Evaluation of potential carcinogenicity of steroidal alkaloids from Veratrum album L. by the DC polarography method.
The presented work is devoted to the study of polarographic reduction in the series of 13 alkaloids isolated from various parts of Veratrum album subsp. lobelianum. The used compounds were evaluated from the point of view of their potential carcinogenicity in anhydrous N,N-dimethylformamide (DMF) by the method of DC polarography. All compounds were reduced during an one two-electron irreversible step. Their potential carcinogenicity characterized by a parameter tg alpha value determined in the presence of alpha-lipoic acid ranged from the highest value 0.257 obtained for the solanidane skeleton containing rubijervine to the value 0.070 for jervine. The tg alpha value determined for rubijervine (0.257) is comparable with the tg alpha of naphto-(2',1',2,3)fluoranthene (0.270)-compound classified by IARC as possible carcinogen for human. The tg alpha values determined for other alkaloids were relatively low and they do not indicate any possible carcinogenic activity.


Prodrug approaches for the development of a long-acting drug delivery systems.
Long-acting formulations are designed to reduce dosing frequency and simplify dosing schedules by providing an extended duration of action. One approach to obtain long-acting formulations is to combine long-acting prodrugs (LA-prodrug) with existing or emerging drug delivery technologies (DDS). The design criteria for long-acting prodrugs are distinct from conventional prodrug strategies that alter absorption, distribution, metabolism, and excretion (ADME) parameters. Our review focuses on long-acting prodrug delivery systems (LA-prodrug DDS), which is a subcategory of long-acting formulations where prodrug design enables DDS formulation to achieve an extended duration of action that is greater than the parent drug. Here, we define LA-prodrugs as the conjugation of an active pharmaceutical ingredient (API) to a promoiety group via a cleavable covalent linker, where both the promoiety and linker are selected to enable formulation and administration from a drug delivery system (DDS) to achieve an extended duration of action. These LA-prodrug DDS results in an extended interval where the API is within a therapeutic range without necessarily altering ADME as is typical of conventional prodrugs. The conversion of the LA-prodrug to the API is dependent on linker cleavage, which can occur before or after release from the DDS. The requirement for linker cleavage provides an additional tool to prolong release from these LA-prodrug DDS. In addition, the physicochemical properties of drugs can be tuned by promoiety selection for a particular DDS. Conjugation with promoieties that are carriers or amenable to assembly into carriers can also provide access to formulations designed for extending duration of action. LA-prodrugs have been applied to a wide variety of drug delivery strategies and are categorized in this review by promoiety size and complexity. Small molecule promoieties (typically MW < 1000 Da) have been used to improve encapsulation or partitioning as well as broaden APIs for use with traditional long-acting formulations such as solid drug dispersions. Macromolecular promoieties (typically MW > 1000 Da) have been applied to hydrogels, nanoparticles, micelles, dendrimers, and polymerized prodrug monomers. The resulting LA-prodrug DDS enable extended duration of action for active pharmaceuticals across a wide range of applications, with target release timescales spanning days to years.


Ultrastructural evaluation of gingival connective tissue in hereditary gingival fibromatosis.
To describe the ultrastructural features of hereditary gingival fibromatosis (HGF) in affected family members and compare microscopic findings with normal gingival (NG) tissue. Gingival tissue samples from nine patients with HGF from five unrelated families were evaluated by transmission electron microscopy. Nine NG tissue samples were used for comparison. Areas containing collagen fibrils forming loops and folds were observed in both groups, whereas oxytalan fibers were frequently identified in the HGF group. The diameter of collagen fibrils and the interfibrillar space among them were more uniform in the NG group than in the HGF group. Fibroblasts were the most common cells found in both the HGF and NG groups and exhibited enlarged, rough endoplasmic reticulum, mitochondria with well-preserved crests, conspicuous nucleoli, and euchromatic chromatin. Other cells, such as mast cells, plasma cells, and macrophages, were also observed. HGF tissues had ultrastructural characteristics that were very similar to those of NG tissues. Oxytalan fibers were observed more frequently in the HGF samples than in the NG samples. Other studies of HGF in patients from different families should be performed to better understand the pathogenesis of this hereditary condition.


Correction to: Practical guidance for the evaluation of disease progression and the decision to change treatment in patients with advanced gastric cancer receiving chemotherapy.
The article Practical guidance for the evaluation of disease progression.


The self-management and transition manual "ALL YOU NEED IS LOVE" for adolescents with chronic kidney disease.
To examine the effects of "ALL YOU NEED IS LOVE", a novel six-week, self-directed patient education manual designed to improve chronic kidney disease knowledge/self-management, health care transition readiness, self-advocacy, and mindfulness skills among adolescents with chronic kidney disease. We enrolled 49 adolescents aged 11-17 years (mean age 14.7 ± 1.9; 53.1% males) from a university hospital kidney center who had CKD stages 3 and greater, who were randomly assigned into the "ALL YOU NEED IS LOVE" patient education only group (n = 31) or the "ALL YOU NEED IS LOVE" patient education plus mindfulness training group (n = 18). Participants completed Qualtrics surveys at baseline, post-intervention, and three-month follow-up. The survey included measures of outcome variables (i.e., self-management/transition readiness, patient self-advocacy, and mindfulness), and an additional demographic questionnaire was included in the baseline survey. Multilevel model analyses were used to examine the effects of group and time on the outcome variables. Multilevel model analyses showed an overall significant time effect across all outcome variables in both groups. However, the group effects were not statistically significant across the outcome variables. Both interventions significantly increased participants' self-management/HCT readiness, self-advocacy, and mindfulness over time. Mindfulness training may not bring additional benefits to the "ALL YOU NEED IS LOVE" education manual. Pediatric nurses and clinicians may utilize the self-directed "ALL YOU NEED IS LOVE" manual to increase self-management/transition readiness, self-advocacy, and mindfulness among adolescents with chronic kidney disease.


The cortico-basal ganglia integrative network: the role of the thalamus.
The thalamus is a critical component of the frontal cortical-basal ganglia-thalamic circuits that mediate motivation and emotional drive, planning and cognition for the development and expression of goal-directed behaviors. Each functional region of the frontal cortex is connected with specific areas of each basal ganglia (BG) structure and of the thalamus. In addition, the thalamus sends a massive, topographically organized projection directly to the striatum. Tract-tracing and physiological experiments have indicated a general topographic organization of the cortical-BG-thalamic loops and supported a model of BG function based on parallel and segregated pathways. However, the learning and execution of appropriate behavioral responses require integration of inputs related to emotional, cognitive, and motor cortical functions. Our recent data indicate that integration may occur via non-reciprocal connections between the striatum and substantia nigra and within "hot spots" of convergence between corticostriatal projections from different functional regions. Similarly, integration may exist in the thalamus. There are non-reciprocal connections between the thalamus and cortex via thalamocortical projections that terminate in the superficial and deep cortical layers. These terminals can influence different functional cortical areas that, in turn, project to the striatum and back to the thalamus. In addition, a non-reciprocal corticothalamic projection terminates in thalamic regions that are parts of other circuits. Finally, 'hot spots' of convergence between terminals from different cortical regions may also occur in the thalamus as is seen in the striatum. Thus, via several different pathways, the thalamus may serve as an important center of integration of networks that underlie the ability to modulate behaviors.


Mast cell stabilisers.
Mast cells play a critical role in type 1 hypersensitivity reactions. Indeed, mast cell mediators are implicated in many different conditions including allergic rhinitis, conjunctivitis, asthma, psoriasis, mastocytosis and the progression of many different cancers. Thus, there is intense interest in the development of agents which prevent mast cell mediator release or which inhibit the actions of such mediators once released into the environment of the cell. Much progress into the design of new agents has been made since the initial discovery of the mast cell stabilising properties of khellin from Ammi visnaga and the clinical approval of cromolyn sodium. This review critically examines the progress that has been made in the intervening years from the design of new agents that target a specific signalling event in the mast cell degranulation pathway to those agents which have been developed where the precise mechanism of action remains elusive. Particular emphasis is also placed on clinically used drugs for other indications that stabilise mast cells and how this additional action may be harnessed for their clinical use in disease processes where mast cells are implicated.


High efficiency, high temperature separations on silica based monolithic columns.
The effect of temperature on separation using reversed-phase monolithic columns has been investigated using a nano-LC pumping system for gradient separation of tryptic peptides with MS detection. A goal of this study was to find optimal conditions for high-speed separations. The chromatographic performance of the columns was evaluated by peak capacity and peak capacity per time unit. Column lengths ranging from 20 to 100 cm and intermediate gradient times from 10 to 30 min were investigated to assess the potential of these columns in a final step separation, e.g. after fractionation or specific sample preparation. Flow rates from 250 to 2000 nL/min and temperatures from 20 to 120°C were investigated. Temperature had a significant effect on fast separations, and a flow rate of 2000 nL/min and a temperature of 80°C gave the highest peak capacity per time unit. These settings produced 70% more protein identifications in a biological sample compared to a conventional packed column. Alternatively, an equal amount of protein identifications was obtained with a 40% reduction in run time compared to the conventional packed column.


Optimization of perfusion CT protocol for imaging of extracranial head and neck tumors.
The in vivo assessment of physiological processes associated with microcirculation in the head and neck tissue by means of perfusion computed tomography is widely used in the management of patients with head and neck tumors. However, there is no systematic consideration of the total acquisition duration and placement of the scans. A simulation study for optimizing perfusion studies of extracranial head and neck tumors, with considerations of reducing radiation dose while maintaining accuracy of the perfusion parameters, is demonstrated here. The suggested that dual-phase optimized protocols may provide reliable estimations of the permeability surface area product as well as blood flow and volume without additional radiation burden and serious patient discomfort. These optimized protocols can potentially be useful in the clinical setting of examining patients with extracranial head and neck tumors.


The low-toxicity versions of LPS, MPL adjuvant and RC529, are efficient adjuvants for CD4+ T cells.
Lipopolysaccharide (LPS) has long been known to enhance innate and adaptive immune responses; however, its extreme toxicity precludes its use in clinical settings. The combined toxicity and adjuvanticity of LPS have contributed to the view that immunological adjuvants need to be highly inflammatory to be maximally effective. Here, we compared the effects of LPS with its less-toxic derivatives, monophosphoryl lipid A (MPL) and a chemical mimetic, RC529, on CD4+ T cell clonal expansion, long-term survival, and T helper cell type 1 (Th1) differentiation. We found that LPS, MPL, and RC529 had similar effects on CD4+ T cell clonal expansion, cell division, and ex vivo survival. Analysis of the ability of activated CD4+ T cells to produce interferon-gamma following a 21-day immunization and challenge protocol with LPS and MPL resulted in similar Th1 differentiation. In contrast, we found that LPS was more effective in promoting long-term CD4+ T cell responses, as we recovered nearly sixfold more cells following immunization/challenge as compared with treatment with MPL. Our results indicate that low-inflammation adjuvants, such as MPL and RC529, are capable of enhancing short-term CD4+ T cell clonal expansion and Th1 differentiation, but inflammatory signaling aids in the long-term retention of antigen-specific T cells.


Unblocked segments in obstetric epidural blocks. The influence of previous regional block on obstetric patients receiving lumbar epidural analgesia during labour.
Two hundred and six patients were studied to discover if previous regional analgesia influenced the incidence of unblocked segments and unilateral analgesia during lumbar epidural block in labour. Previous lumbar epidural or subarachnoid or sacral epidural blocks alone did not appear to influence the incidence of unilateral block or of unblocked segments. The practice of using epidural analgesia to facilitate the insertion of cervical circumsuture during pregnancy did not appear to be contra-indicated.


Effect of etchant variability on shear bond strength of all ceramic restorations - an in vitro study.
The present study was carried out to evaluate the effect of pre-luting surface treatments by 3 different etchant used at 3 different etching periods and their effect on shear bond strength of IPS Empress 2 luted to tooth by dual cure resin cement. Fifty samples of ceramic were divided into four groups as group I control group: No surface treatment, group II: Etched with Hydrofluoric (HF) acid (4.9%), group III: Etched with Ammonium bifluoride acid (9.4%) and group IV: Etched with Phosphoric acid (37%). Group II, III and IV were further divided into 3 Subgroups; namely A, B and C according to the etching periods (20, 60 and 120 s) respectively. The shear bond strength was determined by using a Universal testing Machine. The morphological changes of the surface treated ceramic samples prior to luting to tooth and mode of the fracture failure after shear bond test were observed by using a Scanning Electron Microscope. The mean shear bond strength was highest when IPS Empress 2 ceramic samples were surface treated using 4.9% Hydrofluoric acid gel and 9.4% Ammonium bifluoride acid for 120 s. The least mean shear bond strength was noticed in case of control group, where no surface treatment was done and samples treated by 37% Phosphoric acid. Thus it could be concluded, that Ammonium bifluoride could be an appropriate alternative to be used instead of HF acid. 120 s etching showed highest bond strength values for HF acid (4.9%) and Ammonium bifluoride (9.4%).


Health outcomes, health services utilization, and costs consequences of medicare uninsurance among migrants in Canada: a systematic review.
Medically uninsured groups, many of them migrants, reportedly delay using healthcare services due to costs and often face preventable health consequences. This systematic review sought to assess quantitative evidence on health outcomes, health services use, and health care costs among uninsured migrant populations in Canada. OVID MEDLINE, Embase, Global Health, EconLit, and grey literature were searched to identify relevant literature published up until March 2021. The Cochrane Risk of Bias in Non-randomized Studies - of Interventions (ROBINS-I) tool was used to assess the quality of studies. Ten studies were included. Data showed that there are differences among insured and uninsured groups in reported health outcomes and health services use. No quantitative studies on economic costs were captured. Our findings indicate a need to review policies regarding accessible and affordable health care for migrants. Increasing funding to community health centers may improve service utilization and health outcomes among this population.


Effects of melatonin-pretreated adipose-derived mesenchymal stem cells (MSC) in an animal model of spinal cord injury.
One of the most serious nervous system diseases is spinal cord injury(SCI), which is increasing for various reasons. Although no definitive treatment has yet been identified for SCI, one possible treatment is adipose-derived stem cells(ADSCs). However, a key issue in transplantation is improving cells' survival and function in the target tissue. Melatonin(MT) hormone with antioxidant properties can prolong cell survival and improve cell function. This study investigates the pre-conditioning of ADSCs with melatonin for enhancing the engraftment and neurological function of rats undergoing SCI. 42 male Sprague-Dawley rats were divided into six groups, including Control, Sham, Model, Vehicle, and Lesion treatments A and B. After acquiring white adipose tissue, stem cells were evaluated by flow cytometry. SCI was then applied in Model, Vehicle, A, and B groups. Group A and B received ADSCs and ADSCs + melatonin, respectively, 1 week after SCI, but the vehicle received only an intravenous injection for simulation; The other groups were recruited for the behavioral test. Immunohistochemistry(IHC) was used to assess the engraftment and differentiation of ADSCs in the SCI site. Basso, Beattie, and Bresnahan's score was used to evaluate motor function between the six groups. Histological studies and cell count confirmed ADSCs implantation at the injury site, which was higher in the MT-ADSCs (P < 0.001). IHC revealed the differentiation of ADSCs and MT-ADSCs into neurons, astrocytes, and oligodendrocyte lineage cells, which were higher in MT-ADSCs. Functional improvement was observed in SCI + ADSCs and SCI + MT-ADSCs groups. The pre-conditioning of ADSCs with melatonin positively affects engraftment and neuronal differentiation in SCI but does not impact performance improvement compared to the ADSCs.


Oral co-administration of live attenuated Salmonella enterica serovar Typhimurium expressing chicken interferon-α and interleukin-18 enhances the alleviation of clinical signs caused by respiratory infection with avian influenza virus H9N2.
The combined use of cytokines has shown synergistic and/or additive effects in controlling several viral infections of livestock animals. However, little is known concerning the practical use of chicken cytokine combinations to control avian diseases. Here, we investigated the antiviral efficacy of oral co-administration of chicken interferon-α (chIFN-α) and chicken interleukin-18 (chIL-18) using attenuated Salmonella enterica serovar Typhimurium in chickens infected with avian influenza virus (AIV) H9N2. Our results demonstrate that oral co-administration of S. enterica serovar Typhimurium expressing chIFN-α and chIL-18 produced a greater alleviation of clinical signs caused by respiratory infection with AIV H9N2 in chickens, when compared to administration of S. enterica serovar Typhimurium expressing either chIFN-α or chIL-18 alone. Mortality, clinical symptom severity, and feed and water intake were used to access treatment effectiveness. This enhancement of antiviral immunity was further confirmed by evidence of reduced rectal shedding and decreased replication of AIV H9N2 in several different tissues of challenged chickens including trachea, lung, cecal tonsil, and brain. Furthermore, oral co-administration of chIFN-α and chIL-18 more efficiently modulated the immune responses of chickens against AIV H9N2 by enhancing both humoral and Th1-biased cell-mediated immunity, compared to single administration of either construct. Therefore, our results suggest that the combined administration of two chicken cytokines, chIFN-α and chIL-18, using attenuated S. enterica serovar Typhimurium as an oral carrier, provides an effective means for controlling respiratory disease caused by AIV H9N2 infection.


Serum cholesterol concentration before and after streptokinase in acute myocardial infarction.
To determine if serum cholesterol concentration should be measured before or after streptokinase therapy within the first 24 h of myocardial infarction. Prospective study of patients receiving streptokinase therapy for acute myocardial infarction (AMI). Coronary care unit of a district general hospital. Thirty-one patients (26 men aged 38-74 years, mean 60 years) admitted with a definite diagnosis of myocardial infarction. Streptokinase therapy given intravenously at a mean of 5 h (range 1.5-15 h) after the onset of chest pain. Serum cholesterol concentration just prior to, and 11.5 h (range 4-20.5 h) after streptokinase administration. There was a significant mean fall of 0.4 mmol l-1 (P = 0.002, 95% CI = 0.2-0.6) in serum cholesterol concentration from a pre-streptokinase concentration of 7.0 (range 5.3-9.9) to a post-streptokinase concentration of 6.6 (range 4.9-9.9). In the patients who showed a fall in cholesterol concentration, the magnitude of fall correlated with the baseline cholesterol concentration (r = 0.66, P < 0.01) but not with peak cardiac enzyme activities (r = 0.05, P > 0.2 for aspartate aminotransferase; r = 0.10, P > 0.2 for lactate dehydrogenase), time from onset of chest pain to post-streptokinase measurement (r = 0.27, P > 0.2) or time from streptokinase administration to post-streptokinase measurement (r = 0.01, P > 0.2). Serum cholesterol concentration may be underestimated when measured after streptokinase therapy, particularly when the true basal value is high. Further management of this risk factor may be based more accurately on its measurement before than after streptokinase therapy within the first 24 h of AMI.


Novel purification system for 6xHis-tagged proteins by magnetic affinity separation.
We have developed a novel nickel-silica matrix for the generation of magnetic beads for metal-ion affinity chromatography. In contrast to magnetic Ni-NTA agarose beads, the novel particle type (SiMAC) consists of a magnetic core and a nickel-silica composite matrix with the nickel ions tightly integrated in the silica. This results in a much higher number of chelating groups compared with Ni-NTA agarose beads. With the SiMAC beads, greatly improved purification of histidine-tagged proteins from crude bacterial extracts was achieved. The yield was at least twice as high as with conventional materials, the method is faster, since the coupling step is omitted and there is no need for handling toxic Ni(2+) salts.


The serotonin 5-HT7 receptor agonist LP-44 microinjected into the dorsal raphe nucleus suppresses REM sleep in the rat.
The effects of LP-44, a selective 5-HT7 receptor agonist, and of SB-269970, a selective 5-HT7 receptor antagonist, on spontaneous sleep were studied in adult rats implanted for chronic sleep recordings. The 5-HT7 receptor ligands were microinjected directly into the dorsal raphe nucleus (DRN) during the light period of the 12-h light/12-h dark cycle. Infusion of LP-44 (1.25-5.0 mM) into the DRN induced a significant reduction of rapid-eye-movement sleep (REMS) and of the number of REM periods. Similar effects were observed after the direct administration into the DRN of SB-269970 (0.5-1.0 mM). Pretreatment with a dose of SB-269970 (0.5 mM) that significantly affects sleep variables antagonized the LP-44 (2.5 mM)-induced suppression of REMS and of the number of REM periods. It is proposed that the suppression of REMS after microinjection of LP-44 into the DRN is related, at least in part, to the activation of GABAergic neurons in the DRN that contribute to long projections that reach, among others, the laterodorsal and pedunculopontine tegmental nuclei involved in the promotion of REMS.


An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology.
Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins >150kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a 'resolution revolution', owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM.


Ailanthus altissima leaf extract mediated green production of zinc oxide (ZnO) nanoparticles for antibacterial and antioxidant activity.
Fabricating zinc oxide nanoparticles (ZnO-NPs) from plant extracts is a cost-effective, safe, and environmentally friendly alternative to established chemical procedures. This study was aimed at the environmentally friendly fabrication of ZnO-NPs from plant extract. An additional objective was to investigate the antibacterial and antioxidant activity of these biosynthesized ZnO-NPs. ZnO-NPs were fabricated using the leaf extract of Ailanthus altissima, as an eco-friendly approach. The physicochemical properties of ZnO-NPs were explored using UV-visible spectroscopy, scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectrometry. The bio-fabricated ZnO-NPs were examined for bactericidal activity against pathogenic bacteria (gram-negative and gram-positive) using the agar well diffusion technique. The antioxidant efficiency of ZnO-NPs was assessed using a DPPH assay. A surface Plasmon peak was recorded at 327 nm, showing the existence of ZnO-NPs in the reaction solution of plant extract and zinc sulfate hexahydrate salt. These nanoparticles were predominantly spherical and capped by different functional groups of biomolecules. Furthermore, ZnO-NPs showed a dose-dependent antibacterial and antioxidant activity. At 20 mg/mL ZnO-NPs, the maximum bactericidal potential of ZnO-NPs was reported against Staphylococcus aureus (201.2 mm). ZnO-NPs have an IC50 value of 78.23 µg/mL, indicating that they are an effective antioxidant. This research presents an environmentally acceptable method for producing spherical ZnO-NPs with high antibacterial and antioxidant activities. These bio-fabricated ZnO-NPs could be a good option for applications in medicine and the healthcare industry.


Cow's milk allergy in multiple sclerosis patients.
Exposure to some environmental agent such as different nutrition and contact with allergens may have a role in developing multiple sclerosis (MS). The present study was aimed to evaluate the cow's milk allergy (CMA) in MS patients compared to healthy controls. Between March 2012 and July 2012, 48 MS patients were selected and compared with 48 healthy subjectsto assess the frequency of CMA in MS patients compared to healthy control. Cow's milk specific immunoglobin E (IgE) was determined by Immuno CAP. Sex and the frequency of CMA were compared between study groups by Chi-square test. Total of 96 subjects were assessed (22% male and 78% female). The mean age of the study subjects was 30.8 ± 6.6 years. Mean age of case and control groups was 30.7 (±6.9) versus 30.9 ± 6.3, respectively (P value = 0.83). There were no detection of cow's milk specific IgE in serum of MS patients and healthy subjects. There was no difference between MS and healthy subjects regarding CMA.


Treatment of hypotension in newborns.
Systemic hypotension is a common complication of preterm birth affecting approximately one-third of very low-birthweight infants. There is considerable variation between neonatal units in the reported prevalence of hypotension, the threshold for therapeutic intervention and the nature of any cardiovascular support offered. Systemic hypotension is associated with adverse long-term neurodevelopmental outcome. The majority of preterm infants with hypotension have a normal or high left ventricular output, with low systemic vascular resistance often associated with a haemodynamically significant ductal shunt. Historically, volume expansion, dopamine and dobutamine have been the agents most commonly used to treat hypotension. Some hypotensive preterm infants have low cortisol levels, and corticosteroids are being used increasingly to prevent or treat hypotension in these babies.


Lymphocyte reactivity in patients with cancer of the oral cavity measured by sister chromatid differential staining.
Lymphocyte reactivity in phytohemagglutinin-stimulated peripheral lymphocyte cultures from 22 patients with squamous cell carcinoma of the oral cavity and 29 age- and sex-matched controls was measured by the bromodeoxyuridine-Giemsa method for demonstrating sister chromatid differential staining. One hundred metaphases from each donor were counted, and the proportions of cells in the first, second, and third cycles were scored to estimate the in vitro mitogen responsiveness and cell division rates of the lymphocytes. The in vitro reactivity of peripheral lymphocytes of cancer patients was impaired in comparison with that of the controls, and the degree of reactivity of lymphocytes in cancer patients was not related to the size (greatest diameter) of the primary tumor. Results are discussed in relation to previously existing data.


Student experiences and mentor views of the use of simulation for learning.
This paper presents the findings of a two phase mixed methods study. Phase 1 investigated whether simulation could support the development of a range of clinical skills amongst pre-registration adult and children's nursing students. The second phase of the study gathered mentors' views and experiences of the use of simulation in the preparation of students for practice. Commissioned by the Nursing and Midwifery Council, the United Kingdom (UK) professional body, the study is reported as one of 13 pilot sites using designated practice hours for simulation. The commission resulted from a call to review the current pre-registration nursing curriculum that includes 4600 equally divided theory and practice hours delivered across the programme. Phase 1 included a sample of 69 adult and children's pre-registration students from years one and three of their programme, studying at one UK University. The group attended five simulation sessions including basic life support, manual handling, infection control, clinical decision making and managing violence and aggression. Students completed pre- and post-tests in basic life support and manual handling, and vignettes and objective structured clinical examinations (OSCEs) covering the five areas of simulation. Phase 2 included interviews with six mentors who were supervising students involved in the study. Simulation was positively received by both students and mentors as it was apparent that it offered scope for interdisciplinary learning that could be broadened to inter-professional applications. The study also identified that the use of simulation could provide scope for collaborative working between education providers and clinical staff.


Integrated transcriptomic and quantitative proteomic analysis identifies potential RNA sensors that respond to the Ag85A DNA vaccine.
DNA vaccine has emerged as a promising approach with potential for Tuberculosis (TB) prevention in adults. However, the mechanism behind DNA vaccines is still largely unknown. Utilizing the CRISPR/Cas9 technique, we engineered Ag85A mutated dendritic cells (Ag85A-M-DCs) in which the Ag85A mRNA derived from Mycobacterium tuberculosis was expressed but not the corresponding protein. Control cells (Ag85A-DCs) expressed both Ag85A mRNA and protein. To better understand the mechanism of antigen presentation following DNA vaccination, integrated transcriptomic and proteomic analysis of dendritic cells (DCs), Ag85A-DCs, and Ag85A-M-DCs were performed. A total of 723, 278, and 933 differentially expressed genes (DEGs), and 209, 134, and 509 differentially expressed proteins (DEPs) were identified between Ag85A-M-DCs and DCs, Ag85A-DCs and DCs, and Ag85A-M-DCs and Ag85A-DCs, respectively. Integration analysis detected 59, 15, and 64 associated DEGs/DEPs with the same expression trend between Ag85A-M-DCs and DCs, Ag85A-DCs and DCs, and Ag85A-M-DCs and Ag85A-DCs, respectively. KEGG pathway analysis showed that chemokine signaling pathway and MAPK signaling pathway were enriched in all three pairs of comparisons. The protein and protein interaction network revealed that ANXA1 was in the top 10 high-degree hub genes closely related to other genes in all three pairs of comparisons. The results indicated that Ag85A DNA vaccine might transmit immunogenicity information and induce immune responses by activating chemokine signaling pathway and MAPK signaling pathway. ANXA1 may serve as a key target molecule of the Ag85A vaccine with additional potential for TB prevention.


[Spatial organization of musculature in the Himasthla elongata cercaria (Trematoda: Echinostomatidae)].
Somatic muscles (body-wall and "parenchyma" musculature), muscles of suckers, alimentary tract and excretory bladder of Himasthla elongata cercaria were investigated using fluorescent phalloidin labelling and confocal microscopy. The arrangement of body-wall muscles differs between the certain parts of cercarial body and appears to be the most complicated in the collar district. Among the body-wall musculature, we described U-shaped muscles, which have never been found previously in trematodes. Muscles of oral and ventral suckers are grouped into 6-7 independent layers. In some of those layers, they are arranged bilaterally, which contradicts the tradition to consider the sucker as radially symmetric.


Human Milk, Infant Formula, and Other Milks Fed to Infants and Toddlers in the United States, NHANES 2007-2018.
For the first time, the 2020-2025 Dietary Guidelines for Americans provide specific guidance regarding the types of foods and beverages that should be offered in the first 2 years of life. Milk, in various forms (eg, human milk, infant formula, and cow's milk) contributes a large proportion of key nutrients to the diets of infants and toddlers in the United States. The aim of this study was to determine the types of milk (human milk, infant formula, and other milk) fed to US infants and toddlers in the past 12 years and to describe trends over time. This was a cross-sectional analysis of 2-day, 24-hour dietary recalls. Data from the 2007-2018 National Health and Nutrition Examination Survey were used for these analyses. Infants and toddlers aged 0 through 23.9 months with 2 days of dietary recall data (n = 3,079) were included. The main outcome was proportion of infants and toddlers fed different milk types. Survey-adjusted weighted percentages were used to report sociodemographic characteristics and the proportion of subjects fed each milk type category by age group and survey cycles. Binary and multinomial logistic regressions were used to assess differences in subject characteristics by age groups. Sociodemographic characteristics did not differ by age group. The proportion of infants aged 0 to <6 months fed infant formula only was 60.2% in 2007-2012 and 44.8% in 2013-2018. The proportion of infants aged 6 to <12 months fed partially hydrolyzed infant formula only was 7.3% in 2007-2012 and 13.1% in 2013-2018. In toddlers (>12 months old), cow's milk was the predominant milk type in both 2007-2012 and 2013-2018. The percentage of infants fed any human milk increased over the past decade. Unsweetened cow's milk was the most predominate milk type consumed among toddlers.


Porous molecularly imprinted monolithic capillary column for on-line extraction coupled to high-performance liquid chromatography for trace analysis of antimicrobials in food samples.
A novel porous molecularly imprinted monolithic capillary column (MIMCC) based on ternary porogen was synthesized by in situ technique with sulfaquinoxaline as the template molecule. The characteristics of the MIMCC were investigated by scanning electron microscopy, infrared spectrum, thermogravimetric analysis and solvent resistance test. The saturated adsorption amount of sulfaquinoxaline on MIMCC was 2.7 times over that on the non-imprinted monolithic capillary column (NIMCC). The MIMCC also exhibited good enrichment ability to its analogs and the enrichment factors were 46-211 for five antimicrobials. High permeability and imprinting factors as well as good stability, reproducibility and long lifetime were obtained. An on-line method based on MIMCC solid-phase microextraction coupled with high-performance liquid chromatography was developed for the determination of trace antimicrobials in complex samples. The good linearity for sulfametoxydiazine, sulamethoxazole and sulfaquinoxaline was 0.05-10 µg/L, the limits of detection (LODs) were 10.0-14.0 ng/L. The linear range for mequindox and quinocetone were 0.10-10.0 µg/L, the LODs were 20.0-27.0 ng/L respectively. The recoveries were 71.0-108.2% with relative standard deviation of 1.6-8.5%, correspondingly. The results showed that MIMCC could effectively enrich antimicrobials from complex matrices. The on-line method based on MIMCC and HPLC was selective, sensitive and convenient for trace determination of antimicrobials in complex samples.


Mechanistic insights into the role of serum-glucocorticoid kinase 1 in diabetic nephropathy: A systematic review.
Aberrant expression of serum-glucocorticoid kinase 1 (SGK1) contributes to the pathogenesis of multiple disorders, including diabetes, hypertension, obesity, fibrosis, and metabolic syndrome. SGK1 variant is expressed in the presence of insulin and several growth factors, eventually modulating various ion channels, carrier proteins, and transcription factors. SGK1 also regulates the enzymatic activity of Na+ K+ ATPase, glycogen synthase kinase-3, ubiquitin ligase Nedd4-2, and phosphomannose mutase impacting cell cycle regulation, neuroexcitation, and apoptosis. Ample evidence supports the crucial role of aberrant SGK1 expression in hyperglycemia-mediated secondary organ damage. Diabetic nephropathy (DN), a dreadful microvascular complication of diabetes, is the leading cause of end-stage renal failures with high morbidity and mortality rate. The complex pathogenesis of DN encompasses several influencing factors, including transcriptional factors, inflammatory markers, cytokines, epigenetic modulators, and abnormal enzymatic activities. SGK1 plays a pivotal role by controlling various physiological functions associated with the occurrence and progression of DN; therefore, targeting SGK1 may favorably influence the clinical outcome in patients with DN. This review aimed to provide mechanistic insights into SGK1 regulated DN pathogenesis and summarize the evidence supporting the therapeutic potential of SGK1 inhibition and its consequences on human health.


The pathogenesis of microcephaly resulting from congenital infections: why is my baby's head so small?
The emergence of Zika-virus-associated congenital microcephaly has engendered renewed interest in the pathogenesis of microcephaly induced by infectious agents. Three of the original "TORCH" agents are associated with an appreciable incidence of congenital microcephaly: cytomegalovirus, rubella virus, and Toxoplasma gondii. The pathology of congenital microcephaly is characterized by neurotropic infectious agents that involve the fetal nervous system, leading to brain destruction with calcifications, microcephaly, sensorineural hearing loss, and ophthalmologic abnormalities. The inflammatory reaction induced by these four agents has an important role in pathogenesis. The potential role of "strain differences" in pathogenesis of microcephaly by these four pathogens is examined. Specific epidemiologic factors, such as first and early second trimester maternal infection, and the manifestations of congenital infection in the infant, shed some light on the pathogenesis. Immune aspects of normal pregnancy and their role in congenital infections is examined. In this review, we integrate all these findings to create a unified hypothesis of the pathogenesis of congenital microcephaly induced by these infectious agents.


Biochemical research into psychosis.
A strategy is presented for biological psychosis research with neuroleptics acting as a point of crystallisation like antidepressants do in biological depression research. The neuroleptics chlorpromazine, haloperidol and oxypertine were studied, and it was found that they influence central catecholamine (CA) metabolism in man. An increased central dopamine (DA) turnover was found to occur in psychotic disorders, mostly in the form of motor agitation. As the first of a planned series of studies, chlorpromazine with presumed ability to reduce both DA-ergic and noradrenaline (NA)-ergic transmission and oxypertine as a more selective blocker of NA-ergic transmission were selected for comparison. The overall therapeutic effect of oxypertine was inferior to that of chlorpromazine, whereas oxypertine proved more effective in cases where loss of initiative was predominant. On the other hand, chlorpromazine exerted a more marked influence on extrapyramidal motor functions than oxypertine. In chronic psychotic disorders with inertia, oxypertine thus seems to be a neuroleptic which is strong enough to prevent exacerbation of delusions and hallucinations while at the same time increasing the level of motivation. These findings were in accordance with our predictions. The comparative study is illustrative of the practical significance of the research approach in this study: The biochemical action profile of a neuroleptic seems to be a more reliable indicator of its clinical action than does its chemical structure.


Evaluation of the performance of osteoporosis/fracture screening models to identify high-risk women for osteoporosis: Bushehr elderly health (BEH) program.
This study aimed to evaluate the performance of valid risk assessment models developed for osteoporosis/ fracture screening to identify women in need of bone density measurement in a population of Iranian elderly women. This study was performed using the data of Bushehr Elderly Health (BEH) program, a population-based cohort study of elderly population aged ≥ 60 years. Seven osteoporosis risk assessment tools, including Osteoporosis Risk Assessment Instrument (ORAI), Malaysian Osteoporosis Screening Tool (MOST), Osteoporosis Prescreening Risk Assessment (OPERA), Osteoporosis Prescreening Model for Iranian Postmenopausal women (OPMIP), Osteoporosis Index of Risk (OSIRIS), and Osteoporosis Self-Assessment Tool for Asians (OSTA), as well as Fracture Risk Assessment Tool (FRAX) were included in the study. By using osteoporosis definition based on BMD results, the performance measurement criteria of diagnostic tests such as sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and Youden index for each model were calculated and the models were compared. A total of 1237 female participants with the mean age of 69.1 ± 6.3 years were included. Overall, 733 (59%) participants had osteoporosis, and about 80% had no history of fracture. The sensitivity of the seven models ranged from 16.7% (OSIRIS) to 100% (ORAI and MOST) at their recommended cut-off points. Moreover,their specificity ranged from 0.0% (ORAI and MOST) to 78.9% (OSTA). The FRAX and OPERA had the optimal performance with the Youden index of 0.237 and 0.226, respectively. Moreover, after combining these models, the sensitivity of them increased to 85.4%. We found that the FRAX (model with 11 simple variables) and OPERA (model with 5 simple variables) had the best performance. By combining the models, the performance of each was improved. Further studies are needed to adopt the model and to find the best cut-off point in the Iranian postmenopausal women.


Temporal effects of two interferential current applications on peripheral circulation in children with hemiplegic cerebral palsy.
To determine the short-term effects of sympathetic and peripheral stimulation of interferential current (IFC) on blood flow (BF) in children with hemiplegic cerebral palsy (CP). Thirty children with hemiplegic CP, ranging from 8 to 12 years old, were randomly divided into three groups (10 children/group). The first group received sympathetic stimulation of IFC, the second group received peripheral stimulation of IFC, and the third group (control) received placebo peripheral stimulation of IFC. A frequency of 80-100 Hz at an intensity of 10-20 mA was applied for 20 min. Blood volume pulse (BVP) amplitude was measured before IFC application using a plethysmography sensor at the big toe immediately after and 15 min poststimulation. The data were statistically analyzed and compared. There were statistically significant differences in BVP amplitude among the three time intervals in both the sympathetic and peripheral groups (P < 0.05) with no difference in the control group (P = 0.995). There was a significant increase in BVP amplitude immediately after stimulation compared with before stimulation in both the sympathetic and peripheral groups (P = 0.0001). However, differences between the sympathetic and peripheral groups at the three measured periods were statistically nonsignificant (P > 0.05). Both IFC applications had a proper effect on improving BF in children with hemiplegic CP with no difference in efficacy between sympathetic and peripheral stimulation.


Catecholamine depletion by reserpine blocks the anxiolytic actions of ethanol in the rat.
Neurochemical investigations of the anti-anxiety action of ethanol demonstrate that increased dopaminergic, noradrenergic, and serotonergic activity mediates the anxiolytic actions of ethanol. Results of studies with animals and human beings also confirm an involvement of the sympathetic nervous system in the behavioral actions of ethanol. Because enhanced sympathetic activity increases the release of norepinephrine from sympathetic terminals, the interruption of normal sympathetic activity might disrupt the anxiolytic action of ethanol. The present study examined the effect of chemical sympathectomy by reserpine pretreatment on the subsequent anxiolytic action of ethanol. Seventy-one, female, Sprague-Dawley rats were administered reserpine (0 or 5 mg/kg), followed 17-19 h later by ethanol (0, 0.5, or 1.0 g/kg). Animals were then tested in the elevated plus maze. Compared with saline pretreatment, which did not attenuate the anxiolytic actions of a 1.0-g/kg dose of ethanol, reserpine pretreatment completely blocked the anxiolytic action of a 1.0-g/kg dose of ethanol. Reserpine, by itself, did not possess anxiolytic or anxiogenic actions. Because the anxiolytic action of ethanol involves increased catecholamine activity and the depletion of norepinephrine and dopamine from sympathetic nerve terminals by reserpine blocked the anxiolytic action of ethanol, it is concluded that the anxiolytic action of ethanol requires the presence of normal catecholamine activity. We suggest that prostaglandin activity normally evoked by enhanced sympathetic nervous system activity might be involved in the anxiolytic action of ethanol.


Overweight and Obesity: Prevalence, Consequences, and Causes of a Growing Public Health Problem.
This review considers a variety of perspectives on overweight and obesity (OW/obesity), including measurement and classification; prevalence and changes in prevalence in recent years; genetic, biological, medical, individual, and social correlates of OW/obesity; and treatment approaches. Despite increased attention, OW/obesity is escalating in prevalence worldwide, and the causes are exceedingly complex. A range of innovative studies, including basic research on gut microflora, dietary composition, pharmacologic interventions, and surgical procedures, is generating findings with potential for future prevention and treatment of OW/obesity. Social system changes such as school programs and the awareness of the roles of personal, family, health provider, and cultural experiences related to OW/obesity have also gained traction for vital prevention and treatment efforts over the past decade.


Characterisation of non-coding genetic variation in histamine receptors using AnNCR-SNP.
Almost 90 % of disease-associated genetic variants found using genome wide association studies (GWAS) are located in non-coding regions of the genome. Such variants can affect phenotype by altering important regulatory elements such as promoters, enhancers or repressors, leading to changes in gene expression and consequently disease, such as thyroid cancer and allergic diseases. A number of allergy and atopy related diseases such as asthma and atopic dermatitis are related to histamine receptors; however, these diseases are not fully characterized at the molecular level. Moreover, candidate gene based studies of common variants known as single nucleotide polymorphism (SNPs) located in the coding regions of these receptors have given mixed results. It is important to complement these approaches by identifying and characterising non-coding variants in order to further elucidate the role of these receptors in disease. Here we present an analysis of histamine receptor genes using the tool AnNCR-SNP to characterise variants in non-coding genomic regions. AnNCR-SNP combines bioinformatics and experimental data sets from various sources to predict the effects of genetic variation on gene expression regulation. We find many SNPs located in areas of open chromatin, overlapping with transcription factor binding sites and associated with changes in gene expression in expression quantitative trait loci (eQTL) experiments. Here we present the results as a catalogue of non-coding variation in histamine receptor genes to aid histamine researchers in identifying putative functional SNPs found in GWAS for further validation, and to help select variants for candidate gene studies.


Postoperative stability of bioresorbable plates made of 85:15 poly (L-lactide-co-glycolide) in Le Fort I osteotomy.
Recently, RapidSorb plates (DePuy Synthes) made of 85.15 poly (L-lactide-co-glycolide) have been used for orthognathic surgery; however, reports regarding their effectiveness are limited. We aimed to compare the postoperative stability of RapidSorb plates, RapidSorb combined with titanium (MOJ plates), and MOJ plates in patients who underwent Le Fort I osteotomy at Tokyo Medical and Dental University Hospital. The use of RapidSorb in the maxilla is a load-sharing application and therefore constitutes an approved indication. Discrepancies in the maxillary positions were measured using postoperative computed tomography data at 1 week and 1 year using the centroid method 3-dimensionally. Treatment with RapidSorb alone showed a more vertical discrepancy in the maxilla treatment with MOJ and RapidSorb+MOJ. The RapidSorb4 group was subdivided into 2 groups (under and over 1.0-mm) based on the change in the maxillary centroid. The bone gap at the lateral border of the piriform aperture was significantly larger in the over-1.0-mm group than in the 1.0-mm group. The fixation of RapidSorb alone is not appropriate in load-bearing and unstable applications but is not contraindicated for load-sharing indications. Fixation with RapidSorb combined with MOJ was clinically effective, with results similar to titanium plate-only fixation regarding postoperative stability.


Recurrent aspiration in children: lipid-laden alveolar macrophage quantitation.
We prospectively evaluated bronchial washings from 45 patients aged 1 month to 25 years (mean 3.3 years) for the presence of lipid-laden macrophages. By grading the amount of intracellular oil-red-O-positive material, we determined a semiquantitative lipid-laden macrophage index for each patient. All patients observed to be definite aspirators (n = 22) had indexes greater than or equal to 86 (mean +/- SD, 139 +/- 46). All patients with no clinical suspicion of aspiration (n = 23) had indexes less than or equal to 72 (mean +/- SD, 21 +/- 20). Our results suggest that simply sighting lipid-laden macrophages in a nonspecific finding, but quantitation of these cells may be a very good test for recurrent aspiration of food substances in children. This test appears to be more sensitive than radiographic studies.


Patients' experiences of continuity in the care of type 2 diabetes: a focus group study in primary care.
Continuity of care is fundamental to general practice and type 2 diabetes is a common chronic disease with major health and social impacts. Nevertheless continuity, as experienced by patients with type 2 diabetes, remains a neglected area. To explore perceptions and experiences of continuity of care in general practice from the perspectives of patients with type 2 diabetes, focusing on the advantages and disadvantages of different types of continuity. Focus groups with patients. Seven practices with different organisational structures in Leeds, UK. Seventy-nine patients with type 2 diabetes were recruited. Focus group interviews were conducted with 79 patients with type 2 diabetes from seven practices in Leeds, UK. Patients experienced three different types of continuity: relational (or longitudinal) continuity, cross-boundary (or team) continuity, and continuity of information. Patients' perceptions of continuity were influenced by several factors including a personal relationship between themselves and their healthcare professional, their own beliefs and behaviours, presence of diabetes, and the systems and structures of general practices. Patients identified the advantages and disadvantages of two types of continuity. Relational or longitudinal continuity was important in providing psychosocial care, but with a risk of misdiagnosis. The advantages of cross-boundary or team continuity were to provide physical care, whereas the main disadvantages were the absence of personal care and patient confusion. Perceptions of continuity by patients with type 2 diabetes were influenced by several factors; they perceived several advantages and disadvantages associated with different types of continuity. Patients might expect certain healthcare benefits by following certain types of continuity.


Work of IEEE P1157 Medical Data Interchange committee.
The Institute of Electrical and Electronics Engineers (IEEE) has been working for several years to develop medical system communication standards, under sponsorship of the Engineering in Medicine and Biology Society (EMBS). The work of the IEEE P1157 Medical Data Interchange committee is surveyed in this paper. By 1990 the IEEE P1157 Medical Data Interchange committee plans to have a draft standard ready for ballot, with an initial focus on the interface transactions needed to support a patient care information system. Eventually, the IEEE P1157 Medical Data Interchange committee will develop standards with a wider scope, including the transfer of electronic medical records between different vendor systems.


Evaluation and improvement of different methods for the enrichment of antigen-binding cells of chickens.
Antigen-binding cells were enriched using bovine serum albumin immunized chickens as an experimental model. A comparison of density centrifugation, adherence to derivatized surfaces (polystyrol and gelatin) and affinity chromatography revealed that optimal enrichment was obtained using the latter procedure. Polyacrylamide was superior to agarose.


Hemangiosarcoma of the thyroid gland. Histological diagnosis problems.
A bleeding tumor of the thyroid gland in a 69-year-old woman is presented. Histologically, it was a hemangiosarcoma with proliferation and vessel neoformation: vascular-like clefts or distinct capillaries with tumoral endo- and perithelial structures were present. With reference to the published data, problems of differential diagnosis in the undifferentiated and giant cell carcinomas of the thyroid gland are discussed.


Force-velocity relationship and maximal power on a cycle ergometer. Correlation with the height of a vertical jump.
The force-velocity relationship on a Monark ergometer and the vertical jump height have been studied in 152 subjects practicing different athletic activities (sprint and endurance running, cycling on track and/or road, soccer, rugby, tennis and hockey) at an average or an elite level. There was an approximately linear relationship between braking force and peak velocity for velocities between 100 and 200 rev.min-1. The highest indices of force P0, velocity V0 and maximal anaerobic power (Wmax) were observed in the power athletes. There was a significant relationship between vertical jump height and Wmax related to body mass.


Clinical determination of bone quality: is ultrasound an answer?
Progress in clinical characterization of bone relies on developing a means to clinically assess all of the important determinants of bone quality, specifically, the intrinsic material properties of a bone (stiffness and brittleness) versus the macroscopic structural properties [apparent mass density (g/cc), structural shape and distribution of cortical mass, trabecular architecture, extent of unrepaired microdamage, and defects associated with the accelerated remodeling in early menopause]. Ultrasound devices currently measure parameters related to either of only two basic properties: bone ultrasound attenuation (BUA) or the apparent velocity of wave propagation (AVU). Theory and repeated corroboration in the laboratory have shown that the velocity of sound in solids such as bone has a quantitative relationship to the elastic modulus (or stiffness) and mass density. Although no comparable physical model exists for BUA, growing in vitro and in vivo empirical evidence shows a relationship to stiffness and mass density as well. Therefore, the question of ultrasound's ability to provide additional, clinically useful information about bone quality reduces to this: Does bone quality depend significantly on bone stiffness and does stiffness depend on factors other than bone mass alone? Clinical study results provide mounting evidence of ultrasound's abilities. (1) Numerous studies compare either velocity or BUA with BMC or BMD. The correlation coefficients vary widely between studies, even when repeated by the same investigators and laboratories. Two studies demonstrated this by comparing groups of subjects who are indistinguishable by BMD at the lumbar spine, but whose mean AVU readings are significantly different.(ABSTRACT TRUNCATED AT 250 WORDS)


Improved synthesis and in vitro characterization of chitosan-thioethylamidine conjugate.
The aim of this study was to establish improved reaction conditions for the synthesis of chitosan-thioethylamidine (Ch-TEA) conjugate and to evaluate the properties of the obtained Ch-TEA conjugate. The influence of different factors on the coupling reaction, such as concentration of chitosan solution, employment of reducing agent and deprotection of S-acetyl groups, was evaluated. The cohesive properties and stability of the obtained conjugate were evaluated by disintegration test and by oxidation experiments, respectively. The adhesive properties of Ch-TEA conjugate were evaluated in vitro on freshly excised porcine mucosa via tensile studies and the rotating cylinder method. The permeation-enhancing effect of Ch-TEA conjugate was evaluated in Ussing-type chambers by using rhodamine 123 as model compound. The resulting conjugate displayed 225 micromol immobilized free thiol groups and 102 micromol disulfide bonds per gram polymer. The degree of modification depends mostly on the chitosan concentration employed and the deprotection of S-acetyl groups with hydroxylamine. During oxidation studies the amount of thiol groups decreased by 61%. Disintegration studies of tablets comprising Ch-TEA demonstrated stability for 48 h. In tensile studies, the total work of adhesion of the conjugate was determined to be 5.1-fold increased in comparison to unmodified chitosan. Results from the rotating cylinder method showed more than a 13-fold increase in the adhesion time of thiolated chitosan versus unmodified chitosan. The apparent permeability coefficient (Papp) of the system 0.5% (w/v) Ch-TEA conjugate with 5% (w/v) glutathione was calculated to be 5.35 x 10(-8)cm/s, while the Papp value of the system 0.5% (w/v) unmodified chitosan was 1.73 x 10(-8)cm/s. These features should render Ch-TEA useful as an excipient for various drug delivery systems.


Chemicals of concern in personal care products used by women of color in three communities of California.
Personal care products (PCPs) may contain chemicals associated with adverse health effects. Prior studies found differences in product use by race/ethnicity and suggest some women are disproportionately exposed to chemicals of concern (CoCs). We quantified chemicals linked to cancer, reproductive or developmental harm, or endocrine disruption in PCPs used by women of color. We documented PCPs in stores frequented by Black, Latina, and Vietnamese women in their communities in California and CoCs on ingredient labels of 546 unique hair, skin, makeup, nail, deodorant/perfume, and intimate care products. Community partners chose 31 products for a combined targeted and suspect screen (National Institute of Standards and Technology mass spectral library search) two-dimensional gas chromatography time-of-flight mass spectrometry (GCxGC-TOFMS) analysis to detect chemicals not on ingredient labels. We found that 65% of labels included CoCs, and 74% of labels had undisclosed ingredients listed as "fragrance." The most prevalent chemicals were parabens, cyclosiloxanes, and formaldehyde releasers. GCxGC-TOFMS found additional CoCs, including fragrances, solvents, preservatives, ultraviolet filters, and contaminants. These findings contribute to awareness of potentially hazardous chemicals in PCPs, can help estimate disparities in chemical exposure, and complement research on health inequities due to chemical exposures from various contributors. This study is one of the first detailed assessments of chemicals of concern found in various types of PCPs used by several racial/ethnic groups. We found that over half of the 546 products selected by community partners as marketed to and/or used by them contained ingredients linked to cancer, reproductive or developmental harm, or endocrine disruption. Laboratory analysis identified additional chemicals in a subset of products, including unlabeled fragrance chemicals and contaminants. Elucidating exposures to chemicals in PCPs is important for risk assessment and health inequity research.


Connarus favosus Planch.: An inhibitor of the hemorrhagic activity of Bothrops atrox venom and a potential antioxidant and antibacterial agent.
The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to treat snakebites. To investigate the potential of the aqueous extract of Connarus favosus (AECf) to inhibit hemorrhagic and phospholipase A2 activities induced by Bothrops atrox venom (BaV) and to determine the antioxidant and antimicrobial potentials of the extract. AECf was analyzed phytochemically for phenolics (condensed tannins and hydrolyzable tannins) by colorimetry. Antioxidant activity was evaluated by quantitative assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Fe(3+)/phenanthroline. Antimicrobial activity was evaluated by the minimal inhibitory concentration test, and cytotoxicity was evaluated using human fibroblast cells (MRC-5). Inhibition of BaV-induced hemorrhagic activity was assessed after oral administration of the extract using pre-treatment, post-treatment and combined (BA plus AECf) treatment protocols. Inhibition of indirect hemolysis caused by phospholipase A2 (PLA2) was investigated in vitro. Interaction between AECf and BaV was investigated by SDS-PAGE electrophoresis, Western blot (Wb) and zymography. The phytochemical profile of AECf revealed ten secondary metabolite classes, and colorimetry showed high total phenolic and total (condensed and hydrolyzable) tannin content. AECf exhibited high antioxidant and antimicrobial potentials. The IC50 for the cytotoxic effect was 51.91 (46.86-57.50)µg/mL. Inhibition of BaV-induced hemorrhagic activity was significant in all the protocols, and inhibition of PLA2 activity was significant with the two highest concentrations. The BaV/AECf mixture produced the same bands as BaV by itself in SDS-PAGE and Wb although the bands were much fainter. Zymography confirmed the proteolytic activity of BaV, but when the venom was pre-incubated with AECf this activity was blocked. AECf was effective in reducing BaV-induced hemorrhagic activity when administered by the same route as that used in folk medicine and exhibited antioxidant and antimicrobial potentials.


Mechanism of non-contact ACL injury: OREF Clinical Research Award 2021.
Anterior cruciate ligament (ACL) ruptures significantly impact athletes in terms of return to play and loss of long-term quality of life. Before the onset of this study, understanding the mechanism of ACL injury was limited. Thus, the primary focus of this manuscript is to describe our multi-faceted approach to uncovering the mechanism of noncontact ACL injury (NC-ACLI) with the goal of developing preventive strategies. The initial qualitative analysis of ACL injury events revealed most (70%) injuries involve minimal to no contact and occurr during landing or deceleration maneuvers in team sports with a minor perturbation before the injury that may disrupt the neuromuscular system leading to poor body dynamics. A series of quantitative videotape studies demonstrated differences in leg and trunk positions at the time of NC-ACLI in comparison to control subjects. Analysis of the faulty dynamics provoking NC-ACLI, especially the flat-footed landing component, supports the theory that an axial compressive force is the critical factor responsible for NC-ACLI. Our magnetic resonance imaging study demonstrated the NC-ACLI position was associated with a higher tibial slope, and joint contact occurring on the flat, anterior portion of the lateral femoral condyle versus the round, posterior aspect. Both anatomic conditions favor sliding (pivot shift) over rolling in the presence of an axial compressive force. Subsequent cadaveric studies supported axial compressive forces as the primary component of NC-ACLI. Both a strong eccentric quadriceps contraction and knee abduction moments may increase the compressive force at the joint thereby lowering the axial threshold to injury. This manuscript summarizes the NC-ACLI mechanism portion of the 2021 OREF Clinical Research Award.


Evolutionary insights into photoregulation of the cell cycle in the green lineage.
Plant growth depends solely on light energy, which drives photosynthesis. Thus, linking growth control to light signals during certain developmental events, such as seed or spore germination and organ formation, is a crucial feature that plants evolved to use energy efficiently. How light controls the cell cycle depends on growth habitats, body plans (unicellular vs. multicellular), and photosensors. For example, the photosensors mediating light signaling to promote cell division appear to differ between green algae and land plants. In this review, we focus on cell-cycle regulation by light and discuss the transition of its molecular mechanisms during evolution. Recent advances show that light-dependent cell-cycle control involves global changes in transcription of cell-cycle genes, and is mediated by auxin and cytokinin.


Outpatient Robotic Unilateral Extravesical Ureteral Reimplantation in the Pediatric Population: Short-Term Assessment of Safety.
Robotic extravesical ureteral reimplantation has been established as a viable option for surgical management of vesicoureteral reflux. Typically this procedure is associated with a hospital stay for routine postoperative care. We assessed the short-term safety of robotic unilateral extravesical ureteral reimplantation as a scheduled outpatient procedure in a pediatric population. We retrospectively studied a cohort of patients who underwent robotic extravesical ureteral reimplantation between June 2012 and January 2018. No regional blocks were performed. Patients were discharged from the postanesthesia care unit as part of a scheduled outpatient procedure without an extended stay. Postoperative outcomes included 30-day emergency room visits, readmissions to the hospital and Clavien-Dindo grade I to V complications. Four male and 23 female patients were identified. Median age was 85 months (range 27 to 210) and median weight was 26 kg (13 to 97). Median robotic console time was 140 minutes (range 84 to 257). No patient required a hospital stay for management of pain. Two patients (9%) required unplanned antibiotic therapy postoperatively for bacterial cystitis and pneumonia (Clavien-Dindo grade II complications). The patient with pneumonia was diagnosed during a subsequent emergency room visit. One patient was rehospitalized on postoperative day 4 because of constipation. No Clavien-Dindo grade III or higher complication was observed in any patient. Robotic unilateral extravesical ureteral reimplantation is safe as an outpatient procedure in the pediatric population. Further evaluation is warranted to assess its short and long-term outcomes on a larger scale.


Ontogeny of the metencephalic, mesencephalic and diencephalic content of catecholamines as measured by high performance liquid chromatography with electrochemical detection.
Developmental changes in norepinephrine (NE), dopamine (DA) and epinephrine (E) contents of the rat metencephalon, mesencephalon and diencephalon, have been measured by high performance liquid chromatography with electrochemical detection, from fetal stages (E15 or E17 to E21) to postnatal days (P0 to P30) and compared to the adult levels. The data show a biphasic pattern in NE changes of the three brain areas, with a first increase in the late prenatal period, followed by a further development from P0 to P18, thus reaching the adult levels. A similar pattern of development is found for the mesencephalic and diencephalic DA contents. The E levels of the diencephalon are very low in comparison to the NE and DA concentrations, but present a gradual increase from E17 to P18. The results correlate with the development of catecholamine systems in brain area as measured by other methodological approaches.


Infection in atopic dermatitis.
Atopic dermatitis is a chronic inflammatory skin disease commonly seen in children, but it also occurs in adults. This skin disease is often triggered by bacterial, fungal, or viral skin infections. The mechanisms resulting in this increased propensity for skin infections have been an area of active investigation. Recent studies suggest that the skin of patients with atopic dermatitis has increased avidity for binding to Staphylococcus aureus and is deficient in its ability to generate antimicrobial peptides needed to eradicate infectious agents. The current review will examine recent literature on the etiology and pathogenesis of infection in atopic dermatitis. These observations are likely to have importance implications for management of atopic dermatitis.


Murine monoclonal antibodies against a unique determinant of erythrocytes, related to Rh and U antigens: expression on normal and malignant erythrocyte precursors and Rhnull red cells.
Three murine monoclonal antibodies (Mabs) MB-2D10, LA-18.18 and LA-23.40 were prepared. They reacted with red cells of all common and most rare blood-group phenotypes, with the exception of those of the RhnullU negative and RhmodU negative phenotypes. So far, only a single example of an alloantibody (Duclos or anti-Rh38) of a similar specificity has been found. Serological studies indicated that the Mabs were probably not directed against an antigenic determinant of Rh polypeptides, the LWab glycoprotein or glycophorin B, all structures absent from or aberrantly expressed on Rhnull red cells. The antigen was found to be erythrocyte-specific, and was also present on pro-erythroblasts, erythroblasts and malignant erythroblastoid cells but not on erythroid progenitors in the bone marrow. The Mabs were found to block each other in an immune rosette method and are thus probably directed against the same epitope or against neighbouring epitopes on the same structure. In immunochemical studies, MB-2D10 precipitated the 30-32 kDa Rh polypeptides from red cell membranes and a protein or proteins which formed diffuse and overlapping bands in SDS-polyacrylamide gel electrophoresis, with Mrs of 40-200 kDa (probably the Rh-related glycoproteins). Under certain experimental conditions glycophorin B appeared to be coprecipitated. The 2D10 structure, detected by the Mabs, seems to be part of a complex of proteins and/or glycoproteins, which includes Rh polypeptides, the LWab glycoprotein and glycoproteins recognized by various Mabs with Rh-related specificities. In the red cell membrane, the complex may be associated with glycophorin B.


White matter hyperintensities are not associated with cognitive decline in early Parkinson's disease - The DeNoPa cohort.
Small vessel disease and white matter hyperintensities (WMH) as its surrogate marker are known to predict cognitive decline in the elderly. However, the influence of vascular lesions on cognitive impairment in Parkinson's disease (PD) has been discussed controversial so far. The Aim of this study was to evaluate the predictive role of volume and location of white matter hyperintensities (WMH) on cognitive decline in de novo PD patients. 108 diagnosed drug-naïve PD patients (64 ± 9 years, 38% women) from the DeNoPa Cohort underwent extensive neuropsychological testing with re-testing in 24-month later. Movement Disorder Society criteria for the classification of mild cognitive impairment (MCI) and dementia in PD were applied. Participants that declined from normal cognition or MCI at baseline to MCI or dementia at 24-month follow-up (FU) or from MCI to dementia at 24-month FU were defined as "converters". Subjects with stable cognitive level or improved cognitive status were classified as "non-converters". Magnetic resonance imaging (MRI) was performed, and the extent of WMH was assessed as global volume and as WMH load within cholinergic pathways using the Cholinergic Pathways Hyperintensities Scale. We compared Parkinson's disease subjects with age-matched, neurologically healthy controls. At total of 29 (27%) patients met the criteria for MCI at baseline, whereas 79 (73%) patients had no cognitive impairment. During the 24-month FU 33 patients showed cognitive decline ("converter") compared to 75 "non-converters". Multivariable logistic regression revealed no significant differences between "cognitively impaired" and "cognitively non-impaired" patients and participants of the control group at baseline or between "converter" and "non-converter" regarding the extent of WMH globally or within cholinergic pathways. We could not identify global or localized WMH load as predictive markers of cognitive decline in de novo PD patients indicating that cerebral small vessel disease is not a critical modifier of cognitive function in early PD.


Regulatory potential of COUP-TFs in development: stem/progenitor cells.
The formation of complex organisms is highly dependent on the differentiation of specialized mature cells from common stem/progenitor cells. The orphan nuclear receptors chicken ovalbumin upstream promoter transcription factors (COUP-TFs) are broadly, but not ubiquitously, expressed in multiple tissues throughout embryonic development and COUP-TFs are indispensible for proper organogenesis. Recently, growing evidence suggests a critical role of COUP-TFs in multiple aspects of stem/progenitor cell biology. In this review, we highlight the progress of COUP-TFs function and its underlying mechanism in driving stem/progenitor cell self-renewal, lineage specification, differentiation, maintenance, and cell identity in diverse tissue types. These studies provide novel insights into future clinical utilities of COUP-TFs in stem cell based therapies and in the management of diseases.


Balanitis circumscripta plasmacellularis: case report with ultrastructural study.
A typical case of balanitis circumscripta plasmacellularis is described in a 74-year-old man. Electron microscopy revealed a dense infiltrate composed mainly of plasma cells and macrophages; the former showed considerable rough endoplasmic reticulum and phagolysosomes, while the latter contained variable amounts of sideromes. A few mast cells, eosinophils and extravasated erythrocytes were also present. Connective tissue was scarce; no elastic fibres and viral particles were seen.


Integrated bioinformatics-cheminformatics approach toward locating pseudo-potential antiviral marine alkaloids against SARS-CoV-2-Mpro.
The emergence of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) with the most contagious variants, alpha (B.1.1.7), beta (B.1.351), delta (B.1.617.2), and Omicron (B.1.1.529) has continuously added a higher number of morbidity and mortality, globally. The present integrated bioinformatics-cheminformatics approach was employed to locate potent antiviral marine alkaloids that could be used against SARS-CoV-2. Initially, 57 antiviral marine alkaloids and two repurposing drugs were selected from an extensive literature review. Then, the putative target enzyme SARS-CoV-2 main protease (SARS-CoV-2-Mpro) was retrieved from the protein data bank and carried out a virtual screening-cum-molecular docking study with all candidates using PyRx 0.8 and AutoDock 4.2 software. Further, the molecular dynamics (MD) simulation of the two most potential alkaloids and a drug docking complex at 100 ns (with two ligand topology files from PRODRG and ATB server, separately), the molecular mechanics/Poisson-Boltzmann surface area (MM/PBSA) free energy, and contributions of entropy were investigated. Then, the physicochemical-toxicity-pharmacokinetics-drug-likeness profiles, the frontier molecular orbitals energies (highest occupied molecular orbital, lowest unoccupied molecular orbital, and ΔE), and structural-activity relationship were assessed and analyzed. Based on binding energy, 8-hydroxymanzamine (-10.5 kcal/mol) and manzamine A (-10.1 kcal/mol) from all alkaloids with darunavir (-7.9 kcal/mol) and lopinavir (-7.4 kcal/mol) against SARS-CoV-2-Mpro were recorded. The MD simulation (RMSD, RMSF, Rg, H-bond, MM/PBSA binding energy) illustrated that the 8-hydroxymanzamine exhibits a static thermodynamic feature than the other two complexes. The predicted physicochemical, toxicity, pharmacokinetics, and drug-likeness profiles also revealed that the 8-hydroxymanzamine could be used as a potential lead candidate individually and/or synergistically with darunavir or lopinavir to combat SARS-CoV-2 infection after some pharmacological validation.


Seeing beyond monitors-Critical care nurses' multiple skills in patient observation: Descriptive qualitative study.
The aim of this study was to provide a comprehensive description of multiple skills in patient observation in critical care nursing. Data from semi-structured interviews were analysed using thematic analysis. Experienced critical care nurses (n=20) from three intensive care units in two university hospitals in Finland. Patient observation skills consist of: information gaining skills, information processing skills, decision-making skills and co-operation skills. The first three skills are integrated in the patient observation process, in which gaining information is a prerequisite for processing information that precedes making decisions. Co-operation has a special role as it occurs throughout the process. This study provided a comprehensive description of patient observation skills related to the three-phased patient observation process. The findings contribute to clarifying this part of the competence. The description of patient observation skills may be applied in both clinical practice and education as it may serve as a framework for orientation, ensuring clinical skills and designing learning environments. Based on this study, patient observation skills can be recommended to be included in critical care nursing education, orientation and as a part of critical care nurses' competence evaluation.


Clinical and radiological findings in Pallister-Killian syndrome.
Pallister-Killian syndrome (PKS) is a potentially lethal disorder with facial dysmorphism, pigmentary skin anomalies, developmental delay and major visceral anomalies, such as diaphragmatic hernia, anorectal malformation, and congenital heart disease. PKS is causally associated with mosaic tetrasomy of chromosome 12p. A routine chromosome analysis in peripheral lymphocytes usually fails to detect the mosaic state. A prompt diagnosis rests on clinical awareness and a subsequent chromosome or molecular analysis in fibroblasts, buccal mucosal cells, or bone marrow cells. We report here on three infants with PKS. One infant had aortic dilatation, a previously unreported association in PKS. More importantly, all infants showed a recognizable, though mild, pattern of skeletal changes mainly affecting axial bones, including delayed ossification of the vertebral bodies and pubic bones, flared anterior ribs, and broad metaphyses of the long bones, particularly of the femora. These skeletal changes should be considered as a useful diagnostic sign in PKS. Awareness of the axial skeletal alterations can be helpful in prompting clinicians to search for mosaic tetrasomy 12p and perform chromosomal analysis in appropriate tissue types.


Does pericapsular nerve group block have limited analgesia at the initial post-operative period? Systematic review and meta-analysis.
Perioperative pain management related to hip surgical procedures is challenging. Pericapsular nerve group (PENG) block is a novel technique that is assumed to provide better analgesia in addition to its post-operative motor-sparing effect. In this review, we aim to assess the safety and efficacy of PENG block on pain management in patients undergoing hip surgeries. Fifteen clinical trials with a total of 837 patients were included. Pain scores favored PENG group when compared to FICB group (fascia iliaca compartment block) or analgesics-only group, but the difference could only be detected in the early post-operative period. Afterward, it seems to lose its superiority, and no difference could be detected. In addition, cumulative opioid consumption favored PENG group at 24 h but not at 48 h. Regarding patients' satisfaction, our analysis showed results favoring PENG group, but there was no difference in time to first opioid or length of hospital stay. The incidence of vomiting was lower in PENG, but there was no difference in the incidence of nausea, pruritis, and dizziness. PENG provides better analgesia and lower opioid consumption in the initial post-operative period. Current evidence is not enough, and further high-quality randomized controlled trials with larger sample sizes are required.


StepWatch stride counting: accuracy, precision, and prediction of energy expenditure in children.
Accuracy and precision of the StepWatch (SW) was tested in typically developing youth (TDY) with the prediction of oxygen consumption derived from stride counts. Ten TDY (5 boys, 5 girls) with an average age 14.1 years (SD=2.2) enrolled. Participants underwent treadmill cardiopulmonary exercise testing (CPET) wearing two SW devices at 1, 2, 3, and 4 miles per hour (mph). Average sign-corrected disagreement for strides counted between the 2 monitors was 0.4 (0.52), 0.2 (0.42), 0.1 (0.32), and 0.1 (0.32) for 1, 2, 3, and 4 mph, respectively. The ratio of SW counts to observed manual counts averaged 100.03% with ICC=0.995. Oxygen consumption equations were derived from resting and walking VO(2), age, gender, and stride counts. Excellent accuracy and precision was documented for treadmill walking speeds up to 4 mph while VO(2) per number of strides taken appears to decrease with age and is lower in girls.


The Impact of Radiation-Induced DNA Damage on cGAS-STING-Mediated Immune Responses to Cancer.
Radiotherapy is a major modality used to combat a wide range of cancers. Classical radiobiology principles categorize ionizing radiation (IR) as a direct cytocidal therapeutic agent against cancer; however, there is an emerging appreciation for additional antitumor immune responses generated by this modality. A more nuanced understanding of the immunological pathways induced by radiation could inform optimal therapeutic combinations to harness radiation-induced antitumor immunity and improve treatment outcomes of cancers refractory to current radiotherapy regimens. Here, we summarize how radiation-induced DNA damage leads to the activation of a cytosolic DNA sensing pathway mediated by cyclic GMP-AMP (cGAMP) synthase (cGAS) and stimulator of interferon genes (STING). The activation of cGAS-STING initiates innate immune signaling that facilitates adaptive immune responses to destroy cancer. In this way, cGAS-STING signaling bridges the DNA damaging capacity of IR with the activation of CD8+ cytotoxic T cell-mediated destruction of cancer-highlighting a molecular pathway radiotherapy can exploit to induce antitumor immune responses. In the context of radiotherapy, we further report on factors that enhance or inhibit cGAS-STING signaling, deleterious effects associated with cGAS-STING activation, and promising therapeutic candidates being investigated in combination with IR to bolster immune activation through engaging STING-signaling. A clearer understanding of how IR activates cGAS-STING signaling will inform immune-based treatment strategies to maximize the antitumor efficacy of radiotherapy, improving therapeutic outcomes.


Eruptions and related clinical course among 296 hospitalized adults with confirmed COVID-19.
Limited information exists on mucocutaneous disease and its relation to course of COVID-19. To estimate prevalence of mucocutaneous findings, characterize morphologic patterns, and describe relationship to course in hospitalized adults with COVID-19. Prospective cohort study at 2 tertiary hospitals (Northwell Health) between May 11, 2020 and June 15, 2020. Among 296 hospitalized adults with COVID-19, 35 (11.8%) had at least 1 disease-related eruption. Patterns included ulcer (13/35, 37.1%), purpura (9/35, 25.7%), necrosis (5/35, 14.3%), nonspecific erythema (4/35, 11.4%), morbilliform eruption (4/35, 11.4%), pernio-like lesions (4/35, 11.4%), and vesicles (1/35, 2.9%). Patterns also showed anatomic site specificity. A greater proportion of patients with mucocutaneous findings used mechanical ventilation (61% vs 30%), used vasopressors (77% vs 33%), initiated dialysis (31% vs 9%), had thrombosis (17% vs 11%), and had in-hospital mortality (34% vs 12%) compared with those without mucocutaneous findings. Patients with mucocutaneous disease were more likely to use mechanical ventilation (adjusted prevalence ratio, 1.98; 95% confidence interval, 1.37-2.86); P < .001). Differences for other outcomes were attenuated after covariate adjustment and did not reach statistical significance. Skin biopsies were not performed. Distinct mucocutaneous patterns were identified in hospitalized adults with COVID-19. Mucocutaneous disease may be linked to more severe clinical course.


Augmented dissemination of antibiotic resistance elicited by non-antibiotic factors.
The emergence and rapid spread of antibiotic resistance seriously compromise the clinical efficacy of current antibiotic therapies, representing a serious public health threat worldwide. Generally, drug-susceptible bacteria can acquire antibiotic resistance through genetic mutation or gene transfer, among which horizontal gene transfer (HGT) plays a dominant role. It is widely acknowledged that the sub-inhibitory concentrations of antibiotics are the key drivers in promoting the transmission of antibiotic resistance. However, accumulating evidence in recent years has shown that in addition to antibiotics, non-antibiotics can also accelerate the horizontal transfer of antibiotic resistance genes (ARGs). Nevertheless, the roles and potential mechanisms of non-antibiotic factors in the transmission of ARGs remain largely underestimated. In this review, we depict the four pathways of HGT and their differences, including conjugation, transformation, transduction and vesiduction. We summarize non-antibiotic factors accounting for the enhanced horizontal transfer of ARGs and their underlying molecular mechanisms. Finally, we discuss the limitations and implications of current studies.


Interaction of parvalbumin of pike II with calcium and terbium ions.
Fluorimetric titrations of parvalbumin II (pI 4.2) of pike (Pike II) with Ca2+ and Tb3+ show the CD and EF binding sites to be non-equivalent. The intrinsic binding constants of the strong and the weak sites obtained for Ca2+ are: KsCa = 1.6.10(8) M-1; KwCa = 6.6.10(5) M-1. Differences of the order of 100% were encountered between the Tb3+ binding constants obtained with four different versions of titration. Their average values are: KsTb = 1.9.10(11) M-1; KwTb = 1.0.10(7) M-1. The distances of the strong and the weak sites from the singular Tyr-48, rs = 9.5 A and r2 = 11.5 A, were derived from Förster-type energy transfer and proved compatible with the X-ray structure of parvalbumin III (pI 4.2) of carp (CarpIII). From the distances, it is suggested that CD is the strong and EF the weak metal-binding site of PikeII. Tb3+ was shown by CD spectroscopy to have the same structural effect on PikeII as Ca2+. Removal of the metal ions from PikeII results in a decrease of helix content as monitored by CD spectroscopy. This decrease is larger than that in CarpIII. A concomitant decrease of the fluorescence quantum yield at nearly constant decay time is indicative of mainly static quenching, probably by the non-coordinating carboxylate groups. The maximum helix content is almost completely reestablished upon binding of the first metal ion. However, small changes of the energy transfer in PikeII with one terbium ion bound to the strong site indicate fine structural rearrangements of the strong binding site when Ca2+ is bound to the weak one.


Intravascular Ultrasound in Chronic Total Occlusion Percutaneous Coronary Intervention: Solving Ambiguity and Improving Durability.
Chronic total occlusions remain among the most technically challenging lesions to treat percutaneously. Limitations of 2-dimensional angiography may further hinder successful treatment of these lesions. Intrasvascular ultrasound has a key role in percutaneous recanalization for a chronic total occlusion by providing key lesion characteristics, facilitating guidewire crossing, elucidating the intraplaque or extralaque path of the guidewire, optimizing lesion preparation, guiding stenting and identifying suboptimal results. Live visualization of the guidewire during crossing may reduce extraplaque wire tracking. This review describes the practical uses of intravascular imaging for commonly encountered scenarios when treating chronic total occlusions.


Two-dimensional, J-resolved spectroscopic imaging of GABA at 4 Tesla in the human brain.
A method for measuring brain gamma-amino butyric acid (GABA) levels is presented that combines 2D J-resolved magnetic resonance spectroscopy (J-MRS) techniques with chemical-shift imaging (2D-JMRSI) at 4 Tesla (T). The results of phantom and in vivo experiments agree well in demonstrating that the (2)CH(2) GABA resonance situated at 2.97 ppm can be resolved from the neighboring creatine (Cr) resonance at 3.03 ppm and quantified. Single-voxel, J-resolved standard and metabolite-nulled in vivo experiments on six healthy subjects reveal a broad component from the underlying macromolecules (MM) that resonates at and around 3.00 ppm, which is estimated to contribute approximately 15% to the J-resolved GABA resonance in this large voxel at a repetition time (TR) of 4.5 s. With our 2D-JMSRI at 1.25 s TR, the macromolecule resonance contribution to our GABA measurements is approximated to be 12%. Six healthy human subjects underwent scanning at 4T with this sequence, yielding a global brain GABA concentration of 0.76 +/- 0.20 mM after correction for 12% macromolecule contribution.


Vascular organization of the catfish gill filament.
Light and scanning electron microscopic observations were made on methyl-methacrylate corrosion casts of the blood vessels in the gills of channel catfish (Ictalurus punctatus). The vasculature of the gill filament can be divided into three distinct pathways: 1. the well-known respiratory circulation which includes the afferent filamental artery (AF), afferent lamellar arteriole (AL), lamella (L), efferent lamellar arteriole (EL) and efferent filamental artery (EF), 2. a nutritive pathway from the EF through small nutritive capillaries (NC) and into one of several filamental veins (FV), and 3. an interlamellar circulation in which small prelamellar arterio-venous anastomoses (PAVA) connect the AL into a series of organized vascular spaces (interlamellar vessels, ILV's) that underlie the interlamellar filamental epithelium. Several sinus like spaces associated with AF, EF and the filamental cartilagenous support were also observed. The physiological significance of these vascular pathways is discussed.


Abnormal segregation of alleles and haplotypes at the polymorphic site of the PRNP gene within promoter and intron 1 regions in Polish Holstein-Friesian cattle.
Allele and haplotype segregation at the polymorphic sites within the promoter (23indel) and intron 1 (12indel) regions of the PRNP gene was analyzed in Polish Holstein-Friesian cattle. More 23del/del homozygotes and fewer 23ins/ins homozygotes than expected were observed in the offspring of ♂ 23ins/del × ♀ 23ins/del parents. In the offspring of ♂ 23ins/del × ♀ 23del/del parents and ♂ 23del/del × ♀ 23ins/del parents, a trend toward more 23del/del animals and fewer 23ins/del animals than expected was noted. At the 12indel polymorphic site, the only trend found was one toward fewer 12ins/ins genotypes and more 12ins/del and 12del/del genotypes than expected in the offspring of ♂ 12ins/del × ♀ 12ins/del parents. An analysis of haplotype segregation revealed more 23del-12del/23del-12del diplotypes and fewer 23ins-12ins/23ins-12ins diplotypes at the significance threshold than expected in the offspring of ♂ 23ins-12ins/23del-12del × ♀ 23ins-12ins/23del-12del parents.


Cytidine-containing tails robustly enhance and prolong protein production of synthetic mRNA in cell and in vivo.
Synthetic mRNAs are rising rapidly as alternative therapeutic agents for delivery of proteins. However, the practical use of synthetic mRNAs has been restricted by their low cellular stability as well as poor protein production efficiency. The key roles of poly(A) tail on mRNA biology inspire us to explore the optimization of tail sequence to overcome the aforementioned limitations. Here, the systematic substitution of non-A nucleotides in the tails revealed that cytidine-containing tails can substantially enhance the protein production rate and duration of synthetic mRNAs both in vitro and in vivo. Such C-containing tails shield synthetic mRNAs from deadenylase CCR4-NOT transcription complex, as the catalytic CNOT proteins, especially CNOT6L and CNOT7, have lower efficiency in trimming of cytidine. Consistently, these enhancement effects of C-containing tails were observed on all synthetic mRNAs tested and were independent of transfection reagents and cell types. As the C-containing tails can be used along with other mRNA enhancement technologies to synergically boost protein production, we believe that these tails can be broadly used on synthetic mRNAs to directly promote their clinical applications.