Differences in the metabolism of MPTP in the rodent and primate parallel differences in sensitivity to its neurotoxic effects.
Primates and rodents show marked differences in sensitivity to the neurotoxic effects of MPTP. We and others have previously shown that the toxic effects of MPTP on nigrostriatal cells are dependent on the oxidative metabolism of MPTP to the quaternary species MPP+. We have therefore compared the distribution and metabolism of MPTP in the monkey and several rodent species. Three major differences have been identified: 1) the primate, but not the rodents, showed a persistently high concentration of MPTP metabolites in the caudate nucleus compared to other brain regions; 2) the rodent brains cleared MPTP and its metabolites much more rapidly than did the monkey, and; 3) the predominant metabolite retained by the monkey brain was MPP+, while MPP+ cannot be detected in rodent brains for more than a few hours after injection. The persistence of MPP+ in the primate brain may explain the heightened toxicity of MPTP in this species.


A Kynar piezoelectric film respirometer.
This paper describes a new technique for measuring respiratory gas flows and volumes using polyvinylidene fluoride film.


Apolipoprotein E: from lipid transport to neurobiology.
Apolipoprotein (apo) E has a storied history as a lipid transport protein. The integral association between cholesterol homeostasis and lipoprotein clearance from circulation are intimately related to apoE's function as a ligand for cell-surface receptors of the low-density lipoprotein receptor family. The receptor binding properties of apoE are strongly influenced by isoform specific amino acid differences as well as the lipidation state of the protein. As understanding of apoE as a structural component of circulating plasma lipoproteins has evolved, exciting developments in neurobiology have revitalized interest in apoE. The strong and enduring correlation between the apoE4 isoform and age of onset and increased risk of Alzheimer's disease has catapulted apoE to the forefront of neurobiology. Using genetic tools generated for study of apoE lipoprotein metabolism, transgenic "knock-in" and gene-disrupted mice are now favored models for study of its role in a variety of neurodegenerative diseases. Key structural knowledge of apoE and isoform-specific differences is driving research activity designed to elucidate how a single amino acid change can manifest such profoundly significant pathological consequences. This review describes apoE through a lens of structure-based knowledge that leads to hypotheses that attempt to explain the functions of apoE and isoform-specific effects relating to disease mechanism.


Effects of MCI-154, a novel cardiotonic agent, on mean circulatory filling pressure in anesthetized dogs.
The effects of MCI-154, a novel cardiotonic agent, on mean circulatory filling pressure (an index of total body venous tone), total peripheral resistance and the heart were examined in anesthetized dogs. The bolus injection of MCI-154 (10-100 micrograms/kg i.v.) caused a dose-dependent decrease in mean circulatory filling pressure and resistance to venous return. MCI-154 also decreased the mean blood pressure and total peripheral resistance, and increased cardiac output and heart rate. Right atrial pressure was reduced only by the lowest dose (10 micrograms/kg i.v.) of MCI-154. These hemodynamic effects of MCI-154, except those on mean circulatory pressure and resistance to venous return, reached a maximum with 30 micrograms/kg of the drug. Nitroglycerin (50 micrograms/kg i.v.), a venodilator, decreased mean circulatory filling pressure, resistance to venous return, mean blood pressure and total peripheral resistance, and increased heart rate. However, unlike MCI-154, nitroglycerin did not alter cardiac output and right atrial pressure. These results suggest that the venodilator effect of MCI-154, as well as the positive inotropic and vasodilator effects, could potentially benefit patients with congestive heart failure.


Adhesion formation and prevention after peritoneal injury and repair in the rabbit.
Fifteen rabbits were used to assess peritoneal healing and adhesion formation after suturing and stapling peritoneal edges or excising, cauterizing and abrading areas of peritoneum. Two weeks after peritoneal injury, the amount of adhesions formation was noted. Resection of peritoneal tissue with natural rehealing was preferable to reapproximation of free peritoneal edges with either staples or sutures.


Enhanced CO2 capture efficiency applying amine-based nano magnetite/sulfinol-M nano solvents at high pressures.
Recently, extensive researches were performed on carbon dioxide (CO2) capture using nano solvents. In this study, three different modified magnetic nanoparticles (NPs), including Fe3O4-Proline (Fe3O4-P), Fe3O4-Lysine (Fe3O4-L), and Fe3O4@SiO2-NH2 NPs were produced to alter and enhance the properties of Fe3O4 NPs for CO2 capture in sulfinol-M as a base solvent. CO2 capture experiments were carried out in a high-pressure batch apparatus in which CO2 absorbed into sulfinol/nano-sulfinol. The influences of NP type, NP loading, and gas phase pressure on CO2 capture were studied. Based on the obtained results, Fe3O4-L and Fe3O4@SiO2-NH2 NPs are notably capable on increasing the CO2 capture capacity of sulfinol-M, and they improved CO2 capture up to 6.3% and 13.36% as compared with the base fluid. Moreover, experiments indicated that all nanosolutions have higher CO2 capture efficiency at higher pressures. Eventually, the effect of NPs stability on CO2 capture was investigated and discussed.


A new approach in the kinetics of biological transport. The potential of reversible inhibition studies.
Kinetic equations are derived for reversible inhibition of both active and facilitated transport systems for seven common experimental arrangements. It is shown that the unique features of transport kinetics may be exploited to give new kinds of information. It is also shown that that the familiar rules of enzyme kinetics, though often applied to transport, can be seriously misleading. The analysis leads to the following general conclusions: (1) A competitive mechanism frequently gives rise to non-competitive kinetics, depending on the experimental design, but a non-competitive mechanism never produces competitive kinetics. (2) Inhibition studies on exchange diffusion at equilibrium in non-active systems or in the final steady state in active systems are the only unambiguous kinetic tests to distinguish competitive from non-competitive mechanisms. (3) Substrate analogs that are bound to the carrier and transported are readily distinguished by inhibition kinetics from those not transported, even though both may rapidly enter the cell by another route. (4) Even in non-active systems competitive inhibitors commonly have far different affinities for the substrate sites on the two membranes faces: where sufficient non-polarity allows their penetration into the cell, inhibition kinetics readily establish such sideness in their action. (5) Inhibition kinetics of the mixed competitive and non-competitive type result from moderately asymmetrical binding of inhibitor at the substrate site. (6) Asymmetry is a necessary feature of active transport: hence studies of inhibition kinetics should provide important insights into its mechanism.


M2-polarized macrophages contribute to neovasculogenesis, leading to relapse of oral cancer following radiation.
Despite the fact that radiation is one of the standard therapies in the treatment of patients with oral cancer, tumours can recur even in the early stages of the disease, negatively impacting prognosis and quality of life. We previously found that CD11b(+) bone marrow-derived cells (BMDCs) were recruited into human glioblastoma multiforme (GBM), leading to re-organization of the vasculature and tumour regrowth. However, it is not yet known how these cells contribute to tumour vascularization. In the present study, we investigated the role of infiltrating CD11b(+) myeloid cells in the vascularization and recurrence of oral squamous cell carcinoma (OSCC). In a xenograft mouse model, local irradiation caused vascular damage and hypoxia in the tumour and increased infiltration of CD11b(+) myeloid cells. These infiltrating cells showed characteristics of M2 macrophages (M2Mφs) and are associated with the promotion of vascularization. M2Mφs promoted tumour progression in recurrence after irradiation compared to non-irradiated tumours. In addition, we found that CD11b(+) myeloid cells, as well as CD206(+) M2Mφs, are increased during recurrence after radiotherapy in human OSCC specimens. Our findings may lead to the development of potential clinical biomarkers or treatment targets in irradiated OSCC patients.


Dysfunctional astrocytic regulation of glutamate transmission in a rat model of depression.
Depression is usually associated with alterations in the monoaminergic system. However, new evidences suggest the involvement of the glutamatergic system in the aetiology of depression. Here we explored the glutamatergic system in a rat model of depression (i.e., the flinders sensitive line (FSL)) to reveal the mechanism underlying the emotional and cognitive aspects associated with the disease. We showed a dramatically elevated level of baseline glutamatergic synaptic transmission by whole-cell recordings as well as impairment in long-term potentiation induced by high-frequency stimulation in hippocampal slices from FSL rats compared with Sprague-Dawley rats. At behavioural level, FSL rats displayed recognition memory impairment in the novel object recognition test. Enantioselective chromatography analysis revealed lower levels of D-serine in the hippocampus of FSL rats and both synaptic plasticity and memory impairments were restored by administration of D-serine. We also observed dysfunctional astrocytic glutamate regulation including downregulation of the glia glutamate transporter GLAST as shown by western blot. One possibility is that the dysfunctional astrocytic glutamate reuptake triggers a succession of events, including the reduction of D-serine production as a safety mechanism to avoid NMDA receptor overactivation, which in turn causes the synaptic plasticity and memory impairments observed. These findings open up new brain targets for the development of more potent and efficient antidepressant drugs.


Exercise is the Most Important Medicine for COVID-19.
COVID-19 infection and long COVID affect multiple organ systems, including the respiratory, cardiovascular, renal, digestive, neuroendocrine, musculoskeletal systems, and sensory organs. Exerkines, released during exercise, have a potent crosstalk effect between multiple body systems. This review describes the evidence of how exerkines can mitigate the effects of COVID-19 in each organ system that the virus affects. The evidence presented in the review suggests that exercise should be considered a first-line strategy in the prevention and treatment of COVID-19 infection and long COVID disease.


Electron microscopical demonstration of glucose-6-phosphatase in native cryostat sections fixed with glutaraldehyde through semipermeable membranes.
The cytochemical demonstration of glucose-6-phosphatase (G6Pase) activity in native cryostat sections fixed with glutaraldehyde through semipermeable membranes is superior to conventional methods with regard to exact localization and lack of inactivation and diffusion of the enzyme, together with simultaneous excellent preservation of the tissue fine structure. In rat liver not only hepatocytes but also many bile duct epithelia and endothelia of arterioles and venules show a marked G6Pase activity in the membranes of the endoplasmic reticulum including the nuclear envelope.


HLA-A*02:934, identified in National bone marrow donor registry named after Vasya Perevoshchikov, Russian Federation.
HLA-A*02:934 has one nucleotide change from HLA-A*02:01:01:01 at nucleotide 308.


Cardiac auscultation via simulation: a survey of the approach of UK medical schools.
A decline in clinical skills of medical students and junior doctors is well documented. We aim to determine how the 32 UK medical schools utilise simulated heart sounds to develop medical students' cardiac auscultation skills. Representatives of all 32 UK medical schools were contacted with a survey questionnaire. Data analysis was carried out using SPSS. Continuous variables e.g. teaching group size were described using median and interquartile range (IQR). 27 Medical schools use a form of simulated heart sounds as a teaching method (2 representatives were unsure, 3 did not respond). This teaching is mandatory in 17 schools. Simulation-based teaching tends to be offered during 3rd year of medical training [median = 3rd year, interquartile range (IQR) 2-3]. Seven medical schools offered simulation teaching more than once. The median number of students in each session was 7.5 [IQR = 5.5-9.5]. One medical school reported that they were unsure how best to implement the heart sound simulation into the medical undergraduate curriculum. The results of our survey of all the UK medical schools suggest that heart sound simulation are used mainly as an introduction to heart murmurs rather than a tool for repetitive practice, complementing clinical experience. Most medical schools do not measure the impact of such teaching on clinical examination.


Nosocomial bacterial pneumonia: an overview.
Nosocomial pneumonia accounts for 10% to 20% of all nosocomial infections and represents one of the most serious complications of hospitalization. This review focuses on the etiology, pathogenesis, and prevention of nosocomial pneumonia, with emphasis on infection control procedures to prevent and minimize its occurrence.


Genetic counseling access for parents of newborns who screen positive for cystic fibrosis: Consensus guidelines.
A risk associated with cystic fibrosis newborn screening (CFNBS) is parental misunderstanding of genetic information generated by the over 6600 positive screens reported annually in the United States. CFNBS algorithms incorporating DNA analysis can generate genetic information that requires clinical interpretation and has significance for the newborn, parents, and other relatives. Engagement between CF care centers and trained genetic counseling providers, such as licensed and/or certified genetic counselors (GCs), is variable and limited in providing information to CFNBS positive (CFNBS+) families. Using a modified Delphi process, a workgroup of CFNBS experts developed recommendation statements for engagement of genetic counseling services in CF care centers where CFNBS + diagnostic evaluations are performed. Statements were assessed over three rounds of surveys, one face-to-face meeting, and through public feedback. Seventeen statements achieved >80% consensus (range: 82%-100%). The workgroup affirmed prior CFF policy statements recommending genetic counseling for parents of infants with CFNBS+. The remaining statements addressed infrastructure and logistics of genetic counseling services, including defining appropriate training for genetic counseling providers and counseling content, establishing a path to equal access to genetic counseling providers across CF care centers, and setting a standard for client-centered CFNBS genetic counseling that is respectful of diverse patient needs and autonomy. Implementation of client-centered genetic counseling for CFNBS+ families in CF care centers by providers with expertise in both CF and genetic counseling will require efforts to further define core concepts, enhance the education of providers, and develop opportunities for access via telemedicine.


Characterisation of trocar associated gas leaks during laparoscopic surgery.
During laparoscopy, the abdominal cavity is insufflated with carbon dioxide (CO2) that could become contaminated with viruses and surgical smoke. Medical staff is potentially exposed when this gas leaks into the operating room through the instruments and past trocar valves. No detailed studies currently exist that have quantified these leakage pathways. Therefore, the goal of this study was to quantify the gas leakages through trocars and instruments, during minimally invasive procedures. A model of the surgical environment was created, consisting of a rigid container with an interface for airtight clamping of laparoscopic equipment such as trocars and surgical instruments. The model was insufflated to 15 mm Hg using a pressure generator and a pneumotachograph measured the equipment gas leak. A protocol of several use cases was designed to simulate the motions and forces the surgeon exerts on the trocar during surgery. Twenty-three individual trocars and twenty-six laparoscopic instruments were measured for leakage under the different conditions of the protocol. Trocar leakages varied between 0 L/min and more than 30 L/min, the instruments revealed a range of leakages between 0 L/min and 5.5 L/min. The results showed that leakage performance varied widely between trocars and instruments and that the performance and location of the valves influenced trocar leakage. We propose trocar redesigns to overcome specific causes of gas leaks. Moreover, an international testing standard for CO2 leakage for all new trocars and instruments is needed so surgical teams can avoid this potential health hazard when selecting new equipment.


Change in the location of the equator and recessed muscles in young rabbit eyes.
To study the changes in the location of the equator and the new insertion of extraocular muscle after recession surgery in a rabbit model. An experimental study was performed in ten eyes of five rabbits. Eyes were divided into two groups according to the amount of recession. In right eyes, 5 mm superior recti (SR) muscle recession, approximately 2 mm posterior to the equator, was performed (5 mm recession group), while in the left eyes, 3 mm recession was performed to the location of the equator (3 mm recession group). We measured the distance of the equator from the limbus, as well as the distance between the superior rectus insertion and the equator. The preoperative measurements were compared with the results 2 months after the surgery. The change in location of the superior rectus and the equator was compared between the two eyes. The recessed SR muscle did not show any significant change in location in both groups (p = 0.18 and 0.16 respectively). However, the location of the equator of rabbit eye showed movement of about 1 mm posterior to the initial location with the growth of the eyeballs (p = 0.04 and 0.03, respectively). The location of the equator moved posteriorly at 2 months postoperatively in young rabbit model while the insertion of the recessed SR did not show any significant movement.


Restoration of Dystrophin Expression in Mdx-Derived Muscle Progenitor Cells Using CRISPR/Cas9 System and Homology-Directed Repair Technology.
Duchenne muscular dystrophy (DMD) is a progressive myopathy caused by mutations in genes encoding dystrophin proteins that ultimately lead to depletion of myogenic progenitor cells (MPCs). Several approaches have been used to correctly express the dystrophin gene in induced pluripotent stem cells (iPSCs), including deletion of mutated exon 23 (ΔEx23) by clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated gene 9 (Cas9)-mediated gene editing technology. However, this approach is labor-intensive due to individual colony picking and genotyping to verify allelic modification. Here, we present a protocol to restore the function of the dystrophin gene by using homology-directed repair (HDR)-based CRISPR/Cas9 and inducing myogenic program of reprogrammed iPSCs from Mdx mice by inducible muscle-specific transcription factor MyoD.


Over expression of CD44V8-10 in urinary exfoliated cells as an independent prognostic predictor in patients with urothelial cancer.
CD44 is a widely expressed cell surface adhesion molecule, of which various isoforms arise from alternative RNA splicing mechanisms. Over expression of specific CD44 splice variants, namely CD44v8-10, is evident in various malignant tumors and is considered to be associated with disease progression. In this study, we investigated whether the transcriptional level of CD44v8-10 relative to that of the standard CD44 isoform would predict the extent and prognosis of urothelial cancer. The CD44v8-10- to -standard CD44 ratio was measured in the tissue (40 urothelial cancer specimens and corresponding normal urinary tissue) and spontaneously voided urine samples of 150 patients with urothelial cancer and 50 with benign urological disease by reverse transcriptase-polymerase chain reaction using the set of primers capable of amplifying all CD44 splice variant isoforms. Initially any CD44 variant isoforms were barely detectable in normal urinary tissues, whereas CD44v8-10 was predominantly expressed in most urothelial cancer specimens. Furthermore, the CD44v8-10- to -standard CD44 ratio in urothelial cancer was closely associated with tumor progression. We then compared the ratio in urothelial cancer tissue and urinary exfoliated cells, and noted a linear and significant correlation of these 2 values in the same patients. Therefore, we investigated whether the CD44v8-10- to -standard CD44 ratio in urinary exfoliated cells would predict the prognosis and disease progression. The mean ratio in the urinary exfoliated cells of patients with invasive urothelial cancer was significantly higher than in those with superficial urothelial cancer. Of the patients with superficial bladder cancer disease-free survival rate of those with an elevated versus a normal ratio was significantly lower. Moreover, of the patients with advanced urothelial carcinoma who underwent complete resection disease-free survival of those with an elevated CD44v8-10- to -standard CD44 ratio was significantly lower than that of patients with a normal ratio. These results indicate that CD44v8-10 is strongly expressed in tumor tissue and evident at high levels in urinary exfoliated cells of patients with invasive versus superficial urothelial cancer. An elevated CD44v8-10- to -standard CD44 ratio in urinary exfoliated cells may serve as a novel prognostic predictor and indicator of disease extent in patients with urothelial cancer.


The impact of WALANT anesthesia and office-based settings on patient satisfaction after carpal tunnel release: A patient reported outcome study.
Wide awake local anesthesia with no tourniquet (WALANT) and office-based procedures are used in hand surgery. There are limited literature data on patient satisfaction when comparing methods of anesthesia and location of surgery. We conducted a retrospective single-surgeon study with patient reported satisfaction in three groups. We compared patient impressions of anesthesia type; WALANT vs locoregional anesthesia plus sedation. We also compared satisfaction in three surgery settings; office surgery vs hospital ambulatory minor procedure room vs main operating room. Group 1 office surgery patients had ultrasound guided surgery with WALANT. Group 2 main operating room surgery patients also had ultrasound guided surgery with WALANT. Group 3 main operating room patients had endoscopic surgery with sedation and a tourniquet. Each group had 30 patients with a minimum follow up of 2 months. We measured overall satisfaction, satisfaction with the organization of care, satisfaction with the administration of anesthesia, and satisfaction with the quality of anesthesia. We also collected secondary data on the resolution of the neuropathic symptoms. Procedures performed in an office-based setting showed higher rates of patient satisfaction when compared to the ambulatory day surgery setting. WALANT anesthesia also showed significantly higher rates of patient satisfaction on a numerical analog scale when compared to sedation based on, irrespective of the surgical setting. All patients had resolution of their neuropathic symptoms regardless of the technique performed. We found that carpal tunnel releases performed in an office-based setting produces superior patient satisfaction. WALANT anesthesia also provides improved patient satisfaction when compared to sedation and monitoring techniques, irrespective of the surgical setting and location. Carpal tunnel release with WALANT in an office-based setting is better for patient comfort and satisfaction, with no evidence of lesser clinical outcomes at a short term follow-up. III.


The effect of frying on glycidyl esters content in palm oil.
The changes in palm oil, as affected by frying temperature, and content of the glycidyl esters (GEs) were studied. Potato chips were fried intermittently in palm oil, which was heated for 8 h daily over five consecutive days. Frying was conducted at three frying temperatures: 150, 165 and 180 °C. Thermo-oxidative alterations of the oil were measured by acid and anisidine values, changes in fatty acid composition, total polar components, polar fraction composition and colour components formation. Content of GE was measured by liquid chromatography-mass spectrometry. Results showed that amount of products of hydrolysis, oxidation and polymerization (excluding decrease of degree of unsaturation) increased significantly as a function of frying temperature and time. Between GEs of fatty acids the most abundant were esters of palmitic and oleic acids. With increasing temperature and frying time, the content of GE decreased. The extent of GE decrease was correlated with degree of oil degradation.


Incidence of Stroke in the Aboriginal and Non-Aboriginal Populations of Australia: A Data Linkage Study.
Most estimates of stroke incidence among Aboriginal and Torres Strait Islander (hereinafter Aboriginal) Australians are confined to single regions and include small sample sizes. We aimed to measure and compare stroke incidence in Aboriginal and non-Aboriginal residents across central and western Australia. Whole-population multijurisdictional person-linked data from hospital and death datasets were used to identify stroke admissions and stroke-related deaths (2001-2015) in Western Australia, South Australia, and the Northern Territory. Fatal (including out-of-hospital deaths) and nonfatal incident (first-ever) strokes in patients aged 20-84 years were identified during the 4-year study period (2012-2015), using a 10-year lookback period to exclude people with prior stroke. Incidence rates per 100 000 population/year were estimated for Aboriginal and non-Aboriginal populations, age-standardized to the World Health Organization World Standard population. In a population of 3 223 711 people (3.7% Aboriginal), 11 740 incident (first-ever) strokes (20.6% regional/remote location of residence; 15.6% fatal) were identified from 2012 to 2015, 675 (5.7%) in Aboriginal people (73.6% regional/remote; 17.0% fatal). Median age of Aboriginal cases (54.5 years; 50.1% female) was 16 years younger than non-Aboriginal cases (70.3 years; 44.1% female; P<0.001), with significantly greater prevalence of comorbidities. Age-standardized stroke incidence in Aboriginal people (192/100 000 [95% CI, 177-208]) was 2.9-fold greater than in non-Aboriginal people (66/100 000 [95% CI, 65-68]) aged 20-84 years; fatal incidence was 4.2-fold greater (38/100 000 [95% CI, 31-46] versus 9/100 000 [95% CI, 9-10]). Disparities were particularly apparent at younger ages (20-54 years), where age-standardized stroke incidence was 4.3-fold greater in Aboriginal people (90/100 000 [95% CI, 81-100]) than non-Aboriginal people (21/100 000 [95% CI, 20-22]). Stroke occurred more commonly, and at younger ages, in Aboriginal than non-Aboriginal populations. Greater prevalence of baseline comorbidities was present in the younger Aboriginal population. Improved primary prevention is required. To optimize stroke prevention, interventions should include culturally appropriate community-based health promotion and integrated support for nonmetropolitan health services.


[Preparation of strong cation exchange packings based on monodisperse hydrophilic non-porous resins and their application for fast separation of proteins].
Monodisperse, 3.0 microm non-porous hydrophilic poly (glycidylmethacrylate-co-ethylenedimethacrylate) particles were prepared by an one-step swelling and polymerization method. The particles were modified to be a strong cation exchange (SCX) stationary phase for high performance liquid chromatography (HPLC) in the following steps. First, the particles were completely hydrolyzed. Second, the hydrolyzed particles were treated with epichlorhydrin followed by another hydrolysis of the newly introduced epoxide groups. Third, the particles were reacted with chlorosulfonic acid. The SCX stationary phase was evaluated in light of the ion exchange property, separability and hydrophilicity on the separation and retention of proteins in detail. Four proteins were quickly separated in 1.0 min with linear gradient elution using the synthesized SCX stationary phase. It was found that it followed ion exchange chromatographic (IEC) retention mechanism. The SCX resin was used for the fast purification of lysozyme from egg white and cytochrome-C from pig heart in 3.0 min with only one step. The results obtained were satisfactory.


Effects of microcurrent stimulation on hyaline cartilage repair in immature male rats (Rattus norvegicus).
In this study, we investigate the effects of microcurrent stimulation on the repair process of xiphoid cartilage in 45-days-old rats. Twenty male rats were divided into a control group and a treated group. A 3-mm defect was then created with a punch in anesthetized animals. In the treated group, animals were submitted to daily applications of a biphasic square pulse microgalvanic continuous electrical current during 5 min. In each application, it was used a frequency of 0.3 Hz and intensity of 20 μA. The animals were sacrificed at 7, 21 and 35 days after injury for structural analysis. Basophilia increased gradually in control animals during the experimental period. In treated animals, newly formed cartilage was observed on days 21 and 35. No statistically significant differences in birefringent collagen fibers were seen between groups at any of the time points. Treated animals presented a statistically larger number of chondroblasts. Calcification points were observed in treated animals on day 35. Ultrastructural analysis revealed differences in cell and matrix characteristics between the two groups. Chondrocyte-like cells were seen in control animals only after 35 days, whereas they were present in treated animals as early as by day 21. The number of cuprolinic blue-stained proteoglycans was statistically higher in treated animals on days 21 and 35. We conclude that microcurrent stimulation accelerates the cartilage repair in non-articular site from prepuberal animals.


Bone ingrowth into polymer coated porous synthetic coralline hydroxyapatite.
Porous hydroxyapatite, converted hydrothermally from the calcium carbon exoskeleton of the coral genus Goniopora (CHAG), has been shown to be effective as a scaffold for bone ingrowth (2,3,5-7,9). However, the large pores in the material resulted in low compressive strengths. In a previous study, we found that microcoating the internal surfaces of CHAG with dilactic-polyactic acid (DL-PLA) improved significantly its compressive properties. The objective of this study was to determine the effect of PLA microcoating on bone ingrowth into CHAG plugs. Plugs of thick- (3:1 chloroform to DL-PLA by weight), medium- (10:1), and thin- (30:1) coated as well as uncoated CHAG were implanted transcortically in the proximal third of the diaphysis of the rabbit tibia. Specimens were harvested at 3, 12, and 24 weeks for mechanical testing and contralaterally for histological and histomorphometric assessment. At 12 weeks, uncoated CHAG plugs developed an average ultimate interface shear stress of 26.7 MPa, compared with 17 MPa for 30:1 and 8 MPa for 10:1 and 3:1 coated specimens. At 24 weeks, there were no significant differences in shear stress among any of the specimens. Histomorphometric assessments showed that the ratio of area fractions of new bone to area fractions of new bone and void space increased from 68-70% for 3:1 and 10:1 coated specimens at 3 weeks, and to 85.5-89.5% at 24 weeks. In comparison, uncoated and 30:1 specimens had area fraction ratios of about 82% at 3 weeks and 93% at 24 weeks. Histologic sections demonstrated direct apposition of new bone to both the coating and the hydroxyapatite as well as degradation of the coating.


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Combinations of ivermectin with proteasome inhibitors induce synergistic lethality in multiple myeloma.
Multiple myeloma (MM) is an incurable malignancy of plasma cells. Ivermectin is a US Food and Drug Administration-approved drug for antiparasitic use. Here, we showed that ivermectin exerted anti-MM effects and significantly synergized with proteasome inhibitors in vitro and in vivo. Ivermectin alone exhibited mild anti-MM activity in vitro. Further investigation suggested that ivermectin inhibited proteasome activity in the nucleus by repressing the nuclear import of proteasome subunits, such as PSMB5-7 and PSMA3-4. Therefore, ivermectin treatment caused the accumulation of ubiquitylated proteins and the activation of the UPR pathway in MM cells. Furthermore, ivermectin treatment caused DNA damage and DNA damage response (DDR) signaling pathway activation in MM cells. Ivermectin and bortezomib exhibited synergized anti-MM activity in vitro. The dual-drug treatment resulted in synergistic inhibition of proteasome activity and increased DNA damage. An in vivo study using a human MM cell line xenograft mouse model showed that ivermectin and bortezomib efficiently repressed MM tumor growth in vivo, while the dual-drug treatment was well tolerated by experimental animals. Overall, our results demonstrated that ivermectin alone or cotreated with bortezomib might be promising in MM treatment.


What proportion of people with hip and knee osteoarthritis meet physical activity guidelines? A systematic review and meta-analysis.
To determine the proportion of people with hip and knee osteoarthritis that meet physical activity guidelines recommended for adults and older adults. Systematic review with meta-analysis of studies measuring physical activity of participants with hip and knee osteoarthritis using an activity monitor. Physical activity levels were calculated using the mean average [95% confidence interval (CI)] weighted according to sample size. Meta-analyses determined the proportion of people meeting physical activity guidelines and recommendations of (1) ≥150 min per week of moderate to vigorous physical activity (MVPA) in bouts of ≥10 min; (2) ≥150 min per week of MVPA in absence of bouts; (3) ≥10,000 steps per day and ≥7000 steps per day. The Grades of Research, Assessment, Development and Evaluation (GRADE) approach was used to determine the quality of the evidence. For knee osteoarthritis, 21 studies involving 3266 participants averaged 50 min per week (95% CI = 46, 55) of MVPA when measured in bouts of ≥10 min, 131 min per week (95% CI = 125, 137) of MVPA, and 7753 daily steps (95% CI = 7582, 7924). Proportion meta-analyses provided high quality evidence that 13% (95% CI = 7, 20) completed ≥150 min per week of MVPA in bouts of ≥10 min, low quality evidence that 41% (95% CI = 23, 61) completed ≥150 min per week of MVPA in absence of bouts, moderate quality evidence that 19% (95% CI = 8, 33) completed ≥10,000 steps per day, and low quality evidence that 48% (95% CI = 31, 65) completed ≥7000 steps per day. For hip osteoarthritis, 11 studies involving 325 participants averaged 160 min per week (95% CI = 114, 216) of MVPA when measured in bouts of ≥10 min, 189 min per week (95% CI = 166, 212) of MVPA, and 8174 daily steps (95% CI = 7670, 8678). Proportion meta-analyses provided low quality evidence that 58% (95% CI = 18, 92) completed ≥150 min per week of MVPA in absence of bouts, low quality evidence that 30% (95% CI = 13, 50) completed ≥10,000 steps per day, and low quality evidence that 60% (95% CI = 47, 73) completed ≥7000 steps per day. A small to moderate proportion of people with knee and hip osteoarthritis met physical activity guidelines and recommended daily steps. Future research should establish the effects of increasing physical activity in this population to meet the current physical activity guidelines.


Adrenal cytochrome P-45011 beta-proteoliposomes catalyzing aldosterone synthesis: preparation and characterization.
Purified cytochrome P-45011 beta from bovine adrenocortical mitochondria was successfully incorporated into the liposome membranes composed of phosphatidylcholine, phosphatidylethanolamine and cardiolipin at a molar ratio of 2:2:1. The incorporation of P-45011 beta into the liposome membranes was ascertained by the Ficoll density gradient centrifugation and the protein refractoriness to trypsin digestion. The prepared proteoliposomes containing P-45011 beta and phospholipid at a molar ratio of 1:3000 were unilamellar vesicles of about 40 nm in average diameter. The P-45011 beta embedded in the liposome membranes was found to be more stable than the detergent-solubilized form. The reconstituted system containing the P-45011 beta-proteoliposomes, adrenodoxin and NADPH-adrenodoxin reductase showed catalytic activities not only for the hydroxylation of 11-deoxycorticosterone at 11 beta- and 18-positions but also for its conversion into aldosterone with a turnover number of 2.3 nmol/min per nmol of P-45011 beta. A successive reaction without the intermediates leaving from the enzyme was suggested for the P-45011 beta-mediated conversion of 11-deoxycorticosterone to aldosterone following the result that the formation of aldosterone was linear with respect to time without the lag phase; this was confirmed by the result that radioactivity in aldosterone from 3H-labeled 11-deoxycorticosterone was scarcely decreased by the addition of unlabeled intermediates to the reactions system.


Indications for superselective intra-arterial cisplatin infusion and concomitant radiotherapy in cases of hypopharyngeal cancer.
We retrospectively assessed the indications for superselective intra-arterial infusion of cisplatin with concomitant radiotherapy (RADPLAT) in patients with hypopharyngeal cancer (HPC). Between April 2000 and March 2013, 41 previously untreated patients received superselective intra-arterial infusion of cisplatin (100-120mg/m(2) per week) with simultaneous intravenous infusions of thiosulfate to neutralize cisplatin toxicity and conventional radiotherapy (65-70Gy). During the median follow-up period of 5.5 years, a statistically significant difference in the 5-year overall survival was noted between patients with N0-1 (n=14) and N2b-3 disease (n=27). One-half of deaths were observed to be the result of distant metastasis. The 5-year local control and overall survival were significantly better in patients with unilateral than in those with bilateral primary tumors. All the patients with T4b disease (n=3) died of disease within 2 years. Indications for RADPLAT in patients with HPC were defined as patients with unilateral tumors staged as T3-4a and N0-1.


Activity of monkey primary somatosensory cortical neurons changes prior to active movement.
Changes in the discharge rates of monkey primary somatosensory cortical neurons were recorded during the performance of wrist flexion and extension. Neurons with activity changes that occurred early before movement onset were often found in areas 3a, 1 and 2, but rarely in area 3b. Based on timing considerations, these observations suggest that somatosensory cortical neurons receive central as well as peripheral inputs that modulate their activity and that may be related to changes in tactile threshold before movement.


Differential coupling between muscarinic receptors and G-proteins in regions of the rat brain.
The coupling of muscarinic receptors to G-proteins in various regions of the rat brain was assessed by measuring carbachol-stimulated, low-Km GTPase. The inhibition of carbachol-stimulated GTPase by the M1-selective antagonist pirenzepine was compared to the affinity of pirenzepine for various nuclei within the regions as measured autoradiographically. The rank order of potency of carbachol for stimulating GTPase in various brain regions was similar to that for binding to receptors in those areas. The maximal specific activity (efficacy) of carbachol-stimulated GTPase varied independently of the distribution of total receptors or receptor subtypes. The overall potency of pirenzepine for inhibiting carbachol-stimulated GTPase was not correlated with the overall affinity of pirenzepine for muscarinic receptors in the regions. Comparing results in various brain regions, the data suggest that there are differences in the efficiency of coupling between muscarinic receptors and G-proteins. For example, the pons-medulla appeared to have a small population of pirenzepine-sensitive (M1 or M4) receptors that were coupled very efficiently to G-proteins, whereas in the hippocampus all muscarinic receptors, most of which are pirenzepine-sensitive, appeared to be weakly coupled to G-proteins. It is suggested that variable interactions between receptors and G-proteins may be an important factor in the overall coupling between receptor occupancy and cellular responses to acetylcholine as well as other hormones and transmitters.


Inosine stimulates chemotaxis, Ca2+-transients and actin polymerization in immature human dendritic cells via a pertussis toxin-sensitive mechanism independent of adenosine receptors.
Inosine is an endogenous purine nucleoside, which is formed by adenosine deaminidase during adenosine breakdown and is released into the extracellular space from the sympathetic nervous system or injured cells. Here, we studied the biological activity of inosine on human dendritic cells (DC), which are specialized antigen presenting cells characterized by their ability to migrate from the blood to peripheral tissues, and then to secondary lymphoid organs where they initiate adaptive immune responses. In immature DC, inosine concentration-dependently stimulated Ca(2+)-transients, actin polymerization, and chemotaxis. Experiments with adenosine receptor antagonists and pertussis toxin (PTX) as well as desensitization studies suggested that the activity of inosine was mediated by a G protein-coupled receptor pathway independent of adenosine receptors. DC, induced to mature by lipopolysaccharide, lost their ability to respond towards inosine with these activities. Moreover, inosine did neither influence membrane expression of CD54, CD80, CD83, CD86, HLA-DR, and MHC class I molecules nor modulated secretion of interleukin (IL)-12, IL-10, and tumor necrosis factor alpha in immature and lipopolysaccharide-matured DC. In aggregate, our study indicates that inosine may be involved in the trafficking control system of immature DC, and mediates its chemotactic activity by a PTX-sensitive mechanism independent of adenosine receptors.


Human skin volatiles: a review.
Odors emitted by human skin are of great interest to biologists in many fields; applications range from forensic studies to diagnostic tools, the design of perfumes and deodorants, and the ecology of blood-sucking insect vectors of human disease. Numerous studies have investigated the chemical composition of skin odors, and various sampling methods have been used for this purpose. The literature shows that the chemical profile of skin volatiles varies greatly among studies, and the use of different sampling procedures is probably responsible for some of these variations. To our knowledge, this is the first review focused on human skin volatile compounds. We detail the different sampling techniques, each with its own set of advantages and disadvantages, which have been used for the collection of skin odors from different parts of the human body. We present the main skin volatile compounds found in these studies, with particular emphasis on the most frequently studied body regions, axillae, hands, and feet. We propose future directions for promising experimental studies on odors from human skin, particularly in relation to the chemical ecology of blood-sucking insects.


Creatine kinase B-subunit activity in human serum. II. Evaluation of s-ck b-subunit activity in the diagnosis of acute myocardial infarction.
In a group of 113 consecutive patients taken into a coronary care unit on suspicion of acute myocardial infarction, blood samples were taken every 6 h and the following enzyme activities were measured: creatine kinase (S-CK), aspartate aminotransferase (S-ASAT), alanine aminotransferase (S-ALAT) and lactate dehydrogenase (S-LD). All measurements were made according to the Recommendations of the Scandinavian Committee on Enzymes. On all patients S-CK B subunit activity was determined by immunoinhibition with a specific anti CK M-subunit inhibitory antibody. At peak values of the respective total enzyme activities CK and LD isoenzymes were further qualitatively estimated by electrophoresis. The data indicate that even serial determinations of total CK, ASAT, ALAT and LD activities in serum do not provide the information required for a conclusive diagnosis of myocardial infarction in the individual case. In contrast, the positive predictive value (PV) of S-CK B was found to be 1.0 and the negative predictive value was 0.98. S-CK MB showed a PV pos. of 1.0 and also a PV neg. of 1.0. Electrophoretic determination of S-LD isoenzymes was slightly poorer with a PV pos. of 0.96 and PV neg. of 0.98. S-CK, total activity with nearly 9 per cent false positives had a positive predictive value of only 0.91, but a negative one of 1.0.


Performance and costs of multiple screening strategies for type 2 diabetes: two population-based studies in Shanghai, China.
To compare the performance and the costs of various assumed screening strategies for type 2 diabetes mellitus (T2DM) among Chinese adults, and identify an optimal one for the population. Two multistage-sampling surveys were conducted in Shanghai, China, in 2009 and 2017. All participants were interviewed, had anthropometry, measured fasting plasma glucose (FPG), hemoglobin A1c (A1c) and/or postprandial glucose. The 1999 WHO diagnostic criteria was used to identify undiagnosed T2DM. A previously developed Chinese risk assessment system and a specific risk assessment system developed in this study were applied to calculate diabetes risk score (DRS) 1 and 2. Optimal screening strategies were selected based on the sensitivity, Youden index and the costs using the 2009 survey data as the training set and the 2017 survey data as the validation set. A twofold cross-validation was also performed. Of numerous assumed strategies, FPG ≥5.6 mmol/L alone performed well (Youden index of 71.8%) and cost least (US$18.4 for each case detected), followed by the strategy of DRS2 ≥8 combining with FPG ≥5.6 mmol/L (Youden index of 71.7% and US$20.2 per case detected) and the strategy of DRS1 ≥17 combining with FPG ≥5.6 mmol/L (Youden index of 72.0% and US$21.6 per case detected). However, FPG alone resulted in more subjects requiring oral glucose tolerance test (OGTT) than did combining with DRS. The strategy of FPG ≥5.6 mmol/L combining with A1c ≥4.7% achieved a Youden index of 72.1%, but had a cost as high as US$48.8 for each case identified. Twofold cross-validation also supported the use of FPG alone, but with an optimal cut-off of 6.1 mmol/L. Our results support the use of FPG alone in T2DM screening in Chinese adults. DRS may be used combining with FPG in populations with available electronic health records to reduce the number of OGTT and save costs of screening.


Molecular and pathological studies on natural cases of bovine theileriosis.
The present communication is a part of study conducted on 32 cases of bovine lymphadenopathies. Out of which, six cases of bovine theileriosis were diagnosed on the basis of peripheral blood smear examination and gross lesions in lymph nodes and visceral organs. Nested PCR using two primer sets N516/N517 and 14136/294 was conducted on samples collected from affected lymph nodes confirms Theileria annulata infection in five out of six cases. Sequencing analysis of amplified product showed 92, 94 and 93 % homology of isolate TH1_Bovine, TH2_Bovine and THEN_Bovine for T. annulata with T. annulata Tamilnadu and Pant Nagar. An upregulation of Th2 cytokines in the cases of theileriosis was observed as the level of TNF-α in individual animals varies from higher value (1028 pg/100 μg protein) to as low as 500 pg/100 μg protein. An increase in level of Th2 cytokines IL-4 and IL-10 was also observed. The present study concluded that pathological studies and cytokine analysis of lymph nodes are of paramount importance in disease diagnosis and associated immune response of the animal with lymphadenopathies.


A histological examination of the uterovaginal sperm storage glands in the domestic hen following an insemination with variable semen dosages.
The uterovaginal sperm storage glands of the domestic hen were examined histologically and scored for filling rates 24 hr after an insemination. Two semen dosages (.023 and .047 ml) were employed and a comparison was made between the large and small semen dose. No significant differences were found between the two groups, suggesting that only a specific number of spermatozoa are capable of entering the uterovaginal sperm storage glands during a given time interval.


Tilorone-induced lysosomal storage mimicking the features of mucopolysaccharidosis and of lipidosis in rat liver.
This ultrastructural and histochemical study deals with the lysosomal storage phenomena occurring in the rat liver after repeated oral administration of tilorone, an agent with anti-tumor and anti-viral activities. In the sinusoidal endothelium and in Kupffer cells, the lysosomes were changed into large vacuoles which contained material with the histochemical characteristics of acid glycosaminoglycans. The alterations closely resembled those previously observed in the splenic red pulp of tilorone-treated rats. In hepatocytes, the lysosomes were converted into large multilamellated inclusions indicating storage of polar lipids. The results show that, in the rat liver, tilorone induces cellular alterations mimicking those of inherited mucopolysaccharidoses and lipidoses. After discontinuing drug treatment the two storage phenomena gradually faded at different rates: The lipidosis disappeared within 2 to 4 weeks, whilst mucopolysaccharidosis-like changes were still found 15 weeks after drug withdrawal. The occurrence of lipidosis is not surprising, since by its molecular structure tilorone can be regarded as belonging to the group of amphiphilic cationic drugs which often have this side effect. Much more surprising is the occurrence of mucopolysaccharidosis-like alterations. The exact biochemical identification of the polyanionic storage material and the molecular mechanisms responsible for this drug side effect remain to be established.


Segregated regulatory CD39+CD4+ T cell function: TGF-β-producing Foxp3- and IL-10-producing Foxp3+ cells are interdependent for protection against collagen-induced arthritis.
Oral immunization with a Salmonella vaccine vector expressing enterotoxigenic Escherichia coli colonization factor Ag I (CFA/I) can protect against collagen-induced arthritis (CIA) by dampening IL-17 and IFN-γ via enhanced IL-4, IL-10, and TGF-β. To identify the responsible regulatory CD4(+) T cells making the host refractory to CIA, Salmonella-CFA/I induced CD39(+)CD4(+) T cells with enhanced apyrase activity relative to Salmonella vector-immunized mice. Adoptive transfer of vaccine-induced CD39(+)CD4(+) T cells into CIA mice conferred complete protection, whereas CD39(-)CD4(+) T cells did not. Subsequent analysis of vaccinated Foxp3-GFP mice revealed the CD39(+) T cells were composed of Foxp3-GFP(-) and Foxp3-GFP(+) subpopulations. Although each adoptively transferred Salmonella-CFA/I-induced Foxp3(-) and Foxp3(+)CD39(+)CD4(+) T cells could protect against CIA, each subset was not as efficacious as total CD39(+)CD4(+) T cells, suggesting their interdependence for optimal protection. Cytokine analysis revealed Foxp3(-) CD39(+)CD4(+) T cells produced TGF-β, and Foxp3(+)CD39(+)CD4(+) T cells produced IL-10, showing a segregation of function. Moreover, donor Foxp3-GFP(-) CD4(+) T cells converted to Foxp3-GFP(+) CD39(+)CD4(+) T cells in the recipients, showing plasticity of these regulatory T cells. TGF-β was found to be essential for protection because in vivo TGF-β neutralization reversed activation of CREB and reduced the development of CD39(+)CD4(+) T cells. Thus, CD39 apyrase-expressing CD4(+) T cells stimulated by Salmonella-CFA/I are composed of TGF-β-producing Foxp3(-) CD39(+)CD4(+) T cells and support the stimulation of IL-10-producing Foxp3(+) CD39(+)CD4(+) T cells.


Bioactive pyrrole-based compounds with target selectivity.
The discovery of novel synthetic compounds with drug-like properties is an ongoing challenge in medicinal chemistry. Natural products have inspired the synthesis of compounds for pharmaceutical application, most of which are based on N-heterocyclic motifs. Among these, the pyrrole ring is one of the most explored heterocycles in drug discovery programs for several therapeutic areas, confirmed by the high number of pyrrole-based drugs reaching the market. In the present review, we focused on pyrrole and its hetero-fused derivatives with anticancer, antimicrobial, and antiviral activities, reported in the literature between 2015 and 2019, for which a specific target was identified, being responsible for their biological activity. It emerges that the powerful pharmaceutical and pharmacological features provided by the pyrrole nucleus as pharmacophore unit of many drugs are still recognized by medicinal chemists.


Impairment of thermoregulatory cooling behavior by single cortical spreading depression in the rat.
Effects of single waves of unilateral cortical spreading depression (CSD) on skin-cooling operant behavior were studied in the rat with and without unilateral lesion in the preoptic and anterior hypothalamus (PO/AH). The skin-cooling behavior was severely suppressed for 6 min, with a resulting rise in rectal temperature (Tre), when CSD entered the frontal cortex contralateral to the PO/AH lesion. However, CSD ipsilateral to the PO/AH lesion slightly suppressed the skin-cooling behavior for 4 min with small rise in Tre. In the PO/AH intact rat, while 0.9% NaCl did not affect the skin-cooling behavior, unilateral CSD slightly suppressed it for 3 min almost in the same way as that of the ipsilateral CSD trial in the PO/AH lesioned rat. The results add further evidence to support the involvement of the frontal cortex in the central control of thermoregulation.


Effects of adrenalectomy and adrenal steroids on norepinephrine synthesis and monamine oxidase activity.
Rat heart norepinephrine (NE) tunover was increased 6--10 days after bilateral adrenalectomy. This increase was prevented by administration of deoxycorticosterone acetate (DOCA) but not by either hydrocortisone or corticosterone. Blood pressure decreased following adrenalectomy. This decrease was prevented by DOCA, hydrocortisone and corticosterone. Monoamine oxidase (MAO) activity increased in the heart but not in the liver following adrenalectomy. DOCA prevented the increase in heart MAO activity whereas hydrocortisone and corticosterone were ineffective. In intact animals, heart and liver MAO activity were not changed by 5 days of cold exposure, a procedure which increases NE turnover. It is suggested that the increase in heart NE turnover may be related to the increase in MAO activity seen after adrenalectomy.


Effect of prolactin on the calcium binding and/or transport of ejaculated and epididymal human spermatozoa.
The prolactin concentration in human seminal plasma and in human epididymal cauda fluid was assessed by radioimmunoassay. The prolactin concentration in cauda plasma was found to be similar to that found in male blood serum (5.5 to 9.1 ng/ml) and significantly lower than the concentration of the hormone found in seminal plasma obtained either from euspermic (48 +/- 12 ng/ml) or from vasectomized volunteers (50 +/- 10.2 ng/ml) (P less than 0.001). Calcium binding and/or transport in ejaculated spermatozoa were found to be little (0.29 +/- 0.08 nmoles/10(8) cells) and dependent on a quickly saturable process. The addition of 200 ng of human prolactin/ml induced a 60% increase in this parameter, while 50 ng of prolactin/ml were ineffective. Epididymal human spermatozoa differ from ejaculated sperm cells in showing greater, time-dependent, calcium binding and/or transport under basal conditions (2.00 +/- 0.35 nmoles/10(8) cells/hour), and in being more susceptible to the stimulating action of prolactin (4.4 +/- 0.68 nmoles/10(8) cells/hour in the presence of 50 ng of prolactin/ml).


Ascorbate-and iron-driven redox activity of Dp44mT and Emodin facilitates peroxidation of micelles and bicelles.
Iron (Fe)-induced oxidative stress leads to reactive oxygen species that damage biomembranes, with this mechanism being involved in the activity of some anti-cancer chemotherapeutics. Herein, we compared the effect of the ligand, di-2-pyridylketone 4,4-dimethyl-3-thiosemicarbazone (Dp44mT), or the potential ligand, Emodin, on Fe-catalyzed lipid peroxidation in cell membrane models (micelles and bicelles). These studies were performed in the presence of hydrogen peroxide (H2O2) and the absence or presence of ascorbate. In the absence of ascorbate, Fe(II)/Emodin mixtures incubated with H2O2 demonstrated slight pro-oxidant properties on micelles versus Fe(II) alone, while the Fe(III)-Dp44mT complex exhibited marked antioxidant properties. Examining more physiologically relevant phospholipid-containing bicelles, the Fe(II)- and Fe(III)-Dp44mT complexes demonstrated antioxidant activity without ascorbate. Upon adding ascorbate, there was a significant increase in the peroxidation of micelles and bicelles in the presence of unchelated Fe(II) and H2O2. The addition of ascorbate to Fe(III)-Dp44mT substantially increased the peroxidation of micelles and bicelles, with the Fe(III)-Dp44mT complex being reduced by ascorbate to the Fe(II) state, explaining the increased reactivity. Electron paramagnetic resonance spectroscopy demonstrated ascorbyl radical anion generation after mixing ascorbate and Emodin, with signal intensity being enhanced by H2O2. This finding suggested Emodin semiquinone radical formation that could play a role in its reactivity via ascorbate-driven redox cycling. Examining cultured melanoma cells in vitro, ascorbate at pharmacological levels enhanced the anti-proliferative activity of Dp44mT and Emodin. Ascorbate-driven redox cycling of Dp44mT and Emodin promotes their anti-proliferative activity.


Fuels and chemicals from hemicellulose sugars.
Industrial processes of lignocellulosic material have made use of only the hexose component of the cellulose fraction. Pentoses and some minor hexoses present in the hemicellulose fraction, which may represent as much as 40% of lignocellulosic biomass, have in most cases been wasted. The lack of good methods for utilization of hemicellulose sugars is a key obstacle hindering the development of lignocellulose-based ethanol and other biofuels. In this chapter, we focus on the utilization of hemicellulose sugars, the structure of hemicellulose and its hydrolysis, and the biochemistry and process technology involved in their conversion to valuable fuels and chemicals.


Post-mortem analysis of lactate concentration in diabetics and metformin poisonings.
Lactate is produced in carbohydrate metabolism under anaerobic conditions. Lactic acidosis occurs when the production of lactate exceeds its removal. In post-mortem (PM) context, the lactic acidosis is difficult to interpret due to unknown pathophysiological factors prior to death and PM changes that may affect the lactate levels. We evaluated 1865 medico-legal autopsy cases where the quantitation of glucose, lactate, and ketone bodies was performed as a part of the cause of death (CoD) investigation. Lactate was shown to ascend in a logarithmic manner as the PM interval increased until a plateau was achieved approximately after 8-10 days PM, and the elevation was caused mainly by PM changes. The lactate level was higher than the mean in cases where the CoD was diabetes mellitus type 2 (DM2) or metformin poisoning. Although there was a correlation between metformin and lactate levels, our findings suggest the DM2 and its complications were the cause for elevated lactate levels rather than metformin, since the lactate levels were similar in DM2-associated deaths where no metformin was detected. Elevated lactate levels in PM samples rather referred to metabolic disturbances often caused by DM2. An assay to detect D-lactate in PM samples was described.


Extended analysis of intratumoral heterogeneity of primary osteosarcoma tissue using 3D-in-vivo-tumor-model.
Osteosarcomas are a rare, heterogeneous and malignant group of bone tumors that have a high potential for metastasis and aggressive growth patterns. Treatment of metastasized osteosarcoma is often insufficient and research is compromised by problems encountered when culturing cells or analyzing genetic alterations due to the high level of intratumoral and intertumoral heterogeneity. The chick chorioallantoic membrane (CAM) model, a 3D-in-vivo-tumor-model, could potentially facilitate the investigation of osteosarcoma heterogeneity at an individual and highly specified level. Objective was to establish the grafting and transplantation of different primary osteosarcoma tissue parts onto several consecutive CAMs for tumor profiling and investigation of osteosarcoma heterogeneity. Various parts of primary osteosarcoma tissue were grafted onto CAMs and were transplanted onto another CAM for five to seven consecutive times, enabling further experimental analyzes. Primary osteosarcoma tissue parts exhibited satisfactory growth patterns and displayed angiogenic development on the CAM. It was possible to graft and transplant different tumor parts several times while the tissue viability was still high and tumor profiling was performed. Primary osteosarcoma tissue grew on several different CAMs for an extended time period and neovascularization of serial transplanted tumor parts was observed, improving the versatility of the 3D-in-vivo-tumor-model.


Disparities in Health Care for Hispanic Patients in Physical Medicine and Rehabilitation in the United States: A Narrative Review.
In the United States, the number of people with Hispanic backgrounds is rapidly increasing. This growth, along with other factors, may put them at increased risk for healthcare disparities. Thus, physical medicine and rehabilitation should evaluate disparities in health services through the lens of the evidence base. To our knowledge, this is the first review describing the literature on healthcare disparities for Hispanic patients in the field of physical medicine and rehabilitation. This review explores healthcare utilization, risk factors, access, and health conditions specific to Hispanic populations. Articles used in this narrative review were collected for a 10-yr span (2009-2018) from online databases.Our findings highlight disparities across common conditions and injuries in the physical medicine and rehabilitation literature, including poststroke rehabilitation, hip fracture treatment and rehabilitation, spinal cord injury, and traumatic brain injury. Our review suggests that Hispanic patients may be at risk for worse outcomes after these conditions, because of low access to rehabilitation services and disparities in the referral process. Similar results are reported for pain management and Parkinson disease. As we strive to provide optimal care to an increasingly diverse patient population, researchers and clinicians must consider effects of race and ethnicity on access to and utilization of rehabilitation services.


Effects of pH, humic substances and animal interactions on survival and physiological status of Asellus aquaticus L. and Gammarus pulex (L.) : A field experiment.
The mortality and physiological status (body water content) of Asellus aquaticus (Isopoda) and Gammarus pulex (Amphipoda) were measured after 25 days exposure in 20 natural streams with a pH range of 4.3-7.5 and a colour range of 8-280 mg Pt L-1. In addition, the effects of keeping the animals as single species or together were studied. The response of Gammarus to low pH was an increased mortality and lower physiological status of surviving individuals in streams with a pH lower than 6.0. In Asellus the physiological status was correlated with pH, while the mortality was not pH dependent. The effects of humus on the physiological status of Asellus was significant when fitted to a second order polynomial function. The influence of humus can, however, be regarded as small relative to pH. The interactions between the species could be described as asymmetric under optimal conditions of high pH and low humus concentrations, where the presence of Gammarus decreased the survival and physiological status of Asellus. Acid stress did not seem to reverse the direction of this asymmetry, but the presence of Gammarus improved the physiological status of Asellus at pH lower than 6.0. Since the presence of Asellus did not increase the mortality or decrease the physiological status of Gammarus, this could be explained by Asellus feeding on Gammarus that died from physiological stress solely. This mechanism suggests that food quality, and thus effects of diffuse competition, can be important for the ability to withstand acid stress. The results, though, give no support for the hypothesis that competition from Asellus is important for the disappearance of Gammarus during the acidification of streams.


Patient-centered goal setting in very early supported discharge with continued rehabilitation after stroke.
To examine patients' perception of performance and satisfaction with the activities in their set goals before and after very early supported discharge (VESD) with continued rehabilitation. A descriptive cohort study with data extracted from a randomized controlled trial. Sixty-nine patient allocated to the intervention group were eligible. Before discharge, the patients were asked to set rehabilitation goals, and they were asked to rate the performance and satisfaction of their set goals. At discharge from the rehabilitation, the patients were asked to re-evaluate their experience and satisfaction with the goal performance. One hundred and forty goals were registered. At 81.5% of the set goals, the patients estimated that they performed the task better at discharge than at enrolment and at 86.5% of the set goals the patients were more satisfied with the performance at discharge than at enrolment. Patients with mild to moderate stroke, undergoing a VESD after stroke, reported high performance level for their set goals and were satisfied with their performance execution. Further research is needed to investigate whether the goal should be set preferably at home or at hospital before discharge.Implications for rehabilitationMany of the patients can formulate achievable goals with their rehabilitation after stroke.Patients ongoing rehabilitation after stroke are satisfied with their performance of the set goals.As part of patient-centered care, stroke patients should be given the opportunity to formulate their own goals with their rehabilitation.Short hospital times and fast planning of goal-meetings, seems to influence patient goal setting in early discharge rehabilitation.


5-HT7 receptors expressed in the mouse parafacial region are not required for respiratory chemosensitivity.
A brainstem homeostatic system senses CO2 /H+ to regulate ventilation, blood gases and acid-base balance. Neurons of the retrotrapezoid nucleus (RTN) and medullary raphe are both implicated in this mechanism as respiratory chemosensors, but recent pharmacological work suggested that the CO2 /H+ sensitivity of RTN neurons is mediated indirectly, by raphe-derived serotonin acting on 5-HT7 receptors. To investigate this further, we characterized Htr7 transcript expression in phenotypically identified RTN neurons using multiplex single cell qRT-PCR and RNAscope. Although present in multiple neurons in the parafacial region of the ventrolateral medulla, Htr7 expression was undetectable in most RTN neurons (Nmb+ /Phox2b+ ) concentrated in the densely packed cell group ventrolateral to the facial nucleus. Where detected, Htr7 expression was modest and often associated with RTN neurons that extend dorsolaterally to partially encircle the facial nucleus. These dorsolateral Nmb+ /Htr7+ neurons tended to express Nmb at high levels and the intrinsic RTN proton detectors Gpr4 and Kcnk5 at low levels. In mouse brainstem slices, CO2 -stimulated firing in RTN neurons was mostly unaffected by a 5-HT7 receptor antagonist, SB269970 (n = 11/13). At the whole animal level, microinjection of SB269970 into the RTN of conscious mice blocked respiratory stimulation by co-injected LP-44, a 5-HT7 receptor agonist, but had no effect on CO2 -stimulated breathing in those same mice. We conclude that Htr7 is expressed by a minor subset of RTN neurons with a molecular profile distinct from the established chemoreceptors and that 5-HT7 receptors have negligible effects on CO2 -evoked firing activity in RTN neurons or on CO2 -stimulated breathing in mice. KEY POINTS: Neurons of the retrotrapezoid nucleus (RTN) are intrinsic CO2 /H+ chemosensors and serve as an integrative excitatory hub for control of breathing. Serotonin can activate RTN neurons, in part via 5-HT7 receptors, and those effects have been implicated in conferring an indirect CO2 sensitivity. Multiple single cell molecular approaches revealed low levels of 5-HT7 receptor transcript expression restricted to a limited population of RTN neurons. Pharmacological experiments showed that 5-HT7 receptors in RTN are not required for CO2 /H+ -stimulation of RTN neuronal activity or CO2 -stimulated breathing. These data do not support a role for 5-HT7 receptors in respiratory chemosensitivity mediated by RTN neurons.


The role of microRNAs in glaucoma.
In this review, we aim to provide a comprehensive summary of the various microRNAs (miRNAs) shown to be involved in glaucoma and intraocular pressure regulation. miRNAs are short, single-stranded, and noncoding RNAs that regulate gene expression in a number of physiological conditions and human diseases, including glaucoma. Numerous miRNAs display differential expression in glaucoma-affected tissues, such as aqueous humor, tears, trabecular meshwork, and retina analyzed from patients and animal models, suggesting their potential involvement in glaucoma pathogenesis. Several studies summarized here have also investigated the challenge of delivering intact miRNAs to target tissues in order to develop miRNA-based glaucoma therapies. We extend these reports by conducting an additional layer of analysis that integrates the interaction between glaucoma-related miRNAs and glaucoma-associated genes. We conclude with a comprehensive discussion of the therapeutic potential of miRNAs, the cellular pathways that link these miRNAs together, and the most promising miRNAs for future glaucoma research.


Live-cell imaging combined with immunofluorescence, RNA, or DNA FISH to study the nuclear dynamics and expression of the X-inactivation center.
Differentiation of embryonic stem cells is accompanied by changes of gene expression and chromatin and chromosome dynamics. One of the most impressive examples for these changes is inactivation of one of the two X chromosomes occurring upon differentiation of mouse female embryonic stem cells. With a few exceptions, these events have been mainly studied in fixed cells. In order to better understand the dynamics, kinetics, and order of events during differentiation, one needs to employ live-cell imaging techniques. Here, we describe a combination of live-cell imaging with techniques that can be used in fixed cells (e.g., RNA FISH) to correlate locus dynamics or subnuclear localization with, e.g., gene expression. To study locus dynamics in female ES cells, we generated cell lines containing TetO arrays in the X-inactivation center, the locus on the X chromosome regulating X-inactivation, which can be visualized upon expression of TetR fused to fluorescent proteins. We will use this system to elaborate on how to generate ES cell lines for live-cell imaging of locus dynamics, how to culture ES cells prior to live-cell imaging, and to describe typical live-cell imaging conditions for ES cells using different microscopes. Furthermore, we will explain how RNA, DNA FISH, or immunofluorescence can be applied following live-cell imaging to correlate gene expression with locus dynamics.


Minimally invasive vs. open approach to the simultaneous treatment of colorectal tumors with synchronous liver metastasis: a single center, propensity-score matched analysis from Mayo clinic.
Open combined resections of colorectal primary tumors and synchronous liver metastases have become common in selected cases. However, evidences favoring a minimally invasive (MIS) approach are still limited. The aim of this study is to evaluate the outcomes of MIS vs. open synchronous liver and colorectal resections. 384 cases of synchronous colorectal and liver resections performed at one institution were identified during the study period. MIS vs open approach were compared after a propensity score matching; surgical outcomes were analyzed. MIS cases featured longer operative time (399 vs 300 min, p < 0.001), fewer blood loss (200 vs 500 ml, p = 0.003), and shorter hospitalization (median LOS 4 vs 6 days, p = 0.001). No difference was observed between the two groups for use of Pringle maneuver (p = 0.083), intraoperative blood transfusion (p = 0.061), achievement of negative colorectal (p = 0.176) and liver margins (p = 1.000), postoperative complications (p = 1.000) and significant (Clavien-Dindo ≥ 3a) complications (p = 0.817), delay of adjuvant therapy due to complications (p = 0.555), 30- and 90-day mortality. Synchronous colorectal and liver metastases resections via a minimally-invasive approach in high-volume centers with appropriate expertise result in significantly lower blood loss and length of stay despite longer operative time in comparison to open, with no oncological inferiority.


31P-NMR assay of phosphatidylcholine and phosphatidylethanolamine in AL721.
A 31P-NMR based method has been developed for the assay of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in AL721. The assay is based on the comparison of NMR signal intensities of the phosphatidyl moiety of PC and PE to the signal of triphenylphosphate (TPP), the internal standard. The assay is specific and reliable for the quantitation of phospholipid mixtures. Its uncertainty, due primarily to low intensity of 31P signals, was estimated at less than 6%.


Anti-sclerostin antibodies and abaloparatide have additive effects when used as a countermeasure against disuse osteopenia in female rats.
Prolonged disuse and substantial mechanical unloading are particularly damaging to skeletal integrity. Preclinical studies in rodents and clinical studies have highlighted the need for potent bone anabolic drugs to counteract disuse-induced osteopenia. The aim of present study was to compare the efficacy of romosozumab (Scl-Ab) and abaloparatide (ABL), alone or in combination, to prevent botulinum toxin (BTX) induced bone loss in a rat model. Eighty female Wistar rats were divided into the following six groups: 1. Baseline (n = 12); 2. Control (Ctrl) (n = 12); 3. BTX (n = 12); 4. BTX + Scl-Ab (n = 16); 5. BTX + ABL (n = 12); and 6. BTX + Scl-Ab + ABL (n = 16). Disuse was achieved by injecting 4 IU BTX into the hind limb musculature at study start. Scl-Ab (25 mg/kg) was injected s.c. twice weekly, while ABL (80 μg/kg) was injected s.c. five days a week for four weeks. Hind limb disuse dramatically decreased muscle mass and skeletal integrity and deteriorated the cortical morphology and trabecular microstructure. Treatment with Scl-Ab alone prevented most of the adverse cortical and trabecular effects of disuse, while ABL monotherapy mainly attenuated the disuse-induced loss of femoral areal bone mineral density (aBMD). Moreover, the combination of Scl-Ab and ABL not only counteracted most of the negative skeletal effects of unloading, but also increased aBMD (+10% and +20%), epiphyseal trabecular bone volume fraction (BV/TV) (+25% and +73%), and metaphyseal bone strength (+18% and +30%) significantly above that of Scl-Ab or ABL monotherapy, respectively. The potent and additive osteoanabolic effect of Scl-Ab and ABL, when given in combination, is highly intriguing and underlines that an osteoanabolic bone gain can be maximized by utilizing two pharmaceuticals targeting different cellular signaling pathways. From a clinical perspective, a combination treatment may be warranted in patients where the osteoanabolic effect of either monotherapy is not sufficient, or if a dose-reduction is required due to adverse effects.


A novel approach for enumeration of extracellular vesicles from crude and purified cell culture samples.
The interest in extracellular vesicles (EVs) has been increased in recent years due to their potential application in diagnosis and therapy of severe diseases. The versatile fields of application due to the numerous possible cargos and the targeted delivery system make them a promising biopharmaceutical product. However, their broad size range as well as varied surface protein content result in challenges for the purification, characterization, and quantification. In this study a novel method, based on high-resolution flow cytometry, was examined for the enumeration of EVs in purified as well as crude process samples. In addition to quantification, samples were characterized by dynamic light scattering, zeta potential measurement, and analytical size exclusion chromatography. It has been demonstrated that EVs were successfully enumerated with the novel method, offering great benefits for development and monitoring of EV processes.


Good practice in continence services.
The Department of Health, England, issued new guidance on continence services earlier this year. Sue Thomas examines the guidelines with a view to the future of continence care.


Young Sca-1+ bone marrow stem cell-derived exosomes preserve visual function via the miR-150-5p/MEKK3/JNK/c-Jun pathway to reduce M1 microglial polarization.
Polarization of microglia, the resident retinal immune cells, plays important roles in mediating both injury and repair responses post-retinal ischemia-reperfusion (I/R) injury, which is one of the main pathological mechanisms behind ganglion cell apoptosis. Aging could perturb microglial balances, resulting in lowered post-I/R retinal repair. Young bone marrow (BM) stem cell antigen 1-positive (Sca-1+) cells have been demonstrated to have higher reparative capabilities post-I/R retinal injury when transplanted into old mice, where they were able to home and differentiate into retinal microglia. Exosomes were enriched from young Sca-1+ or Sca-1- cells, and injected into the vitreous humor of old mice post-retinal I/R. Bioinformatics analyses, including miRNA sequencing, was used to analyze exosome contents, which was confirmed by RT-qPCR. Western blot was then performed to examine expression levels of inflammatory factors and underlying signaling pathway proteins, while immunofluorescence staining was used to examine the extent of pro-inflammatory M1 microglial polarization. Fluoro-Gold labelling was then utilized to identify viable ganglion cells, while H&E staining was used to examine retinal morphology post-I/R and exosome treatment. Sca-1+ exosome-injected mice yielded better visual functional preservation and lowered inflammatory factors, compared to Sca-1-, at days 1, 3, and 7 days post-I/R. miRNA sequencing found that Sca-1+ exosomes had higher miR-150-5p levels, compared to Sca-1- exosomes, which was confirmed by RT-qPCR. Mechanistic analysis found that miR-150-5p from Sca-1+ exosomes repressed the mitogen-activated protein kinase kinase kinase 3 (MEKK3)/JNK/c-Jun axis, leading to IL-6 and TNF-α downregulation, and subsequently reduced microglial polarization, all of which contributes to reduced ganglion cell apoptosis and preservation of proper retinal morphology. This study elucidates a potential new therapeutic approach for neuroprotection against I/R injury, via delivering miR-150-5p-enriched Sca-1+ exosomes, which targets the miR-150-5p/MEKK3/JNK/c-Jun axis, thereby serving as a cell-free remedy for treating retinal I/R injury and preserving visual functioning.


Effects of shear stress on endothelial cell haptotaxis on micropatterned surfaces.
Endothelial cell (EC) migration plays a critical role in vascular remodeling. Here we investigated the interactions between haptotaxis (induced by extracellular matrix gradient) and mechanotaxis (induced by mechanical forces) during EC migration. A micropatterning technique was used to generate step changes of collagen surface density. Due to haptotaxis, ECs developed focal adhesions and migrated into the area with higher surface density of collagen. Different levels of fluid shear stress were applied on ECs in the direction perpendicular to collagen strips. Shear stress at 2 dyn/cm2 did not affect haptotaxis, while shear stress at 3 dyn/cm2 or higher was sufficient to drive the migration of most ECs in the flow direction and against haptotaxis. Immunostaining revealed the increase of focal adhesions and lamellipodial protrusion in the direction of flow. These results suggest that shear stress beyond a certain threshold can be a predominant factor to determine the direction of EC migration.


Spindle cell reaction to nontuberculous mycobacteriosis in AIDS mimicking a spindle cell neoplasm. Evidence for dual histiocytic and fibroblast-like characteristics of spindle cells.
We report 5 patients with AIDS who had an unusual spindle cell proliferation in the lymph nodes and skin caused by nontuberculous mycobacteriosis. The spindle cell proliferation in these tissues may mimic a spindle cell neoplasm and pose a diagnostic problem if an infectious aetiology is not suspected. The fibroblast-like spindle cells contained numerous acid fast bacilli. They were strongly positive for antibody markers of monocyte/macrophage and leukocyte derivation: Leu M3, Mo-9, T-200, and HLA-DR, and variably positive for alpha-1 anti-chymotrypsin and lysozyme. Ultrastructurally these spindle cells were predominantly fibroblast-like with poorly developed features of macrophages. These results reveal the dual macrophage and fibroblastic character of the spindle cells and probably imply a functional differentiation rather than a histogenetic one.


A new method for estimating race/ethnicity and associated disparities where administrative records lack self-reported race/ethnicity.
To efficiently estimate race/ethnicity using administrative records to facilitate health care organizations' efforts to address disparities when self-reported race/ethnicity data are unavailable. Surname, geocoded residential address, and self-reported race/ethnicity from 1,973,362 enrollees of a national health plan. We compare the accuracy of a Bayesian approach to combining surname and geocoded information to estimate race/ethnicity to two other indirect methods: a non-Bayesian method that combines surname and geocoded information and geocoded information alone. We assess accuracy with respect to estimating (1) individual race/ethnicity and (2) overall racial/ethnic prevalence in a population. The Bayesian approach was 74 percent more efficient than geocoding alone in estimating individual race/ethnicity and 56 percent more efficient in estimating the prevalence of racial/ethnic groups, outperforming the non-Bayesian hybrid on both measures. The non-Bayesian hybrid was more efficient than geocoding alone in estimating individual race/ethnicity but less efficient with respect to prevalence (p<.05 for all differences). The Bayesian Surname and Geocoding (BSG) method presented here efficiently integrates administrative data, substantially improving upon what is possible with a single source or from other hybrid methods; it offers a powerful tool that can help health care organizations address disparities until self-reported race/ethnicity data are available.


Epidemiologic Opportunities and Challenges in Studying Environmental Risk Factors for Rheumatic Diseases.
Most rheumatic diseases have a stronger environmental than hereditary etiology. This article summarizes the key environmental risk factors for rheumatic diseases, the data sources that generated these findings, and the key pitfalls with existing research that every rheumatology clinician should know. Emerging research opportunities hold promise to revolutionize this field, and soon.


Focused ultrasound-mediated blood-brain barrier opening combined with magnetic targeting cytomembrane based biomimetic microbubbles for glioblastoma therapy.
Glioblastoma is the most common type of brain tumor. Due to the presence of the blood-brain barrier, the effects of chemotherapy have been unsatisfactory. The combination of focused ultrasound and microbubbles to reversibly open the blood-brain barrier is now considered a key factor in improving treatment outcomes of glioblastoma. In this study, we developed bionic drug delivery microbubbles, which in combination with focused ultrasound had an obvious inhibitory effect on glioblastoma. We extracted the brain microvascular cell membranes, combined them with lipid components, and loaded them with superparamagnetic iron oxide and doxorubicin to prepare biomimetic drug delivery microbubbles (FeDOX@cellMBs). We demonstrated that FeDOX@cellMBs retained the intrinsic properties of loading, such as magnetic properties and drug toxicity, both in vitro and in vivo. FeDOX@cellMBs exhibited good tumor targeting and uptake under the combined action of magnetic and focused ultrasound. Importantly, the FeDOX@cellMBs demonstrated excellent internal stability and effectively inhibited tumor growth in orthotopic glioblastoma mice. Finally, organ H&E staining confirmed that FeDOX@cellMBs were safe for use. In conclusion, FeDOX@cellMBs successfully penetrated the blood-brain barrier and effectively inhibited glioblastoma growth under the combined effects of focused ultrasound and magnetic stimulation. These results provide a new approach for the treatment of glioblastoma, with implications for future clinical translation.


T-lymphocyte reactivity during the menstrual cycle in women.
Reactivity of blood lymphocytes to nonspecific mitogenic stimulation with phytohemagglutinin (PHA) was measured in nine healthy, regularly cycling women at three phases of their menstrual cycles corresponding to peak levels of estradiol (midfollicular phase), peak levels of progesterone (midluteal phase), and the lowest levels of both hormones (menstrual phase). Sampling points were verified by radioimmunoassay of estrogen, progesterone, luteinizing hormone, and follicle-stimulating hormone. There were significant increases in reactivity associated with an increasing concentration of PHA and with autologous plasma vs AB plasma. However, no differences were found in reactivity to PHA over the three menstrual cycle phases and correlational analyses indicated no relationship between counts and any of the hormones measured.


A cytopathological approach to diagnosing intrathoracic lymphadenopathy using aspirates obtained by the transbronchial needle aspiration method.
Transbronchial needle aspiration (TBNA) is an effective, safe and cost-effective technique that allows for sampling of the mediastinal lymph node and peribronchial lesions. It is used in bronchogenic carcinoma staging, peribronchial and submucosal lesions, diagnosis of sarcoidosis and tuberculosis, differentiating submucosal invasion, and in diagnosing mediastinal masses. From our experience at the University of Abant Izzet Baysal and from a review of the literature, we discuss the adequacy and the differential diagnosis of aspiration material obtained by TBNA and cytopathological-histopathological evaluation in intrathoracic lymphadenopathies to increase the success rate of the TBNA method.


Pleistocene foxes (Vulpes, Canidae, Carnivora) from the Taurida Cave, Crimea.
A mandible fragment and four isolated teeth of the fossil foxes, Vulpes alopecoides (Del Campana, 1913), Vulpes cf. vulpes (Linnaeus, 1758) and Vulpes sp., are described from the Early Pleistocene locality of Taurida cave (Crimea, Late Villafranchian, 1.8-1.5 Ma). Based on the size and morphological features, a mandible fragment and two M1 are attributed to V. alopecoides. The structure of m1 of Vulpes cf. vulpes is similar to that of V. vulpes and V. alopecoides, but the size greatly exceeds the size limits for the latter species. Although m1 of Vulpes sp. fits the size of the lower carnassials of V. alopecoides, its morphology is definitely unique among the Early Pleistocene representatives of the genus Vulpes.


[Quantitative determination of amino acids from blood-stained filter papers using high-performance thin layer chromatography and a TLC-scanner (author's transl)].
A method is described to determine amino acids by extraction from pieces of bloodstained and dried filter papers and following dansylation and separation by High-Performance Thin Layer Chromatography (HPTLC). Quantitative determination is possible by means of a TLC-Scanner measuring the fluorescent light of the dansylated amino acids. The possible use of this procedure as screening test is discussed.


Cocktail-like gradient gelatin/hyaluronic acid bioimplant for enhancing tendon-bone healing in fatty-infiltrated rotator cuff injury models.
The regeneration of enthesis tissue (native tendon-bone interface) at the post-surgically repaired rotator cuff remains a challenge for clinicians, especially with the emergence of degenerative affection such as fatty infiltration that exacerbate poor tendon-bone healing. In this study, we proposed a cocktail-like hydrogel with a four-layer structure (BMSCs+gNC@GH) for enhancing fatty infiltrated tendon-bone healing. As collagen and hyaluronic acid are the main biomacromolecules that constitute the extracellular matrix of enthesis tissue, this hydrogel was composed of UV-curable gelatin/hyaluronic acid (GelMA/HAMA) dual network gel (GH) with nanoclay (NC) and stem cells loaded. The results showed that NC exhibited a cocktail-like gradient distribution in GH, which effectively mimicked the structure of native enthesis and supported the long-term culture and encapsulation of BMSCs. What's more, the gradient variation of NC provided a biological signal for promoting gradient osteogenic differentiation of cells. Based on the in vivo results, BMSCs+gNC@GH effectively promoted fibrocartilage layer regeneration at the tendon-bone interface and inhibited fatty infiltration. Therefore, BMSCs+gNC@GH group exhibited better biomechanical properties. Thus, this cocktail-like implant may be a promising tissue-engineered scaffold for tendon-bone healing, and it provides a new idea for the development of scaffolds with the function of inhibiting degeneration.


The perfectionistic patient.
Family members, residents, and care givers often react negatively when perfection is demanded by the geriatric patient. This type of reaction presents a challenge to the nurse providing care. When a geriatric patient is perfectionistic, it is important to step back, to observe and assess, and to plan appropriate strategies that will facilitate the patient's adjustment. When the behavior is related to situational stressors, it may be appropriate to respond to the stressors and ignore the behavior. When the behavior results from a feeling of loss of control and powerlessness, intervention should be directed in that direction. In any case, actions of the nurse should respond to the patient's need for self-esteem and improved ability to cope with the environment. Responding to the perfectionistic geriatric patient presents a particular challenge to the nurse in long-term care, and the nurse's strategic intervention can significantly increase the patient's ability to adjust to the ongoing changes that accompany the aging process.


Controlled growth of NiCo₂O₄ nanorods and ultrathin nanosheets on carbon nanofibers for high-performance supercapacitors.
Two one-dimensional hierarchical hybrid nanostructures composed of NiCo2O4 nanorods and ultrathin nanosheets on carbon nanofibers (CNFs) are controllably synthesized through facile solution methods combined with a simple thermal treatment. The structure of NiCo2O4 can be easily controlled to be nanorods or nanosheets by using different additives in the synthesis. These two different nanostructures are evaluated as electrodes for high performance supercapacitors, in view of their apparent advantages, such as high electroactive surface area, ultrathin and porous features, robust mechanical strength, shorter ion and electron transport path. Their electrochemical performance is systematically studied, and both of these two hierarchical hybrid nanostructures exhibit high capacitance and excellent cycling stability. The remarkable electrochemical performance will undoubtedly make these hybrid structures attractive for high-performance supercapacitors with high power and energy densities.


Structural analysis of the dual agonism at GLP-1R and GCGR.
Glucagon-like peptide-1 receptor (GLP-1R) and glucagon receptor (GCGR), two members of class B1 G protein-coupled receptors, play important roles in glucose homeostasis and energy metabolism. They share a high degree of sequence homology but have different functionalities. Unimolecular dual agonists of both receptors developed recently displayed better clinical efficacies than that of monotherapy. To study the underlying molecular mechanisms, we determined high-resolution cryo-electron microscopy structures of GLP-1R or GCGR in complex with heterotrimeric Gs protein and three GLP-1R/GCGR dual agonists including peptide 15, MEDI0382 (cotadutide) and SAR425899 with variable activating profiles at GLP-1R versus GCGR. Compared with related structures reported previously and supported by our published pharmacological data, key residues responsible for ligand recognition and dual agonism were identified. Analyses of peptide conformational features revealed a difference in side chain orientations within the first three residues, indicating that distinct engagements in the deep binding pocket are required to achieve receptor selectivity. The middle region recognizes extracellular loop 1 (ECL1), ECL2, and the top of transmembrane helix 1 (TM1) resulting in specific conformational changes of both ligand and receptor, especially the dual agonists reshaped ECL1 conformation of GLP-1R relative to that of GCGR, suggesting an important role of ECL1 interaction in executing dual agonism. Structural investigation of lipid modification showed a better interaction between lipid moiety of MEDI0382 and TM1-TM2 cleft, in line with its increased potency at GCGR than SAR425899. Together, the results provide insightful information for the design and development of improved therapeutics targeting these two receptors simultaneously.


Identification of the novel HLA-A*24:567N allele in a Chinese individual.
HLA-A*24:567N differs from HLA-A*24:02:01:01 by a single nucleotide deletion at position 149.


A single oral dose of celecoxib-loaded solid lipid nanoparticles for treatment of different developmental stages of experimental schistosomiasis mansoni.
Schistosomiasis, a neglected tropical parasitic disease, is associated with severe pathology, mortality and economic loss. The treatment and control of schistosomiasis depends mainly on a single dose of praziquantel (PZQ). Drug repurposing and nanomedicine attract great attention to improve anti-schistosomal therapy. Previously, we reported that celecoxib (CELE), the non-steroidal anti-inflammatory drug, showed potent anti-schistosomal efficacy in an oral dose of 20 mg/kg/day for five days against different developmental stages of Schistosoma mansoni (S. mansoni) infection in mice. The aim of the current study was to shorten the duration of CELE treatment to reach an effective single oral dose against different developmental stages of S. mansoni infection using solid lipid nanoparticles (SLNs) as nano-carriers. The latter enhance the solubility, bioavailability and drug delivery and hence can decrease the frequency of administration which is of great clinical value. CELE-loaded SLNs showed good colloidal properties, high entrapment efficiency and drug loading, sustained biphasic release pattern with excellent storage stability. The used regimen was efficient against different developmental stages of S. mansoni infection with the most pronounced effect against the juvenile stage where the worm load, the hepatic egg count and the intestinal egg count were reduced by 86.39%, 91.45% and 90.11%, respectively. Meanwhile, when targeting the invasive and the adult stages, it induced reduction in the worm load by 73.55% and 78.22%, the hepatic egg count by 69.99% and 75.39% and the intestinal egg count by 77.57% and 79.89%, respectively. Additionally, CELE-loaded SLNs caused extensive tegumental damage of adult worms and marked improvement in the liver pathology.