Towards an integrated blood pressure self-monitoring solution for stroke/TIA in Ireland: a mixed methods feasibility study for the TASMIN5S IRL randomised controlled trial.

BACKGROUND: Optimising blood pressure (BP) control is one of the most important modifiable risk factors in preventing subsequent stroke where the risk increases by one-third for every 10 mmHg rise in systolic BP. This study evaluated the feasibility and potential effectiveness of blood pressure self-monitoring with planned medication titration, to inform a definitive trial of the intervention, in patients with a previous stroke or transient ischaemic attack (TIA). METHODS: Patients with a history of stroke/TIA and sub-optimal BP control were invited to take part in a mixed methods feasibility study for a randomised controlled trial. Those meeting the inclusion criteria with systolic BP >130 mmHg were randomised to a self-monitoring intervention group or usual care group. The intervention involved self-monitoring BP twice a day for 3 days within a 7-day period, every month, following text message reminders. Treatment escalation, based on a pre-agreed plan by the general practitioner (GP) and patient, was initiated according to the results of these readings. Semi-structured interviews were carried out with patients and clinicians and analysed thematically. RESULTS: Of those identified, 47% (32/68) attended for assessment. Of those assessed, 15 were eligible for recruitment and were consented and randomised to the intervention or control group on a 2:1 basis. Of those randomised, 93% (14/15) completed the study and there were no adverse events. Systolic BP was lower in the intervention group at 3 months. Participants found the intervention acceptable and easy to use. GPs found it easy to incorporate into their practice activity without increasing workload. CONCLUSIONS: TASMIN5S, an integrated blood pressure self-monitoring intervention in patients with a previous stroke/TIA, is feasible and safe to deliver in primary care. A pre-agreed three-step medication titration plan was easily implemented, increased patient involvement in their care, and had no adverse effects. This feasibility study provides important information to inform a definitive trial to determine the potential effectiveness of the intervention in patients post-stroke or TIA. TRIAL REGISTRATION: ISRCTN57946500 . Registered on 12/08/2019.

Direct-write 3D printing and characterization of a GelMA-based biomaterial for intracorporeal tissue.

We develop and characterize a biomaterial formulation and robotic methods tailored for intracorporeal tissue engineering (TE) via direct-write (DW) 3D printing. Intracorporeal TE is defined as the biofabrication of 3D TE scaffolds inside of a living patient, in a minimally invasive manner. A biomaterial for intracorporeal TE requires to be 3D printable and crosslinkable via mechanisms that are safe to native tissues and feasible at physiological temperature (37 °C). The cell-laden biomaterial (bioink) preparation and bioprinting methods must support cell viability. Additionally, the biomaterial and bioprinting method must enable the spatially accurate intracorporeal 3D delivery of the biomaterial, and the biomaterial must adhere to or integrate into the native tissue. Current biomaterial formulations do not meet all the presumed intracorporeal DW TE requirements. We demonstrate that a specific formulation of gelatin methacryloyl (GelMA)/Laponite®/methylcellulose (GLM) biomaterial system can be 3D printed at physiological temperature and crosslinked using visible light to construct 3D TE scaffolds with clinically relevant dimensions and consistent structures. Cell viability of 71%-77% and consistent mechanical properties over 21 d are reported. Rheological modifiers, Laponite® and methylcellulose, extend the degradation time of the scaffolds. The DW modality enables the piercing of the soft tissue and over-extrusion of the biomaterial into the tissue, creating a novel interlocking mechanism with soft, hydrated native tissue mimics and animal muscle with a 3.5-4 fold increase in the biomaterial/tissue adhesion strength compared to printing on top of the tissue. The developed GLM biomaterial and robotic interlocking mechanism pave the way towards intracorporeal TE.

Resveratrol supplementation significantly influences obesity measures: a systematic review and dose-response meta-analysis of randomized controlled trials.

This study aimed to summarize earlier randomized controlled trials on the effects of resveratrol supplementation on body weight (BW), body mass index (BMI), waist circumference (WC) and fat mass (FM). We searched PubMed, SCOPUS, Cochrane Library and Google Scholar from inception to April 2018 using relevant keywords. All clinical trials investigating the effects of resveratrol supplementation on BW, BMI, WC and FM in adults were included. Overall, 28 trials were included. Pooled effect sizes suggested a significant effect of resveratrol administration on weight (weighted mean differences [WMD]: -0.51 kg, 95% confidence interval [CI]: -0.94 to -0.09; I2  = 50.3%, P = 0.02), BMI (WMD: -0.17 kg m-2 , 95% CI: -0.32, -0.03; I2  = 49.6%, P = 0.02) and WC (WMD: -0.79 cm, 95% CI: -1.39, -0.2; I2  = 13.4%, P = 0.009), respectively. However, no significant effect of resveratrol supplementation on FM was found (WMD: -0.36%, 95% CI: -0.88, 0.15; I2  = 0.0%, P = 0.16). Findings from subgroup analysis revealed a significant reduction in BW and BMI in trials using resveratrol at the dosage of <500 mg d-1 , those with long-term interventions (≥3 month), and performed on people with obesity. Taken together, the data suggest that resveratrol supplementation has beneficial effects to reduce BW, BMI and WC, but not FM.

Squishy nanotraps: hybrid cellulose nanocrystal-zirconium metallogels for controlled trapping of biomacromolecules.

A brick-and-mortar-like ultrasoft nanocomposite metallogel is formed by crosslinking cellulose nanocrystals (CNC) with ammonium zirconium carbonate (AZC) to trap and reconfigure dextran, a model biomacromolecule. The bricks (CNC) reinforce the metallogel, compete with dextran in reacting with AZC, and decouple long-time dextran dynamics from network formation, while the mortar (AZC) imparts bimodality to the dextran diffusion.

Higher circulating levels of chemokine CXCL10 in patients with breast cancer: Evaluation of the influences of tumor stage and chemokine gene polymorphism.

BACKGROUND: The CXCL10 receptor, CXCR3, is preferentially expressed on Th1 and NK cells. Therefore, CXCL10 acts as a chemoattractant for these cells. OBJECTIVE: The aim was to evaluate the CXCL10 levels and a single nucleotide polymorphism (SNP), rs4508917, in chemokine gene, in patients with breast cancer (BC). METHODS: A total of 200 subjects including 100 women with BC and 100 healthy women were enrolled into study. The serum CXCL10 levels were measured by ELISA and the SNP rs4508917 was determined by polymerase chain reaction-restriction length polymorphism (PCR-RFLP). RESULTS: The CXCL10 levels were significantly higher in patients than control group (P< 0.0001). There was also significant difference between tumor stages regarding the CXCL10 levels (P< 0.0001). The frequencies of GG genotype and G allele at rs4508917 were significantly higher in patients than controls (P< 0.0001). The CXCL10 levels were higher in patients with GG genotype whereas they were lower in healthy subjects having GG genotype as compared with those having AA genotype at rs4508917 (P< 0.001). CONCLUSION: Higher CXCL10 levels in patients with BC represent that the chemokine may contributes in tumor development. The rs4508917 may play a role in the susceptibility to BC. Different association was also observed between rs4508917 and CXCL10 levels in patients with BC and healthy subjects.

Probiotic Yogurt Culture Bifidobacterium Animalis Subsp. Lactis BB-12 and Lactobacillus Acidophilus LA-5 Modulate the Cytokine Secretion by Peripheral Blood Mononuclear Cells from Patients with Ulcerative Colitis.

BACKGROUND: There are some evidences for the immunomodulation disorders in the response to intestinal microbiota in inflammatory bowel disease. Yogurt is a fermented milk product made with a starter culture consisting of different probiotics which could be colonized in intestine. However, the role of probiotics in the aetiopathogenesis of ulcerative colitis (UC) has not been clarified. To determine how the immune system responds to these bacteria this study was planned. METHODS: Bifidobacterium lactis BB-12 (B. lactis) and Lactobacillus acidophilus LA-5 (L. acidophilus) were cultivated on MRS broth. PBMCs of 36 UC patients were separated by Ficoll-Hypaque centrifugation and co-cultured with different concentrations of UV killed bacteria in RPMI-1 640 plus 10% FCS for 48/72 h. IL-10, TGF-ß, IFN-γ and TNF-α were measured in supernatant of PBMCs by ELISA. RESULTS: Both bacteria significantly augmented IL-10, TGF-ß, IFN-γ and TNF-α compared to control (p<0.001). The secretion levels of IL-10 and TGF-ß by B. lactis- compared to L. acidophilus-stimulated PBMCs were significantly higher (p<0.05, p<0.01 respectively). The secretion levels of TNF-α and IFN-γ by PBMCs after 72 h were significantly lower compared to 48 h stimulation by B. lactis (p<0.001, p<0.035 respectively). CONCLUSION: These data show that both probiotics may trigger the pro- and anti-inflammatory immune response of UC patients. It seems that IL-10/TGF-ß uprising by B. lactis could be the reason of TNF-α/IFN-γ reduction. Therefore albeit B. lactis still stimulates the effector Th cells but because of more stimulatory effect on Tregs, it could be a good potential therapeutic candidate for further investigation.

Circulating levels of interleukin-35 in patients with multiple sclerosis: evaluation of the influences of FOXP3 gene polymorphism and treatment program.

The regulatory T (Treg) cells play a major role in the control of the autoimmunity and inflammation, and IL-35 has been described as an immunosuppressive cytokine that is mainly produced by CD4(+)FOXP3(+) Treg cells. The aim of this study was to evaluate the serum levels of IL-35 and a single nucleotide polymorphism (SNP), rs3761548, in FOXP3 gene in patients with multiple sclerosis. The blood samples were collected from 140 multiple sclerosis (MS) patients (including 51 untreated and 89 treated patients) and 140 healthy subjects as a control group. The serum levels of IL-35 were measured by ELISA. The DNA was analyzed for SNP rs3761548 in FOXP3 gene using SSP-PCR. There was no significant difference between untreated MS patients and control group regarding the mean serum levels of IL-35, although this parameter was higher in untreated patients. However, the mean serum level of IL-35 in treated MS patients was significantly higher than that in the control group (P < 0.008). The mean serum levels of IL-35 in patients who were treated with interferon-ß, methylprednisolone, or with the both interferon-ß and methylprednisolone were significantly higher than that in the healthy group (P < 0.01, P < 0.01, and P < 0.2, respectively). The frequencies of AA and AC genotypes at rs3761548 in the FOXP3 gene were significantly higher in MS group as compared with healthy subjects (P < 0.05). The frequency of CC genotype at rs3761548 was significantly lower in the MS group in comparison with healthy control subjects (P < 0.001). Moreover, the frequency of A allele was significantly higher whereas the frequency of C allele was significantly lower in MS patients in comparison to healthy subjects (P < 0.001). The mean serum level of IL-35 was significantly lower in MS patients or healthy subjects with AA genotype as compared with those with CC genotype at rs3761548 in FOXP3 gene (P < 0.01 and P < 0.001, respectively). These results showed higher serum levels of IL-35 in treated MS patients representing that the benefit effects of treatment may in part performed through the upregulation of the IL-35 production. The SNP rs3761548 may influence the susceptibility to MS disease and the serum levels of IL-35.

Augmenting the expression of NKp44 molecule and the natural killer activity in peripheral blood mononuclear cells from patients with malignant colorectal carcinoma.

OBJECTIVE: NKp44 and NKG2D are of the main NK activating receptors involved in recognition and killing of tumors. Here we studied the stimulatory effects of PHA and/or K562 cell line on induction of NKp44 and NKG2D expression and the NK activity of PBMCs from patients with colorectal carcinoma (CRC). MATERIALS AND METHODS: Peripheral blood samples were collected from 10 patients with CRC. The peripheral blood mononuclear cells (PBMCs) from each patient received a single stimulation with PHA or double stimulation with PHA and irradiated K562 cell line (iK562). The expression of CD56, NKG2D and NKp44 were detected by flowcytometry. The NK activity of PBMCs against a colorectal carcinoma cell line named as SW742 was determined with 51Cr-release assay. RESULTS: Double stimulation of PBMCs with PHA+iK562 significantly augmented the number CD56(+) cells compared to PHA alone and non-stimulated PBMCs (P<0.000, P<0.0000; respectively). A single stimulation of PBMCs with PHA resulted in an enhancement in NKG2D and NKp44 expression from 16.6±3.3% (for non-stimulated PBMCs) to 42±5.6% and 48.1±3.8% respectively (p<0.05). Double stimulation of PBMCs augmented the NKp44 expression significantly in comparison with single stimulation with PHA (73.6±12%, p<0.05). Double stimulation of PBMCs significantly enhanced the NK activity against SW742 target cells compared to single stimulation with PHA (p<0.05). DISCUSSION AND CONCLUSION: Our results demonstrated that the mitogen and iK562 exposure to PBMCs can significantly improve NK activity which is co-related to the higher expression of NKp44 and NKG2D. These data may help to improve cancer immunotherapy protocols.

Elimination of apoptotic spermatozoa by magnetic-activated cell sorting improves the fertilization rate of couples treated with ICSI procedure.

Couples with unexplained infertility (UI) tend to have low fertilization rates using current in vitro fertilization procedures. The aim of this study was to evaluate whether elimination of apoptotic spermatozoa could increase the likelihood of pregnancy by intra-cytoplasmic sperm injection (ICSI). A total of 74 couples with UI were divided into two groups including the control group (n = 37) for which spermatozoa prepared by density gradient centrifugation were injected into oocytes and the study group (n = 37) for which spermatozoa was further processed by magnetic-activated cell sorting to eliminate apoptotic spermatozoa, then injected into oocytes. Fertilization, cleavage, pregnancy and birth rates were analyzed in two groups. The fertilization rate was significantly higher in the study group compared with the control group (73.41% vs. 61.11%; p = 0.03). On day 3, the number of eight blastomeric non-fragmented embryos per oocytes was also significantly higher in study group as compared with controls (45.05% vs. 34.16%; p = 0.049). The pregnancy and birth rates were 43.24 and 40.5% in study group and 35.11 and 27% in control group respectively. Statistical analyses showed that the differences in the pregnancy and live-birth rates between study and control groups were not significant (p = 0.37 and 0.16 respectively). These results demonstrate that non-apoptotic spermatozoa display higher fertilization potential and embryo quality following ICSI.

In vitro modulation of natural killer activity of human peripheral blood mononuclear cells against prostate tumor cell line.

OBJECTIVE: Natural killer (NK) cells have long been known to be involved in the recognition and lysis of tumor cells but the mechanisms contributing to deficient NK activity in patients with cancer remains unclear. Manipulation of them is likely essential to the success of cancer immunotherapy protocols although optimal stimulation and maintenance of NK activity remains elusive. Here we studied the stimulatory effects of PHA and K562, on NK activity of human peripheral blood mononuclear cells (PBMCs). MATERIALS AND METHODS: The NK activity of PBMCs against DU-145 was determined with 51Cr-release assay. The PBMCs were stimulated with PHA, either on one occasion or repetitively on three occasions (1-PHA-PBMC or 3-PHA-PBMC, respectively), and were incubated with irradiated K562 (iK562). The expression of CD56, NKG2D and MICA/B were detected on PBMCs and cell lines with flow cytometry. RESULTS: PHA stimulation increased the proportion of CD56+ cells and upregulated NKG2D expression on 1-PHA-PBMC and 3-PHA-PBMC, but co-incubation with iK562 decreased NKG2D expression on 1-PHA-PBMC without change of NKG2D expression on the 3-PHA-PBMC. NK activity of 1-PHA-PBMC appeared to decrease with co-incubation with iK562 compared to a significant increase in activity of 3-PHA-PBMC. A similar increase in interferon-γ (IFN-γ) secretion from 3-PHA-PBMC was demonstrated compared to 1-PHA-PBMC. DISCUSSION AND CONCLUSION: Our results demonstrated that varying the mitogen exposure to PBMCs affect the influence of iK562 on NK activity. This effect appeared to be unrelated to the subsequent expression of NKG2D or IFN-γ secretion. These results may be beneficial in the development of future cancer immunotherapy protocols.

Are natural killer cells distributed in relationship to nerve fibers in the pregnant mouse uterus?

Specialized lymphocytes, called uterine Natural Killer (uNK) cells, appear in human and rodent uteri and become abundant at implantation sites during decidualization and early pregnancy. The hallmark of human uNK cells is intense expression of CD56, a neural cell adhesion glycoprotein (NCAM-1) while mature (granulated) mouse uNK cells express asialoGM1, a brain ganglioside. Murine uNK cells initiate the normal structural changes induced in maternal spiral arteries by pregnancy but regulation of their recruitment, localization and activation is incompletely understood. To address whether uNK cell distribution is co-localized with nerve fiber distribution, sections of gestation day (gd) 6-12 implantation sites from C57BL/6 (B6) mice were studied. Nerve fibers reactive with antibodies to pan neurofilament 150 kD or with tyrosine hydroxylase, an enzyme restricted to sympathetic fibers, were present the walls of branches from the uterine artery in the mesentery. Reactivity was lost as the vessels crossed the myometrium and entered endometrium/decidua. Periodic Acid Schiffs reactive uNK cells were absent from the mesentery and enriched in decidua basalis where they transcribed NCAM-1 and associated with non-innervated segments of the uterine arteries, including spiral arteries. These data suggest that the localization and activation of mature uNK cells are unlikely to be neurotransmitter regulated.

Effect of surfactant HLB and different formulation variables on the properties of poly-D,L-lactide microspheres of naltrexone prepared by double emulsion technique.

The aim of this work was to investigate the role of HLB of emulsifier as well as volume of the internal aqueous phase (W(1)) and presence of salt in the external aqueous phase (W(2)) on the morphology, size and encapsulation efficiency of poly(D,L-lactide) microspheres containing naltrexone HCl. PLA microparticles containing naltrexone HCl, an effective opiate antagonist, were prepared by a water-in-oil-in-water emulsification-solvent evaporation procedure. One of the five different emulsifiers: span 80, span 20, tween 85, tween 80 and tween 20, with HLB values from 4-17 were added to W(1). Presence of emulsifier in W(1) resulted in smaller particles with a more dense and uniform internal structure. Incorporation of span 80 (HLB 4.3, suitable for W/O emulsions) yield the highest encapsulation efficiency. Increasing the HLB value to 8 or 11 (span 20 or tween 85) decreased the efficiency of naltrexone HCl-loading. HLB values higher than 15 (tween 80 or tween 20) increased encapsulation efficiency unexpectedly, which could be attributed to migration of these emulsifiers to the O/W(2) interface and modifying the surface properties of microparticles. Increasing the internal water phase volume from 0.2-1.8 ml resulted in larger particle size with poor encapsulation efficiency. Addition of 10% w/w NaCl to the W(2) changed the surface morphology of microspheres from a porous form to a smooth surface. It was shown that, by selecting the appropriate HLB value of emulsifier in W(1), addition of salt to W(2) and controlling the volume of W(1), one can control the encapsulation efficiency, size and morphology of microspheres.

Soluble HLA class I molecules in malignant pleural and peritoneal effusions and its possible role on NK and LAK cytotoxicity.

PURPOSE: To examine the amount of sHLA-I in malignant pleural and peritoneal effusions and its possible role in natural immune defense. METHODS: Three groups of patients (75 patients with malignancy, 21 with infection, and 27 with other diseases) were studied for sHLA-I value using an ELISA method. Cytolytic activity of freshly isolated pleural and peritoneal effusion-associated lymphoid (EAL) cells from 14 of cases with malignancy were examined and compared to that of ten non-cancerous patients. EAL cells were co-cultured with the autologous cell-free effusions immediately after collection and 3 days after incubation with IL-2. RESULTS: The mean value of sHLA-I in effusions was 1.01+/-1.36 micro g/ml, 0.97+/-1.20 micro g/ml, and 0.49+/-0.45 micro g/ml, respectively. Despite higher mean sHLA-I levels in malignant and infected patients, no significant difference between these groups was observed ( P >0.05). Generally, the amount of sHLA-I in peritoneal effusions was higher than that for pleural effusions, but the difference was not significant. There were also no statistical differences in the sHLA-I levels between sub-groups of patients with malignancy. EAL cells' killing activity in malignant and infected effusions was 68.15+/-11.73 and 78.28+/-14.41, respectively ( P=0.08). No correlation between sHLA-I level and NK activity of EAL cells from the patients was found. Almost all malignant cases after exposure to cell-free effusions displayed an increase in NK activity (from 68.66+/-11.13 to 74.2+/-12.39, P=0.042) and a decrease in LAK activity (74.5+/-18.30 vs 67.72+/-16.46, P=0.040). Whereas, the same experiment performed for non-malignant effusions showed a decrease in both NK activity and LAK activity. Changes in NK and LAK activity were not correlated with the amount of sHLA-I in the effusions. CONCLUSION: The presence of sHLA-I, particularly in malignant effusions, suggests a role for these molecules in tumor immunity in the peritoneal or plural environment; however, at least with these group of patients, sHLA-I appears not to be a unique determining factor on EAL cells' killing activity.