Antemortem inhibition of phosphoinositide 3-kinase causes changes in meat quality traits in broiler chickens.

Phosphoinositide 3-kinase (PI3K) is an enzyme that mediates endocrinological responses, such as intracellular signaling of insulin and growth factors, and plays important roles in muscle homeostasis and growth. In this study, the effect of antemortem PI3K activity on meat quality traits was investigated using broiler chickens whose PI3K was inhibited pharmacologically. Breast and thigh muscles were harvested from broilers treated with the PI3K inhibitor wortmannin, and meat quality traits were evaluated by determination of color, water-holding capacity, and breaking strength. The pH and concentrations of glycogen and free amino acids were also investigated as determinants of the chemical properties of meat. The results indicated that antemortem PI3K inhibition by wortmannin modified breast muscle color with lower L∗ values (P < 0.05) and b∗ values (P < 0.05) and higher a∗ values (P < 0.05). Antemortem PI3K inhibition also increased the water-holding capacity of breast muscles (P < 0.05), although breaking strength was not much affected. In addition, antemortem PI3K inhibition increased the concentrations of free amino acids in breast muscles, especially arginine (P < 0.05) and glutamic acid (P < 0.05). Similar effects were observed in thigh muscles. Lower glycogen levels at sacrifice (P < 0.05) and the resultant higher pH during the postmortem period (P < 0.05) were associated with PI3K inhibition-induced changes in meat quality traits. The wortmannin-treated muscles shared certain features with dark, firm, and dry meat which is a common abnormal meat. These findings suggest that antemortem PI3K activity contributes to meat quality traits and is involved in the molecular mechanism of the production of meat quality abnormalities.

Homodigital artery flap reconstruction for fingertip amputation: a comparative study of the oblique triangular neurovascular advancement flap and the reverse digital artery island flap.

This fingertip reconstruction study retrospectively compared sensory recovery and active range of motion outcomes in neurovascular island advancement and reverse digital artery island flaps. Seventeen oblique triangular flaps and 14 reverse digital artery island flaps were performed for nail bed level fingertip amputations (Ishikawa subzone II). There was no significant difference between the two procedures in the Semmes-Weinstein monofilament test and range of motion results. For static and moving two-point discrimination tests, however, those with a reverse digital artery island flap required a longer period for sensory recovery compared to those with an oblique triangular advancement flap. This trend equilibrated at 12 months after surgery showing no significant difference in both static and moving two-point discrimination tests between the procedures.

Cytokine mediated immune responses in the Japanese pufferfish (Takifugu rubripes) administered with heat-killed Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the Mongolian dairy product.

The important role played by cytokines in host innate immunity and the interaction of subsets of immune and inflammatory cells through cytokines offer avenues for immune interventions. We investigated 16 cytokine gene responses in the Japanese pufferfish, Takifugu rubripes orally treated with a heat-killed lactic acid bacterium (LAB), Lactobacillus paracasei spp. paracasei (strain 06TCa22) (Lpp) isolated from a Mongolian dairy product at 1mgg(-1)body weightd(-1) for 3days. Additionally, we assessed superoxide anion production (SAP) and phagocytic activity (PA) of head kidney cells and resistance to Vibrio harveyi infection in treated fish. Significant up-regulation of pro-inflammatory (IL-1ß, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immunity inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intra-cellular pathogen killing (I-IFN-1 and IFN-γ), anti-inflammatory (IL-10) and peripheral T cell expansion and survival controlling (IL-2, IL-7, IL-15, IL-21 and TGF-ß1) cytokines was observed in the treated fish. Furthermore, significantly increased SAP, PA and pathogen resistance were observed in the treated fish compared to untreated fish. Our results indicate the enhancement of cytokine mediated immunity in T. rubripes by the use of the heat-killed Lpp as a potential immunostimulant and would be of great use in immunomodulation trials with the possibility to monitor positive immune response.

Elevated cytokine responses to Vibrio harveyi infection in the Japanese pufferfish (Takifugu rubripes) treated with Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the Mongolian dairy product.

With the aim of evaluating the effect of a Mongolian dairy product derived Lactobacillus paracasei spp. paracasei (strain 06TCa22) (Lpp) on the cytokine-mediated immune responses to Vibrio harveyi infection, we examined 16 cytokine expressions in the Japanese pufferfish, Takifugu rubripes. Fish were orally treated with the heat-killed Lpp at 1 mg g(-1) body weight d(-1) for 3 days. At 24 h posttreatment, fish were infected by an intramuscular injection of 0.1 mL V. harveyi bacterial suspension (10(8) cfu mL(-1)). Additionally, superoxide anion production (SAP) and phagocytic activity (PA) of head kidney cells were assessed during 120 h postinfection period. Significant up-regulation of pro-inflammatory (IL-1ß, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immune inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intra-cellular pathogen killing (I-IFN-1 and IFN-γ), anti-inflammatory (IL-10) and lymphocyte agonistic (IL-2, IL-7, IL-15, IL-21 and TGF-ß1) cytokines was observed in the treated fish compared to control ones during the pathogen infection. Furthermore, significantly increased SAP and PA (P < 0.01; 0.05) were recorded in the treated fish compared to untreated fish. These results suggest the beneficial role of Lpp in enhancement of cytokine-mediated immunity in the Japanese pufferfish against V. harveyi infection and application of this product as a potential fish immunostimulant.

Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells induced with heat-killed probiotics isolated from the Mongolian dairy products.

Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney (HK) cells to heat-killed lactic acid bacteria probiotics isolated from the Mongolian dairy products were investigated by transcriptomic examination. The HK cells were incubated with two heat-killed bacteria, namely Lactobacillus paracasei spp. paracasei (strain 06TCa22) and L. plantarum (strain 06CC2) and the responses of 16 cytokine genes at 0 (control), 1, 4, 8, 12, 24 and 48 h post-stimulation were assayed by multiplex RT-PCR analysis (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter, Inc.). The 16 genes included in the assay were pro-inflammatory cytokines (IL-1ß, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immune regulators (IL-12p35, IL-12p40 and IL-18), antiviral (I-IFN-1 and IFN-γ) and other regulatory (IL-2, IL-7, IL-15, IL-21, IL-10 and TGF-ß1) cytokines. Despite the differences in the transcriptional profiles, expression of all the cytokines tested here was significantly elevated by both the probiotic bacterial stimulants compared with the unstimulated control. Therefore, this in vitro study has demonstrated the modulation of cytokine defense mechanisms in the HK cells by the two heat-killed probiotics indicating their potentiality as novel immunostimulants to fish. However, strain-dependent varied expression of important cytokines (cell-mediated immune regulators, antiviral and anti-inflammatory cytokines) suggests better efficacy of L. paracasei spp. paracasei strain as fish immunostimulant. Further in vivo studies to elucidate the cytokine regulation networks will validate our present observations. A careful evaluation of ant-inflammatory properties may be undertaken using single strain to affirm the immunostimulatory capability. Moreover, application timings and frequency to assess the longevity of immunostimulant effects and to make the application cost-effective need to be evaluated before any practical use in aquaculture.

Large magnetoresistance through a single molecule due to a spin-split hybridized orbital.

Using organic materials in spintronic devices raises a lot of expectation for future applications due to their flexibility, low cost, long spin lifetime, and easy functionalization. However, the interfacial hybridization and spin polarization between the organic layer and the ferromagnetic electrodes still has to be understood at the molecular scale. Coupling state-of-the-art spin-polarized scanning tunneling spectroscopy and spin-resolved ab initio calculations, we give the first experimental evidence of the spin splitting of a molecular orbital on a single non magnetic C(60) molecule in contact with a magnetic material, namely, the Cr(001) surface. This hybridized molecular state is responsible for an inversion of sign of the tunneling magnetoresistance depending on energy. This result opens the way to spin filtering through molecular orbitals.

Upsizing the femoral component increases patellofemoral contact force in total knee replacement.

In posterior stabilised total knee replacement (TKR) a larger femoral component is sometimes selected to manage the increased flexion gap caused by resection of the posterior cruciate ligament. However, concerns remain regarding the adverse effect of the increased anteroposterior dimensions of the femoral component on the patellofemoral (PF) joint. Meanwhile, the gender-specific femoral component has a narrower and thinner anterior flange and is expected to reduce the PF contact force. PF contact forces were measured at 90°, 120°, 130° and 140° of flexion using the NexGen Legacy Posterior Stabilized (LPS)-Flex Fixed Bearing Knee system using Standard, Upsized and Gender femoral components during TKR. Increasing the size of the femoral component significantly increased mean PF forces at 120°, 130° and 140° of flexion (p = 0.005, p < 0.001 and p < 0.001, respectively). No difference was found in contact force between the Gender and the Standard components. Among the patients who had overhang of the Standard component, mean contact forces with the Gender component were slightly lower than those of the Standard component, but no statistical difference was found at 90°, 120°, 130° or 140° of flexion (p = 0.689, 0.615, 0.253 and 0.248, respectively). Upsized femoral components would increase PF forces in deep knee flexion. Gender-specific implants would not reduce PF forces.

Enlarged post-operative posterior condyle tightens extension gap in total knee arthroplasty.

We investigated whether the extension gap in total knee replacement (TKR) would be changed when the femoral component was inserted. The extension gap was measured with and without the femoral component in place in 80 patients with varus osteoarthritis undergoing posterior-stabilised TKR. The effect of a post-operative increase in the size of the femoral posterior condyles was also evaluated. The results showed that placement of the femoral component significantly reduced the medial and lateral extension gaps by means of 1.0 mm and 0.9 mm, respectively (p < 0.0001). The extension gap was reduced when a larger femoral component was selected relative to the thickness of the resected posterior condyle. When the post-operative posterior lateral condyle was larger than that pre-operatively, 17 of 41 knees (41%) showed a decrease in the extension gap of > 2.0 mm. When a specially made femoral trial component with a posterior condyle enlarged by 4 mm was tested, the medial and lateral extension gaps decreased further by means of 2.1 mm and 2.8 mm, respectively. If the thickness of the posterior condyle is expected to be larger than that pre-operatively, it should be recognised that the extension gap is likely to be altered. This should be taken into consideration when preparing the extension gap.

Spin-polarized scanning tunneling microscopy and spectroscopy study of chromium on a Cr(001) surface.

Several tens of chromium layers were deposited at 250 °C on a Cr(001) surface and investigated by spin-polarized scanning tunneling microscopy (SP-STM), Auger electron spectroscopy (AES) and scanning tunneling spectroscopy (STS). Chromium is found to grow with a mound-like morphology resulting from the stacking of several monolayers which do not uniformly cover the whole surface of the substrate. The terminal plane consists of an irregular array of Cr islands with lateral sizes smaller than 20 × 20 nm(2). Combined AES and STS measurements reveal the presence of a significant amount of segregants prior to and after deposition. A detailed investigation of the surface shows that it consists of two types of patches. Thanks to STS measurements, the two types of area have been identified as being either chromium pure or segregant rich. SP-STM experiments have evidenced that the antiferromagnetic layer coupling remains in the chromium mounds after deposition and is not significantly affected by the presence of the segregants.

Pancreatitis-associated protein-I and pancreatitis-associated protein-III expression in a rat model of kainic acid-induced seizure.

The pancreatitis-associated protein (PAP) family (also known as the regenerating gene (Reg) family) is a group of 16 kDa secretory proteins structurally classified as the calcium dependent-type lectin superfamily. Some PAP family members are expressed in neurons following peripheral nerve injury and traumatic brain injury. To determine whether PAP family members are expressed in non-traumatic brain injury, expressions were analyzed following kainic acid (KA)-induced seizure. PAP-I (also known as Reg2 in rat and RegIII-beta in mouse) and pancreatitis associated protein-III (PAP-III; RegIII-gamma in mouse) messenger ribonucleic acid (mRNA) was transiently expressed in some restricted areas, such as the hippocampus and parahippocampal area; expression was observed immediately at a maximal level 1 day after seizure. Expression disappeared within 3 days after seizure. In situ hybridization (ISH) and immunohistochemistry revealed neuronal PAP-I and PAP-III expression in the hippocampal dentate gyrus, perirhinal and entorhinal cortices, and the posterior cortical nucleus of the amygdala. The number of PAP-III mRNA-positive neurons was significantly greater than PAP-I mRNA-positive neurons. The majority of positive neurons co-localized with c-Jun, but not with glutamic acid decarboxylase (GAD). These results may suggest that PAP-I and PAP-III induction in non-GABAergic neurons would protect neurons against damage following seizure.

Possible contribution of hyalocytes to idiopathic epiretinal membrane formation and its contraction.

AIM: To address the cellular components and the contractile mechanisms of the idiopathic epiretinal membrane (ERM). METHODS: Ten surgically removed ERMs were fixed in 4% paraformaldehyde and analysed by whole-mount immunohistochemistry with anti-glial fibrillar acidic protein (GFAP) and alpha smooth-muscle actin (alphaSMA) antibodies. Type I collagen gel contraction assay, an established wound-healing assay in vitro, was performed using cultured bovine hyalocytes or normal human astrocytes (NHA) to evaluate the contractile property of the cells in the presence of tissue growth factor (TGF)-beta2. The expression of alphaSMA was also analysed by western blot analysis to examine myofibroblastic transdifferentiation of the cells. Vitreous-induced collagen gel contraction was also evaluated. RESULTS: All membranes were composed of alphaSMA immunopositive cells in contracted foci and GFAP immunopositive cells in the periphery. No apparent double positive cells were observed in any membranes examined. Cultured hyalocytes showed overexpression of alphaSMA and hypercontraction of collagen gels in response to TGF-beta2, while glial cells showed marginal change. The vitreous from ERM patients also caused overexpression of alphaSMA and hypercontraction of the gels embedding hyalocytes, which were almost completely inhibited in the presence of anti-TGF-beta2 neutralising antibody. CONCLUSIONS: Hyalocytes might be one of the critical components of ERM mediating its contractile property through the effect of TGF-beta2 in the vitreous fluid.

Residual internal limiting membrane in epiretinal membrane surgery.

BACKGROUND/AIM: To examine the degree of the residual internal limiting membrane (ILM) after epiretinal membrane (ERM) peeling. METHODS: Sixty-one eyes of 59 patients with ERM were enrolled. After ERM peeling, residual ILM was visualised with Brilliant Blue G (BBG). The residual ILM pattern was divided into three groups: (1) residual type (ILM mostly remained), (2) half type (approximately half of ILM remained), (3) no residual type (ILM mostly removed with ERM). If ILM remained, residual ILM was removed in all cases and histologically examined using the flat mount method in 10 cases. The correlation between the degree of ERM evaluated by preoperative best-corrected visual acuity (BCVA) and residual ILM pattern was also examined. RESULTS: Twenty-eight eyes (45.9%) were of the residual type. Three eyes (4.9%) were of the half type, and 30 eyes (49.2%) were of no residual type. The mean preoperative BCVA showed no significant correlation with the residual ILM pattern. Flat mount immunohistochemistry revealed many remnant cells, both glial fibrillar acidic protein positive and negative, on residual ILMs in all specimens examined. No recurrence that needed surgical treatment was observed. CONCLUSION: Residual ILM with remnant cells seems to be frequent after ERM removal. Intraoperative staining with BBG may be helpful in determining the extent of ILM removal.

Vascular endothelial growth factor expression by hyalocytes and its regulation by glucocorticoid.

AIM: Tumour necrosis factor-alpha (TNF-alpha) is one of the major inflammatory cytokines involved in the pathogenesis of various vitreoretinal diseases. The authors investigated the effect of hypoxia, TNF-alpha and dexamethasone on vascular endothelial growth factor (VEGF) expression by cultured hyalocytes. METHODS: Hyalocytes were isolated from bovine vitreous. Hypoxic and TNF-alpha-dependent effects on cultured hyalocytes were investigated using several assays to determine VEGF protein expression, hypoxia-inducible factor (HIF)-1alpha protein levels, HIF-1alpha-DNA-binding ability and VEGF mRNA stability. The effects of dexamethasone on VEGF expression and its intracellular signalling under hypoxic or TNF-alpha stimulated conditions were also examined. RESULTS: Hypoxic conditions and TNF-alpha stimulation induce VEGF expression in hyalocytes. These stimuli also stabilise HIF-1alpha protein and increase its DNA-binding ability. Dexamethasone significantly inhibits both HIF-1alpha protein levels and HIF-1alpha-DNA-binding activity, and also decreases the hypoxic- and TNF-alpha -dependent induction of VEGF expression in hyalocyte. However, dexamethasone has no significant effect on the stability of VEGF mRNA. CONCLUSIONS: Hyalocytes may be involved in various vitreoretinal diseases by increasing HIF-1alpha protein stability and HIF-1alpha-DNA binding, and thus increasing VEGF production under pathological conditions. Dexamethasone seems to be capable of inhibiting hypoxic and TNF-alpha dependent VEGF production, presumably via its inhibitory effects on HIF-1alpha protein levels and its DNA-binding activity.

Angiotensin-I converting enzyme inhibitory peptide derived from porcine skeletal muscle myosin and its antihypertensive activity in spontaneously hypertensive rats.

Crude myosin light chain was extracted from Japanese domestic pork loin and digested with pepsin. Antihypertensive peptide was isolated from this digest as a measure of its inhibitory activity for angiotensin-I converting enzyme (ACE). Through isolation with some chromatographies, a single active fraction was isolated, and it was detected as an octapeptide, Val-Lys-Lys-Val-Leu-Gly-Asn-Pro, from 47th to 54th positions of myosin light chain. The 50% inhibitory concentration of this peptide was 28.5 muM. Kinetic evaluation showed that this peptide was a noncompetitive inhibitor, but it was slowly hydrolyzed by ACE. At the dose of 10 mg/kg, this peptide showed antihypertensive activity after a maximum of 3 h of administration and was estimated as a temporally effective hypotensor.

Morphologic evaluation of the caudal end of the inferior petrosal sinus using 3D rotational venography.

BACKGROUND AND PURPOSE: The inferior petrosal sinus (IPS) is the main transvenous access route used to examine or treat lesions involving the cavernous sinus. To carry out these procedures successfully, one must have a detailed knowledge of the anatomy of the venous system around the junction of the IPS and the internal jugular vein (IJV). MATERIALS AND METHODS: Eighty-three sides in 63 patients (26 men, 37 women; mean, 56.5 years of age) were examined by using 3D rotational venography (3DRV). RESULT: The drainage patterns of the IPS could be classified into the following 6 types, with emphasis on the level of IPS-IJV junction: type A, the IPS drains into the jugular bulb in 1/83 sides (1.2%); type B, the IPS drains into the IJV at the level of the extracranial opening of the hypoglossal canal in 29/83 sides (34.9%); type C, the IPS drains into the lower extracranial IJV in 31/83 sides (37.3%); type D, the IPS forms a plexus and has multiple junctions to the IJV near the jugular foramen in 5/83 sides (6.0%); type E, the IPS drains directly into the vertebral venous plexus (VVP) with no connection to the IJV in 3/83 sides (3.6%); and type F, the IPS is absent in 14/83 sides (16.9%). Each type is also characterized by the way of anastomosis with the VVP. CONCLUSION: This classification seemed to be rational from the embryologic viewpoint, and it may be useful in establishing treatment strategies that involve endovascular manipulation via the IPS.

Effects of a humidity-stabilizing sheet on the color and K value of beef stored at cold temperatures.

The effects of a humidity-stabilizing sheet containing glycerol, on the color and K value of beef stored at cold temperatures were investigated in this study. A drip-absorbing sheet seems to be effective for the preservation of meat quality, while a humidity-stabilizing sheet containing glycerol prevents the increase of metmyoglobin in cold stored beef. Maintaining samples wrapped in humidity-stabilizing sheets containing glycerol at low temperature for 7days was a functional method for conserving the concentration of inosine monophosphate. Beef samples wrapped in sheets containing glycerol had lower K values than samples wrapped in sheets not containing glycerol. When the surface of the meat starts to desiccate, the humidity-stabilizing sheet releases moisture that has been absorbed from the beef in the early stages of storage. Thus, glycerol could potentially play a role as a humidity-stabilizing controller and color preservative. This research suggests that a humidity-stabilizing sheet containing glycerol is a useful moisture controller and prevents deterioration of meat quality, discoloration, and desiccation.

N-methyl-D-aspartate receptors play important roles in acquisition and expression of the eyeblink conditioned response in glutamate receptor subunit delta2 mutant mice.

Classical eyeblink conditioning has been known to depend critically on the cerebellum. Apparently consistent with this, glutamate receptor subunit delta2 null mutant mice, which have serious morphological and functional deficiencies in the cerebellar cortex, are severely impaired in delay paradigm. However, these mutant mice successfully learn in trace paradigm, even in '0-trace paradigm,' in which the unconditioned stimulus starts just after the conditioned stimulus terminates. Our previous studies revealed that the hippocampus and the muscarinic acetylcholine receptors play crucial roles in 0-trace paradigm in glutamate receptor subunit delta2 null mutant mice unlike in wild-type mice, suggesting a large contribution of the forebrain to 0-trace conditioning in this type of mutant mice. In the present study, we investigated the role of N-methyl-D-aspartate receptors in 0-trace eyeblink conditioning in glutamate receptor subunit delta2 null mutant mice. Mice were injected intraperitoneally with the noncompetitive N-methyl-d-aspartate receptor antagonist (+)MK-801 (0.1mg/kg) or saline, and conditioned with 350-ms tone conditioned stimulus followed by 100-ms periorbital shock unconditioned stimulus. Glutamate receptor subunit delta2 null mutant mice that received (+)MK-801 injection exhibited a severe impairment in acquisition of the conditioned response, compared with the saline-injected glutamate receptor subunit delta2 null mutant mice. In contrast, wild-type mice were not impaired in acquisition of 0-trace conditioned response by (+)MK-801 injection. After the injection solution was changed from (+)MK-801 to saline, glutamate receptor subunit delta2 null mutant mice showed a rapid and partial recovery of performance of the conditioned response. On the other hand, when the injection solution was changed from saline to (+)MK-801, glutamate receptor subunit delta2 null mutant mice showed a marked impairment in expression of the pre-acquired conditioned response, whereas impairment of the expression was small in wild-type mice. Injection of (+)MK-801 had no significant effects on spontaneous eyeblink frequency or startle eyeblink frequency to the tone conditioned stimulus in either glutamate receptor subunit delta2 null mutant mice or wild-type mice. These results suggest that N-methyl-D-aspartate receptors play critical roles both in acquisition and expression of the conditioned response in 0-trace eyeblink conditioning in glutamate receptor subunit delta2 null mutant mice.

Effect of NaCl on lipid peroxidation-derived aldehyde, 4-hydroxy-2-nonenal formation in minced pork and beef.

Pork and beef containing NaCl were stored at 0 °C and changes in 4-hydroxynonenal (HNE) content were analyzed after 0, 3, 7 and 10 days. As an index of the lipid peroxidation level, malonaldehyde (MA) content was also analyzed. In pork, HNE content in samples with 2% NaCl increased (P<0.05) after 7 and 10 days of storage. In beef, HNE content in all samples increased during storage. The HNE content in samples with NaCl was higher than those of the control, after 7 and 10 days of storage, although the difference was not always significant. Judging from MA content, NaCl may act as a pro-oxidant in pork and beef. HNE may accumulate in pork and beef in which lipid peroxidation is progress.