrosome amplification in tumor progression, and mechanistic view of how centrosomes are amplified in cells through focusing on loss of p53 and aberrant activities of CDK2-cyclins will be discussed.


MRI Study on the Relationship Between Nerve and Blood Vessel in Unilateral Angiogenic Trigeminal Neuralgia.
Trigeminal neuralgia (TN) is a common cranial nerve disease. To investigate the relationship between the trigeminal nerve and the responsible blood vessel in patients with unilateral vascular trigeminal neuralgia (VTN). Thirty patients with unilateral VTN were confirmed by microvascular decompression. Among the 30 patients, the responsible blood vessels were present in 30 cases on the affected side and 17 cases on the uninfected side (1). The location of the intersection of the trigeminal nerve and the responsible blood vessel: the affected side is located 2/5 behind the trigeminal nerve cisternal segment; the healthy side is located 3/5 anterior to the cisternal segment (2). Symptomatic vessels were located within the cistern between the origin and 2/5ths of the cistern length, and non-symptomatic vessels were located beyond the 2/5ths location (3). Direction of intersection: on the affected side, the responsible vessel was located inside and above the trigeminal nerve in 27 cases, (27/30, 90%), and outside and below the trigeminal nerve in 3 cases (3/30, 10%). On the unaffected side, the responsible vessel was located inside and above the nerve in 16 cases (16/17, 94%) and outside and below the nerve in 1 case (1/17, 5.8%) (4). Intersection form: 3 cases (3/30, 10%) on the affected side, the responsible blood vessel contacted the trigeminal nerve, in 26 cases (26/30, 86%) the responsible blood vessel compressed the trigeminal nerve, and in 1 case (1/30, 5%) the responsible blood vessel caused the trigeminal nerve to be twisted; 8 cases (8/17, 47%) of the contralateral side contacted the trigeminal nerve with the responsible blood vessel, and in 9 cases (9/17, 53%) the responsible blood vessel compressed the trigeminal nerve. Patients with unilateral VTN have differences in the location and form of the intersection of the trigeminal nerve and the responsible vessel on the affected side and the contralateral side.


Effect of immunosuppressive agents and sunscreens on UV carcinogenesis in the hairless mouse.
The effect of two immunosuppressive agents, azathioprine and cyclophosphamide, with and without UVB sunscreen protection on UV-induced skin carcinogenesis was studied in the albino hairless mouse. In a daily treatment regime spanning 9 weeks, groups of mice were immunosuppressed with either drug, and were exposed to minimally erythemal doses of a light source simulating the UV portion of the solar spectrum. The accumulated UV exposure alone induced skin tumours in 77% of mice. Azathioprine, but not cyclophosphamide, significantly enhanced the incidence of UV tumorigenesis. Photoprotection by topical application of one of two commonly used UVB sunscreens, 2-ethyl-hexyl-p-methoxycinnamate (2-EHMC) or octyl-N-dimethyl-p-aminobenzoate (o-PABA), reduced the UV tumour incidence to zero in immunologically normal mice and to 8-15% in immunosuppressed mice. Unexpressed latent tumour initiations were revealed in all sunscreen-protected groups by the subsequent application of a tumour promoter, croton oil. In immunologically normal mice 2-EHMC had allowed initiations in 39% of UV-irradiated mice, and o-PABA in 16.5%. However, in UV-irradiated mice immunosuppressed with azathioprine there had been initiations in 78% of mice protected with 2-EHMC and 65% of mice protected with o-PABA. Photoprotected mice immunosuppressed with cyclophosphamide did not show the same increase in UV-initiations (22% with 2-EHMC, 23% with o-PABA). These results provide evidence that azathioprine increases the susceptibility of the skin to UV carcinogenesis. However, UVB sunscreens afford effective protection from overt tumour expression in the absence of a tumour promoter.


Triglycerides and systolic blood pressure negatively mediate the direct relationship of vitamin D status to resting energy expenditure: A cross sectional analysis.
We determined whether individual components of metabolic syndrome (MetS) mediated the direct association of vitamin D status (25OHD) on resting energy expenditure (REE). Multiple linear regression determined predictors of REE from data on 180 men and women from two ethnic groups. We then modelled a mediation pathway through components of MetS on the direct association between 25OHD and REE. The mediation modelling used the PROCESS SPSS Macro (version 4.0) based on 5000 bootstrapped samples, with the adjustment for different sets of covariates. REE was significantly predicted by age, fat mass (FM), fat free mass (FFM), ethnicity, inverse ln insulin, 25OHD, triglycerides (TG), systolic blood pressure (SBP) and, to some extent, by time of REE measurements (p < 0.094). Adjustment for all these covariates, resulted in a negative indirect mediation effect of TG [β coefficient (bootstrapped SE): 0.95 (0.519); bootstrapped 95% CI: 2.172, -0.165; p < 0.05] and a concurrent negative mediation of SBP [β coefficient (bootstrapped SE): 0.72(0.484); bootstrapped 95% CI: 1.851, -0.011; p < 0.05]. There remained a positive direct pathway from 25OHD to REE [β coefficient (S.E): 4.715 (2.129); p = 0.028], however the total effect of 25OHD was dampened [β coefficient (S.E): 3.04 (2.126); p = 0.154]. Independent of insulin sensitivity, a negative mediation by TG and SBP dampened the overall effect of 25OHD on REE.


Comparing administered and market-based water allocation systems through a consistent agent-based modeling framework.
Water allocation can be undertaken through administered systems (AS), market-based systems (MS), or a combination of the two. The debate on the performance of the two systems has lasted for decades but still calls for attention in both research and practice. This paper compares water users' behavior under AS and MS through a consistent agent-based modeling framework for water allocation analysis that incorporates variables particular to both MS (e.g., water trade and trading prices) and AS (water use violations and penalties/subsidies). Analogous to the economic theory of water markets under MS, the theory of rational violation justifies the exchange of entitled water under AS through the use of cross-subsidies. Under water stress conditions, a unique water allocation equilibrium can be achieved by following a simple bargaining rule that does not depend upon initial market prices under MS, or initial economic incentives under AS. The modeling analysis shows that the behavior of water users (agents) depends on transaction, or administrative, costs, as well as their autonomy. Reducing transaction costs under MS or administrative costs under AS will mitigate the effect that equity constraints (originating with primary water allocation) have on the system's total net economic benefits. Moreover, hydrologic uncertainty is shown to increase market prices under MS and penalties/subsidies under AS and, in most cases, also increases transaction, or administrative, costs.


Carotenoid composition and photon-use efficiency of photosynthesis inGossypium hirsutum L. grown under conditions of slightly suboptimum leaf temperatures and high levels of irradiance.
Cotton (Gossypium hirsutum L. var. DP 61) was grown at different temperatures during 12-h light periods, with either 1800-2000 μmol photons m-2 s-1 (high photon flux density, PFD) or 1000-1100 μmol m-2 s-1 (medium PFD) incident on the plants. Night temperature was 25°C in all experiments. Growth was less when leaf temperatures were below 30°C during illumination, the effect being greater in plants grown with high PFD (Winter and Königer 1991). Leaf pigment composition and the photon-use efficiency of photosynthesis were analysed to assess whether plants grown with high PFD and suboptimal temperatures experienced a higher degree of high irradiance stress during development than those grown with medium PFD. The chlorophyll content per unit area was 3-4 times less, and the content of total carotenoids about 2 times less, with the proportion of the three xanthophylls zeaxanthin + antheraxanthin + violaxanthin being greater in leaves grown at 20-21°C than in leaves grown at 33-34°C. In leaves from plants grown at 21°C and 1800-2000 μmol photons m-2 s-1, zeaxanthin accounted for as much as 34% of total carotenoids in the middle of the photoperiod, the highest level recorded in this study. This finding is consistent with a protective role of zeaxanthin under conditions of excess light. At the lower temperatures, the photochemical efficiency of photosystem II, measured as the ratio of variable to maximum fluorescence yield (F V/F M) after 12-h dark adaptation, was 0.76 in medium PFD plants and 0.75 in high PFD plants compared with 0.83 and 0.79, respectively, at the higher temperatures. The photon-use efficiency of O2 evolution (ϕ) based on absorbed light between 630 and 700nm, decreased with decrease in temperature from 0.102 to 0.07 under conditions of high PFD, but remained above 0.1 at medium PFD. Owing to compensatory reactions in these long-term growth experiments, sustained differences inF V/F M and ϕ were much less pronounced than the differences in chlorophyll content and dry matter, particularly in plants which had developed at high PFD and low temperature. In fact, in these plants, which exhibited pronounced photobleaching, a largely functional photosynthetic apparatus was still maintained in cells adjacent to the lower leaf surfaces. This was indicated by measurements of photon use efficiencies of photosynthetic O2 evolution with leaves illuminated first at the upper, and then at the lower surface.


Myelopathy in systemic lupus erythematosus: clinical, laboratory, radiological and progression findings in a cohort of 1,193 patients.
To describe clinical, laboratory, radiological and progression characteristics of myelopathy in systemic lupus erythematosus (SLE). A retrospective analysis was performed on a cohort of 1193 patients with SLE (ACR criteria) in order to identify patients with myelopathy (neuropsychiatric ACR). Disease activity was assessed by the SLE activity index (SLEDAI) on the date of the event and functional capacity was assessed by the Expanded Disability Status Scale (EDSS) at the last visit. We identified 14 (1.2%) patients with myelopathy. All were women with a mean age of 30±11.5 years. Myelopathy occurred at the diagnosis of SLE in four (28%) patients; and nine (64%) patients had another type of neuropsychiatric manifestation associated. Neurological recurrence was observed in one (7%) patient. Disease activity was observed in 2 (14%) patients. Cerebrospinal fluid presented pleocytosis on 7 (53%) patients; antiphospholipid antibodies were positive in 5 (45%). Magnetic resonance imaging (MRI) showed T2 hyperintensity with a predominance of longitudinal involvement in 6 (86%) patients. Most were treated with intravenous corticosteroids and cyclophosphamide. No patient had full recovery and four (36%) had high EDSS scores. Three (21%) patients died from sepsis early in the course of their myelopathy, during or after immunosuppressive therapy. Myelopathy occurred in 14 (1.2%) of the patients in our cohort and this may be the first manifestation of the disease occurring independently of systemic disease activity. Although rare, myelopathy shows great morbidity and mortality, can be recurrent and MRI is critical for diagnosis.


Structural analysis of a pectic polysaccharide from the leaves of Diospyros kaki.
A pectic polysaccharide DL-2A with a molar mass of 8.5 x 10(5), was obtained from the boiling water extract of Diospyros kaki leaves. It had [alpha]20D -21.8 degrees (c 0.22, H2O) and consisted of rhamnose, arabinose, galactose, xylose and galacturonic acid units in the molar ratio of 0.4:3.4:2.4:1.0:0.8, along with traces of glucuronic acid. About 16.7% of galacturonic acid existed as the methyl ester. A combination of linkage analyses, periodate oxidation, partial acid hydrolysis, selective lithium-degraded reaction, ESIMS, 1H- and 13C- NMR spectral analyses revealed its structural features. It was found that DL-2A possessed an alpha-(1-->4)-galacturonan backbone with some insertions of alpha-1,2-Rhap residues. The side-chains of arabino-3,6-galactan were attached to the backbone via O-4 of Rhap residues and O-3 of GalAp residues, while 4-linked xylose residues (forming short linear chains) were directly linked to O-4 of rhamnose residues, not as part of the xylogalacturonan. These novel structural features enlarge the knowledge on the fine structure of pectic substances in the plant kingdom.


Vaccine adjuvants alter TCR-based selection thresholds.
How T cell receptor (TCR) specificity evolves in vivo after protein vaccination is central to the development of helper T (Th) cell function. Most models of clonal selection in the Th cell compartment favor TCR affinity-based thresholds. Here, we demonstrated that depot-forming vaccine adjuvants did not require Toll-like receptor (TLR) agonists to induce clonal dominance in antigen-specific Th cell responses. However, readily dispersible adjuvants using TLR-9 and TLR-4 agonists skewed TCR repertoire usage by increasing TCR selection thresholds and enhancing antigen-specific clonal expansion. In this manner, vaccine adjuvants control the local accumulation of Th cells expressing TCR with the highest peptide MHC class II binding. Clonal composition was altered by mechanisms that blocked the local propagation of clonotypes independently of antigen dose and not as a consequence of interclonal competition. This capacity of adjuvants to modify antigen-specific Th cell clonal composition has fundamental implications for the design of future protein subunit vaccines.


Hydroxyl radicals release in rat striatum involves metabotropic glutamate receptors.
Disruption of glutamate homeostasis frequently leads to oxidative stress and to the release of hydroxyl radicals (radical OH). Here, we investigated, via a microdialysis approach, the possible involvement of metabotropic glutamate receptors in the glutamate-induced release of hydroxyl radicals in adult rat striatum. Glutamate was applied at low amount, resulting in a moderate release that was not inhibited by dizocilpine (MK-801), a specific NMDA receptor antagonist. (RS)-alpha-methyl-4-carboxyphenylglycine (MCPG), a broad spectrum metabotropic antagonist, that does not exert any effect on the basal release of radical OH suppressed their response to glutamate. (+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (t-ACPD), a non-selective metabotropic glutamate receptors agonist, promoted an radical OH release almost similar to that observed after glutamate, which was similarly impaired by co-infusion with MCPG. By contrast, infusion of (RS)-3,5-dihydroxyphenylglycine (DHPG), a more specific group I metabotropic glutamate receptors agonist, did not result in any appreciable radical OH response. Thus, beside NMDA receptors, some metabotropic glutamate receptors may also be involved in the glutamate-induced release of hydroxyl radicals.


A novel DNA adjuvant, N3, enhances mucosal and systemic immune responses induced by HIV-1 DNA and peptide immunizations.
The study was designed to evaluate a novel cationic lipid DNA adjuvant (N3) and its function for HIV-1gp160/rev DNA plasmid delivered intranasally. The primary N3/HIV-DNA plasmid immunizations were boosted intranasally with a gp41 peptide in a anionic L3 adjuvant. This novel prime-boost strategy of mucosal immunization provided a broad HIV-1 envelope specific immunity, and recognition of viruses of subtypes A, B and C. Intranasal N3-adjuvanted gp160/rev DNA prime followed by one L3-peptide boosting immunization, induced broadly neutralizing antibodies against HIV-1 in the mucosa and systemically. The needle-free intranasal prime-boost strategy using two different adjuvant formulations reduced significantly the dose of DNA needed.


Isolated multiple bilateral echogenic papillary muscles: A unique sonographic feature of trisomy 13.
Echogenic papillary muscles are noted in 30% of fetuses with trisomy 13. All reported fetuses with trisomy 13 and echogenic papillary muscles have exhibited additional abnormal sonographic findings. A 21.7-week fetus demonstrated three papillary echogenicities in each cardiac ventricle during ultrasound examination. Chromosomal analysis of amniocytes showed the karyotype 47,XX,+13. Multiple bilateral papillary muscles may provide the only sonographic sign of fetal trisomy 13.


Weight management in youth with rapid-onset obesity with hypothalamic dysregulation, hypoventilation, autonomic dysregulation, and neural crest tumor (ROHHAD-NET): literature search and case report.
Rapid-onset obesity with hypothalamic dysfunction, hypoventilation, autonomic dysregulation, and neural endocrine tumor (ROHHAD-NET) syndrome is a youth-onset constellation of symptoms including rapid-onset obesity with hypothalamic dysfunction, hypoventilation, and autonomic dysregulation. Despite growing understanding of the clinical classification of this syndrome there is limited investigation into treatment of the rapid-onset obesity which can be progressive and life-limiting. The purpose of this case report is to describe the clinical timeline and treatment of severe obesity in a patient with of ROHHAD-NET and propose recommendations for the treatment of associated obesity. We present the case of a 10-year-old female with a clinical presentation consistent with ROHHAD-NET who achieved clinically meaningful weight loss with a combination of lifestyle modification and anti-obesity pharmacotherapies. We report on the use of three separate pharmacological agents and ultimately the referral for bariatric surgery. Given that early-onset obesity and hypoventilation are life-limiting components of this condition, early recognition and treatment are essential to improve health outcomes.


Effect of double antibiotic and calcium hydroxide pastes on dislodgement resistance of an epoxy resin-based and two calcium silicate-based root canal sealers.
The purpose of the present study was to determine the dislodgement resistance of AH Plus, MTA Fillapex, and Total Fill BC sealer to root canal dentin walls following placement of calcium hydroxide (CH) or double antibiotic paste (DAP) medicaments. Root canals of 90 single-rooted human mandibular premolar teeth were instrumented with Reciproc rotary instruments to a size R50. The teeth were randomly divided into two experimental groups: those receiving an intracanal medicament with either CH or DAP and a control group (n = 30). After 3 weeks, the medicaments were removed by irrigation with 5 mL of 2.5 % NaOCl, 5 mL 17 % EDTA, and 5 mL distilled water under sonic agitation. Each group was then subdivided into three subgroups (n = 10), and the canals were filled with either AH Plus, MTA Fillapex, or Total Fill BC sealer. After 1 week, a push-out test was applied to the specimens and the results were analyzed using one-way analysis of variance (ANOVA) and post hoc Tamhane's tests. Regardless of the type of intracanal medicament used, Total Fill BC Sealer and MTA Fillapex showed the highest and lowest bond strength, respectively (P < 0.05). The use or absence of medicaments did not significantly affect the dislodgment resistance of root canal fillings (P > 0.05). Prior CH placement improved dislodgement resistance of all sealers without statistical difference (P > 0.05). Adhesive failure between core and sealer was the most frequent failure mode. Prior application of CH or DAP did not significantly affect the adhesion of the AH Plus, MTA Fillapex, and Total Fill BC Sealers. There is a little information about the influence of antibiotic medicaments on the bond strength of root canal sealer. These findings suggest that the use of DAP does not affect the adhesion strength of AH Plus, MTA Fillapex, and Total Fill BC Sealers.


Rapid changes of mitochondrial Ca2+ revealed by specifically targeted recombinant aequorin.
Introduction of Ca2+ indicators (photoproteins, fluorescent dyes) that can be trapped in the cytosolic compartment of living cells has yielded major advances in our knowledge of Ca2+ homeostasis. Ca2+ however regulates functions not only in the cytosol but also within various organelles where indicators have not yet been specifically targeted. Here we present a novel procedure by which the free Ca2+ concentration of mitochondria, [Ca2+]m, can be monitored continuously at rest and during stimulation. The complementary DNA for the Ca2+ sensitive photoprotein aequorin was fused in frame with that encoding a mitochondrial presequence. The hybrid cDNA was transfected into bovine endothelial cells and stable clones were obtained expressing variable amounts of mitochondrially targeted apoaequorin. The functional photoprotein could be reconstituted in intact cells by incubation with purified coelenterazine and [Ca2+]m could thus be monitored in situ. This allowed the unprecedented direct demonstration that agonist-stimulated elevations of cytosolic free Ca2+, [Ca2+]i, (measured in parallel with Fura-2) evoke rapid and transient increases of [Ca2+]m, which can be prevented by pretreatment with a mitochondrial uncoupler. The possibility of targeting aequorin to cellular organelles not only offers a new and powerful method for studying aspects of Ca2+ homeostasis that up to now could not be directly approached, but might also be used in the future as a tool to report in situ a variety of apparently unrelated phenomena of wide biological interest.


Comparison of utilizing modified hold-relax, muscle energy technique, and instrument-assisted soft tissue mobilization on hamstring muscle length in healthy athletes: Randomized controlled trial.
The hamstring muscle shortness is the primary risk factor for sports-related injuries. Numerous treatments are available for lengthening of hamstring muscle. The main purpose of this study was to compare the immediate effect of modified hold-relax, muscle energy technique (MET), and instrument assisted soft tissue mobilization-Graston techniques (IASTM-GT) on length of hamstring muscle in young healthy athletes. 60 athletes comprising of 29 females and 31 males were recruited in the present study. Participants were allocated to 3 groups of IASTM-GT (N = 20, 13 male, 7 female), Modified Hold-relax (N = 20, 8 male, 12 female), and MET (N = 20, 7 male, 13 female). Active knee extension and passive straight leg raising (SLR), and toe touch test were performed before and immediately after the intervention by a blinded assessor. For the comparison of dependent variables across time, 3*2 repeated measure ANOVA was utilized. Interaction of group by time was significant for passive SLR (P < 0.001). Interaction of group by time was not significant for active knee extension (P = 0.17). The results showed that dependent variables increased significantly in all groups. The effect size (Cohen's d) in the groups of IASTM-GT, modified Hold-relax, and MET was 1.7, 3.17, and 3.12, respectively. Although the measures were improved in all groups, it seems that IASTM-GT can be used as a safe and efficient treatment, which can be a suitable candidate alongside modified hold-relax and MET for increasing the hamstrings muscle length in healthy athletes.


Serum glycoproteins in the liver diseases. IV. Alpha-1 acid glycoprotein level in liver cirrhosis.
Circulating alpha-1 acid glycoprotein level in cirrhotic patients was determined by radioimmunoassay, and was compared to the ones in normal subjects and chronic active hepatitis with sublobular necrosis. Serum alpha-1 acid glycoprotein levels in liver cirrhosis (p less than 0.001) and chronic active hepatitis with sublobular necrosis (p less than 0.02) were significantly reduced comparing to the normal subjects, although any statistically significant difference was not observed between the formers. In liver cirrhosis, thie serum alpha-1 acid glycoprotein level correlated negatively with serum albumin concentration but neither with serum alpha-1 globulin fraction nor with Indocyanine green clearance rate.


In vivo enantioselectivity of epibatidine interaction with calcium-modulating drugs.
Several recent studies have implicated the involvement of calcium ion fluxes in the response to nicotine on a cellular level. The influence of a calcium agonist (BAY K 8644) and calcium blockers on (+)- and (-)-epibatidine-induced antinociception was investigated in mice using the tail-flick test. The effect of (+)-epibatidine but not (-)-epibatidine was significantly increased by BAY K 8644 pretreatment. This potentiation by BAY K 8644 was blocked by pretreating the animals with nifedipine at 2 mg/kg. The calcium channel antagonist, nimodipine, reduced significantly the antinociception induced by (+)-epibatidine but not (-)-epibatidine (12 microg/kg, s.c.). Similar results were also observed with the calcium chelator EGTA. Thus, epibatidine isomers pharmacological effect was differentially modulated by calcium signaling process secondary to receptor activation.


Production of surfactant protein C in the baculovirus expression system: the information required for correct folding and palmitoylation of SP-C is contained within the mature sequence.
Surfactant protein C (SP-C) is synthesized in the alveolar type II cells of the lung as a 21 kDa propeptide which is proteolytically processed to a 4.2 kDa mature active form. The main function of this extremely hydrophobic protein is to enhance lipid insertion into the air/liquid interface in the lung upon inhalation. This is necessary to maintain a relatively low surface tension at this interface during breathing. In this report we describe the production of mature human SP-C in the baculovirus expression system. The recombinant protein contains a secondary structure with a high alpha-helical content (73%), comparable to native SP-C, as determined by circular dichroism and attenuated total reflection Fourier transform infrared analysis. The expressed protein is a mixture of dipalmitoylated (15%) and non-palmitoylated SP-C. This suggests that the information required for palmitoylation is contained within the sequence of the mature protein. The activity of the protein to insert phospholipids into a preformed monolayer of lipids at an air/liquid interface was determined with a captive bubble surfactometer. Recombinant SP-C significantly reduced the surface tension at the air/liquid interface during dynamic expansion and compression. We conclude that correctly folded, dipalmitoylated and active SP-C can be expressed in the baculovirus expression system. Our results may facilitate investigations into the relation between structure and function of SP-C and into protein palmitoylation in general.


Androgen and androgen-metabolizing enzymes in metastasized lymph nodes of breast cancer.
Androgen receptor and androgen metabolizing enzymes, 17β-hydroxysteroid dehydrogenase type 5 (17βHSD5) and 5α-reductase1 (5α1), are frequently detected in primary tumor of breast cancer, but their status in metastatic lymph nodes has not been examined. The biological role of androgen in breast cancer and its metastatic process also remain unknown. In this study, we used immunohistochemistry to localize the expression of androgen receptor, 17βHSD5, and 5α1 in primary tumors and paired metastatic lymph nodes and correlated the findings with clinicopathologic factors of individual patients. Approximately 70% of primary tumors and paired metastatic lymph nodes expressed androgen receptor, with significant correlation between both lesions. However, 17βHSD5 and 5α1 immunoreactivity was decreased in metastatic lymph nodes. Alone or in tandem with androgen receptor, 5α1 was associated with significantly lower Ki-67 index, lower pathologic grade, and higher estrogen receptor positivity, but androgen receptor/5α1 double positivity in lymph nodes was associated with larger lymph node metastasis and higher TNM stage. In conclusion, androgen receptor immunoreactivity remained stable during the process of metastasis, whereas androgen-metabolizing enzymes decreased. Although results of our study and previous reports imply additional roles of androgen metabolism in the metastasis process, especially conversion by 5α1, there may be divergence between its effects on primary tumor and those in metastatic lymph nodes.


Mitochondrial oxidative stress and dysfunction in rat brain induced by carbofuran exposure.
Repeated low-dose exposure to carbofuran exerts its neurotoxic effects by non-cholinergic mechanisms. Emerging evidence indicates that oxidative stress plays an important role in carbofuran neurotoxicity after sub-chronic exposure. The purpose of the present study is to evaluate the role of mitochondrial oxidative stress and dysfunction as a primary event responsible for neurotoxic effects observed after sub-chronic carbofuran exposure. Carbofuran was administered to rats at a dose of 1 mg/kg orally for a period of 28 days. There was a significant inhibition in the activity of acetylcholinesterase (66.6%) in brain samples after 28 days of carbofuran exposure. Mitochondrial respiratory chain functions were assessed in terms of MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) reduction and activity of succinate dehydrogenase in isolated mitochondria. It was observed that carbofuran exposure significantly inhibited MTT reduction (31%) and succinate dehydrogenase activity (57%). This was accompanied by decrease in low-molecular weight thiols (66.6%) and total thiols (37.4%) and an increase in lipid peroxidation (43.7%) in the mitochondria isolated from carbofuran-exposed rat brain. The changes in mitochondrial oxidative stress and functions were associated with impaired cognitive and motor functions in the animals exposed to carbofuran as compared to the control animals. Based on these results, it is clear that carbofuran exerts its neurotoxicity by impairing mitochondrial functions leading to oxidative stress and neurobehavioral deficits.


Structural diversity and dynamics of microtubules and polymorphic tubulin assemblies.
Tubulin, the main protein of microtubules (MTs), has the potency of forming a variety of other assembly products in vitro: rings, ring-crystals, C- and S-shaped ribbons, 10 nm fibres, hoops, sheets, heaped sheets, MT doublets, MT triplets, double-wall MTs, microtubules, curled ribbons, and paracrystals. The supramolecular subunits of all of them are the protofilaments which might be arranged either parallel to the axis (e.g., in MTs, ribbons) or curved (e.g., in hoops, microtubules). There is strong evidence that in the second case the protofilaments have an inside-out orientation compared to MTs. All assembly products mentioned are described structurally and their relevance to the in vivo situation is considered. Moreover, MTs and the other assemblies undergo permanent changes. These dynamics occurring in both individual assemblies and assembly populations are discussed from the structural point of view.


Stem Cells and Organs-on-chips: New Promising Technologies for Human Infertility Treatment.
Having biological children remains an unattainable dream for most couples with reproductive failure or gonadal dysgenesis. The combination of stem cells with gene editing technology and organ-on-a-chip models provides a unique opportunity for infertile patients with impaired gametogenesis caused by congenital disorders in sex development or cancer survivors. But how will these technologies overcome human infertility? This review discusses the regenerative mechanisms, applications, and advantages of different types of stem cells for restoring gametogenesis in infertile patients, as well as major challenges that must be overcome before clinical application. The importance and limitations of in vitro generation of gametes from patient-specific human-induced pluripotent stem cells (hiPSCs) will be discussed in the context of human reproduction. The potential role of organ-on-a-chip models that can direct differentiation of hiPSC-derived primordial germ cell-like cells to gametes and other reproductive organoids is also explored. These rapidly evolving technologies provide prospects for improving fertility to individuals and couples who experience reproductive failure.


Economic and disease burden of breast cancer associated with suboptimal breastfeeding practices in Mexico.
Exclusive breastfeeding and longer breastfeeding reduce women's breast cancer risk but Mexico has one of the lowest breastfeeding rates worldwide. We estimated the lifetime economic and disease burden of breast cancer in Mexico if 95% of parous women breastfeed each child exclusively for 6 months and continue breastfeeding for over a year. We used a static microsimulation model with a cost-of-illness approach to simulate a cohort of Mexican women. We estimated breast cancer incidence, premature mortality, disability-adjusted life years (DALYs), medical costs, and income losses due to breast cancer and extrapolated the results to 1.116 million Mexican women of age 15 in 2012. Costs were expressed in 2015 US dollars and discounted at a 3% annual rate. We estimated that 2,186 premature deaths (95% CI 2,123-2,248), 9,936 breast cancer cases (95% CI 9,651-10,220), 45,109 DALYs (95% CI 43,000-47,217), and $245 million USD (95% CI 234-256) in medical costs and income losses owing to breast cancer could be saved over a cohort's lifetime. Medical costs account for 80% of the economic burden; income losses and opportunity costs for caregivers account for 15 and 5%, respectively. In Mexico, the burden of breast cancer due to suboptimal breastfeeding in women is high in terms of morbidity, premature mortality, and the economic costs for the health sector and society.


Deciphering the TCR Repertoire to Solve the COVID-19 Mystery.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected several millions and killed more than quarter of a million worldwide to date. Important questions have remained unanswered: why some patients develop severe disease, while others do not; and what roles do genetic variabilities play in the individual immune response to this viral infection. Here, we discuss the critical role T cells play in the orchestration of the antiviral response underlying the pathogenesis of the disease, COVID-19. We highlight the scientific rationale for comprehensive and longitudinal TCR analyses in COVID-19 and reason that analyzing TCR repertoire in COVID-19 patients would reveal important findings that may explain the outcome disparity observed in these patients. Finally, we provide a framework describing the different strategies, the advantages, and the challenges involved in obtaining useful TCR repertoire data to advance our fight against COVID-19.


Electromyographic activity in the immobilized shoulder girdle musculature during ipsilateral kinetic chain exercises.
To quantify the electromyographic activity in the shoulder girdle musculature during ipsilateral kinetic chain exercises performed in a shoulder immobilizer in asymptomatic men. Descriptive. Motion analysis laboratory at a tertiary care center. Five asymptomatic male volunteers, ages 24 to 32 years. Fine-wire (supraspinatus, infraspinatus, upper subscapularis) and surface (deltoids, trapezii, biceps, serratus anterior) electrodes recorded electromyographic activity from each muscle during a split-stance cross-body rotation (twisting to the opposite side at high, mid, and low levels), split stance attempted ipsilateral floor touch, and attempted overhead reach. All movements were initiated from the immobilized scapula and were tested with and without a combined step. Mean peak normalized (percentage of maximum voluntary contraction [%MVC]) electromyographic activity of each muscle during each exercise. For all exercises, biceps and infraspinatus activity remained low (<10% MVC), whereas upper subscapularis activity was moderate to very high (29%-68% MVC). Supraspinatus activity was low (<20% MVC) for all motions except the attempted overhead reach (23% MVC). Serratus electromyographic activity was less than 20% of MVC for all motions and was most responsive to added stepping (23%-136% MVC without stepping vs 24%-199% MVC with stepping). Cross-body rotation at lower heights progressively increased serratus activity while decreasing supraspinatus, upper trapezius, and anterior deltoid activity. Based on these electromyographic data, selected kinetic chain exercises could potentially be implemented during periods of shoulder immobilization. All exercises examined could potentially be safe after superior labral anteroposterior repair, but not after subscapularis repair. All exercises, with the exception of the attempted overhead reach, could potentially be safe after supraspinatus repair, with or without concomitant infraspinatus repair. Early activation of the serratus anterior could potentially be achieved by performing cross-body rotations, particularly at lower heights.