Enhancement of interleukin-2 production by bovine peripheral blood mononuclear cells treated with the combination of anti-programmed death-ligand 1 and cytotoxic T lymphocyte antigen 4 chimeric monoclonal antibodies

Our previous studies demonstrate the therapeutic efficacy against bovine diseases of an anti-bovine programmed death-ligand 1 (PD-L1) chimeric antibody. In humans, PD-1 and PD-L1 antibodies are more effective when combined with an antibody targeting cytotoxic T lymphocyte antigen 4 (CTLA-4) and these combination therapies are therefore clinically used. Here we generated an anti-bovine CTLA-4 chimeric antibody (chAb) to enhance the therapeutic efficacy of the PD-L1 antibody. We further analyzed the effects of dual blockade of CTLA-4 and PD-1 pathways on T-cell responses. The established anti-bovine CTLA-4 chAb showed comparable blocking activity on the binding of bovine CTLA-4 to CD80 and CD86 as the anti-bovine CTLA-4 mouse monoclonal antibody. Anti-bovine CTLA-4 chAb also significantly increased IL-2 production from bovine peripheral blood mononuclear cells (PBMCs). Further, the combination of anti-CTLA-4 chAb with anti-PD-L1 chAb significantly upregulated IL-2 production by PBMCs. These results suggest that the combination of antibodies have higher potential to enhance immune responses against pathogens compared with single administration.     

Congenital syringohydromyelia in a crossbred (Holstein-Friesian × Japanese Black) beef calf

A 5-day-old male crossbred beef calf presented with a well-coordinated bilateral hopping gait of the hind limbs. Postmortem CT showed a poorly defined oval-shaped region at the L3–L4 spinal segments, which had high signal intensity on T2 weighted postmortem MRI images. On pathological examination, we identified a large cystic cavity filled with a large amount of cerebrospinal fluid on the cut surface of the spinal region. Histopathological examination revealed that the spinal cord parenchyma was compressed by the cystic structure, and the cystic cavity was lined with a thin layer of discrete ependymal cells, indicating syringohydromyelia. This is the first reported case of a Holstein-Friesian × Japanese Black crossbred calf with solitary syringohydromyelia. Our findings suggest that myelodysplasia with cystic cavities can be suspected by CT, without the need for MRI.     

Evaluation of adverse events in small‐breed dogs treated with maropitant and a single dose of doxorubicin

BackgroundThe recommended doxorubicin (DOX) dose for small dogs is 1 mg/kg. Recent data suggest that DOX‐induced gastrointestinal (GI) toxicosis can be reduced with maropitant treatment.ObjectivesTo investigate the incidence of adverse events (AEs) in small‐breed dogs administered a single 25 mg/m² DOX followed by administration of maropitant (DOX25). The primary aim was to assess myelo‐ and GI toxicoses for 2 weeks after DOX administration. The secondary aim was to compare the incidence and grades of AEs found in the DOX25 group with a historical control group (DOX 1 mg/kg without administration of antiemetic or antidiarrheal medications).AnimalsNineteen small‐breed tumor‐bearing dogs.MethodsA prospective, observational study of tumor‐bearing dogs, weighing 5 to 10 kg, administered a single 25 mg/m² dose of DOX IV, followed by administration of maropitant for the next 5 days.ResultsInappetence, vomiting, and diarrhea were found in 7/19, 2/19, and 6/19 of the DOX25 dogs, respectively. Neutropenia and thrombocytopenia was 12/19 and 3/19, respectively. Most AEs were grades 1 and 2, except for grades 3 and 4 inappetence and neutropenia in 3 and 4 dogs, respectively. Furthermore, febrile neutropenia occurred in 3/19 dogs in the DOX25 group. All AEs between the DOX25 and historical control groups were not significantly different.Conclusions and Clinical ImportanceVomiting and diarrhea were deemed acceptable with 25 mg/m² DOX followed by maropitant treatment in 5 to 10 kg dogs; however, additional supportive care might be needed for dogs with inappetence and neutropenia.     

Enhancement of saccharification by overexpression of poplar cellulase in sengon

Lignocellulosic material from trees has great potential to form the basis of the second generation for bioethanol production because trees produce most of the biomass on the earth. We modified the wall structure of sengon (Paraserianthes falcataria) through overexpression of poplar cellulase in the cell walls. The overexpression did not decrease cellulose content but caused a decrease in xyloglucan bound to the walls. The level of saccharification and successive ethanol production was increased in the wood of the transgenic sengon overexpressing poplar cellulase compared with that of the wild type plant, and even after delignification of the wood. We propose that a xyloglucan intercalated into cellulose microfibrils could be one of the recalcitrant components in the saccharification of lignocelluloses.     

Proteomic analysis of the G-layer in poplar tension wood

Angiosperm trees bend their stems by forming tension wood at the upper side of leaning stems. Most tension wood has a cellulose-rich G-layer in the innermost surface of the fiber cell wall. Strong tensile stress is considered to occur in the G-layer. This study undertook to identify the proteins involved in G-layer formation and function through a proteomic analysis of G-layer-localized protein. G-layers of poplar were loosened by sonication and isolated as doughnut-shaped pieces of thinly sliced transverse sections. The proteins, once extracted with urea/detergent solution, were separated by two-dimensional polyacrylamide gel electrophoresis, and 110 spots were subjected to liquid chromatography tandem mass spectrometry (LC/MS/MS). A database search for these spots’ mass spectrum patterns identified 72 proteins. In addition, all peptide digestion mixtures of G-layer proteins were separated by strong cation exchange chromatography and 39 proteins were identified using LC/MS/MS analysis. Proteins involved in wall formation, such as lignin biosynthesis-related protein, xyloglucan endotransglucosylase, and fasciclin-like arabinogalactan protein, were notably detected in the G-layer.     

Effect of pressure on organic acid production from Japanese beech treated in supercritical water

The effect of pressure as described by density was studied on organic acid production from Japanese beech (Fagus crenata) treated in supercritical water. At a reaction temperature of 380°C, the maximum yield of organic acids was 35% at a pressure of 30 MPa (density of water: 0.53 g/ml) for 1 min in a batch-type system. Furthermore, the yield of organic acids decreased with increasing reaction pressure. It was also found that fragmented products from sugars such as methylglyoxal and glycolaldehyde could be more easily converted to organic acids than dehydrated products such as furfural and 5-hydroxymethyl furfural. This result suggests that organic acids can be mainly derived from fragmented products. Part of this article was presented at the 55th Annual Meeting of the Japan Wood Research Society (2005), and at the 17th Annual Meeting of Japan Institute of Energy (2007)     

Genetic identification and morphology of naturally spawned eggs of the Japanese eel Anguilla japonica collected in the western North Pacific

Eggs of the Japanese eel Anguilla japonica collected in the western North Pacific were identified by onboard species-specific polymerase chain reaction (PCR) and DNA nucleotide sequencing after the cruise. Fish eggs of various species were collected by large plankton net tows at 12 stations along the southern part of the West Mariana Ridge on 19–25 May 2009. A total of 43 fish eggs were distinguished morphologically as possibly being of A. japonica. Thirty-one of those were analyzed by PCR, which included 15 eggs collected at 12°50–55′N, 141°15–20′E (in 5 tows) that showed positive results. The 16S ribosomal RNA (rRNA) gene sequences of eggs determined after the cruise indicated that 31 A. japonica eggs had been collected. The remaining eggs were of mesopelagic eel species (Serrivomeridae and Derichthyidae), or unidentified species. The morphological characteristics of the A. japonica eggs were consistent with those of artificially spawned eggs, except they had a slightly larger diameter. The egg diameter range did not overlap with those of mesopelagic eels of the Serrivomeridae, which often spawn in the same area as A. japonica. These results suggest that egg diameter and embryo shape can be used to morphologically identify naturally spawned A. japonica eggs.     

Occurrence of heavy metals in the sediments of Uranouchi Inlet,Kochi prefecture,Japan

The present status of contamination by heavy metals, and the impact of cage culture on sediments at the Uranouchi Inlet, Kochi Prefecture, Japan, were investigated from two stations influenced by intensive aquaculture and a control station, during May–July 2006. The moisture content of the sediments at the aquaculture stations was over 65%, and the organic matter was always over 100 mg/g dry wt. In contrast, the highest moisture content and organic matter at the control station was 45.5% and 62.9 mg/g dry wt, respectively. Concentrations of zinc (Zn) (178 ± 4.8 mg/kg dry wt) and copper (Cu) (125 ± 1.2 mg/kg dry wt) were highest at the aquaculture stations. Lead (Pb) was highest (50.7 ± 0.77 mg/kg dry wt) at the aquaculture station though it was as high as 33.2 ± 0.77 mg/kg dry wt at the control station. One-way ANOVA showed that the differences in concentrations of Zn and Cu in sediments from the aquaculture and control stations were highly significant (P = 0.01), whereas Pb showed no such trend. Occurrence of a large fraction of labile Zn (56.1%) and Cu (40.3%) in these sediments warrants attention. Although factors other than metals may explain the distribution observed, the information presented here may be useful in predicting long-term effects of heavy metal contamination from aquaculture in the marine environment.     

Cellulose esters as compatibilizers in wood/poly(lactic acid) composite

One of the important issues relevant to wood/plastic composite molding is the compatibility between hydrophilic wood and hydrophobic plastic. Polyolefins modified with maleic anhydride, which have been frequently used for wood and polyolefin composites, are not effective for poly(lactic acid) composites. Because compounds with both hydrophilic and hydrophobic groups are potential compatibilizers, cellulose esters of several carboxylic acids, RCOO-cellulose (R = CH₃, C₂H₅, C₃H₇, C₄H₉, C₅H₁₁, C₁₁H₂₃), were synthesized and their effects as compatibilizers of wood/poly(lactic acid) composites (WPLC) were examined in this study. The mechanical properties of WPLC were improved with a small amount of added cellulose esters, especially cellulose butyrate or cellulose valerate. The relevant effect of added cellulose esters on the thermal properties of wood/PLA composite was analyzed by measuring dynamic viscoelasticity. Part of this report was presented at the 56th Annual Meeting of Japan Wood Research Society, Akita, August 2006 and the 9th International Conference on Frontiers of Polymers and Advanced Materials, Cracow, Poland, July, 2007