CD4<Superscript>+</Superscript>FOXP3<Superscript>+</Superscript> T regulatory cells decrease and CD3<Superscript>+</Superscript>CD8<Superscript>+</Superscript> T cells recruitment in TILs from melanoma metastases after electrochemotherapy

Electrochemotherapy (ECT) represents an effective local treatment for skin unresectable melanoma metastases with high overall objective response rate. ECT is based on the combination of anti-neoplastic drugs administration and cancer cells electroporation. Whether ECT can also activate the immune system is a matter of debate, however a significant recruitment of dendritic cells in melanoma treated metastases has been described. Herein we investigated immediate and late effects of ECT treatment on T cell subsets in ECT-treated lesions by fluorescent immunohistochemistry. Biopsies from melanoma patients (n = 10) were taken before ECT (t0), at d1 and d14 from treatment. At t0, CD3⁺CD4⁺ T cells were the most represented T cells, well detected in the perilesional dermis, particularly at tumour margin, while CD3⁺CD8⁺ T cells were less represented. CD4⁺FOXP3⁺ T regulatory (Treg) cells were present in the perilesional dermis and within the lesion. ECT induced a significant decrease of CD4⁺FOXP3⁺ Treg cells percentage in the perilesional dermis, observed at d1 and at d14 (p < 0.001). CD3⁺CD8⁺ T cells frequency significantly increased at d14 from treatment in the perilesional dermis (p < 0.001). Furthermore calreticulin translocation to the plasma membrane, a hallmark of immunogenic cell death, was observed in metastatic cells after ECT. The data reported here confirm that ECT induces a local response, with a lymphoid infiltrate characterized by CD4⁺FOXP3⁺ Treg cells decrease and CD3⁺CD8⁺ T cells recruitment in the treated lesions. These results might contribute to design novel combinational therapeutic approaches with ECT and immunotherapy in order to generate a systemic long-lasting anti-melanoma immunity.     

Surface antigen profiles of leukocytes and melanoma cells in lymph node metastases are associated with survival in AJCC stage III melanoma patients

There is an urgent need to identify more accurate prognostic biomarkers in melanoma patients, particularly in those with metastatic disease. This study aimed to identify melanoma and leukocyte surface antigens predictive of survival in a prospective series of AJCC stage IIIb/c melanoma patients (n = 29). Live cell suspensions were prepared from melanoma metastases within lymph nodes (LN). The suspensions were immuno-magnetically separated into CD45⁺ (leukocyte) and CD45^? (non-hematopoietic, enriched melanoma cell) fractions. Surface antigens on CD45^? and CD45⁺ cell populations were profiled using DotScan? microarrays (Medsaic Pty. Ltd.) and showed differential abundance levels for 52 and 78 antigens respectively. Associations of the surface profiles with clinicopathologic and outcome data (median follow-up 35.4 months post LN resection) were sought using univariate (log-rank test) and multivariate (Wald’s test; modelled with patient’s age, gender and AJCC staging at LN recurrence) survival models. CD9 (p = 0.036), CD39 (p = 0.004) and CD55 (p = 0.005) on CD45⁺ leukocytes were independently associated with distant metastasis-free survival using multivariate analysis. Leukocytes with high CD39 levels were also significantly associated with increased overall survival (OS) in multivariate analysis (p = 0.016). LNs containing leukocytes expressing CD11b (p = 0.025), CD49d (p = 0.043) and CD79b (p = 0.044) were associated with reduced OS on univariate analysis. For enriched melanoma cells (CD45^? cell populations), 11 surface antigens were significantly correlated with the disease-free interval (DFI) between diagnosis of culprit primary melanoma and LN metastasis resection. Nine antigens on CD45⁺ leukocytes also correlated with DFI. Following validation in independent datasets, surface markers identified here should enable more accurate determination of prognosis in stage III melanoma patients and provide better risk stratification of patients entering clinical trials.     

Electrochemotherapy is effective in the treatment of rat bone metastases

Bone metastases impair general health status, quality of life and survival of patients. Electrochemotherapy (ECT), which combines electroporation (EP) and the administration of anticancer drugs, has been recently introduced into clinical practice for the local treatment of solid tumours. In the present study, the ability of EP with bleomycin (Bleo) to induce MRMT-1 rat breast cancer cell death was investigated in vitro. Then, an in vivo model for bone metastases was set up by the inoculation of MRMT-1 cells in rat proximal tibia. 7 days after tumour induction the animals were treated with Bleo, EP, Bleo followed by EP (ECT), or left untreated. ECT eliminated the tumour in 6 out of 8 (75 %) treated metastases. Radiological evaluation showed that the Honore score in ECT-treated animals was significantly lower when compared with the other groups (p < 0.0005) and not significantly different from healthy controls. Bone morphology in ECT-treated animals, evaluated by histological and microtomographical analyses, showed intact cortical and trabecular bone structure with new bone apposition. Histomorphometric evaluation showed that ECT-treated metastases had significantly higher bone volume, trabecular number, trabecular thickness and bone mineral density compared with those of untreated metastases (respectively p < 0.0005 for BV/TV, Tb.N and BMD; p < 0.05 for Tb.Th) or metastases treated with Bleo (p < 0.05 for BV/TV, Tb.N, p < 0.005 for BMD) or EP (p < 0.005 for BV/TV, Tb.N; p < 0.0005 for BMD). These findings suggest that early ECT treatment of bone metastases is minimally invasive, safe and effective, thus providing pre-clinical evidence for its use in the treatment of human bone metastases.     

Mother-child bonding at 1 year; associations with symptoms of postnatal depression and bonding in the first few weeks

Some mothers experience neutral or negative feelings toward their new infant. This study examined the association between symptoms of postnatal depression and mother–infant bonding and the persistence of these feelings over the first year. Bonding was assessed using the Mother–Infant Bonding Scale (MIBQ), at four times postnatal, “early weeks” (1–4 weeks), 9 weeks, 16 weeks and 1 year, in 50 depressed, Edinburgh Postnatal Depression scale (EPDS) ≥13 at 4 weeks post natal, and 29 non-depressed mothers. A significant association between the EPDS score at 4 weeks and bonding score at 1–4 weeks, 9 weeks, and at 1 year postnatal, χ ²(1)?=?9.85, p?<?0.01, 5.44, p?<?0.05 and 5.21, p?<?0.05, respectively, was found, with a trend at 16 weeks. There was a strong association between bonding in the early weeks and all later time points χ ²(1)?=?17.26, p?<?0.001, 7.89, p?<?0.01 and 13.69, p?<?0.001, respectively. Regression showed early bonding rather than early depression was the major predictor of bonding at 1 year. Women who are depressed postnatally can fail to bond well with their baby and this can persist for a year. Early identification and intervention for poor bonding is indicated.     

Imbalance of circulating dendritic cell subsets in chronic obstructive pulmonary disease

Dendritic cells (DCs) play an unsettled role in chronic obstructive pulmonary disease (COPD) pathogenesis. Two main blood subsets, myeloid (m) and plasmacytoid (p) DCs, have been identified in humans. Phenotype and frequency of circulating DC subsets were assessed by multi-parametric flow cytometry in 28 COPD patients and 30 healthy controls (15 never smokers and 15 smokers). Proportion and absolute number of pDCs were significantly reduced in COPD patients in comparison with never smokers (p < 0.001 and p < 0.003) along with a marked increase of the mDC/pDC ratio (p < 0.001). Analysis of peripheral lymphocyte subsets showed that the naive/memory T cell ratio was significantly reduced in COPD patients in comparison with never smokers (p < 0.001). Similar perturbations in the distribution of DCs and T cells also occurred in control smokers. This study is the first report of an imbalance of blood DCs in COPD. Influence of smoking and clinical relevance of these findings are discussed.     

Potential role of plasmacytoid dendritic cells for FOXP3 regulatory T cell development in human colorectal cancer and tumor draining lymph node

FOXP3⁺ regulatory T cells (Tregs) play an important role in the maintenance of tumor immunity tolerance. Compared with conventional myeloid dentritic cells (mDCs), plasmacytoid dendritic cells (pDCs) exhibit poor immunostimulatory ability, and their interaction with T cells often promotes the development of Tregs. The aim of this study was to determine FOXP3⁺ Tregs and CD123⁺pDCs infiltration in colorectal cancer and tumor draining lymph node (TDLN), and to evaluate the clinical significance and relationship between pDCs infiltration and Tregs development in the CRC tolerogenic milieu. An immunohistochemical assay was conducted to assess FOXP3⁺Tregs and CD123⁺pDCs infiltration in tumor tissue and in metastatic-free TDLN (mfTDLN) and metastatic TDLN (mTDLN). The results showed that FOXP3⁺ Tregs infiltration was more frequent in tumor tissue than in adjacent normal mucosa (P < 0.001). FOXP3⁺Tregs infiltration was associated with advanced TNM stage and lymph node metastasis (P < 0.01 and P < 0.01 for TNM stage and lymph node metastasis, respectively). Different from FOXP3⁺Tregs, CD123⁺pDCs frequencies were lower in most CRC tumor tissues, whereas the positive rate of CD123 expression in CRC was significantly higher than in adjacent normal mucosa tissue (P < 0.01). Compared to mfTDLN, mTDLN was significantly enriched in FOXP3⁺ Tregs (P < 0.01) and increased in pDC/mDC ratio (P < 0.01). The statistical analysis demonstrated a significant correlation in both Tregs and pDC/mDC ratio in mTDLN. These results suggest that there are more FOXP3⁺ Tregs with a stronger prognostic significance which might promote tumor tolerance, and that CD123⁺pDCs might contribute to Tregs development in the CRC tolerogenic milieu.     

Haematological and acute-phase response of dogs with experimental skin <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> infection to treatment with antibiotic and parthenolide

Changes in white blood cells, leukogram patterns, the positive acute-phase protein (APP) fibrinogen and negative APPs (albumin and arylesterase) were monitored to evaluate their potential as sensitive indicators throughout the course of therapy in canine skin Pseudomonas aeruginosa infection. The study was performed on 15 male mixed-breed dogs, divided in three groups of 5 dogs each. Dogs from group A were injected subcutaneously with P. aeruginosa bacterial culture (1?×?10⁸ CFU/mL) at a dose of 0.3 mL/kg and treated with enrofloxacin (5 mg/kg, s.c.) on post infection hour 48 for 10 consecutive days. Dogs from group B were infected and treated with a combination of enrofloxacin (at above-mentioned dose and intervals) and parthenolide (feverfew extract 90 mg, 0.7 % parthenolide). The schedule consisted of daily oral intake of two capsules of feverfew beginning on post infection hour 4 and continued for 6 days. The control group C included healthy dogs, injected s.c. with 0.3 mL/kg physiological saline. The haematological indices and APPs were assayed before infection and on 4th, 24th, 48th and 72nd hours and on 7th, 10th and 14th days after infection. Infected and antibiotic-treated dogs responded with significant leukocytosis, left shift, eosinopaenia and lymphopaenia between hours 24 and 72. In this group, fibrinogen increased substantially by post infection hours 24 (p?<?0.01 vs 0 h; p?<?0.05 vs group C), 48 (p?<?0.001 vs 0 h; p?<?0.05 vs group C) and 72 (p?<?0.001 vs 0 h; p?<?0.01 vs group C) while albumin reduction was marked by hours 48 (p?<?0.05 vs 0 h) and 72 (p?<?0.05 vs 0 h; p?<?0.001 vs group C) and day 7 (p?<?0.01 vs 0 h; p?<?0.001 vs group C). The combination of enrofloxacin and parthenolide modified, at a significant extent, the deviations in studied parameters except for eosinophil percentage, which persisted low.     

Can muscle shortening alone, explain the energy cost of muscle contraction in vivo?

Purpose

Decreased whole-body energy cost of running has been associated with an increased Achilles tendon stiffness. It is usually assumed that this lower energy cost can be attributed to less muscle fascicle shortening with a stiffer tendon. Increased fiber shortening is an important determinant of muscle energetics in vitro. However, other factors, like increased muscle activation may be important when considering whole muscle energetics in vivo.

Methods

To determine the effects of a small additional muscle shortening on skeletal muscle energy requirement, 19 subjects performed 30 plantarflexions on two separate occasions: isometric (ISO) and isokinetic (KIN, 6.98 rad s^–1), each with a target of 50 % of maximum isometric torque. Medial gastrocnemius muscle fascicle length (FL) was measured by ultrasound and rate of oxyhemoglobin (HbO₂) desaturation was measured during blood flow occlusion using near-infrared spectroscopy.

Results

KIN resulted in significantly greater muscle shortening (23.8 ± 1.3 mm) than ISO (18.3 ± 1.0 mm, p < 0.001, mean ± SEM), and greater shortening velocity (KIN = 2.5 ± 0.3 FL s^–1, ISO = 1.1 ± 0.1 FL s^–1, p < 0.001). Rate of HbO₂ desaturation was 19 ± 7 %, greater in KIN than ISO (p < 0.01), despite 19 ± 2 % lower mean torque (p < 0.001) and 9.8 ± 1.6 Nm s lower mean impulse per contraction (p < 0.001) in KIN compared to ISO. Root mean square for EMG was significantly greater (p < 0.05) during KIN (73 ± 3 %) than during ISO (63 ± 2 %).

Conclusion

These results illustrate that muscle energy requirement is greater when muscle fascicle shortening and/or velocity of shortening is increased, and suggest that greater activation contributes to that increased energy requirement.     

Bevacizumab and cetuximab with conventional chemotherapy reduced pancreatic tumor weight in mouse pancreatic cancer xenografts

Pancreatic cancer remains the fourth leading cause of cancer-related death in the USA with a 5-year survival rate of 5 %. The effects of epidermal growth factor receptor and vascular endothelial growth factor A blockade with chemotherapy on pancreatic tumor growth were examined. Mice bearing human PANC-1 cell xenografts were divided into three groups: T-CR (gemcitabine, cisplatin, and 5-fluorouracil), T-TR (cetuximab, bevacizumab, gemcitabine, cisplatin, and 5-fluorouracil), and vehicle control (T). The therapies were administered via intraperitoneal injections every 4 days for seven cycles from 7 weeks after cancer cell implantation. Mice treated with T-TR had significant reductions in tumor weight as compared to the control group (p < 0.05). Although mice in the T-CR group experienced a significant reduction in body weight gain, serum albumin, and gastrocnemius muscle mass (p < 0.05), no such reductions were observed in the T-TR group. Mice treated with T-TR had slightly increased CD11c⁺ DC and CD49b⁺ NK cell levels in the spleen (p < 0.05) and significantly lower tumor VEGF expression (p < 0.05). Tumor carcinoembryonic antigen expression was significantly reduced in both treatment groups (p < 0.05). Thus, addition of bevacizumab and cetuximab to gemcitabine, cisplatin, and fluorouracil may represent an effective treatment option for pancreatic cancer that warrants further study.     

The order effect of combined endurance and strength loadings on force and hormone responses: effects of prolonged training

Purpose

To examine acute responses and recovery of force and serum hormones to combined endurance and strength loadings utilizing different orders of exercises before and after training.

Methods

Physically active men were matched to an order sequence of endurance followed by strength (E + S, n = 12) or strength followed by endurance (S + E, n = 17). The subjects performed one experimental loading consisting of steady-state cycling and a leg press protocol before and after 24 weeks of order-specific combined training.

Results

No between-group difference in acute reductions of force was observed at week 0 (E + S ?23 %, p < 0.001; S + E ?22 %, p < 0.01) and 24 (E + S ?25 %, p < 0.001; S + E ?27 %, p < 0.001) and recovery in force was completed after 24 h in both groups at week 0 and 24. Concentrations of growth hormone (22-kDa) increased post-acute loading at week 0 (E + S, +57 fold, p < 0.05; S + E, +300 fold, p < 0.001; between-groups p < 0.001) and 24 (E + S, +80 fold, p < 0.01; S + E, +340 fold, p < 0.05; between-groups p < 0.05). No significant acute responses in concentrations of testosterone were observed at week 0 or 24. However, at week 0 testosterone was reduced during recovery following the E + S loading only (24 h ?23 %, p < 0.01; 48 h ?21 %, p < 0.001; between-groups at 24 and 48 h, p < 0.05), but was no longer observed after training. 1RM strength improved similarly in E + S (13 %, p < 0.001) and S + E (17 %, p < 0.001).

Conclusions

This study showed an order effect (E + S vs. S + E) in concentrations of testosterone during 2 days of recovery at week 0, which was diminished after 24 weeks of training. The initial difference in testosterone concentrations during recovery did not seem to be associated with strength development.     

Low BRAF and NRAS expression levels are associated with clinical benefit from DTIC therapy and prognosis in metastatic melanoma

Metastatic melanoma is characterized by a poor response to chemotherapy. Furthermore, there is a lack of established predictive and prognostic markers. In this single institution study, we correlated mutation status and expression levels of BRAF and NRAS to dacarbazine (DTIC) treatment response as well as progression-free and overall survival in a cohort of 85 patients diagnosed with advanced melanoma. Neither BRAF nor NRAS mutation status correlated to treatment response. However, patients with tumors harboring NRAS mutations had a shorter overall survival (p < 0.001) compared to patients with tumors wild-type for NRAS. Patients having a clinical benefit (objective response or stable disease at 3 months) on DTIC therapy had lower BRAF and NRAS expression levels compared to patients progressing on therapy (p = 0.037 and 0.003, respectively). For BRAF expression, this association was stronger among patients with tumors wild-type for BRAF (p = 0.005). Further, low BRAF as well as NRAS expression levels were associated with a longer progression-free survival in the total population (p = 0.004 and <0.001, respectively). Contrasting low NRAS expression levels, which were associated with improved overall survival in the total population (p = 0.01), low BRAF levels were associated with improved overall survival only among patients with tumors wild-type for BRAF (p = 0.013). These findings indicate that BRAF and NRAS expression levels may influence responses to DTIC as well as prognosis in patients with advanced melanoma.     

Contributory Anti-Inflammatory Effects of Mesenchymal Stem Cells,Not Conditioned Media,On Ovalbumin-Induced Asthmatic Changes in Male Rats

Our aim in selecting an appropriate cell fraction and conditioned media (CM) was to achieve the suitable candidate for ameliorating long-term chronic asthmatic changes of respiratory tract. Thirty-six rats were classified into healthy and sensitized groups, which were further divided into three subgroups; rats received systemically 50 μl volume of PBS, CM, or 2?×?10⁶ rat bone marrow-derived mesenchymal stem cells (rBMMSCs). Tracheal responsiveness (TR), immunologic responses, and recruitment of rBMMSCs into the lungs were evaluated. A high degree of TR and total WBC and percentages of eosinophils and neutrophils was significantly recorded in all sensitized groups rather than of controls (p?<?0.001 to p?<?0.05). Concurrently, a significant improvement of TR and eosinophil and neutrophil return toward normal levels was evident in sensitized rats receiving cells as compared to parallel asthmatic animals. Flow cytometric monitoring of lymphocyte subpopulation revealed a decrease in the number of CD3⁺CD4⁺ and concurrent increase in CD3⁺CD8⁺ in all sensitized rats as compared to control (p?<?0.001 to p?<?0.05). Noticeably, no significant modulatory effects of either cell or CM administration were achieved on the CD3⁺CD4⁺ and CD3⁺CD8⁺ populations in non-asthmatic rats. Corroborating our results, the number of CD3⁺CD4⁺ tended to increase (p?<?0.05) which coincided with a decreased manner of CD3⁺CD8⁺ populations as compared to other asthmatic groups (p?<?0.01 to p?<?0.05). Moreover, stem cells could efficiently transmigrate to the lung parenchyma, albeit the dynamic of asthmatic changes stimulated the rate of recruited cells. Our study shed light on superior effects of mesenchymal stem cells, but not CM, in attenuating chronic asthmatic changes in the model of rat.     

Recurring patterns in stationary intervals of abdominal uterine electromyograms during gestation

Abdominal uterine electromyograms (uEMG) studies have focused on uterine contractions to describe the evolution of uterine activity and preterm birth (PTB) prediction. Stationary, non-contracting uEMG has not been studied. The aim of the study was to investigate the recurring patterns in stationary uEMG, their relationship with gestation age and PTB, and PTB predictivity. A public database of 300 (38 PTB) three-channel (S1–S3) uEMG recordings of 30 min, collected between 22 and 35 weeks’ gestation, was used. Motion and labour contraction-free intervals in uEMG were identified as 5-min weak-sense stationarity intervals in 268 (34 PTB) recordings. Sample entropy (SampEn), percentage recurrence (PR), percentage determinism (PD), entropy (ER), and maximum length (L _MAX) of recurrence were calculated and analysed according to the time to delivery and PTB. Random time series were generated by random shuffle (RS) of actual data. Recurrence was present in actual data (p < 0.001) but not RS. In S3, PR (p < 0.005), PD (p < 0.01), ER (p < 0.005), and L _MAX (p < 0.05) were higher, and SampEn lower (p < 0.005) in PTB. Recurrence indices increased (all p < 0.001) and SampEn decreased (p < 0.01) with decreasing time to delivery, suggesting increasingly regular and recurring patterns with gestation progression. All indices predicted PTB with AUC ≥0.62 (p < 0.05). Recurring patterns in stationary non-contracting uEMG were associated with time to delivery but were relatively poor predictors of PTB.     

Changes of FibroScan,APRI, and FIB-4 in chronic hepatitis B patients with significant liver histological changes receiving 3-year entecavir therapy

Noninvasive fibrosis tests have been used widely for evaluation of liver fibrosis in patients with chronic hepatitis B (CHB). We aimed to investigate the influence of antiviral treatment on FibroScan, APRI, and FIB-4 in CHB patients with significant liver histological changes (SLHC) defined as inflammatory grade ≥ A2 and/or fibrosis stage ≥ F2. A total of 104 CHB patients with SLHC at the baseline were included. FibroScan, APRI, and FIB-4 values were compared before and after 3-year entecavir (ETV) treatment. Liver stiffness measurement values decreased significantly after 3-year ETV treatment in cirrhosis group (from 13.6 to 9.6 kPa, p = 0.018), significant fibrosis group (from 8.4 to 5.8 kPa, p = 0.001), and mild fibrosis group (from 5.5 to 4 kPa, p < 0.001). APRI decreased significantly after 3-year ETV treatment in patients with cirrhosis (from 0.80 to 0.25, p < 0.001), patients with significant fibrosis (from 0.54 to 0.24, p < 0.001), and those with mild fibrosis (from 0.35 to 0.23, p < 0.001). FIB-4 decreased significantly after 3-year ETV treatment in patients with cirrhosis (from 1.27 to 0.81, p = 0.007) and significant fibrosis (from 1.12 to 0.78, p < 0.001), while did not decrease significantly in patients with mild fibrosis (from 0.90 to 0.80, p = 0.389). FibroScan, APRI, and FIB-4 values decreased significantly after 3-year ETV treatment in CHB patients, which indicates that these noninvasive fibrosis tests might be useful for monitoring regression of liver fibrosis and assessing treatment efficacy during long-term ETV treatment.     

Cytokine response to acute running in recreationally-active and endurance-trained men

To compare the cytokine response to exhaustive running in recreationally-active (RA) and endurance-trained (ET) men. Eleven RA men (VO_2max 55 ± 7 mL·min^?1·kg^?1) and 10 ET men (VO_2max 68 ± 7 mL·min^?1·kg^?1) followed a controlled diet and refrained from volitional exercise for 8 days. On the fourth day, participants completed 60 min of treadmill running (65 % VO_2max), followed by intermittent running to exhaustion (70 % VO_2max). Fasting blood was obtained at baseline, after 20, 40 and 60 min of exercise, at the end of intermittent exercise, during 2 h of recovery and on four follow-up days (FU1–FU4). Tumour necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-1 receptor antagonist (IL-1ra) and creatine kinase (CK) were measured. Exercise increased the concentrations of all cytokines and CK, but there were no significant differences between groups. IL-1β increased (2.2–2.5-fold, P < 0.001) during exercise, while TNF-α was increased (1.6–2.0-fold, P < 0.001) during exercise and for 2 h post-exercise. IL-6 (71–84-fold, P < 0.001) and IL-1ra (52–64-fold, P < 0.001) were increased throughout exercise and up to FU1, peaking immediately after exercise and at 1.5–2 h post-exercise, respectively. CK concentrations were increased (P < 0.001) throughout exercise and up to FU4, peaking at FU1, but were not associated with changes in any cytokines. Exhaustive running resulted in modest and transient increases in TNF-α and IL-1β, and more marked and prolonged increases in IL-6 and IL-1ra, but improved training status did not affect this response. Increased CK might indicate either exercise-induced muscle cell disruption or increased cell permeability, although neither appears to have contributed to the increased cytokine concentrations.     

Stage-dependent detection of CD14+ and CD16+ cells in the human heart after myocardial infarction

Monocytes are critically involved in cardiovascular wound healing processes. Human monocytes can be classified into two subsets based on the expression of CD14 and CD16. Here, we examined the temporal and spatial distribution of CD14⁺ and CD16⁺ cells after myocardial infarction (MI) in human heart and spleen tissue and correlated it with markers of cardiac repair. Heart samples obtained at autopsy were histologically classified into acute (AMI; n?=?11), subacute (SAMI; n?=?10) and old (OMI; n?=?16) MI, or control myocardium (CONTR; n?=?8). Histochemical analyses revealed marked fibrosis in OMI (p?<?0.001 vs. CONTR). The adhesion molecule CD56 was also strongly expressed in OMI (p?<?0.01 vs. CONTR) and found to correlate with fibrosis (p?<?0.001). The number of capillaries was reduced in OMI (p?<?0.01 vs. CONTR; p?<?0.05 vs. AMI), whereas the hypoxia indicator carbonic anhydrase IX was predominantly expressed in AMI (p?<?0.01 vs. OMI and CONTR) and SAMI (p?<?0.05 vs. OMI and CONTR). The monocyte chemoattractrant osteopontin was also more highly expressed in hearts of SAMI patients (p?<?0.01 vs. CONTR). Numbers of CD14⁺ monocytes were found to correlate with CD16⁺ cells (p?<?0.05) and inversely with fibrosis (p?<?0.05). Regarding a MI-associated release of monocytes from spleen reservoirs, a non-significant reduction of splenic CD14⁺ and CD16⁺ cells was detected in subjects with AMI. In conclusion, disease stage-specific alterations in CD14⁺ and CD16⁺ cells in human heart may contribute to cardiac repair processes following MI.     

Cysteine protease inhibitor of <Emphasis Type="Italic">Schistosoma japonicum</Emphasis> - A parasite-derived negative immunoregulatory factor

Studies have shown that cysteine protease inhibitors from some parasites have immunosuppressive effects on the host. We previously have cloned a novel cysteine protease inhibitor from Schistosoma japonicum and purified its recombinant version (protein named rSj-C). Its possible inhibitory effect on the host immune response has not been described.This study shows that rSj-C inhibits lysosomal cysteine protease of murine dendritic cells (DCs). After DCs were incubated with rSj-C and then with soluble adult worm antigen (AWA) of S. japonicum, the mean fluorescence intensity of MHC class II antigens on the surface of DCs decreased significantly by flow cytometry. These results indirectly proved that rSj-C can suppress exogenous-antigen presentation by DCs. The flow cytometric assay revealed that in comparison with control groups, the proportion of CD4⁺CD25⁺Foxp3⁺ T cells among CD4⁺CD25⁺ T cells of Schistosom-infected mice increased significantly 8 weeks after the infected mice were injected with rSj-C (p ? 0.05). Additionally, the expression levels of cytokines IL-4 and TGF-β produced by T cells increased significantly as compared with these levels in the normal group (p ? 0.05). These results clearly show that the cysteine protease inhibitor from S. japonicum is a new parasite-derived immunosuppressive factor.     

Clinical and laboratory features of invasive community-onset methicillin-resistant Staphylococcus aureus infection: a prospective case–control study

Differences between the features of invasive community-onset methicillin-resistant Staphylococcus aureus (cMRSA) and methicillin-susceptible S. aureus (cMSSA) infections are incompletely understood. Fifty-seven patients with invasive cMRSA infection were prospectively identified at two teaching hospitals; for each cMRSA case, two cases of invasive cMSSA infection acted as controls. The primary outcome was 30-day all-cause mortality. Patients with invasive cMRSA infection were more likely to be Aboriginal (25% vs. 14%, age-adjusted odds ratio [OR] 2.5, p?=?0.037), reside in a long-term care facility and/or have been hospitalised in the previous year (51% vs. 34%, p?=?0.04) and less likely to have endocarditis (2% vs. 12%, p?=?0.02) or require admission to an intensive care unit or high-dependency area (7% vs. 21%, p?=?0.02). All-cause mortality at 30 days was similar in the cMRSA and cMSSA groups (9% vs. 7%, p?=?0.68). Panton–Valentine leukocidin (PVL) genes were detected in a similar proportion of cMRSA and cMSSA isolates (32% vs. 27%, p?=?0.49) and the presence of PVL genes was associated with younger age (35 years vs. 55 years, p?<?0.001), Aboriginal ethnicity (38% vs. 10%, p?<?0.001), skin and soft-tissue infection (54% vs. 19%, p?<?0.001), lower illness severity at presentation (SAPS II score 9 vs. 21, p?=?0.001) and shorter hospitalisation (9 days vs. 24 days, p?<?0.001). Patients with “PVL-positive” and “PVL-negative” S. aureus infection had similar 30-day all-cause mortality (4% vs. 9%, p?=?0.28). Few clinical features differentiated patients with invasive cMRSA infection from those with infection caused by cMSSA. Invasive “PVL-positive” S. aureus infection was associated with less morbidity but similar mortality to “PVL-negative” infection.     

Effect of adiposity on insulin action after acute and chronic resistance exercise in non-diabetic women

Purpose

Obesity may attenuate metabolic health improvements following lifestyle interventions. However, the effect of adiposity on insulin action following resistance exercise in young non-diabetic women is unknown. The purpose of this study was to test the hypothesis that adiposity attenuates improvements in insulin sensitivity and glucose-stimulated insulin secretion (INS_0–60/GLC_0–60) after both acute resistance exercise (ARE) and progressive training (PRT).

Methods

Twenty-six young non-diabetic women (21.2 ± 0.7 years) were randomly assigned to control (C; n = 7; BF 40.1 ± 2.1 %) or exercise groups: normal body fat (NBF; n = 8; BF 29.9 ± 2.3 %) and high body fat (HBF; n = 12; BF 48.2 ± 1.4 %). Acute whole-body exercises were performed at 60 % of 1-RM for three sets of 8–12 repetitions, and PRT was performed 3 days/week for 7 weeks. A 75 g OGTT was conducted before and after ARE and PRT to estimate insulin sensitivity (Matsuda index) and INS_0–60/GLC_0–60. Insulin area under the curve (AUC) was calculated using the trapezoidal model.

Results

ARE had no statistical effect on insulin action across groups. Strength and fat-free mass (via DXA) increased after PRT in both NBF and HBF (p < 0.05), but only HBF women decreased BF (p < 0.01). HBF women were less insulin sensitive at baseline compared to NBF women (p < 0.05). Insulin sensitivity increased 95 % and INS_0–60/GLC_0–60 decreased 32 % following PRT in NBF, but not HBF or C (p < 0.05). After training, enhanced insulin sensitivity was inversely related to decreased INS_0–60/GLC_0–60 (r = ?0.71, p < 0.001), fasting insulin (r = ?0.71, p < 0.001), and insulin AUC (r = ?0.85, p < 0.001).

Conclusion

Seven weeks of PRT increases insulin sensitivity and reduces glucose-stimulated insulin secretion in NBF, but not HBF women. Obesity attenuates exercise-induced improvements in glucose regulation in young non-diabetic women.     

Subcutaneous cholera toxin exposure induces potent CD103+ dermal dendritic cell activation and migration

CD103⁺ dermal dendritic cells (dDCs) are a recently described DC subset of the skin shown to be the principal migratory DCs capable of efficiently cross‐presenting antigens and activating CD8⁺ T cells. Harnessing their activity would promote vaccine efficacy, but it has been unclear how this can be achieved. We tested a panel of adjuvants for their ability to affect dDCs. In comparison to the other adjuvants tested, the capacity of cholera toxin (CT) to induce the migration of dDCs was unique. Within 24 h of CT injection, large numbers of highly activated dDCs (including CD103⁺ dDCs) migrated to the draining lymph nodes and cross‐presented coinjected antigens, potently activating naïve CD8⁺ T cells. Peptide vaccines adjuvanted with CT induced T‐cell responses uniquely characterized by dynamic cytokine responses including the production of IL‐2, and such vaccines were protective in situations reliant on CD8⁺ T‐cell responses, including liver‐stage Plasmodium challenge, or tumor challenge. This study is the first to examine the effects of adjuvants on CD103⁺ dDCs and identifies CT as a prototypical adjuvant for the activation of CD103⁺ dDCs, opening the way to development of vaccines and adjuvants that specifically target dDCs and generate effective CD8⁺ T‐cell responses.