Pre‐synaptic and post‐synaptic neuronal activity supports the axon development of callosal projection neurons during different post‐natal periods in the mouse cerebral cortex

Callosal projection neurons, one of the major types of projection neurons in the mammalian cerebral cortex, require neuronal activity for their axonal projections [H. Mizuno et al. (2007) J. Neurosci., 27, 6760–6770; C. L. Wang et al. (2007) J. Neurosci., 27, 11334–11342]. Here we established a method to label a few callosal axons with enhanced green fluorescent protein in the mouse cerebral cortex and examined the effect of pre‐synaptic/post‐synaptic neuron silencing on the morphology of individual callosal axons. Pre‐synaptic/post‐synaptic neurons were electrically silenced by Kir2.1 potassium channel overexpression. Single axon tracing showed that, after reaching the cortical innervation area, green fluorescent protein‐labeled callosal axons underwent successive developmental stages: axon growth, branching, layer‐specific targeting and arbor formation between post‐natal day (P)5 and P9, and the subsequent elaboration of axon arbors between P9 and P15. Reducing pre‐synaptic neuronal activity disturbed axon growth and branching before P9, as well as arbor elaboration afterwards. In contrast, silencing post‐synaptic neurons disturbed axon arbor elaboration between P9 and P15. Thus, pre‐synaptic neuron silencing affected significantly earlier stages of callosal projection neuron axon development than post‐synaptic neuron silencing. Silencing both pre‐synaptic and post‐synaptic neurons impaired callosal axon projections, suggesting that certain levels of firing activity in pre‐synaptic and post‐synaptic neurons are required for callosal axon development. Our findings provide in‐vivo evidence that pre‐synaptic and post‐synaptic neuronal activities play critical, and presumably differential, roles in axon growth, branching, arbor formation and elaboration during cortical axon development.     

Enhancement of cancer vaccine therapy by systemic delivery of a tumor-targeting Salmonella-based STAT3 shRNA suppresses the growth of established melanoma tumors

Cancer vaccine therapies have only achieved limited success when focusing on effector immunity with the goal of eliciting robust tumor-specific T-cell responses. More recently, there is an emerging understanding that effective immunity can only be achieved by coordinate disruption of tumor-derived immunosuppression. Toward that goal, we have developed a potent Salmonella-based vaccine expressing codon-optimized survivin (CO-SVN), referred to as 3342Max. When used alone as a therapeutic vaccine, 3342Max can attenuate growth of aggressive murine melanomas overexpressing SVN. However, under more immunosuppressive conditions, such as those associated with larger tumor volumes, we found that the vaccine was ineffective. Vaccine efficacy could be rescued if tumor-bearing mice were treated initially with Salmonella encoding a short hairpin RNA (shRNA) targeting the tolerogenic molecule STAT3 (YS1646-shSTAT3). In vaccinated mice, silencing STAT3 increased the proliferation and granzyme B levels of intratumoral CD4(+) and CD8(+) T cells. The combined strategy also increased apoptosis in tumors of treated mice, enhancing tumor-specific killing of tumor targets. Interestingly, mice treated with YS1646-shSTAT3 or 3342Max alone were similarly unsuccessful in rejecting established tumors, whereas the combined regimen was highly potent. Our findings establish that a combined strategy of silencing immunosuppressive molecules followed by vaccination can act synergistically to attenuate tumor growth, and they offer a novel translational direction to improve tumor immunotherapy.     

Antibiotic therapy against acute tonsillopharyngitis in children due to group A beta-hemolytic streptococci: comparison of clinical efficacy, the bactericidal effects, and effects on oral flora between cefditoren pivoxil for 5 days and amoxicillin for 10 days

We compared the clinical efficacy, the bactericidal effects, effect on the oral microbial flora, and adverse reactions between cefditoren pivoxil (CDTR-PI) for 5 days and amoxicillin (AMPC) for 10 days in children with acute group A beta-hemolytic streptococci (GAS) tonsillopharyngitis, and simultaneously examined the emm genotype and drug susceptibility of the isolated GAS. The results showed that the clinical efficacy was 100% for CDTR-PI and 97.9% for AMPC, with no difference between the two groups, and the bacterial elimination rate was 100% in both groups. No serious adverse event was noted in either group. On the other hand, concerning changes in the oral microbial flora between before and after treatment, the amount of bacteria showed no change in the CDTR-PI group (p = 0.5761) but clearly decreased in the AMPC group (p = 0.0049). This indicates that CDTR-PI does not disturb the oral microbial flora compared with AMPC. Also, the emm types determined in the 112 GAS strains isolated in this study were similar to those that have recently been isolated frequently in Japan. Concerning the drug resistance, none of the isolates showed resistance to beta-lactam antibiotics, but 45% of them were resistant to macrolides. The advantages of short-term treatment are considered to include a lower cost, improvement in drug compliance, decrease in the frequency of the occurrence of adverse reactions, decrease in the frequency of the appearance of drug-resistant strains, and alleviation of the psychological burden of patients and their parents. For these reasons, we conclude that CDTR-PI for 5 days is a useful option for the treatment of acute GAS tonsillopharyngitis in children.     

Minimum Five-Year Follow-Up Wear Measurement of Longevity Highly Cross-Linked Polyethylene Cup Against Cobalt-Chromium or Zirconia Heads

We investigated the efficacy of combining highly cross-linked polyethylene with ceramic heads on further reduction in polyethylene wear compared with the combination with cobalt-chromium heads via PolyWare computer-assisted method. A prospective cohort study was performed on 102 cementless total hip arthroplasties using Longevity (Zimmer, Warsaw, Ind) highly cross-linked polyethylene liners. Either 26-mm zirconia heads or 26-mm cobalt-chromium heads were randomly used in 51 hips each. At a mean follow-up of 6.7 years, no significant differences were identified between the groups for total penetration rate and steady-state wear rate. Osteolysis was not observed in any hips in either group. In conclusion, no advantage was seen for the 26-mm zirconia head compared with the 26-mm cobalt-chromium head in this period.     

Role of salivary tumour necrosis factor alpha in HIV-positive patients with oral manifestations

In HIV-1-infected patients, oral manifestations such as recurrent apthous ulcers are often seen. A total of 29 HIV-infected patients were examined to determine salivary tumour necrosis factor alpha (TNFalpha) concentrations by enzyme-linked immunosorbent assay, the amount of HIV-1 RNA copy by Amplicor HIV-1 Monitor test, number of CD4 cells by flow cytometry and oral manifestations by oral examination. TNFalpha concentration was significantly correlated with the amount of HIV-1 RNA, however, not with the number of CD4 cells in HIV-1-infected patients. Further, patients with oral manifestations showed significantly higher concentrations of TNFalpha in saliva and HIV-1 RNA copies in serum than those without oral manifestations. Following recovery from oral ulcers, TNFalpha concentration was decreased by half to 20 times lower than the level of that during ulcer incidence. Our results suggest that salivary TNFalpha is a good indicator for oral manifestations and HIV RNA amounts in HIV-1-infected patients.     

Five-year results of metal-on-metal resurfacing arthroplasty in Asian patients

Clinical results of 50 metal-on-metal resurfacing arthroplasties in 45 Japanese patients were evaluated to a minimum follow-up of 5 years. The predominant diagnosis was developmental dysplasia or dislocation of the hip (70%). One patient died of an unrelated cause and another was lost to follow-up. Two hips received revision surgery, including 1 femoral neck fracture and 1 septic loosening. In the remaining 46 hips, 1 hip showed femoral component loosening. Clinical scores of the 46 hips were satisfactory at the final follow-up. The survival rate at 5 years was 96% when failure was attributed to revision for any reason. Metal-on-metal resurfacing arthroplasty in Japanese patients, who have a different distribution of hip diseases from European and American patients, showed similarly promising early clinical results.     

Primary palmar hyperhidrosis locus maps to 14q11.2-q13

Primary palmar hyperhidrosis (PPH) is a unique disorder of unknown cause. It is characterized by excessive perspiration of the eccrine sweat gland in the palm, sole, and the axilla. It is presumed that PPH results from overactivation of the cholinergic sympathetic nerve or dysfunction of the autonomic nervous system. There have been no genetic studies on the disease. We performed a linkage analysis of 11 families including 42 affected and 40 unaffected members using genome-wide DNA polymorphic markers to identify the disease locus. Diagnosis of their PPH was made by direct inspection, interviewing and measurement of the sweating rate with perspirometer. Consequently, from data of three of the 11 families examined, the combined maximum two-point LOD scores of 3.08 and 3.16 (recombination fraction = 0) were obtained at the D14S283 and D14S264 loci, respectively, on chromosome 14q11.2-q13, under an assumption that two liability conditions depend on age. These regions were ruled out in eight other families. Haplotype analysis of the three families supported that one of the PPH locus is assigned at minimum to about a 6-cM interval between D14S1070 and D14S990 and at maximum to about a 30-cM interval between D14S1070 and D14S70. This is the first report of systemic mapping of the PPH locus.     

Non-destructively differentiating the roles of creep, wear and oxidation in long-term in vivo exposed polyethylene cups

Wear of polyethylene acetabular cups in patients of total hip arthroplasty is routinely deduced from the penetration of the femoral head into the acetabular liner as observed in the radiographs. However, the linear penetration thus measured represents the cumulative contribution of two components, one due to wear, and the other due to creep or irreversible deformation of the polyethylene structure. The erroneous attribution to wear of the entire penetration displacement of the head in the cup might lead to misinterpretation of the actual performance of acetabular cups. The aim of this study was to quantify the head displacement components due to wear and to creep, as they occur in vivo in acetabular cups, and to relate them to the oxidation state of the material by means of advanced Raman spectroscopy procedures. Throughout the investigation, we compared the behaviors on the molecular scale of acetabular cups subjected to different sterilization methods (i.e., γ-irradiation and ethylene oxide treatment).     

The influence of chemical activation on tooth bleaching using 10% carbamide peroxide

The aim of this study was to assess the influence of manganese gluconate, a chemical activator of bleaching agents, at a concentration of 0.01% on the efficiency of a 10% carbamide peroxide-based bleaching agent. Forty bovine incisors were immersed in a 25% instant coffee solution for seven days and randomly divided into two groups. Group 1 was the control group and consisted of 10% carbamide peroxide-based bleaching gel only. Group 2 consisted of 10% carbamide peroxide-based bleaching gel and 0.01% manganese gluconate. Three readings of color were taken using the Vita Easyshade spectrophotometer: the initial reading, a reading at seven days, and a reading at 14 days. Total color variation was calculated by ΔE*Lab. Data were submitted to the statistical t-test (5%), which showed that after seven days group 2 had a significant increase in the degree of tooth bleaching compared with group 1. The mean values (±SD) were 16.33 (±3.95) for group 1 and 19.29 (±4.97) for group 2. However, the results for group 1 and group 2 were similar after 14 days. Adding 0.01% manganese gluconate to 10% carbamide peroxide bleaching gel increased the degree of tooth bleaching after a seven-day treatment and did not influence the resulting shade after 14 days.