A Method for the Autonomous Control of Lower Limb Exo-skeletons for Persons with Paraplegia.
Efforts have recently been reported by several research groups on the development of computer-controlled lower limb orthoses to enable legged locomotion in persons with paraplegia. Such systems must employ a control framework that provides essential movements to the paraplegic user (i.e., sitting, standing, and walking), and ideally enable the user to autonomously command these various movements in a safe, reliable, and intuitive manner. This paper describes a control method that enables a paraplegic user to perform sitting, standing, and walking movements, which are commanded based on postural information measured by the device. The proposed user interface and control structure was implemented on a powered lower limb orthosis, and the system was tested on a paraplegic subject with a T10 complete injury. Experimental data is presented that indicates the ability of the proposed control architecture to provide appropriate user-initiated control of sitting, standing, and walking. The authors also provide a link to a video that qualitatively demonstrates the user's ability to independently control basic movements via the proposed control method.


Phytochemicals, antioxidant, antinociceptive and anti-inflammatory potential of the aqueous extract of Teucrium stocksianum bioss.
Despite availability of a substantial number of potent synthetic drugs, medicinal plants are still playing a key role in the discovery of novel and effective drug molecules. Numerous researchers are focusing on the plant based medicines due to its strong safety profiles. Teucrium species exhibit profound antidiabetic, analgesic and spasmolytic activities. The methanolic extract and essential oil of Teucrium stocksianum possess strong antinociceptive activity. The aim of the current research study was to determine the phytochemicals, antioxidant, analgesic and anti-inflammatory potential of the aqueous extract of Teucrium stocksianum Bioss (AETS). Phytochemical screening was carried out according to standard procedures. The antioxidant potential of the extract was ascertained with the stable organic free radical (2, 2-diphenyl-1-picryl-hydrazyl). Three different pain models, including acetic acid induced writhing, formalin induced paw licking and tail immersion tests were carried out for the determination of antinociceptive potential, while the anti-inflammatory activity was evaluated through carrageenan induced paw edema test in mice. The antinociceptive and anti-inflammatory potentials of AETS were assessed at 100, 200 and 300 mg/kg body weight, while acute toxicity were observed at 1500 mg/kg body weight in various groups of mice. Phytochemical screening has shown the occurrence of flavonoids saponins, reducing sugars, terpenoids and tannins. AETS exhibited profound antioxidant activity and has shown maximum activity (60.06 ± 0.846) at 250 μg/ml. In the three pain models AETS displayed marked dose dependent antinociceptive potential. AETS exhibited 63.5, 67.61 and 64% activity in acetic acid induced, formalin induced paw licking and tail immersion tests respectively. The antinociceptive effect of AETS and reference standard drug Tramadol(R) was significantly reversed by Naloxone, endorsed the central analgesic potential of AETS. Similarly the extract also reversed the paw edema in dose dependent manner. AETS displayed significant (53.81%) anti-inflammatory effects at a dose of 300 mg/kg that persisted till 5(th) h. In acute toxicity test AETS was found safe at 1500 mg/kg body weight. AETS exhibited profound antioxidant activity. The test sample displayed marked antinociceptive potential in all the test procedures, indicating the peripheral and central analgesic effects of AETS. The plant extract also displayed marked anti-inflammatory activity at all test doses.


Trends and Determinants in Breastfeeding among Korean Women: A Nationwide Population-Based Study.
Many efforts have been launched to promote breastfeeding in Korea. However, breastfeeding trends and associated factors with breastfeeding in Korea remain unknown. This study aimed to examine trends and determinants in breastfeeding using the Korea National Health and Nutrition Examination Survey (KNHANES) (2010-2018). We analyzed data from the KNHANES V (2010-2012), VI (2013-2015), and VII (2016-2018). A total of 9232 women aged 19-49 years were included in this study. We performed multivariable logistic regression analyses to investigate breastfeeding prevalence trends and associated factors with breastfeeding. Compared to 2010-2012, the odds ratio associated with breastfeeding during 2013-2015 and 2016-2018 increased to 1.30 (95% confidence interval (CI): 1.11-1.51) and 1.40 (95% CI: 1.21-1.63), respectively. The breastfeeding rate was associated with 40-49 years (OR, 95% CI: 0.47, 0.34-0.64 compared to 19-29 years), richer and poorer income (1.20, 1.03-1.39 in richer group and 1.24, 1.05-1.46 in poorer group compared to richest group), education level (0.74, 0.65-0.86 in ≤12 years of education compared to ≥13 years of education), smoking status (1.77, 1.38-2.28 in non-smoking compared to smoking), and self-rated health (1.40, 1.14-1.70 in good and 1.20, 1.00-1.44 in average compared to bad). Education programs and policies such as the Baby-Friendly Hospital Initiative (BFHI) and mother-friendly workplaces are necessary to increase the rates of breastfeeding in these groups.


Eagle syndrome after a fracture of complete ossified stylohyoid ligament from indirect trauma treated using local steroid injection: A case report.
Stylohyoid complex syndrome is characterized by various cervicopharyngeal symptoms related to the ossification and abnormality of the styloid process, stylohyoid ligament, and the lesser horn of the hyoid bone. Eagle syndrome is the most well-known of the spectra of these diseases. Although surgical treatment is considered effective, conservative treatment may be beneficial if symptoms arise because of inflammation of the soft tissues attached to the styloid process or hyoid bone. A 68-year-old man presented with pain in the right side of the neck and odynophagia after trauma on his philtrum. He was diagnosed with Eagle syndrome elicited by a fracture from indirect trauma. Despite analgesic medication and physiotherapy, the pain had somewhat relieved but persisted for 1 year. Computed tomography revealed complete ossification of the bilateral stylohyoid complex. A fracture was observed in the ampulla on the right side of the neck. One year later, the fracture resolved by complete union. Ultrasonography was performed and abnormal ossification was observed on the right side of the neck. Five milligrams of dexamethasone at a concentration of 1 kg/m was slowly injected into the tender point under ultrasonographic guidance. The patient reported immediate reduction of pain and was satisfied with the resolution. No recurrence was observed during a 6-month follow-up period. Although traumatic fracture of the ossified ligament elicited the syndrome, the results were satisfactory because the origin of the patient's pain was presumed to arise from inflammatory conditions. This case demonstrates that treatment with local steroid injection may be appropriate for patients who present with pain originating from muscles and ligaments.


[Inhibitory effect of the recombinant adenovirus pre-miR-10a on K562 cell proliferation].
To investigate the changes of proliferation and apoptosis in K562 cells over-expressing miR-10a. K562 cells were infected with pAd-pre-miR-10a virus, and then the levels of miR-10a and bcr/abl in transfected K562 cells was detected using RT-PCR and the level of BCR/ABL using Western blotting. The methylthiazolyl tetrazolium (MTT) assay and flow cytometry (FCM) were used to monitor the changes of proliferation and apoptosis of K562 cells after transfection. Compared with the control groups, K562 cells transfected with pAd-pre-miR-10a presented the significantly increased level of miR-10a, the significantly decreased expressions of bcr/abl fusion gene and BCR/ABL fusion protein, the inhibited proliferation ability and the promoted apoptosis (P<0.05). The recombinant adenovirus of pAd-pre-miR-10a can inhibit the proliferation of K562 cells and promote cell apoptosis significantly.


[Residents training in private practice : Conflicts between circumstances, willingness, and reality].
The increasing average age of urologists in private practice and outpatient treatment needs means that there is an increased demand for young urologists to cover outpatient care. Due to the increasing range of treatments, residents lack the mediation of those treatment methods and medical conditions which are necessary to become competent in the complete field of urology. To assess the willingness, the requirements, and the current implementation of residents training in private practice. A 14-item online survey was sent to 1326 urologists in private practice to measure the willingness, the requirements, and the current implementation of residents training. Of the 219 (17 %) respondents, 25 % have already trained residents, 41 % were authorized to provide training to residents, and 87 % have taken the appointment of a trainee into consideration. The main reason for employing a trainee was the enjoyment of the teaching experience. The main reason against employing a trainee was the lack of financial compensation. Urologists in private practice are willing and able to participate in residents' training. An extended integration of urological training into private practice can contribute to ensure the teaching of a wide variety of diseases/procedures and influence structural changes in the field of urology. Residents, private practitioners, and clinicians should search for solutions together with those responsible in the health care system for better integration of private practice into urology residents training.


Population data of 17 short tandem repeat loci in 2923 individuals from the Han population of Nantong in East China.
Nantong is located in mid-eastern China, and the Han population in Nantong may be greatly affected by population admixture between northern and southern Han Chinese populations. In this study, we analyzed 17 autosomal short tandem repeat (STR) loci on 2923 unrelated individuals collected from the Han population of Nantong. No significant deviation from Hardy-Weinberg equilibrium was observed at all STR loci, and the expected heterozygosity ranged from 0.6184 to 0.9187. The combined match probability (CMP) was 3.87 × 10(-21), and the combined power of discrimination (CPD) was 99.999999999999999999613 %. No significant difference of allele frequencies was observed between Nantong and other Han populations at all STR loci, as well as Dai, Mongolian, and Tibetan. Significant differences were only observed between Nantong Han and Uyghur at TH01, as well as Nantong Han and Dong at CSF1PO and FGA. Nantong Han showed significant differences between She, Bouyei, and Miao at multiple STR loci.


Robust neuroprotective effects of intranasally delivered iNOS siRNA encapsulated in gelatin nanoparticles in the postischemic brain.
The therapeutic efficacy of intranasal iNOS siRNA delivery was investigated in the postischemic rat brain after encapsulating on in gelatin nanoparticles (GNPs; diameter 188.0 ± 60.9 nm) cross-linked with 0.0667% glutaraldehyde (GA). Intranasally delivered GNPs were found in extracellular and intracellular compartments of many brain regions, including the olfactory bulb, cerebral cortex, and striatum at 1 hour after infusion and continued to be detected for days. Infarct volumes were markedly suppressed (maximal reduction to 42.1 ± 2.6%) at 2 days after 60 minutes of middle cerebral artery occlusion (MCAO) when iNOS siRNA/GNPs were delivered at 6 hours post-MCAO. In addition, this protective effect was manifested by reductions in neurological and behavioral deficits that were sustained for 2 weeks. Therapeutic potency of iNOS siRNA/GNPs was significantly greater and sustained longer than that of bare siRNA and prolonged and efficient iNOS by iNOS siRNA/GNP is responsible for the robust neuroprotective effect.


Optimal lipid targets for the new era of cardiovascular prevention.
Optimal lipid targets (OLT) should be the goal for all individuals treated in the new era of cardiovascular (CV) disease prevention. Evidence supports that average LDL cholesterol (LDL-C) values in Westernized populations are not optimal. Lessons from nature and science support a physiologic LDL-C target of <70 mg/dL. Clinical trial evidence further supports optimal LDL-C targets, although several critical questions remain unanswered. Using a calculated LDL-C may have limitations in clinical practice. Non-HDL-C cholesterol may be a better predictor of outcomes, and should therefore be provided on all laboratory reports. Specific HDL cholesterol (HDL-C) targets are significantly more complicated. Although a low HDL-C predicts a less favorable outcome independent of LDL-C level, an HDL-C level > 50 mg/dL is associated with lower CV risk. Clinical trials on HDL-C have thus far been disappointing. OLT should be the goal for all individuals as an important part of addressing global CV risk.


The effect of air pollution on deaths, disease burden, and life expectancy across China and its provinces, 1990-2017: an analysis for the Global Burden of Disease Study 2017.
Air pollution is an important public health concern in China, with high levels of exposure to both ambient and household air pollution. To inform action at provincial levels in China, we estimated the exposure to air pollution and its effect on deaths, disease burden, and loss of life expectancy across all provinces in China from 1990 to 2017. In all 33 provinces, autonomous regions, municipalities, and special administrative regions in China, we estimated exposure to air pollution, including ambient particulate matter pollution (defined as the annual gridded concentration of PM2·5), household air pollution (defined as the percentage of households using solid cooking fuels and the corresponding exposure to PM2·5), and ozone pollution (defined as average gridded ozone concentrations). We used the methods of the Global Burden of Diseases, Injuries, and Risk Factors Study 2017 to estimate deaths and disability-adjusted life-years (DALYs) attributable to air pollution, and what the life expectancy would have been if air pollution levels had been less than the minimum level causing health loss. The average annual population-weighted PM2·5 exposure in China was 52·7 μg/m3 (95% uncertainty interval [UI] 41·0-62·8) in 2017, which is 9% lower than in 1990 (57·8 μg/m3, 45·0-67·0). We estimated that 1·24 million (95% UI 1·08-1·40) deaths in China were attributable to air pollution in 2017, including 851 660 (712 002-990 271) from ambient PM2·5 pollution, 271 089 (209 882-346 561) from household air pollution from solid fuels, and 178 187 (67 650-286 229) from ambient ozone pollution. The age-standardised DALY rate attributable to air pollution was 1513·1 per 100 000 in China in 2017, and was higher in males (1839·8 per 100 000) than in females (1198·3 per 100 000). The age-standardised death rate attributable to air pollution decreased by 60·6% (55·7-63·7) for China overall between 1990 and 2017, driven by an 85·4% (83·2-87·3) decline in household air pollution and a 12·0% (1·4-22·1) decline in ambient PM2·5 pollution. 40·0% of DALYs for COPD were attributable to air pollution, as were 35·6% of DALYs for lower respiratory infections, 26·1% for diabetes, 25·8% for lung cancer, 19·5% for ischaemic heart disease, and 12·8% for stroke. We estimated that if the air pollution level in China was below the minimum causing health loss, the average life expectancy would have been 1·25 years greater. The DALY rate per 100 000 attributable to air pollution varied across provinces, ranging from 482·3 (371·1-604·1) in Hong Kong to 1725·6 (720·4-2653·1) in Xinjiang for ambient pollution, and from 18·7 (9·1-34·0) in Shanghai to 1804·5 (1339·5-2270·1) in Tibet for household pollution. Although the overall mortality attributable to air pollution decreased in China between 1990 and 2017, 12 provinces showed an increasing trend during the past 27 years. Pollution from ambient PM2·5 and household burning of solid fuels decreased markedly in recent years in China, after extensive efforts to control emissions. However, PM2·5 concentrations still exceed the WHO Air Quality Guideline for the entire population of China, with 81% living in regions exceeding the WHO Interim Target 1, and air pollution remains an important risk factor. Sustainable development policies should be implemented and enforced to reduce the impact of air pollution on long-term economic development and population health. Bill & Melinda Gates Foundation; and China National Key Research and Development Program.


Microstructure of titanium-cement-lithium disilicate interface in CAD-CAM dental implant crowns: a three-dimensional profilometric analysis.
Peri-implantitis is an infection of the implant surface caused by adhesion of bacteria that generate bone resorption and sometimes even consequent implant loss. Both screw-retained and cemented fixed implants are affected. The purpose of this study is to investigate the morphological defects at the cemented interface between titanium abutment and ceramic crown, comparing different adhesive cements used to fill the marginal gap. Twelve computer-aided design-computer-aided manufacturing dental crowns were cemented to titanium abutments using three different resin composite cements. Sealed margins were polished using grommets with descending diamond particle size. Three groups of four crowns each were made according to the cement used, namely RelyX Unicem (3 M ESPE), Panavia F 2.0 (Kuraray), and NX3 (Nexus Kerr). Samples were analyzed using optical inspection, three-dimensional profilometry, and image analysis, including analysis of variance. Although RelyX showed significantly lower root mean square surface roughness (4.4 ± 1.5 μm) than that of NX3 (7.0 ± 2.9 μm), it showed no significant difference with Panavia (3.7 ± 1.5 μm). The marginal gap was significantly wider in Panavia (149 ± 108 μm) as compared with NX3 (71 ± 45 μm) and Relyx (64 ± 34 μm). For all groups, homogeneous heights of both metal-cement and ceramic-cement gaps were observed. Moreover, all samples showed homogeneity of the margins and absence of instrumental bias, thus validating both procedure and materials. When using the chosen polishing method, RelyX Unicem showed both low roughness and marginal width, and thus the smoothest and more continuous abutment-crown interlayer, promising a low probability of occurrence of peri-implantitis.


Continuous monitoring of jugular bulb oxygen saturation in comatose patients--therapeutic implications.
Comatose patients run a high risk of developing cerebral ischaemia which may considerably influence final outcome. It would therefore be extremely useful if one could monitor cerebral blood flow in these patients. Since there is a close correlation between the arteriovenous difference of oxygen and cerebral blood flow, it was a logical step to place a fiberoptic catheter in the jugular bulb for continuous measurement of cerebrovenous oxygen saturation. We have monitored cerebral oxygenation in 54 patients, comatose because of severe head injury, intracerebral haemorrhage or subarachnoid haemorrhage. Normal jugular venous oxygen saturation (SJVO2) ranges between 60 and 90%. A decline to below 50% is considered indicative of cerebral ischaemia. Spontaneous episodes of desaturation (SJVO2 < 50% for at least 15 min) were frequent during the acute phase of these insults. Many of these desaturation episodes could be attributed to hyperventilation, even though considered moderate. Likewise, insufficient cerebral perfusion pressure and severe vasospasm were found to be important causes of desaturation episodes. In many instances, tailoring of ventilation or induced hypervolaemia and hypertension were capable of reversing these low flow states. The new method of continuous cerebrovenous oximetry is expected to contribute to a better outcome by enabling timely detection and treatment of insufficient cerebral perfusion.


Motion analysis of right ventricular dysfunction under mild or moderate pressure overload caused by acute pulmonary embolism.
Acute pulmonary embolism (APE) is the third most common cause of death in the United States. Appearing as a sudden blockage in a major pulmonary artery, APE may cause mild, moderate or severe right ventricular (RV) overload. Although severe RV overload produces diagnostically obvious RV mechanical failure, little progress has been made in gaining a clinical and biophysical understanding of moderate and mild acute RV overload and its impact on RV functionality. In the research described here, we conducted a pilot study in pigs using echocardiography and observed the following abnormalities in RV functionality under acute mild or moderate RV overload: (i) occurrence of paradoxical septal motion with "waving" dynamics; (ii) decrease in local curvature of the septum (p < 0.01); (iii) lower positive correlation between movement of the RV free wall and movement of the septum (p < 0.05); (iv) slower rate of RV fractional area change (p < 0.05); and (v) decrease in movement stability, particularly in the middle of the septum (p < 0.05).


[Clinical, cytogenetic and immunological studies of 2 cases with B-cell acute lymphocytic leukemia. Comparison between B-ALL and Burkitt's lymphoma reported in Japan].
Two patients with acute lymphocytic leukemia of B-cell phenotype (B-ALL) are described. They were 77-year-old female and 34-year-old male. One patient presented with marked splenomegaly, and the other with rupture of spleen on admission. Leukemic morphology revealed a typical L3 profile by FAB classification system in both cases. Immunologic analysis showed the presence of surface immunoglobulins in both cases, and one phenotype was IgM kappa, whereas the other was IgG kappa. Cytogenetic study revealed the typical translocations (8; 14) in both cases. Following chemotherapy, complete remission was achieved in one case, but the other died 36 days after admission. Including our two cases, we studied 12 cases of B-ALL and 77 cases of Burkitt's lymphoma reported in Japan, investigating the clinical prognosis as well as the biological features. We concluded that there are no significant difference of survival between B-ALL and Burkitt's lymphoma. We estimated it is due to oncogenesis from the same original cell in despite of the difference in main tumor site between two diseases.


Somatosensory evoked potentials in healthy people.
The results of a study of somatosensory evoked potentials recorded in 37 healthy subjects of both sexes are described. Comparison of the results of tests on three age subgroups showed selectivity in the change in latencies and amplitudes of waves of the somatosensory responses depending on the subject's age. Ipsilateral responses show greater variability but a lower amplitude and frequency of appearance of the individual components than contralateral responses. Their latent periods also were longer than those of the contralateral responses.


Nanoencapsulation application to prolong postharvest shelf life.
This review offers our opinion on current and future trends regarding nanoencapsulation interventions to extend postharvest shelf life of stored grains, fruits, and vegetables. Herein, we considered two major factors influencing postharvest shelf life for comments: aerobic food spoilage microorganisms and stored pests. Nanoemulsions, edible/active coatings, and nanopackaging loading essential oils as antimicrobial, antioxidant, or pesticide showed promising results in prolonged shelf life at room/cold storage without compromising quality, organoleptic properties, and postharvest physiology. Trends with nanoencapsulation using plant-based pesticides as agrochemical-free methods to keep produce fresh longer were commented as potential candidates for prolonging the shelf life of stored grains and fruits at the postharvest stage. Research with potential large-scale feasibility is intensive, but safety assessment is required and remains little explored.


Specialized Intercellular Communications via Tunnelling Nanotubes in Acute and Chronic Leukemia.
Effectual cell-to-cell communication is essential to the development and differentiation of organisms, the preservation of tissue tasks, and the synchronization of their different physiological actions, but also to the proliferation and metastasis of tumor cells. Tunneling nanotubes (TNTs) are membrane-enclosed tubular connections between cells that carry a multiplicity of cellular loads, such as exosomes, non-coding RNAs, mitochondria, and proteins, and they have been identified as the main participants in healthy and tumoral cell communication. TNTs have been described in numerous tumors in in vitro, ex vivo, and in vivo models favoring the onset and progression of tumors. Tumor cells utilize TNT-like membranous channels to transfer information between themselves or with the tumoral milieu. As a result, tumor cells attain novel capabilities, such as the increased capacity of metastasis, metabolic plasticity, angiogenic aptitude, and chemoresistance, promoting tumor severity. Here, we review the morphological and operational characteristics of TNTs and their influence on hematologic malignancies' progression and resistance to therapies, focusing on acute and chronic myeloid and acute lymphoid leukemia. Finally, we examine the prospects and challenges for TNTs as a therapeutic approach for hematologic diseases by examining the development of efficient and safe drugs targeting TNTs.


Biosynthesis of non-sulfated high-molecular-weight glycosaminoglycans and specific-sized oligosaccharides.
Non-sulfated forms of glycosaminoglycans (NSGAGs) including hyaluronan, chondroitin and heparosan with high-molecular-weight (HMW) are extensively used biomaterials, while NSGAGs oligosaccharides display strong bioactivities. However, microbial production of HMW-NSGAGs and oligosaccharides with specific size are always challenging. Here, a membrane shield strategy was developed to produce HMW-NSGAGs by recruiting type II NSGAG synthases in Corynebacterium glutamicum. By enhancing precursor supplies and reinforcing cell membrane, the MWs of hyaluronan, heparosan and chondroitin reached 4100 kDa, 3000 kDa and 2400 kDa, respectively. In parallel, a synchronized depolymerization-polymerization strategy was developed by co-expressing NSGAGs synthases and lyases. With cell membrane as a filter, we achieved the direct biosynthesis of NSGAGs tetrasaccharides and disaccharides. The titers of chondroitin, hyaluronan and heparosan tetrasaccharides and disaccharides reached 10.9 g L-1, 12.1 g L-1 and 5.8 g L-1, respectively. The strategies developed here should also be applicable to the biosynthesis of other polysaccharides.


Gene regulatory networks mediating canonical Wnt signal-directed control of pluripotency and differentiation in embryo stem cells.
Canonical Wnt signaling supports the pluripotency of embryonic stem cells (ESCs) but also promotes differentiation of early mammalian cell lineages. To explain these paradoxical observations, we explored the gene regulatory networks at play. Canonical Wnt signaling is intertwined with the pluripotency network comprising Nanog, Oct4, and Sox2 in mouse ESCs. In defined media supporting the derivation and propagation of ESCs, Tcf3 and β-catenin interact with Oct4; Tcf3 binds to Sox motif within Oct-Sox composite motifs that are also bound by Oct4-Sox2 complexes. Furthermore, canonical Wnt signaling upregulates the activity of the Pou5f1 distal enhancer via the Sox motif in ESCs. When viewed in the context of published studies on Tcf3 and β-catenin mutants, our findings suggest Tcf3 counters pluripotency by competition with Sox2 at these sites, and Tcf3 inhibition is blocked by β-catenin entry into this complex. Wnt pathway stimulation also triggers β-catenin association at regulatory elements with classic Lef/Tcf motifs associated with differentiation programs. The failure to activate these targets in the presence of a mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) inhibitor essential for ESC culture suggests MEK/ERK signaling and canonical Wnt signaling combine to promote ESC differentiation.


Differential Mueller matrix imaging of partially depolarizing optically anisotropic biological tissues.
Since recently, a number of innovative polarization-based optical imaging modalities have been introduced and extensively used in various biomedical applications, with an ultimate aim to attain the practical tool for the optical biopsy and functional characterization of biological tissues. The techniques utilize polarization properties of light and Mueller matrix mapping of microscopic images of histological sections of biological tissues or polycrystalline films of biological fluids. The main drawback of currently developed laser polarimetry approaches and Mueller matrix mapping techniques is poor reproducibility of experimental data. This is due to azimuthal dependence of polarization and ellipticity values of most matrix elements to sample orientation in respect to incidence light polarization. Current study aims to generalize the methods of laser polarimetry for diagnosis of partially depolarizing optically anisotropic biological tissues. A method of differential Mueller matrix mapping for reconstruction of linear and circular birefringence and dichroism parameter distributions of partially depolarizing layers of biological tissues of different morphological structure is introduced and practically implemented. The coordinate distributions of the value of the first-order differential matrix elements of histological sections of brain tissue with spatially structured, optically anisotropic fibrillar network, as well as of parenchymatous tissue of the rectum wall with an "islet" polycrystalline structure are determined. Within the statistical analysis of polarization reproduced distributions of the averaged parameters of phase and amplitude anisotropy, the significant sensitivity of the statistical moments of the third and fourth orders to changes in the polycrystalline structure of partially depolarizing layers of biological tissue is observed. The differentiation of female reproductive sphere connective tissue is realized with excellent accuracy. The differential Mueller matrix mapping method for reconstruction of distributions of linear and circular birefringence and dichroism parameters of partially depolarizing layers of biological tissues of different morphological structures is proposed and substantiated. Differential diagnostics of changes in the phase (good balanced accuracy) and amplitude (excellent balanced accuracy) of the anisotropy of the partially depolarizing layers of the vagina wall tissue with prolapse of the genitals is realized. The maximum diagnostic efficiency of the first-order differential matrix method was demonstrated in comparison with the traditional methods of polarization and Mueller matrix mapping of histological sections of light-scattering biological tissues.


Relationship between average muscle fibre conduction velocity and EMG power spectra during isometric contraction, recovery and applied ischemia.
The relationship between muscle fibre conduction velocity (MFCV) and the power spectrum of surface EMGs in 3 human volunteers was studied during isometric contractions at 40% maximum voluntary contraction. In addition, the recovery of these two parameters was measured during short lasting contractions at the same force level every 30 s. The recovery phase was also studied during ischaemia, thereby preventing the recovery of MFCV. The mean MFCV was calculated by the cross-correlation method. The measurements were facilitated by a real-time estimation of the cross-correlation and the MFCV and by a graphic display of the digitised signal. During contraction a nearly linear relation was found between MFCV and the median frequency of the power spectrum (MPF). During recovery this relationship was lost in one subject: MPF restored much faster then MFCV. During recovery under ischemia MFCV did not recover, but MPF recovered partially in all subjects. It is concluded that the shift of the power spectrum to lower frequencies during fatigue cannot be explained by changes in MFCV alone. Central mechanisms also influence the power spectrum and studying the recovery of local muscle fatigue during ischemia may separate these influences from that of MFCV on the power spectrum during fatigue.


Assessment of respiratory system compliance by a flow recording method.
Standard methods for the assessment of the compliance of the respiratory system require recordings of both airway pressure and ventilatory volume. A method based on the recording of respiratory flow only, with no need of measuring airway pressure was reported some years ago. The purpose of the present study was to test a modification of this flow-recording technique. The technique is based on the measurement of inspiratory flow with and without the addition of a source of external compliance added to the inspiratory circuit. The difference in inspiratory flow between these two situations can be used to derive the compliance of the respiratory system. The method was and in patients with acute respiratory failure, and compared with a standard method based on airway pressure and inspiratory gas flow recording. Model experiments showed strong correlation between the new flow-recording method and the standard method (r = 0.99, P < 0.001) with a coefficient of variation of less than 2% with both methods. In patients with acute respiratory failure, the flow and standard methods gave similar results when no end-inspiratory pause was applied (mean difference 5 +/- 40 ml/kPa (x +/- s.d.)). The standard method gave significantly higher compliance values, different from the flow method, when an end-inspiratory pause of 10% was applied (mean difference 53 +/- 2 ml/kPa, P < 0.01). The flow method was not influenced by the use of an end-inspiratory pause.(ABSTRACT TRUNCATED AT 250 WORDS)


Contrast-enhanced versus systematic transrectal ultrasound-guided prostate cancer detection: an overview of techniques and a systematic review.
An overview is given of the current conventional and contrast-specific transrectal ultrasound (TRUS) techniques for prostate imaging that may be used for guiding biopsy. Subsequently, a systematic literature search resulted in inclusion of four studies that directly compared systematic and contrast-enhanced-targeted TRUS-guided biopsy of the prostate. All four studies revealed that contrast-enhanced-targeted TRUS-guided biopsy significantly improved the positive biopsy core rate. In the largest study, the mean Gleason score of the contrast-enhanced-targeted TRUS-guided biopsies was significantly higher than that of the systematic biopsies. Future randomized clinical trials, preferably in a non-academic multi-institutional set-up, will have to be performed to confirm the value of contrast-enhanced-targeted TRUS-guided biopsy in regular clinical practice.


Preclinical studies of non-stick thin film metallic glass-coated syringe needles.
Our objective in this study was to determine the biocompatibility and hemocompatibility of thin film metallic glass (TFMG) and its potential use in hypodermic needles for intramuscular or intravenous injection. Mouse and rabbit models were employed under approval from the Institutional Animal Care and Use Committee (n = 5/group, two groups in total for both animal models). Platelet-rich plasma (PRP) was collected from the whole blood of rabbits (ear vein) without anti-coagulant for use in in vitro coagulation tests. Histological analysis and optical microscopy were used to assess the endothelial structure of the inner lining of veins after being punctured with needles and detained for 3 days. Histological analysis of ear vein sections revealed that the extent of endothelial damage after puncturing with a TFMG-coated needle was 33% less than that produced by bare needles. Our results confirm that the deposition of a thin TFMG layer (e.g., Zr53Cu33Al9Ta5) on the surface of hypodermic needle can have remarkably clinical benefits, including anti-adhesion, reduced invasion, and minimal endothelial damage. Our results also confirm the good biocompatibility and hemocompatibility of the TFMG coatings.


Surviving COVID-19 After Hospital Discharge: Symptom, Functional, and Adverse Outcomes of Home Health Recipients.
Little is known about recovery from coronavirus disease 2019 (COVID-19) after hospital discharge. To describe the home health recovery of patients with COVID-19 and risk factors associated with rehospitalization or death. Retrospective observational cohort. New York City. 1409 patients with COVID-19 admitted to home health care (HHC) between 1 April and 15 June 2020 after hospitalization. Covariates and outcomes were obtained from the mandated OASIS (Outcome and Assessment Information Set). Cox proportional hazards models were used to estimate the hazard ratio (HR) of risk factors associated with rehospitalization or death. After an average of 32 days in HHC, 94% of patients were discharged and most achieved statistically significant improvements in symptoms and function. Activity-of-daily-living dependencies decreased from an average of 6 (95% CI, 5.9 to 6.1) to 1.2 (CI, 1.1 to 1.3). Risk for rehospitalization or death was higher for male patients (HR, 1.45 [CI, 1.04 to 2.03]); White patients (HR, 1.74 [CI, 1.22 to 2.47]); and patients with heart failure (HR, 2.12 [CI, 1.41 to 3.19]), diabetes with complications (HR, 1.71 [CI, 1.17 to 2.52]), 2 or more emergency department visits in the past 6 months (HR, 1.78 [CI, 1.21 to 2.62]), pain daily or all the time (HR, 1.46 [CI, 1.05 to 2.05]), cognitive impairment (HR, 1.49 [CI, 1.04 to 2.13]), or functional dependencies (HR, 1.09 [CI, 1.00 to 1.20]). Eleven patients (1%) died, 137 (10%) were rehospitalized, and 23 (2%) remain on service. Care was provided by 1 home health agency. Information on rehospitalization and death after HHC discharge is not available. Symptom burden and functional dependence were common at the time of HHC admission but improved for most patients. Comorbid conditions of heart failure and diabetes, as well as characteristics present at admission, identified patients at greatest risk for an adverse event. No direct funding.


n-polynomial exponential type p-convex function with some related inequalities and their applications.
In this paper, the idea and its algebraic properties of n-polynomial exponential type p-convex function have been investigated. Authors prove new trapezium type inequality for this new class of functions. We also obtain some refinements of the trapezium type inequality for functions whose first derivative in absolute value at certain power are n-polynomial exponential type p-convex. At the end, some new bounds for special means of different positive real numbers are provided as well. These new results yield us some generalizations of the prior results. Our idea and technique may stimulate further research in different areas of pure and applied sciences.


Shh/Ptch and EGF/ErbB cooperatively regulate branching morphogenesis of fetal mouse submandibular glands.
The hedgehog family includes Sonic hedgehog (Shh), Desert hedgehog, and Indian hedgehog, which are well known as a morphogens that play many important roles during development of numerous organs such as the tongue, pancreas, kidney, cartilage, teeth and salivary glands (SMG). In Shh null mice, abnormal development of the salivary gland is seen after embryonic day 14 (E14). Shh also induced lobule formation and lumen formation in acini-like structures in cultured E14 SMG. In this study, we investigated the relationship between Shh and epidermal growth factor (EGF)/ErbB signaling in developing fetal mouse SMG. Administration of Shh to cultured E13 SMG stimulated branching morphogenesis (BrM) and induced synthesis of mRNAs for EGF ligands and receptors of the ErbB family. Shh also stimulated activation of ErbB signaling system such as ERK1/2. AG1478, a specific inhibitor of ErbB receptors, completely suppressed BrM and activation of EGF/ErbB/ERK1/2 cascade in E13 SMGs cultured with Shh. The expressions of mRNA for Egf in mesenchyme and mRNA for Erbb1, Erbb2 and Erbb3 in epithelium of E13 SMG were specifically induced by administration of Shh. These results show that Shh stimulates BrM of fetal mouse SMG, at least in part, through activation of the EGF/ErbB/ERK1/2 signaling system.


Selection criteria among gastroenterologists and surgeons for laparoscopic antireflux surgery.
Symptomatic gastroesophageal reflux disease (GERD) affects a substantial proportion of the American population. The diagnosis and treatment of GERD has advanced tremendously over the past 30 years. However, there remains a lack of understanding about the differences and advantages that laparoscopic antireflux surgery offers and a lack of agreement on the ideal surgical candidate. The purpose of this study was to determine whether a significant difference exists in the practice habits and selection criteria for surgery between gastroenterologists and laparoscopic surgeons. Surveys were sent to 1,000 randomly selected members of the American Gastroenterological Association (AGA) and to 1,000 randomly selected members of the Society of American Gastrointestinal Endoscopic Surgeons (SAGES). As a result, 20% of the AGA surveys and 33% of the SAGES surveys were completed and returned. The AGA group considered patients whose symptoms are not well controlled, those who have complications of disease, and those who require significant lifestyle changes to control their symptoms as the best candidates for surgical evaluation. As a group, gastroenterologists remain somewhat hesitant to refer patients for laparoscopic antireflux surgery. Surgeons considered patients whose symptoms have been well controlled with medical therapy, those who have complications of disease, and those who require significant lifestyle changes to control their symptoms as ideal candidates for fundoplication. A consensus should be reached between surgeons and gastroenterologists in establishing criteria for surgical intervention to manage GERD.


Striatal projections from the lateral and posterior thalamic complexes. An anterograde tracer study in the cat.
Striatal projections from the lateral intermediate (LI) and posterior (Po) thalamic complexes were studied with the anterograde tracers wheat germ agglutinin-horseradish peroxidase and Phaseolus vulgaris leucoagglutinin. Projections to the lateral part of the head and body of the caudate nucleus (CN) and to the putamen (Pu) were found to arise from the ventral parts of the caudal subdivision of the LI besides the well established sources in the intralaminar and ventral thalamic nuclei. No projections to the CN and only a few to the Pu were found to arise from the medial division of the Po. The presence of terminal and intercalated varicosities in the thalamostriatal fibers suggests that they form both terminal and en passant synapses. Thalamostriatal fibers from these thalamic sectors were unevenly distributed within the CN, with patches of either low-density innervation or with no projections at all interspersed within irregular, more densely innervated areas. The former coincided with the acetylcholinesterase-poor striosomes and the latter areas of dense projection with the extrastriosomal matrix.


Controlling Immunity and Inflammation through Integrin-Dependent Regulation of TGF-β.
Integrins promote cell adhesion and migration, but also control local activation of latent transforming growth factor (TGF)-β contained in extracellular matrix or cell-surface reservoirs. Integrin-dependent activation of TGF-β has emerged as a crucial mechanism whereby tissue-borne cells instruct circulating and resident immune cells. Moreover, this regulation has wide pathophysiological implications in wound healing, tissue fibrosis, antibody production, pathogen clearance, inflammation, autoimmunity, cancer, and possibly metabolic disorders including diabetes. Here we review the spatiotemporal control of TGF-β activation by integrins, and its effects on immune cell signaling and function. We discuss the underlying molecular and cell-biological mechanisms, the implications for human health and disease, and possibilities for future therapeutic exploitation.


Neonatal thyroxine treatment: changes in the number of corticotropin-releasing-factor (CRF) and neuropeptide Y (NPY) containing neurons and density of tyrosine hydroxylase positive fibers (TH) in the amygdala correlate with anxiety-related behavior of wistar rats.
Neonatal hyperthyroidism induces persisting alterations in the adult brain, e.g. in spatial learning and hippocampal morphology. In the present study, the relationship between anxiety-related behavior and amygdala morphology was investigated in the adult rat after transient neonatal hyperthyroidism (daily s.c. injections of 7.5 microg L-thyroxine in 0.5 ml 0.9% NaCl solution from postnatal day p1 to p12). The behavioral tests used to study anxiety-related behavior were the motility test, elevated plus-maze and fear-sensitized acoustic startle response. In the amygdala, the number of neurons containing the anxiogenic peptide corticotropin releasing factor (CRF-ir and CRF mRNA) and anxiolytic neuropeptide Y (NPY-ir), the total number of neurons and the density of tyrosine hydroxylase immunoreactive (TH-ir) fibers were quantified. Thyroxine-treated pups presented an accelerated development including opening of eyes and snout elongation as typical signs of hyperthyroidism. Thyroxine-treated adult animals displayed a reduced anxiety in the motility box and elevated plus maze, a reduction in the number of CRF-ir neurons in the central nucleus of the amygdala, as well as an increase in the number of NPY-ir neurons and density of TH-ir fibers in nuclei of the basolateral complex of the amygdala. Moreover, there was a reduction in the total number of neurons in all nuclei of the basolateral complex (despite the higher number of NPY-ir neurons), but not central nucleus of the amygdala. The number of CRF-ir neurons in the central nucleus correlated positively with anxiety-related behavior, and the number of NPY-ir neurons and the density of TH-ir fibers in the basolateral complex correlated inversely with anxiety-related behavior. The findings suggested a shift toward an anxiolytic rather than anxiogenic distribution of peptidergic neurons and fibers in the amygdala at adult age following transient neonatal hyperthyroidism.


Interdisciplinary diagnosis and treatment planning for Class III malocclusion and determining the appropriate anterior tooth positions for individual patients.
Patients presenting with Class III malocclusion often present significant challenges for the orthodontist and restorative clinician. The characteristic anterior crossbite is commonly associated with a maxillo-mandibular skeletal base discrepancy leading to both functional and esthetic issues. Three potential incisal tooth positions are discussed using clinical examples including implants.


Visual Morbidity and Ocular Complications in Presumed Intraocular Tuberculosis: An Analysis of 354 Cases from a Non-Endemic Population.
To describe the visual morbidity associated with intraocular tuberculosis (TB) and to report ocular complications of intraocular TB. Retrospective cohort study of patients diagnosed with intraocular TB at a tertiary referral eye care hospital in central London. Data on baseline and final best-corrected visual acuity (BCVA) along with ocular complications were collected and analyzed. In total, 354 patients with intraocular TB were included in this study. Mean age was 48.45 years (range 19-94) and follow-up was 22.56 ± 7.44 (range 6-30) months post cessation of therapy. Blindness as defined by the World Health Organization was reported in 11 (3.11%) patients at baseline and 13 (3.67%) patients at follow-up. The most common ocular complications were cystoid macular edema (n = 107, 30.5%) and glaucoma (n = 99, 28.12%). Intraocular TB can be associated with significant ocular morbidity. Early recognition, treatment, and regular follow-up of this complex disease may help to reduce visual morbidity and ocular complications.


The ability of stroke volume variation measured by a noninvasive cardiac output monitor to predict fluid responsiveness in mechanically ventilated children.
Continuous noninvasive cardiac output monitoring (NICOM) is a clinically useful tool in the pediatric setting. This study compared the ability of stroke volume variation (SVV) measured by NICOM with that of respiratory variations in the velocity of aortic blood flow (△Vpeak) and central venous pressure (CVP) to predict of fluid responsiveness in mechanically ventilated children after ventricular septal defect repair. The study investigated 26 mechanically ventilated children after the completion of surgery. At 30 min after their arrival in an intensive care unit, a colloid solution of 10 ml/kg was administrated for volume expansion. Hemodynamic variables, including CVP, stroke volume, and △Vpeak in addition to cardiac output and SVV in NICOM were measured before and 10 min after volume expansion. The patients with a stroke volume increase of more than 15 % after volume expansion were defined as responders. The 26 patients in the study consisted of 13 responders and 13 nonresponders. Before volume expansion, △Vpeak and SVV were higher in the responders (both p values <0.001). The areas under the receiver operating characteristic curves of △Vpeak, SVV, and CVP were respectively 0.956 (95 % CI 0.885-1.00), 0.888 (95 % CI 0.764-1.00), and 0.331 (95 % CI 0.123-0.540). This study showed that SVV by NICOM and △Vpeak by echocardiography, but not CVP, reliably predicted fluid responsiveness during mechanical ventilation after ventricular septal defect repair in children.


Antiviral agents.
Potent effective antiviral drugs recently have been licensed for several viral diseases, ushering in a new era in the treatment of viral diseases. Several unique features in the process of a viral infection have been identified as target points for inhibition. The unique steps and the interfering compounds are the subject of this review.


Arabidopsis TANGLED identifies the division plane throughout mitosis and cytokinesis.
In premitotic plant cells, the future division plane is predicted by a cortical ring of microtubules and F-actin called the preprophase band (PPB). The PPB persists throughout prophase, but is disassembled upon nuclear-envelope breakdown as the mitotic spindle forms. Following nuclear division, a cytokinetic phragmoplast forms between the daughter nuclei and expands laterally to attach the new cell wall at the former PPB site. A variety of observations suggest that expanding phragmoplasts are actively guided to the former PPB site, but little is known about how plant cells "remember" this site after PPB disassembly. In premitotic plant cells, Arabidopsis TANGLED fused to YFP (AtTAN::YFP) colocalizes at the future division plane with PPBs. Strikingly, cortical AtTAN::YFP rings persist after PPB disassembly, marking the division plane throughout mitosis and cytokinesis. The AtTAN::YFP ring is relatively broad during preprophase/prophase and mitosis; narrows to become a sharper, more punctate ring during cytokinesis; and then rapidly disassembles upon completion of cytokinesis. The initial recruitment of AtTAN::YFP to the division plane requires microtubules and the kinesins POK1 and POK2, but subsequent maintenance of AtTAN::YFP rings appears to be microtubule independent. Consistent with the localization data, analysis of Arabidopsis tan mutants shows that AtTAN plays a role in guidance of expanding phragmoplasts to the former PPB site. AtTAN is implicated as a component of a cortical guidance cue that remains behind when the PPB is disassembled and directs the expanding phragmoplast to the former PPB site during cytokinesis.


Population fluctuation and damage caused by phytophagous mites on three rubber tree clones.
Our aim was to investigate the population fluctuation and the damage caused by the phytophagous mites Calacarus heveae Feres, Tenuipalpus heveae Baker, and Eutetranychus banksi (McGregor) on clones FX 2784, FX 3864, and MDF 180 in rubber tree crops from southeastern Bahia, Brazil. Moreover, we tested for the influence of climatic variables on occurrence patterns of these species throughout weekly samples performed from October to April. The infestation peaks was between mid-January and late February. The clones FX 2784 and FX 3864 had the highest infestations and more severe damage possibly caused by C. heveae, which was the most frequent and abundant species in all clones. We found that sunlight duration and rainfall were the most important factors for C. heveae while T. heveae was affected by rainfall and temperature. Eutetranychus banksi was only affected by sunlight duration. However, the best models had low goodness of fit. We concluded that the clones FX 2784 and FX 3864 had a higher susceptibility to mite attack, and the association between climatic variables and favorable physiological conditions were determinant for the population increase of the species from January to April.


Synthesis of oligonucleotides using the 2-(levulinyloxymethyl)-5-nitrobenzoyl group for the 5'-position of nucleoside 3'-H-phosphonate and -H-phosphonothioate derivatives.
Synthetic studies on phosphodiester, phosphorothioate, and phosphorodithioate-linked oligonucleotides in terms of 2-(levulinyloxymethyl)-5-nitrobenzoyl (LMNBz) group as the base-labile protecting group for the 5'-hydroxyl groups of nucleoside 3'-H-phosphonate and -H-phosphonothioate derivatives, are described.


Activation of protein kinase C by naturally occurring ether-linked diglycerides.
Recent studies have demonstrated that ether-linked diglycerides are endogenous constituents of biologic tissues and accumulate during agonist stimulation (Daniel, L. W., Waite, M., and Wykle, R. L. (1986) J. Biol. Chem. 261, 9128-9132) and myocardial ischemia (Ford, D. A., and Gross, R. W. (1989) Circ. Res. 64, 173-177). Although protein kinase C previously had been thought to specifically require 1,2-diacyl-sn-glycerol (DAG) molecular species for activation, the present study demonstrates that purified rat brain protein kinase C is activated by naturally occurring ether-linked diglycerides (e.g. 1-O-hexadec-1'-enyl-2-octa-dec-9'-enoyl-sn-glycerol and 1-O-hexadecyl-2-octa-dec-9'-enoyl-sn-glycerol) with a similar dose response curve to that for DAG molecular species. Although in vitro assays demonstrated that DAG could partially activate protein kinase C in the absence of free calcium, activation by ether-linked diglycerides required free calcium concentrations found only in stimulated cells (greater than 1 microM [Ca2+]free). To substantiate these findings the alpha and beta isoforms of protein kinase C from rat brain cortical grey matter were resolved by hydroxylapatite chromatography. Although the beta isoform of protein kinase C was substantially activated by DAG in the absence of free calcium, activation by ether-linked diglycerides had an absolute requirement for physiologic increments in free calcium ion found in stimulated cells. Since ether lipids are localized in specific subcellular membrane compartments, accumulate during several pathophysiologic perturbations and are effective activators of protein kinase C with separate and distinct calcium requirements in comparison to DAG, these results suggest that ether-linked diglycerides are important and potentially specific biologic activators of one or more isoforms of protein kinase C.


Cornu aspersum mucin attenuates indomethacins-induced gastric ulcers in mice via alleviating oxidative stress and inflammation.
In the past three decades, a significant progress has been made in the prevention and treatment of gastric ulcers. The incidence of the disease has decreased, but gastric ulcer is still a medical problem. Currently, the available drugs for gastric ulcer treatment have many side effects; therefore, searching for new and safe therapeutic agents is mandatory. The present study aims to investigate the gastroprotective potential of Cornu aspersum (C. aspersum) mucin against gastric ulcers, and the mechanisms related to oxidative stress and inflammation. C. aspersum mucin was collected from 50 snails. The characteristics of C. aspersum mucin (chemical and microbiological) were evaluated. Mice were pretreated with famotidine and C. aspersum mucin (7.5 and 15 ml/kg b.w.) for 5 days, and then gastric ulcers were induced by indomethacin. Macroscopic examination, biochemical estimations, and Quantitative real-time PCR were carried out. Also, histopathological and immunohistopathological examinations were evaluated. We found that the high dose of the mucin significantly decreased the gastric mucosal malondialdehyde (MDA) and nitric oxide (NO) contents as well as interleukin 1β (IL-1β) and nuclear factor kappa β (NF-ҡB) expression, and inducible nitric oxide synthase (iNOS) immunostaining. It also increased the gastric mucosal GSH and catalase contents as well as hemoxygenase-1 (HO-1) and nuclear factor-erythroid 2-related factor 2 (Nrf2) expressions with regressions in gastric mucosal lesions. In conclusion, C. aspersum mucin could be a potential therapeutic candidate to protect against gastric ulceration.


Detection of differential gene copy number using denaturing high performance liquid chromatography.
Genomic rearrangements leading to deletion or duplication of gene(s) resulting in alterations in gene copy number underlie the molecular lesion in several genetic disorders. Methods currently used to determine gene copy number including real time PCR, southern hybridization, fluorescence in situ hybridization, densitometric scanning of PCR product etc. have certain disadvantages and are also expensive and time consuming. Herein, we describe a simple and rapid method to assess gene copy number using denaturing high performance liquid chromatography (dHPLC). We used X chromosome genes as model to compare the gene copy numbers present on this chromosome in males and females. DNA from these samples were amplified by biplex PCR using primer pairs specific for X chromosome genes only (target gene) and for genes present on both X and Y chromosomes (internal control). Amplified products were analyzed using HPLC under non-denaturing conditions. The ratio of peak areas (target gene/internal control) of the amplified products was approximately twice in female samples than male samples (p < 0.001) demonstrating that the differential gene copy number can be easily detected using this method. This method can potentially be used for diagnostic purpose where the need is to distinguish samples based on the differential gene copy numbers.


Lactate Dehydrogenase A Depletion Alters MyC-CaP Tumor Metabolism, Microenvironment, and CAR T Cell Therapy.
To enhance human prostate-specific membrane antigen (hPSMA)-specific chimeric antigen receptor (CAR) T cell therapy in a hPSMA+ MyC-CaP tumor model, we studied and imaged the effect of lactate dehydrogenase A (LDH-A) depletion on the tumor microenvironment (TME) and tumor progression. Effective LDH-A short hairpin RNA (shRNA) knockdown (KD) was achieved in MyC-CaP:hPSMA+ Renilla luciferase (RLuc)-internal ribosome entry site (IRES)-GFP tumor cells, and changes in tumor cell metabolism and in the TME were monitored. LDH-A downregulation significantly inhibited cell proliferation and subcutaneous tumor growth compared to control cells and tumors. However, total tumor lactate concentration did not differ significantly between LDH-A knockdown and control tumors, reflecting the lower vascularity, blood flow, and clearance of lactate from LDH-A knockdown tumors. Comparing treatment responses of MyC-CaP tumors with LDH-A depletion and/or anti-hPSMA CAR T cells showed that the dominant effect on tumor growth was LDH-A depletion. With anti-hPSMA CAR T cell treatment, tumor growth was significantly slower when combined with tumor LDH-A depletion and compared to control tumor growth (p < 0.0001). The lack of a complete tumor response in our animal model can be explained in part by (1) the lower activity of human CAR T cells against hPSMA-expressing murine tumors in a murine host, and (2) a loss of hPSMA antigen from the tumor cell surface in progressive generations of tumor cells.


Evaluation and management of arrhythmia in the athletic patient.
Athletes may present with palpitations, syncope, or arrest resulting in the diagnosis of arrhythmia, or screening may result in diagnosis of conditions with predisposition to arrhythmia. This chapter focuses on 3 common arrhythmic conditions in athletes-atrial fibrillation, premature ventricular contractions (PVCs), and the athlete with an implanted device. (1) Atrial fibrillation: most studies show that atrial fibrillation is more common in competitive athletes, particularly those participating in long-term endurance sports. Postulated mechanisms include morphologic changes such as atrial dilatation, autonomic changes such as increased vagal tone, or inflammatory changes due to sports participation. Treatment options include long-term antiarrhythmic agents, "pill in the pocket" medications, or radiofrequency ablation, a highly successful procedure in athletes. (2) Premature ventricular contractions: data conflict on whether the incidence of PVCs is increased in highly trained individuals. Very frequent PVCs in athletes, however, can be a manifestation of underlying heart disease, and athletes presenting with PVCs should undergo evaluation. In the absence of underlying heart disease, PVCs do not carry a poor prognosis, and US guidelines do not recommend restriction from sports. (3) Implanted devices: the safety of sports for the athlete with an implanted device is unknown, and current guidelines recommend against participation in vigorous competitive sports, based on postulated risks including failure to defibrillate and risk of injury. Many athletes with defibrillators and pacemakers do participate in sports. Ongoing research will better delineate the risks of sports for the athlete with an implanted device.


Analysis of the claims submitted to the patient insurance (PI) scheme in Sweden during 1975-1979 in obstetrical and gynaecological cases.
The Patient Insurance (PI) scheme in Sweden was instituted on 1 January 1975. From the PI it is possible for a patient to obtain acknowledgement of an injury sustained in connection with medical treatment or operation--and receive financial compensation therefore--without having to prove that the injury was the result of fault or neglect. Special conditions of undertaking are laid down for the guidance of PI assessors which, if fulfilled, will entitle the patient to indemnity from the respective county council (via the PI) for injury sustained in connection with medical care received from county institutions or staff. As the Swedish PI scheme was the first of its kind in the world, an evaluation of its application during the first 5 years seemed justified. The present work constitutes the obstetrical-gynaecological part of that project. The objectives of the study were: to analyse and group according to injury all claims concerning obstetrical or gynaecological intervention submitted to the PI during 1975-79; to evaluate available literature on such injuries in relation to the present findings; to make recommendations for changes in the treatment routines of frequently occurring or serious injuries; and to study the working procedures of the PI, especially regarding its assessment of patient claims. Chapter I sets out the historical background of the PI. The extent of a patient's right to indemnity from the PI is discussed, covering the conditions of undertaking that must be fulfilled on the patient's part, and the PI's obligations. The Patient Injuries Committee is presented, to which injured parties can appeal against decisions of the PI. Chapters II--X. During the period studied, altogether 275 claims concerning obstetrical and gynaecological treatment were submitted to the PI, i.e. 2.5% of all claims received by the PI during that period. Most of the injuries resulted from surgical intervention. The claims have been grouped into six gynaecological and three obstetrical chapters. The reporting frequency to the PI was low during this period and therefore no statistical information can be derived from the analysis. A review of the current literature revealed a similar pattern of severe complications as that found among claims to the PI. In one respect, however, the present study is unique, since complications resulting from certain types of intervention, though performed under so varying circumstances, have never before been compiled and evaluated. The reported injuries were fairly evenly distributed among the 26 counties and the three different sizes of hospital.(ABSTRACT TRUNCATED AT 400 WORDS)


From single to multiple deficit models of developmental disorders.
The emerging etiological model for developmental disorders, like dyslexia, is probabilistic and multifactorial while the prevailing cognitive model has been deterministic and often focused on a single cognitive cause, such as a phonological deficit as the cause of dyslexia. So there is a potential contradiction in our explanatory frameworks for understanding developmental disorders. This paper attempts to resolve this contradiction by presenting a multiple cognitive deficit model of developmental disorders. It describes how this model evolved out of our attempts to understand two comorbidities, those between dyslexia and attention deficit hyperactivity disorder (ADHD) and between dyslexia and speech sound disorder (SSD).


The tigecycline resistance gene tetX has an expensive fitness cost based on increased outer membrane permeability and metabolic burden in Escherichia coli.
Livestock-derived tetX-positive Escherichia coli with tigecycline resistance poses a serious risk to public health. Fitness costs, antibiotic residues, and other tetracycline resistance genes (TRGs) are fundamental in determining the spread of tetX in the environment, but there is a lack of relevant studies. The results of this study showed that both tetO and tetX resulted in reduction in growth and an increased in the metabolic burden of E. coli, but the presence of doxycycline reversed this phenomenon. Moreover, the protection of E. coli growth and metabolism by tetO was superior to that of tetX in the presence of doxycycline, resulting in a much lower competitiveness of tetX-carrying E. coli than tetO-carrying E. coli. The results of RNA-seq showed that the increase in outer membrane proteins (ompC, ompF and ompT) of tetX-carrying E. coli resulted in increased membrane permeability and biofilm formation, which is an important reason for fitness costs. Overall, the increased membrane permeability and metabolic burden of E. coli is the mechanistic basis for the high fitness cost of tetX, and the spread of tetO may limit the spread of tetX. This study provides new insights into the rational use of tetracycline antibiotics to control the spread of tetX.


Electrical signal propagated across acupoints along Foot Taiyang Bladder Meridian in rats.
To investigate the electrical signals propagated along Foot Taiyang Bladder Meridian (BL) in a rat model. The experiments were performed on Dark-Agouti (DA), DA.1U and Sprague Dawley (SD) rats. The antidromic electrical stimulation was applied on the nerve innervating "Pishu" (BL 20) to mimic the acupoint electro-acupuncture (EA). The activities recording from adjacent nerve innervating acupoint "Danshu" (BL 19) or "Weishu" (BL 21) were recorded as indics for acupoint, including the mechanical threshold and discharge rate. After mimic EA on BL 20, C and Aδ units from adjacent BL 19 or BL 21 were sensitized including the decrease in mechanical threshold and increase in discharge rates in DA, DA.1U and SD rats, especially in DA rats. The average discharge rate increased from 2.40±0.26 to 6.06±0.55 and from 1.92±0.42 to 6.17±1.10 impulse/min (P<0.01), and the mechanical threshold decreased from 0.52±0.12 to 0.24±0.05 and from 0.27±0.02 to 0.16±0.01 mmol/L (P<0.01) in C (n=15) and Aδ (n=18) units in DA rats. The net change in discharge rates from C units were 152.5%, 144.7% and 42.4% in DA, DA.1U and SD rats, respectively, among which DA rat's was the highest (P<0.05). In Aδ units, the net change in DA rats were also the highest (221.5%, 139.2% and 49.2% in DA, DA.1U and SD rats). These results showed that mimic acupoint EA activated adjacent acupoints along BL in three rat strains, which might be related to propagated sensation along meridians (PSM). In addition, DA rats were more sensitive and might be a good model animal for PSM research.


Endocrine complications of celiac disease: a case report and review of the literature.
The purpose of this article is to review recent literature regarding endocrine disorders related to celiac disease (CD). We describe a case report and review existing literature on the endocrine manifestations of CD. CD is an autoimmune disorder characterized by intestinal inflammation in response to gluten. CD can cause a wide range of extra-intestinal complications, including endocrine manifestations. Metabolic bone disease including osteoporosis and osteopenia, vitamin D deficiency, secondary hyperparathyroidism and less frequently osteomalacia can be seen. In CD, fracture risk is increased by 30-40%, while risk for hip fracture is approximately doubled. The risk for other endocrine disorders, particularly autoimmune endocrinopathies, is also increased in those with CD compared to the general population. Epidemiologic data indicate the risk for hypothyroidism is 3-4 times higher among those with CD, while risk of type 1 diabetes is greater than double. Risk for primary adrenal insufficiency is a striking 11-fold higher in those with versus without CD, though the absolute risk is low. Fertility is reduced in women with CD before diagnosis by 37% while male fertility in the absence of hypogonadism does not appear to be affected. Other endocrine conditions including hyperthyroidism, ovarian failure, androgen insensitivity, impaired growth and growth hormone deficiency and autoimmune polyendocrine syndromes have also been associated with CD. CD is associated with a wide range of endocrine manifestations.


Active fragments of native streptokinase obtained by partial tryptic degradation.
By partial tryptic digestion of streptokinase, degradation products of the molecular weight range 27 000--42 000 were obtained. Among these, fibrinolysis activating components could be demonstrated.


Capturing the dynamic nature of stress exposure in the Pittsburgh Girls Study.
The science of stress exposure and health in humans has been hampered by differences in operational definitions of exposures and approaches to defining timing, leading to results that lack consistency and specificity. In the present study we aim to empirically derive variability in type, timing and chronicity of stress exposure for Black and White females using prospectively collected data in the Pittsburgh Girls Study (PGS). The PGS is an ongoing 20-year longitudinal, community-based study. In this paper we focused on annual caregiver reports of three domains of stress: subsistence (e.g., resource strain, overcrowding); safety (e.g., community violence, inter-adult aggression), and caregiving (e.g., separation, maternal depression) from early childhood through adolescence. Z-scores were used to conduct a finite mixture model-based latent class trajectory analysis. Model fit was compared using the Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC). We examined differences in timing and chronicity of stress exposure between Black and White girls. Distinct trajectory groups characterized by differential timing and chronicity of stress exposure were observed across all stress domains. Six trajectories characterized subsistence and safety stress, and five characterized caregiving stress. Variability in initial level, chronicity, and magnitude and timing of change was observed within and across domains of stressors. Race differences also varied across the domains: race differences in timing and chronicity were most pronounced for the subsistence and safety domains, whereas Black and White girls had similar levels of exposure to caregiving stress. Substantial variability in timing and chronicity was observed within and across stress domains. Modeling specific domains and dimensions of stress exposure is likely important in testing associations between exposure and health; such specificity may lead to more effective deployment of preventive interventions based on stress exposure.


Antidiabetic activities of aqueous and ethanolic extracts of Piper betle leaves in rats.
Leaves of Piper betle (Piperaceae) possess several bioactivities and are used in traditional medicinal systems. However, its antidiabetic activity has not been scientifically investigated so far. The aim of this study therefore, was to investigate the antidiabetic activity of Piper betle leaves. This was tested in normoglycaemic and strepozotocin (STZ)-induced diabetic rats using oral administration of hot water extract (HWE) and cold ethanolic extract (CEE). In normoglycaemic rats, both HWE and CEE significantly lowered the blood glucose level in a dose-dependent manner. In glucose tolerance test, both extracts markedly reduced the external glucose load. The antidiabetic activity of HWE is comparable to that of CEE. Moreover, HWE failed to inhibit the glucose absorption from the small intestine of rats. Both extracts were found to be non-toxic and well tolerated after following chronic oral administration (no overt signs of toxicity, hepatotoxicity or renotoxicity). However, the weight of the spleen had increased in treated groups possibly indicating lymphoproliferative activity. It is concluded that HWE and CEE of Piper betle leaves possess safe and strong antidiabetic activity.


Heightened condition dependent expression of structural coloration in the faces, but not wings, of male and female flies.
Structurally colored sexual signals are a conspicuous and widespread class of ornament used in mate choice, though the extent to which they encode information on the quality of their bearers is not fully resolved. Theory predicts that signaling traits under strong sexual selection as honest indicators should evolve to be more developmentally integrated and exaggerated than nonsexual traits, thereby leading to heightened condition dependence. Here, we test this prediction through examination of the sexually dimorphic faces and wings of the cursorial fly Lispe cana. Males and females possess structural UV-white and golden faces, respectively, and males present their faces and wings to females during close-range, ground-based courtship displays, thereby creating the opportunity for mutual inspection. Across a field-collected sample of individuals, we found that the appearance of the faces of both sexes scaled positively with individual condition, though along separate axes. Males in better condition expressed brighter faces as modeled according to conspecific flies, whereas condition scaled with facial saturation in females. We found no such relationships for their wing interference pattern nor abdomens, with the latter included as a nonsexual control. Our results suggest that the structurally colored faces, but not the iridescent wings, of male and female L. cana are reliable guides to individual quality and support the broader potential for structural colors as honest signals. They also highlight the potential for mutual mate choice in this system, while arguing for 1 of several alternate signaling roles for wing interferences patterns among the myriad taxa which bear them.


Aspergillus in the Papanicolaou stain: morphology, fluorescence and diagnostic feasibility.
Aspergillus species exhibit a distinct and clear fluorescence in Papanicolaou-stained cytological samples. The Papanicolaou (PAP) stain enhances the autofluorescence of cultured aspergilli and allows better cytological recognition of the fungus by fluorescence microscopy when it is not easily discerned from its surroundings by light microscopy. Morphological properties can be better distinguished and facilitate the differentiation of aspergillus organisms from other filamentous fungi. Neither light nor fluorescence microscopy, the cytological quality nor the presence of phagocytosed hyphae in alveolar macrophages allow distinction between infection and contamination with Aspergillus species. Only the presence of eosinophilic inflammation permits a tentative diagnosis of an Aspergillus infection. In conclusion, PAP fluorescence reduces the need for special stains, is superior to and quicker than other investigative techniques and enhances the sensitivity and specificity of cytological investigation when a rapid and reliable identification of Aspergillus is needed.


Ecological risk-benefit analysis of a wetland development based on risk assessment using "expected loss of biodiversity".
Ecological risk from the development of a wetland is assessed quantitatively by means of a new risk measure, expected loss of biodiversity (ELB). ELB is defined as the weighted sum of the increments in the probabilities of extinction of the species living in the wetland due to its loss. The weighting for a particular species is calculated according to the length of the branch on the phylogenetic tree that will be lost if the species becomes extinct. The length of the branch on the phylogenetic tree is regarded as reflecting the extent of contribution of the species to the taxonomic diversity of the world of living things. The increments in the probabilities of extinction are calculated by a simulation used for making the Red List for vascular plants in Japan. The resulting ELB for the loss of Nakaikemi wetland is 9,200 years. This result is combined with the economic costs for conservation of the wetland to produce a value for the indicator of the "cost per unit of biodiversity saved." Depending on the scenario, the value is 13,000 yen per year-ELB or 110,000 to 420,000 yen per year-ELB (1 US dollar = 110 yen in 1999).


Corrigendum: Enhancing Heat Tolerance of the Little Dogwood Cornus canadensis L. f. with Introduction of a Superoxide Reductase Gene from the Hyperthermophilic Archaeon Pyrococcus furiosus.
[This corrects the article on p. 26 in vol. 7, PMID: 26858741.].


Analogs of a potent maxi-K potassium channel opener with an improved inhibitory profile toward cytochrome P450 isozymes.
Quinolinone 1 is a potent maxi-K potassium channel opener. In an effort to design analogs of 1 with a better inhibitory profile toward the CYP2C9 isozyme, the two acidic sites were chemically modified independently to generate a number of analogs. These analogs were evaluated as maxi-K channel openers in vitro using Xenopus laevis oocytes expressing cloned hSlo maxi-K channels. Compounds 15, 17, and 19 showed potent activity as maxi-K channel openers and were further evaluated for inhibition of the activity of the CYP2C9 isozyme. Compounds 17 and 19 showed diminished inhibitory potency against 2C9 and also against a panel of other more common CYP isozymes.


Multi-strategy assisted chaotic coot-inspired optimization algorithm for medical feature selection: A cervical cancer behavior risk study.
Real-world optimization problems require some advanced metaheuristic algorithms, which functionally sustain a variety of solutions and technically explore the tracking space to find the global optimal solution or optimizer. One such algorithm is the newly developed COOT algorithm that is used to solve complex optimization problems. However, like other swarm intelligence algorithms, the COOT algorithm also faces the issues of low diversity, slow iteration speed, and stagnation in local optimization. In order to ameliorate these deficiencies, an improved population-initialized COOT algorithm named COBHCOOT is developed by integrating chaos map, opposition-based learning strategy and hunting strategy, which are used to accelerate the global convergence speed and boost the exploration efficiency and solution quality of the algorithm. To validate the dominance of the proposed COBHCOOT, it is compared with the original COOT algorithm and the well-known natural heuristic optimization algorithm on the recognized CEC2017 and CEC2019 benchmark suites, respectively. For the 29 CEC2017 problems, COBHCOOT performed the best in 15 (51.72%, 30-Dim), 14 (48.28%, 50-Dim) and 11 (37.93%, 100-Dim) respectively, and for the 10 CEC2019 benchmark functions, COBHCOOT performed the best in 7 of them. Furthermore, the practicability and potential of COBHCOOT are also highlighted by solving two engineering optimization problems and four truss structure optimization problems. Eventually, to examine the validity and performance of COBHCOOT for medical feature selection, eight medical datasets are used as benchmarks to compare with other superior methods in terms of average accuracy and number of features. Particularly, COBHCOOT is applied to the feature selection of cervical cancer behavior risk dataset. The findings testified that COBHCOOT achieves better accuracy with a minimal number of features compared with the comparison methods.


Two cell lineages, myf5 and myf5-independent, participate in mouse skeletal myogenesis.
In skeletal muscle development, the myogenic regulatory factors myf5 and myoD play redundant roles in the specification and maintenance of myoblasts, whereas myf6 has a downstream role in differentiating myocytes and myofibers. It is not clear whether the redundancy between myf5 and myoD is within the same cell lineage or between distinct lineages. Using lineage tracing and conditional cell ablation in mice, we demonstrate the existence of two distinct lineages in myogenesis: a myf5 lineage and a myf5-independent lineage. Ablating the myf5 lineage is compatible with myogenesis sustained by myf5-independent, myoD-expressing myoblasts, whereas ablation of the myf6 lineage leads to an absence of all differentiated myofibers, although early myogenesis appears to be unaffected. We also demonstrate here the existence of a significant myf5 lineage within ribs that has an important role in rib development, suggested by severe rib defects upon ablating the myf5 lineage.


A case report of a rare intramuscular granular cell tumor.
Granular cell tumors (GCTs) were firstly described by Weber in 1854 and 70 years later by Abrikossoff and classified as benign tumors. Originally considered muscle tumors, they have been identified as neural lesions, due to their close association with nerve and to their immunohystochemical characteristics. GCTs are uncommon tumors and they may arise in any part of the body; they have been mainly observed in tongue, chest wall and upper extremities; less frequent sites are larynx, gastrointestinal tract, breast, pituitary stalk and the female anogenital region. Here we report a case of GCT showing an uncommon localization such as the upper third of the right rectus muscle of the abdominal wall. A 45 year-old woman of Caucasian origin presented to the surgeon with a 6-month history of light pain in the upper third of the abdominal wall. Radiological exams (Ultrasonography, Computed Tomography and Contrast magnetic resonance imaging) showed a localized in the right rectus abdominis muscle. After excision, histological and immunohystochemical analysis, with the support of electron microscopy, allowed making diagnosis of granular cell tumor. After fist description by Abrikosoff in 1926 of GCT like mesenchymal tumor of unknown origin, in recent years immunohystochemical techniques definitely demonstrated the histogenetic derivation of GCT from Schwann cells. Granular cell tumors are rare, small, slow-growing, solitary and painless subcutaneous nodules which behave in a benign fashion, but can have a tendency to recur; in rare cases they can metastasize, when they became malignant; there are some clinical and histological criteria to suspect the malignance of this tumor. It is important that clinicians, radiologists and pathologists are aware of the clinical presentation and histopathology of GCT for appropriate management, counselling and follow-up. In our case we had a complete radiological, morphological and immunohystochemical characterization of the lesion and a definitive diagnosis of benignity confirmed by electron microscopy.


Preclinical research strategies for newly approved drugs as reflected in early publication patterns.
The present study aimed to explore early publication patterns (i.e. up to 1 year after obtaining regulatory approval) for newly registered drugs. From the website of the US Food and Drug Administration, all newly approved drugs for 6 calendar years between 1991 and 2011 were identified. Non-clinical original publications for these compounds were retrieved from PubMed and their abstracts analysed for type of study and reported data. This yielded 170 compounds for which 1954 original non-clinical publications were identified, i.e. a median/mean of 5/11.5 publications per compound; however, number of publications per compound varied widely (0-90) and this variation exhibited a non-Gaussian distribution. The earliest non-clinical publication typically was published less than 5 years before regulatory approval and more than 5 years after filing of the primary patent, but some compounds exhibited notable deviations from this pattern. Publications most often reported on efficacy related to the target indication and on potential future indications, with fewer studies addressing mechanisms of action, potency, selectivity, pharmacokinetics and toxic effects. For most compounds, data at the cellular and in vivo level were published, with fewer reports on effects on isolated tissues and even fewer at the molecular level. The preferred species for cellular studies was human, whereas for in vivo studies, it was rats and mice. In 75 % of cases, the lead author of the publication came from an academic institution, and most studies were published in classic pharmacology journals. We conclude that number, timing and scope of early non-clinical publications on newly approved drugs exhibit major variance. Factors potentially associated with such variance are explored in a companion paper.


Analysis of emergency department management of suspected bacterial meningitis.
Previous studies of emergency department management of bacterial meningitis have indicated that there are often long delays before initiation of antibiotics. The purpose of our study was to determine whether these delays were related to specific aspects of patient management. From 1981 through 1988, we retrospectively reviewed the medical records of 122 patients primarily evaluated in the ED and admitted for suspected bacterial meningitis at a university (55) and a community (67) hospital. The median time (interquartile range) from ED registration until initiation of antibiotics (time to antibiotics) was 3.0 hours (1.6 and 4.3 hours, respectively) (total range, 0.5 to 18 hours). The time to antibiotics was not significantly related to the time of ED registration. Ninety percent of the total time to antibiotics occurred after the initial physician encounter. Time to antibiotics was significantly (P less than .00005) longer for patients in whom computed tomography scan and/or laboratory analysis of cerebrospinal fluid preceded initiation of antibiotics compared with patients in whom antibiotic administration was not contingent on the results of these procedures (4.3 [3.2 and 6.0] versus 1.9 [1.2 and 3.4] hours, respectively). Also, time to antibiotics was significantly (P less than .00005) longer for patients in whom antibiotics were initiated on the ward as compared with in the ED (4.5 [3.5 and 6.8] versus 2.2 [1.4 and 3.5] hours, respectively). We conclude that long delays exist in the ED before initiation of antibiotics for cases of suspected bacterial meningitis, and that in general these delays appear to be physician generated and to a great extent potentially avoidable.


Mn2+ Quenching Assay for Store-Operated Calcium Entry.
Store-operated Ca2+ entry (SOCE) pathway plays important roles in many cellular processes, which is largely studied by using fluorescent Ca2+ indicator, Fura-2. Extracellular Mn2+ is able to cross the plasma membrane through SOCE and quenches the fluorescence signals from Fura-2. Thus, the fluorescence quenching rate by Mn2+ composes a convenient assay to monitor the extent of SOCE. This chapter describes an experimental method of Mn2+ quenching assay for both cultured esophageal epithelial and skeletal muscle cells. It also explains how to perform a quantitative analysis of graded SOCE.


Effects of time and temperature during melanging on the volatile profile of dark chocolate.
Chocolate made from small-batch production is known for distinct sensory properties that differentiate its products from large-scale production. Specifically, small-batch processing includes a melanging step, a chocolate refining (a process involving time and temperature to refine texture and flavor) process that occurs in a stone wet-grinder. Chocolatiers understand that melanging is essential to flavor and overall quality, however the influence of melanging on the aroma chemistry of the finished chocolate is anecdotal and largely uncharacterized. Here, we evaluated the effects of time and temperature of melanging on the volatile chemistry of the finished chocolate. Specifically, chocolate aroma was profiled using HS/SPME-GC-MS for three different time and temperature combinations. A total of 88 compounds were annotated by mass spectrometry and included a diverse set of chemical classes such as pyrazines, aldehydes, terpenes, alcohols, esters, and ketones. Analysis of variance (ANOVA), principal component analysis (PCA), and partial least squares analysis (PLS) revealed that the overall aroma profile was influenced by the type of melanging, and time had a greater effect than temperature. Example compounds affected by time include 2-methylpropanal, dimethyl sulfide, and benzaldehyde. Particle size was also measured for each sample. Majority particle size was found to be below 25 microns generally at all time points beyond 8 h. Analysis showed significant p-values for the temperature variable for several compounds, but significant p-values for the time variable were apparent for a greater number of compounds. For compounds which showed dependency on both time and temperature, the p-value for the time variable was much smaller in most cases. Both PCA and OPLS analyses suggested the same trends. These data support that time is a critical factor in determining the aroma chemistry of chocolate and affects a diverse set of known flavor active compounds.


LncRNA XLOC_006390 promotes pancreatic carcinogenesis and glutamate metabolism by stabilizing c-Myc.
The role of lncRNAs in the regulation of glutamate metabolism and metabolic reprogramming of pancreatic cancer (PC) during nutrient deprivation is largely unknown. Our study found that alpha-ketoglutarate (aKG) levels were significantly reduced in the absence of XLOC_006390. We subsequently confirmed that the decrease in aKG was mainly due to the downregulation of glutamate dehydrogenase 1 (GDH1) at the mRNA level. Therefore, we first screened transcription factors targeting the GDH1 gene promoter and confirmed that c-Myc regulates GDH1 transcription. c-Myc binds to the promoter of GDH1 and activates its transcription. Downregulation of GDH1 mRNA levels by XLOC_006390 deletion could be rescued by overexpression of c-Myc. Overexpression of XLOC_006390 promoted the protein stability of c-Myc by blocking its ubiquitination. Clinically, XLOC_006390 was positively correlated with the mRNA level of GDH1, and c-Myc positively regulated GDH1 gene expression, which was tightly associated with PC patient prognosis. The dysregulated lncRNA/c-Myc axis increased glutamate metabolism, promoting PC progression to a higher stage. Therefore, XLOC_006390/c-Myc may be a potential target for PC, and its abnormal activation also indicates the progression of PC.


Patients' perceptions of patient care providers with tattoos and/or body piercings.
This study evaluated patients' perceptions of patient care providers with visible tattoos and/or body piercings. As tattooing and body piercing are increasingly popular, research that informs nursing administrators regarding policies on patient care providers having visible tattoos and body piercings is warranted. A total of 150 hospitalized adult patients compared pictures of male and female patient care providers in uniform with and without tattoos and/or nonearlobe body piercings. Patient care providers with visible tattoos and/or body piercings were not perceived by patients in this study as more caring, confident, reliable, attentive, cooperative, professional, efficient, or approachable than nontattooed or nonpierced providers. Tattooed female providers were perceived as less professional than male providers with similar tattoos. Female providers with piercings were perceived as less confident, professional, efficient, and approachable than nonpierced female providers. Nursing administrators should develop and/or evaluate policies regarding patient care providers with visible tattoos and/or body piercings.


Phase 1 safety trial of a natural product cocktail with antibacterial activity in human volunteers.
New antibiotics are urgently needed to reduce the health burden of antibiotic-resistant bacterial infection. Natural products (NPs) derived from plants and animals are a current focus of research seeking to discover new antibacterial molecules with clinical potential. A cocktail of NPs based on a medieval remedy for eye infection eliminated biofilms of several highly antibiotic-resistant bacterial species in laboratory studies, and had a promising safety profile in vitro and in a mouse model. A necessary prelude to refining this remedy into a defined, synthetic mixture suitable for testing with wound infections is to firstly establish safety when applied to healthy human skin. We aimed to assess skin-related outcomes of the preparation in a sample of healthy volunteers. This prospective, single arm, non-randomised Phase I clinical trial consisted of a single patch test intervention with 48-h follow-up. Volunteers were staff, students and members of the public recruited from the University of Warwick and surrounding locality. Adults aged 18-79 years, with no history of severe immunity-related disease, diabetes, recent infection, or known pregnancy were eligible. A 100 µl application of a filter-sterilised NP mixture, comprising ground garlic, onion, white wine and bovine bile, was applied to skin on the upper arm and covered with a dressing. The primary outcome was skin-related adverse events over 48 h. Digital photographs were captured where bothersome, salve-related events were reported. 109 volunteers, aged 18-77 years, were recruited between June and July 2021. Sample mean age was 37.6 (SD 16.1) years, and 63 (58%) participants were female. Outcome data were obtained for 106/109 (97%); two participants were lost to follow-up and one removed the skin patch after nine hours due to a bothersome garlic odour. Twenty-one (19.8%) participants reported any patch-test related sign or symptom; of these 14 (13.2%) participants reported minor events related to the salve, including itchiness, redness, or garlic odour. No serious events were reported. We found no evidence of serious skin-related adverse events related to the NP preparation.Trial registration: International Standard Randomised Controlled Trial Number (ISRCTN10773579). Date registered: 08/01/2021.


Ex vivo determination of opiate antagonist binding at mu-opioid ([3H]-DAGO) receptors in hypothalamic micropunches from maturing female rats: comparison between SDZ 210-096 and nalmefene.
We have previously shown that the duration of opioid receptor blockade is critical in determining the degree of opioid antagonist effect following peripheral injection of naloxone and naltrexone. In the present work, we have used this ex vivo technique to compare receptor occupancy of a new opiate antagonist, SDZ 210-096 (SDZ), to that of nalmefene (NLM) in maturing female rats. Two doses (SDZ, 5.6 and 50 mg/kg; NLM, 2.5 and 50 mg/kg) were injected subcutaneously into 3 groups of rats (infantile, juvenile and peripubertal). Micropunches from hypothalamic coronal slices (300 microns) were removed at various times post-injection for quantification of mu-opioid receptors with [3H]-DAGO. Acute administration of the lower dose of SDZ inhibited ligand binding almost completely by 3 h but 50% recovery was observed in all age groups by 12 h. In contrast, SDZ 50 mg/kg provided 80-100% antagonism for at least 24 h. Age-related differences in the ability of SDZ to inhibit [3H]-DAGO binding were observed in that hypothalamic mu-opioid receptors were blocked for longer periods in younger rats. Determination of receptor occupation following NLM injection confirmed that it too has prolonged duration of action but a 24 h blockade is not achieved with either dose of this antagonist. Age-related and dose-related changes in receptor occupancy were minimal compared to SDZ. These studies clarify the interaction of these antagonists at hypothalamic mu-opioid receptors and provide information which allows a clearer interpretation of results in experiments involving opioid blockade.


Immunotherapy response-associated Akkermansia: canary in a coal mine?
Gut microbes can shape cancer immunotherapy responses; however, specific taxa have not been tested prospectively as biomarkers. Derosa and colleagues now demonstrate that the presence of Akkermansia muciniphila in baseline stool samples can be associated with improved outcomes and enhanced antitumor immune cell infiltrates in patients with lung cancer treated with immune checkpoint blockade (ICB).