Preliminary study of an oral vaccine against infectious hematopoietic necrosis virus using improved yeast surface display technology

Infectious hematopoietic necrosis virus (IHNV) is a common pathogen that causes severe disease in the salmonid aquaculture industry. Because oral vaccines induce more efficient mucosal immunity than parenteral immunization, an oral vaccine was developed with an improved yeast cell surface display technology to induce an immune response to IHNV. The oral yeast vaccine, designated EBY100/pYD1-bi-G, was delivered orally to rainbow trout (Oncorhynchus mykiss) on days 1 and 32, and the nonspecific and specific immune responses were measured 50 days after the first vaccination. In the hindgut, spleen, and head kidney, the expression of IFN-1 and Mx-1 was significantly upregulated after oral vaccination with EBY100/pYD1-bi-G, and the highest expression of IFN-1 and Mx-1 was observed in the spleen (7.5-fold higher than the control group) and head kidney (3.9-fold higher than the control group), respectively. Several markers of the adaptive immune response (IgM, IgT, CD4, and CD8) were also significantly upregulated, and the highest expression of these markers was observed in the hindgut, suggesting that the mucosal immune response was successfully induced by oral vaccination with EBY100/pYD1-bi-G. Sera from the orally vaccinated rainbow trout showed higher anti-IHNV neutralizing antibody titers (antibody titer 81 ± 4) than the control sera (antibody titer 7 ± 3), and the relative percentage survival after IHNV challenge was 45.8% compared with 2% in the control group. Although the protection afforded by this orally delivered vaccine was lower than that of a DNA vaccine (83%–98%), it is a promising candidate vaccine with which to protect larval fish against IHNV, which are most susceptible to the virus and difficult to inject with a DNA vaccine.     

Histological alterations in gills of Astyanax aff. bimaculatus caused by acute exposition to zinc

Increasing contamination of aquatic ecosystems by metals has caused various morphological, physiological and biochemical changes in aquatic organisms, and the gills of fish are recognized as indicators of environmental quality. In this context, the present work proposed to study the effects of different concentrations of zinc (Zn) in the histology of gills of yellow tail lambari (Astyanax aff. bimaculatus) after acute exposure. Seventy-two adult males of A. aff. bimaculatus were used, the treatments were six concentrations of Zn: 0; 3; 5; 10; 15; and 20 mg/L of water, by 96 h, and gills, muscle and bone fragments were removed. Fragments of gills were fixed and included, sectioned in a rotary microtome and stained with toluidin blue. Fragments of bone, muscle and gills were dehydrated and digested to quantify the absorption of Zn. The median lethal concentration (LC₅₀) 96 h after Zn acute exposure was 10 mg/L of water. Noteworthy, Zn was highly toxic in acute exposure trials starting at the concentration 5 mg/L. The exposure of fish to the metal caused branchial histopathological changes correlated with increasing concentration, caused the death of fish at concentrations of 10, 15 and 20 mg/L. The histological alterations observed in the gills were hyperplasia, lamellar fusion, aneurysm, destruction of the lamellar epithelium, rupture of membrane, deletion of secondary lamellar high, which presented more severity in treatments exposed to the highest concentrations. In conclusion, gills of A. aff. bimaculatus presented profound histological alterations as a result of Zn exposure, and hence, proved to be excellent indicators of environmental contamination.     

Arsenic induced toxicity and histopathological changes in gill and liver tissue of freshwater fish,tilapia (Oreochromis mossambicus)

Acute toxicity of arsenic to tilapia (Oreochromis mossambicus) and its histopathological impacts on gill and liver tissue were evaluated. The median lethal concentration (96 h; LC₅₀) of arsenic (NaAsO₂) was calculated as 28.22 ppm in repeated semi static test method. Fish were exposed to 3 ppm, 28 ppm and 56 ppm concentrations of NaAsO₂ and gill and liver samples were collected after 48 h, 96 h and 192 h of exposure. The changes in gill were characterized by epithelial hyperplasia, epithelial lifting and oedema, lamellar fusion, aneurism, desquamation and necrosis, whereas, the liver tissue showed focal lymphocytic and macrophage infiltration, congestion, vacuolization and shrinkage of hepatocytes, dilation of sinusoids, cloudy swelling, vacuolar degeneration, focal necrosis and nuclear hypertrophy. The result showed that acute arsenic toxicity severely affects the normal behavior and vital organs which is deleterious for the exposed fish.     

Pentraxin 3 as a potential biomarker of acetaminophen-induced liver injury

ObjectiveOverdose of acetaminophen (APAP) can lead to severe liver injury in humans and experimental animals. Pentraxin-3 (PTX-3) is produced and released by several cell types. In this study, we aimed to evaluate whether PTX-3 is a potential biomarker in the identification of APAP-induced liver injury.Materials and methodsThirty adult male Wistar rats were randomly divided into three groups: control, APAP-1 and APAP-2 groups. APAP-1 (1 g/kg) and APAP-2 (2 g/kg) group rats were given APAP by gastric tube. Liver tissues and blood samples were obtained for biochemical and histopathological analysis. Biochemical parameters, plasma and liver PTX-3 levels and degree of liver necrosis were measured in all groups.ResultsAPAP treatments caused necrosis in liver and accompanied by elevated liver PTX-3 levels after 48 h. In APAP-1 and APAP-2 groups when compared with control group (7.5 ± 3.3 ng/mg protein), mean liver PTX-3 concentrations were 14.1 ± 3.0 (p = 0.032) and 28.5 ± 8.2 (p < 0.001) ng/mg protein, respectively. All rats (100%) in the APAP-2 group had the degree 3 liver necrosis. However 10%, 40% and 50% of rats had the degree 1, the degree 2 and the degree 3 liver necrosis in the APAP-1 group, respectively. The degrees of liver necrosis of the APAP-1 and APAP-2 groups were higher than the group of control (p < 0.001 and p < 0.001, respectively).ConclusionsPTX-3 may have a role in the APAP-induced liver injury in the rats. The elevated liver PTX-3 in the APAP-induced hepatic necrosis might be a marker of acute histological liver damage. Further prospective studies are necessary to clarify the prognostic value of liver PTX-3 for prediction of histological hepatic necrosis in the APAP-induced liver injury.     

The use of phospholipase A2 to prepare acellular porcine corneal stroma as a tissue engineering scaffold

This study was to develop a method using phospholipase A₂ (PLA₂) to prepare acellular porcine corneal stroma (APCS) for tissue engineering. The APCS was prepared from native porcine cornea (NPC) that was treated with 200 U/ml PLA₂ and 0.5% sodium deoxycholate (SD). The removal of DNA content, representing decellularization efficiency, reached to 91%, while all hydroxyproline and 80% of glycosaminoglycan were retained in the APCS when compared with NPC. The residual PLA₂ and SD were 0.35 ± 0.04 U/mg dry weight and 4.3 ± 0.8 ng/mg dry weight respectively. The extracts of APCS had no inhibitory effects on proliferation of corneal epithelial and endothelial cells as well as keratocytes. There was no sign of infiltration of neutrophilic leukocytes or leukomonocytes at 2 weeks after subcutaneous implantation of APCS. The prepared APCS displayed similar light transmittance to NPC. There were no significant differences in the areal modulus and curvature variation between APCS and NPC. Rabbit lamellar keratoplasty showed that the grafts of APCS were epithelialized completely in 8 ± 2 days, and their transparency was restored in 84 ± 11 days when the light transmittance of APCS-transplanted corneas displayed no significant difference compared with native corneas. Corneal neovascularization, corneal deformation, and graft degradation were not observed within 12 months.     

Sonographic evaluation of endothelial function in letrozole and tamoxifen users

ObjectivesStudies have shown that women previously treated for breast cancer present fewer cardiovascular events, indicating a possible protective effect of tamoxifen treatment. The effects of these aromatase inhibitors on cardiovascular protection remain controversial. The aim of this study was to compare some cardiovascular risk markers among breast cancer survivors following treatment with tamoxifen group (TMXg), letrozole group (LTZg) or no endocrine treatment group (NETg).MethodsA total of 103 breast cancer survivors: 35 using TMXg, 34 using letrozole group (LTZg) and 34 using no endocrine treatment group (NETg) were evaluated. Ultrasonographic evaluation of brachial artery flow-mediated dilation (FMD), carotid intima–media thickness (IMT) and stiffness index (β); blood total cholesterol, HDL and triglycerides were assessed.ResultsAll three groups presented similar values of HDL and IMT. TMXg showed the lowest total cholesterol (219.29 ± 36.31 mg/dL vs. 250.59 ± 38.37 mg/dL vs. 245.09 ± 35.35 mg/dL; TMXg vs. LTZg vs. NETg, respectively; p < 0.01—ANOVA), the highest triglycerides (139.34 ± 41.82 mg/dL vs. 111.35 ± 28.22 mg/dL vs. 122.09 ± 33.42 mg/dL; p < 0.01), the highest FMD (6.32 ± 2.33% vs. 4.10 ± 2.06% vs. 4.66 ± 2.52%; p < 0.01) and the lowest stiffness index (β) (5.08 ± 1.68 vs. 6.28 ± 1.75 vs. 5.99 ± 1.86; p = 0.01). LTZg did not differ significantly from NETg on any evaluated parameter.ConclusionsWe did not observe any effect of LTZg on the evaluated cardiovascular risk parameters compared to NETg. As such, the observed difference on lipid values, stiffness index (β) and FMD between women receiving tamoxifen and letrozole might be best attributed to the beneficial effect of tamoxifen than to a detrimental effect of letrozole.     

Alzheimer's disease presenilin-1 exon 9 deletion and L250S mutations sensitiZe SH-SY5Y neuroblastoma cells to hyperosmotic stress-induced apoptosis

Mutations in the presenilin-1 (PS1) and presenilin-2 (PS2) genes account for the majority of early-onset familial Alzheimer's disease cases. Recent studies suggest that presenilin gene mutations predispose cells to apoptosis by mechanisms involving altered calcium homeostasis and oxidative damage. In the present study, we determined whether PS1 mutations also sensitize cells to hyperosmotic stress-induced apoptosis. For this, we established SH-SY5Y neuroblastoma cell lines stably transfected with wild-type PS1 or either the PS1 exon 9 deletion (ΔE9) or PS1 L250S mutants. Cultured cells were exposed to an overnight (17 h) serum deprivation, followed by a 30 min treatment with either 20 mM glucose, 10 nM insulin-like growth factor-1 or 20 mM glucose +10 nM insulin-like growth factor-1. Cells were then cultured for a further 3, 6 or 24 h and stained for apoptotic condensed nuclei using propidium iodide. Confirmation that cells were undergoing an active apoptotic process was achieved by labelling of DNA strand breaks using the terminal dUTP nick end labelling (TUNEL) technique. We also determined cell viability using 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. Propidium iodide staining revealed that all cell lines and controls showed an increased number of apoptotic cells appearing with condensed nuclei at 24 h compared with 6 h and 3 h. High glucose-induced hyperosmotic stress resulted in significantly more apoptotic cells in the PS1 ΔE9 and PS1 L250S mutation cell lines at 24 h, compared with the wild-type PS1 lines (P<0.001, ANOVA for both comparisons). Mean values (±S.D.) for the percentage number of apoptotic cells at 24 h following high glucose treatment were 16.1±3.5%, 26.7±5.5% and 31.0±5.7% for the wild-type PS1, PS1 ΔE9 and PS1 L250S lines, respectively. The pro-apoptotic effects of high glucose treatment were reversed by 10 nM insulin-like growth factor-1, although to a lesser extent in the mutation cell lines (5.8±2.4%, 15.2±7.3% and 13.2±2.0% for the wild-type PS1, PS1 ΔE9 (P<0.01 for comparison with wild-type PS1) and PS1 L250S (P<0.01 for comparison with wild-type PS1) transfected lines, respectively. TUNEL labelling of cells at 24 h following treatment gave essentially the same results pattern as obtained using propidium iodide. The percentage number of apoptotic cells with DNA strand breaks (means±S.D.) following high glucose treatment was 15.4±2.6% for the wild-type PS1, 26.8±3.2% for the PS1 ΔE9 (P<0.001 for comparison with wild-type PS1) and 29.7±6.1% for the PS1 L250S transfected lines (P<0.001 for comparison with wild-type PS1). The PS1 ΔE9 and PS1 L250S transfected lines also showed a higher number of apoptotic cells with DNA strand breaks at 24 h following high glucose plus insulin-like growth factor-1 treatment (11.4±2.0% and 14.3±2.8%, respectively), compared with values for the wild-type PS1 lines (8.5±2.4%). These differences were significant (P<0.01) for the comparision of wild-type PS1 and PS1 L250S, but not PS1 ΔE9 lines. The mutation-related increases in number of apoptotic cells at 24 h following high glucose treatment were not accompanied by significant differences in cell viability at this time-point.Our results indicate that PS1 mutations predispose to hyperosmotic stress-induced apoptosis and that the anti-apoptotic effects of insulin-like growth factor-1 are compromised by these mutations. Perturbations of insulin-like growth factor-1 signalling may be involved in PS1 mutation-related apoptotic neuronal cell death in Alzheimer's disease.     

Toxicity of Moringa oleifera seed extract on some hematological and biochemical profiles in a freshwater fish,Cyprinus carpio

The study was carried out to investigate the acute and sublethal toxicity of Moringa oleifera seed extract on hematological and biochemical variables of a freshwater fish Cyprinus carpio under laboratory conditions. The 96 h LC50 value of M. oleifera seed extract to the fish C. carpio was estimated by probit analysis method and was found to be 124.0 mg/L (with 95% confidence limits). For sublethal studies a non lethal dose of 1/10th of 96 h LC50 value (12.40 mg/L) was taken. During acute treatment (96 h), hematological variables like red blood cell count (RBC), hemoglobin (Hb), hematocrit (Hct), and mean corpuscular hemoglobin concentration (MCHC) were significantly (P < 0.05) decreased in fish exposed to seed extract. However a significant (P < 0.05) increase in white blood cell count (WBC), mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) value was observed in the exposed fish during above treatment period when compared to that of the control groups. Biochemical parameters such as plasma protein and glucose levels significantly declined in fish exposed to seed extract while a significant (P < 0.05) increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activity was observed. During sublethal treatment (12.40 mg/L), WBC count, MCV, MCH, plasma glucose, AST, ALT and ALP activities were gradually elevated (P < 0.05) at the end of 7, 14, 21, 28 and 35th days in seed extract exposed fish, whereas plasma protein level declined. However, a biphasic trend was noticed in Hb, Hct, RBC and MCHC levels. This study may provide baseline information about the toxicity of M. oleifera seed extract to C. carpio and to establish safer limit in water purification.     

Contribution of nitric oxide synthase (NOS) in blood–brain barrier disruption during acute focal cerebral ischemia in normal rat

Endogenous level of nitric oxide (NO) is increased in the brain following the stroke, and deactivation of NO synthase has been shown to attenuate its destructive actions in animal stroke models using middle cerebral artery occlusion (MCAO) procedures. However, little is known about the effects of NO in cerebral vascular integrity and edema during acute cerebral ischemia. Here we investigated whether NO plays any role in the progression of blood–brain barrier (BBB) disruption and edema formation in ischemia/reperfusion injury. Intraperitoneal administration of NO substrate l-arginine (300 mg/kg), or NOS inhibitor (l-NAME, 1 mg/kg), was done in normal rats at 20 min before a 60-min MCAO. Mean arterial blood pressures (MAP) and regional cerebral blood flow (rCBF) were continuously recorded during experiment. Neurological deficit score (NDS) was evaluated 12 h after termination of MCAO followed with evaluations of cerebral infarction volume (CIV), edema formation and cerebral vascular permeability (CVP), as determined by the Evans blue dye extravasations (EBE) technique. No significant changes were observed in the values of MAP and rCBF with l-arginine or l-NAME during ischemia or reperfusion periods. There was a 75–85% reduction in rCBF in during MCAO which returned back to its pre-occlusion level during reperfusion. Acute cerebral ischemia with or without l-arginine augmented NDS (4.00 ± 0.44 and 3.00 ± 0.30), in conjunction with increased CIV (518 ± 57 mm³ and 461 ± 65 mm³), provoked edema (3.09 ± 0.45% and 3.30 ± 0.49%), and elevated EBE (8.28 ± 2.04 μg/g and 5.09 ± 1.41 μg/g). Inhibition of NO production by l-NAME significantly improved NDS (1.50 ± 0.22), diminished CIV (248 ± 56 mm³), edema (1.18 ± 0.58%) and EBE (1.37 ± 0.12 μg/g). This study reconfirms the cerebroprotective properties of reduced tissue NO during acute ischemic stroke, and it also validates the deleterious actions of increased NOS activity on the disruption of cerebral microvascular integrity and edema formation of ischemia/reperfusion injuries in normal rat, without changing arterial blood pressure or blood flows to ischemic regions.     

Synaptic degradation of cardiac autonomic nerves in streptozotocin-induced diabetic rats

Background: Cardiac autonomic neuropathy (CAN) is a common complication in type I diabetes mellitus (DM). Nevertheless, the relationship between functional and structural disturbances of cardiac autonomic nerves remains unclear. Methods and results: To clarify this relationship, we studied heart rate variability (HRV) and ultrastructural changes of cardiac autonomic nerves in streptozotocin (STZ)-induced DM in rats. STZ was injected (65 mg/kg intravenous) into the tail vein of male Wistar rats to destroy β cells in the pancreatic islets. After STZ injection, fasting blood sugar (FBS) increased from baseline values of 75 ± 3 mg/dl up to 328 ± 12 mg/dl within 1 week and it reached up to 353 ± 24 mg/dl within 17 weeks. HR in these rats was decreased within 20 days and low HR was maintained for the observation period. TP and HF power started decreasing 20 days after STZ injection, and this decrease progressed throughout the observation period. The L/H power ratio was decreased 80 days after STZ. Electron microscopic findings indicated a depletion of neurotransmitter vesicles and degradation of parasympathetic nerve endings but not of sympathetic ones in the SA node region of the heart in the early stages of DM. In the late stages of DM, the same region showed degradation of both sympathetic and parasympathetic nerve endings. Conclusion: Synaptic degradation in parasympathetic nerves immediately after the onset of DM, and in sympathetic nerves much later in the development of DM is consistent with functional derangements in cardiac autonomic nerve activities assessed by HRV analysis.     

Alpha-mannosidase activity in goats fed with Sida carpinifolia

Human α-mannosidosis results from α-mannosidase deficiency and progressive accumulation of mannose-rich oligosaccharides in lysosomes. Two days before Saanen goats were fed with Sida carpinifolia, α-mannosidase activity in leukocytes was 128±28 nmoles 4-MU/h/mg protein (first trial) and 104±6 nmoles 4-MU/h/mg protein (second trial). At day 5, after the introduction of S. carpinifolia diet, the α-mannosidase activity in leukocytes was significantly increased, both in the first (288±13 nmoles 4-MU/h/mg protein) and in the second trial (303±45 nmoles 4-MU/h/mg protein), and it returned to normal levels 2 days after the withdrawal of the plant from the diet (114±7 nmoles 4-MU/h/mg protein in first trial, and 108±25 nmoles 4-MU/h/mg protein in the second one). Plasma α-mannosidase activity decreased significantly 4 days after animal exposure to the S. carpinifolia diet (769±167 nmoles 4-MU/h/ml) and returned to normal values 10 days after the withdrawal of the plant from the diet (1289±163 nmoles 4-MU/h/ml). Thin-layer chromatography showed an abnormal excretion of oligosaccharides in urine as of day 2 after diet exposure, which persisted until one day after the withdrawal of the plant. Animals presented neurological clinical signs beginning at day 37 (in the first trial) and at day 25 (in the second trial) after being fed with the plant. The results obtained herein suggest that oligosaccharides observed in urine are a result of a decrease in α-mannosidase activity in plasma. S. carpinifolia seems to have other compounds that act on α-mannosidase enzyme in leukocytes in a competitive manner with swainsonine. The increase in α-mannosidase enzyme in leukocytes could be attributed to one of these compounds present in S. carpinifolia.     

Effects of a continuous-combined regimen of low-dose hormone therapy (oestradiol and norethindrone acetate) and tibolone on the quality of life in symptomatic postmenopausal women: A double-blind,randomised study

ObjectiveThis study compared the effects of a continuous-combined regimen of low-dose hormone therapy (LD-HT) versus tibolone and supplemental calcium/vitamin D3 (control) on quality of life (QoL) in symptomatic postmenopausal women.DesignThis study was a prospective, randomised, double-blind, comparative trial with a control group.SettingThe study was conducted in a climacteric outpatient clinic in the University Hospital of Federal University of Juiz de Fora, Brazil.PopulationA total of 174 postmenopausal women under 60 years of age who attended the climacteric outpatient clinic between June 2009 and June 2011 were recruited. These women complained of moderate or intense vasomotor symptoms and exhibited no contraindications for the use of hormone therapy.InterventionsThe patients were randomised into three groups: (1) daily treatment with 2.5 mg tibolone (n = 64), (2) 50 mg calcium carbonate + 200 IU vitamin D3 (Ca/Vit D3, n = 54) or (3) 1 mg oestradiol + 0.5 mg norethindrone acetate (E2/NETA, n = 56) for 12 weeks.Primary outcome measuresThe primary outcome was the evaluation of QoL using the Women's Health Questionnaire (WHQ) in all subjects at baseline and after 4, 8 and 12 weeks of treatment.ResultsA total of 130 women in the following groups completed the study: tibolone (n = 42), Ca/Vit D3 (n = 44) and E2/NETA (n = 44). An improved QoL based on the WHQ was observed at T0 (80.12 ± 14.04, 77.73 ± 15.3, 77.45 ± 15.4) and T12 (57.0 ± 15.5, 55.7 ± 16.7, 58.4 ± 12.6) for the tibolone, E2 + NETA and Ca/Vit D3 groups, respectively (p values <0.05). The three groups exhibited significantly different scores at T12 for sexual behaviour and vasomotor symptoms. The tibolone group exhibited better sexual function compared with the E2/NETA and Ca/Vit D3 groups (4.2 ± 26, 5.6 ± 2.8, 5.4 ± 2.8, respectively, p values <0.05). LD-HT was superior to tibolone and Ca/Vit D3 treatment for improvements in vasomotor symptoms (3.2 ± 1.5, 4.0 ± 1.8, 4.3 ± 2.0, respectively, p values <0.05). Adverse effects were few and mild.ConclusionsAn improved QoL was observed in the three study groups. Tibolone primarily improved sexual function, and E2/NETA exhibited a superior response for vasomotor symptoms.     

Alteration in biochemical parameters during plateletpheresis in healthy donors: A compendious analysis

ObjectivesDuring plateletpheresis, citrate induces hypocalcemia and hypomagnesemia, which are usually transient and self-limiting, but they can lead to significant donor discomfort. The aim of study was to determine the effect of citrate infusion on a multitude of biochemical parameters during plateletpheresis in healthy donors and to correlate changes with adverse donor reactions.MethodsThe study was conducted on 60 healthy plateletpheresis donors. Blood samples were drawn on three occasions, a baseline pre-donation sample, 30 min at start of procedure and 30 min post procedure. Heparinized samples were taken to measure ionized calcium and plain samples to measure serum calcium, serum magnesium, parathyroid hormone, total protein and serum albumin.ResultsThere was statistically significant decline in mean total calcium (9.27 ± 0.66 mg/dl to 8.72 ± 0.87 mg/dl) and ionized calcium (3.8 ± 0.51 mg/dl to 2.9 ± 0.67 mg/dl) from baseline until 30 min after the start of procedure respectively. A significant fall in serum magnesium, total protein and serum albumin was observed. The mean parathyroid hormone showed significant increase from baseline levels till at the completion of procedure (19.94 ± 12.1 pg/ml to 92.08 ± 36.78 pg/ml). If the yield was set constant, there was negative correlation between ACD used and pre-donation platelet count. Majority of adverse donor reactions were hypocalcemic reactions, which were more with Amicus double yield plateletpheresis and were managed with calcium supplementation.ConclusionPlateletpheresis induces marked reduction in serum calcium and magnesium levels. Moreover, increase in parathyroid hormone levels was significant. In addition, decline in total protein and serum albumin may be a concern in donors also participating in plasmapheresis.     

Serum vitamin D status is associated with the presence but not the severity of primary open angle glaucoma

ObjectivesVitamin D is involved in visual health and function. Our objective was to determine whether age-related vitamin D insufficiency was associated with the presence and the severity of primary open angle glaucoma (POAG) in a case-control study of older adults.Study designCase-control study.Main outcome measures. One hundred fifty cases diagnosed with moderate-to-severe POAG (mean, 75.1 ± 8.5 years; 42.0% female) and 164 healthy controls (mean, 73.0 ± 7.9 years; 59.8% female) were included. POAG diagnosis was based on classical diagnostic criteria of optic nerve cupping and/or RNFL thinning, measured with optical coherence tomography. Severe POAG was defined as Humphrey visual field mean deviation (MD) worse than −12 dB. Vitamin D insufficiency was defined as serum 25OHD ≤ 75 nmol/L. Age, gender, mean arterial pressure, vitamin D supplementation, visual acuity, and intraocular pressure were used as potential confounders.ResultsPOAG cases had lower mean serum 25OHD concentration than controls (42.9 ± 25.7 nmol/L versus 49.4 ± 29.5 nmol/L, P = 0.039) and a greater prevalence of vitamin D insufficiency (90.7% versus 82.3%, P = 0.032). Increased mean serum 25OHD concentrations were associated with lower POAG frequency, even after adjustment for potential confounders (OR = 0.89 per 10 nmol/L of 25OHD, P = 0.045). Similarly, vitamin D insufficiency was associated with POAG (OR = 2.09, P = 0.034). Among POAG cases, no 25OHD difference was observed between moderate and severe POAG cases (respectively, 39.2 ± 23.3 nmol/L versus 45.1 ± 26.7 nmol/L, P = 0.188); and no between-group difference regarding the prevalence of vitamin D insufficiency (88.9% versus 94.0%, P = 0.313).ConclusionsDecreased serum 25OHD concentration was associated with POAG. There was no 25OHD difference between moderate and severe POAG.     

Nutrition education in postmenopausal women: Changes in dietary and cardiovascular indices

ObjectiveThe aim of the current study was to examine whether a diet rich in dairy products followed by a nutrition education program for the prevention of osteoporosis could have any adverse effect on certain cardiovascular disease (CVD) risk factors over a 5-month intervention period.MethodsA total sample of 82 women (55–65 years old) was randomized to a dietary intervention group (IG: n = 42), attending biweekly nutrition education program and provided with low-fat, fortified dairy products and to a control group (CG: n = 40). Changes in dietary, biochemical and clinical indices related to CVD were determined at the end of the 5-month intervention period.ResultsThe IG was found to have a higher decrease in the percentage of energy intake derived from total fat and a higher increase in the intake of calcium, phosphorus, magnesium and potassium compared to the CG (p < 0.05). Furthermore, the IG subjects were found to have a lower increase in BMI (0.7 ± 0.1 versus 1.4 ± 0.2 Kg/m², p = 0.011) and systolic blood pressure (SBP) (2.5 ± 2.9 versus 7.8 ± 2.2 mmHg, p = 0.040) and a higher decrease in serum total cholesterol (−5.2 ± 3.3 versus 6.9 ± 5.1 mg/dl, p = 0.042) and LDL-cholesterol levels (−20.0 ± 2.6 versus −12.4 ± 4.2 mg/dl, p = 0.034) compared to the CG.ConclusionsThe findings of the current study indicate that a dietary intervention aiming to minimize the risk for osteoporosis did not have any adverse effects on CVD risk factors. On the contrary, it has induced favourable changes in BMI, serum lipids and SBP.     

Mesenchymal stem cell proliferation and differentiation on an injectable calcium phosphate – Chitosan composite scaffold

Calcium phosphate cement (CPC) can be molded or injected to form a scaffold in situ, has excellent osteoconductivity, and can be resorbed and replaced by new bone. However, its low strength limits CPC to non-stress-bearing repairs. Chitosan could be used to reinforce CPC, but mesenchymal stem cell (MSC) interactions with CPC-chitosan scaffold have not been examined. The objective of this study was to investigate MSC proliferation and osteogenic differentiation on high-strength CPC-chitosan scaffold. MSCs were harvested from rat bone marrow. At CPC powder/liquid (P/L) mass ratio of 2, flexural strength (mean ± sd; n = 5) was (10.0 ± 1.1) MPa for CPC-chitosan, higher than (3.7 ± 0.6) MPa for CPC (p < 0.05). At P/L of 3, strength was (15.7 ± 1.7) MPa for CPC-chitosan, higher than (10.2 ± 1.8) MPa for CPC (p < 0.05). Percentage of live MSCs attaching to scaffolds increased from 85% at 1 day to 99% at 14 days. There were (180 ± 37) cells/mm² on scaffold at 1 day; cells proliferated to (1808 ± 317) cells/mm² at 14 days. SEM showed MSCs with healthy spreading and anchored on nano-apatite crystals via cytoplasmic processes. Alkaline phosphatase activity (ALP) was (557 ± 171) (pNPP mM/min)/(μg DNA) for MSCs on CPC-chitosan, higher than (159 ± 47) on CPC (p < 0.05). Both were higher than (35 ± 32) of baseline ALP for undifferentiated MSCs on tissue-culture plastic (p < 0.05). In summary, CPC-chitosan scaffold had higher strength than CPC. MSC proliferation on CPC-chitosan matched that of the FDA-approved CPC control. MSCs on the scaffolds differentiated down the osteogenic lineage and expressed high levels of bone marker ALP. Hence, the stronger CPC-chitosan scaffold may be useful for stem cell-based bone regeneration in moderate load-bearing maxillofacial and orthopedic applications.     

Adhesion,migration and mechanics of human adipose-tissue-derived stem cells on silk fibroin–chitosan matrix

Silk fibroin–chitosan (SFCS) scaffold is a naturally derived biocompatible matrix with potential reconstructive surgical applications. In this study, human adipose-derived mesenchymal stem cells (ASCs) were seeded on SFCS scaffolds and cell attachment was characterized by fluorescence, confocal, time-lapse, atomic force, and scanning electron microscopy (SEM) studies. Adhesion of ASCs on SFCS was 39.4 ± 4.8% at 15 min, increasing to 92.8 ± 1.5% at 120 min. ASC adhered at regions of architectural complexity and infiltrate into three-dimensional scaffold. Time-lapse confocal studies indicated a mean ASC speed on SFCS of 18.47 ± 2.7 μm h^?1 and a mean persistence time of 41.4 ± 9.3 min over a 2.75 h study period. Cytokinetic and SEM studies demonstrated ASC–ASC interaction via microvillus extensions. The apparent elastic modulus was significantly higher (p < 0.0001) for ASCs seeded on SFCS (69.0 ± 9.0 kPa) than on glass (6.1 ± 0.4 kPa). Also, cytoskeleton F-actin fiber density was higher (p < 0.05) for ASC seeded on SFCS (0.42 ± 0.02 fibers μm^?1) than on glass-seeded controls (0.24 ± 0.03 fibers μm^?1). Hence, SFCS scaffold facilitates mesenchymal stem cell attachment, migration, three-dimensional infiltration, and cell–cell interaction. This study showed the potential use of SFCS as a local carrier for autologous stem cells for reconstructive surgery application.     

Acute influence of restricted ankle dorsiflexion angle on knee joint mechanics during gait

BackgroundRestrictions in range of ankle dorsiflexion (DF) motion can persist following ankle injuries. Ankle DF is necessary during terminal stance of gait, and its restricted range may affect knee joint kinematics and kinetics. The purpose of this study was to investigate the acute influence of varied levels of restricted ankle DF on knee joint sagittal and frontal plane kinematics and kinetics during gait.MethodsThirty healthy volunteers walked with a custom-designed ankle brace that restricted ankle DF. Kinematics and kinetics were collected using a 7-camera motion analysis system and two force plates. Ankle dorsiflexion was restricted in 10-degree increments, allowing for four conditions: Free, light (LR), moderate (MR) and severe restriction (SR). Knee angles and moments were measured during terminal stance.ResultsReal peak ankle DF for Free, LR, MR, and SR were 13.7 ± 4.8°, 11.6 ± 5.0°, 7.5 ± 5.3°, and 4.2 ± 7.2°, respectively. Peak knee extension angles under the same conditions were − 6.7 ± 6.7°, − 5.4 ± 6.4°, − 2.5 ± 7.5°, and 0.6 ± 7.8°, respectively, and the peak knee varus moment was 0.48 ± 0.17 Nm/kg, 0.47 ± 0.17 Nm/kg, 0.53 ± 0.20 Nm/kg, and 0.57 ± 0.20 Nm/kg. The knee varus moment was significantly increased from MR condition with an 8-degree restriction in ankle DF.ConclusionKnee joint kinematics and kinetics in the sagittal and frontal planes were affected by reduced ankle DF during terminal stance of gait. Differences were observed with restriction in ankle DF range of approximately 8°.Level of evidencelevel III     

Oxidative status and chymotrypsin-like activity in right and left ventricle hypertrophy in an experimental model of emphysema

Although cardiac muscle hypertrophy has been studied in association with several diseases, its mechanism in patients with emphysema, in particular in relation to oxidative stress and proteolysis, remains unknown. The role of oxidative stress and proteolysis in right and left ventricle hypertrophy was investigated in hamsters with emphysema induced by 2 different doses of papain (20 mg/mL, E20 and 40 mg/mL, E40). The thickness of the ventricles, total and cardiac weight, lipid peroxidation, carbonyl proteins, total antioxidant capacity (TAC), and proteasomal proteolytic activity were evaluated in the right ventricle (RV) and the left ventricle (LV) of control and emphysema hamsters. RV thickness was increased by 12% in the E20 group and by 29% in the E40 group. Lipid peroxidation measured by chemiluminescence was increased in the E40 group (from 3350.68 ± 392.44 URL/g tissue to 4696.63 ± 1076.70 URL/g tissue, p < 0.05). TAC also increased only in the E40 group. In the LV, chemiluminescence values increased from 4044.77 ± 503.39 to 5517.10 ± 388.27 in the E20 group and to 8169.14 ± 1748.77 URL/g tissue in the E40 group (p < 0.05, both). TAC significantly increased in the E20 and E40 groups. No differences were detected in substances reactive to thiobarbituric acid or carbonyl proteins when comparing ventricles or doses. Chymotrypsin-like proteolytic activity significantly decreased in both groups and ventricles. Emphysema can induce right and left ventricle lipid peroxidation and result in antioxidant mobilization. These data together support the idea that cardiac hypertrophy in response to emphysema is mediated in part by proteolytic pathways with involvement of reactive species.