Cell clusters in intervertebral disc degeneration:an attempted repair mechanism aborted via apoptosis / 대한해부학회지

Cell clusters are a histological hallmark feature of intervertebral disc degeneration. Clusters arise from cell proliferation, are associated with replicative senescence, and remain metabolically, but their precise role in various stages of disc degeneration remain obscure. The aim of this study was therefore to investigate small, medium, and large size cell-clusters. For this purpose, human disc samples were collected from 55 subjects, aged 37–72 years, 21 patients had disc herniation, 10 had degenerated non-herniated discs, and 9 had degenerative scoliosis with spinal curvature <45°.15 non-degenerated control discs were from cadavers. Clusters and matrix changes were investigated with histology, immunohistochemistry, and Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Data obtained were analyzed with spearman rank correlation and ANOVA. Results revealed, small and medium-sized clusters were positive for cell proliferation markers Ki-67 and proliferating cell nuclear antigen (PCNA) in control and slightly degenerated human discs, while large cell clusters were typically more abundant in severely degenerated and herniated discs. Large clusters associated with matrix fissures, proteoglycan loss, matrix metalloproteinase-1 (MMP-1), and Caspase-3. Spatial association findings were reconfirmed with SDS-PAGE that showed presence to these target markers based on its molecular weight.Controls, slightly degenerated discs showed smaller clusters, less proteoglycan loss, MMP-1, and Caspase-3. In conclusion, cell clusters in the early stages of degeneration could be indicative of repair, however sustained loading increases large cell clusters especially around microscopic fissures that accelerates inflammatory catabolism and alters cellular metabolism, thus attempted repair process initiated by cell clusters fails and is aborted at least in part via apoptosis.

Effect of atmospheric carbon dioxide concentration on the cultivation of bovine Mycoplasma species.

Recommendations for bovine mycoplasma culture CO2 concentrations are varied and were not empirically derived. The objective of this study was to determine whether the growth measures of bovine mycoplasma isolates differed when incubated in CO2 concentrations of 10 or 5% or in candle jars (2.7 ± 0.2% CO2). Growth of Mycoplasma bovis (n = 22), Mycoplasma californicum (n = 18), and other Mycoplasma spp. (n = 10) laboratory isolates was evaluated. Isolate suspensions were standardized to approximately 108 cfu/mL and serially diluted in pasteurized whole milk to achieve test suspensions of 102 and 106 cfu/mL. One hundred microliters of each test dilution was spread in duplicate onto the surface of a modified Hayflick's agar plate. Colony growth was enumerated on d 3, 5, and 7 of incubation. A mixed linear model included the fixed effects of CO2 treatment (2.7, 5, or 10%), species, day (3, 5, or 7), and their interactions, with total colony counts as the dependent variable. Carbon dioxide concentration did not significantly affect overall mycoplasma growth differences, but differences between species and day were present. Colony counts (log10 cfu/mL) of M. bovis were 2.6- and 1.6-fold greater than M. californicum and other Mycoplasma spp., respectively. Growth at 7 d of incubation was greater than d 3 and 5 for all species. These findings were confirmed using field isolates (n = 98) from a commercial veterinary diagnostic laboratory. Binary growth responses (yes/no) of the field isolates were not different between CO2 treatments but did differ between species and day of incubation. On average, 57% of all field isolates were detected by 3 d of incubation compared with 93% on d 7. These results suggest that the range of suitable CO2 culture conditions and incubation times for the common mastitis-causing Mycoplasma spp. may be broader than currently recommended.

Empirical and model-based estimates of spatial and temporal variations in net primary productivity in semi-arid grasslands of Northern China.

Spatiotemporal variations in net primary productivity (NPP) reflect the dynamics of water and carbon in the biosphere, and are often closely related to temperature and precipitation. We used the ecosystem model known as the Carnegie-Ames-Stanford Approach (CASA) to estimate NPP of semiarid grassland in northern China counties between 2001 and 2013. Model estimates were strongly linearly correlated with observed values from different counties (slope = 0.76 (p < 0.001), intercept = 34.7 (p < 0.01), R2 = 0.67, RMSE = 35 g C·m-2·year-1, bias = -0.11 g C·m-2·year-1). We also quantified inter-annual changes in NPP over the 13-year study period. NPP varied between 141 and 313 g C·m-2·year-1, with a mean of 240 g C·m-2·year-1. NPP increased from west to east each year, and mean precipitation in each county was significantly positively correlated with NPP-annually, and in summer and autumn. Mean precipitation was positively related to NPP in spring, but not significantly so. Annual and summer temperatures were mostly negatively correlated with NPP, but temperature was positively correlated with spring and autumn NPP. Spatial correlation and partial correlation analyses at the pixel scale confirmed precipitation is a major driver of NPP. Temperature was negatively correlated with NPP in 99% of the regions at the annual scale, but after removing the effect of precipitation, temperature was positively correlated with the NPP in 77% of the regions. Our data show that temperature effects on production depend heavily on recent precipitation. Results reported here have significant and far-reaching implications for natural resource management, given the enormous size of these grasslands and the numbers of people dependent on them.

Pathogenic variant in NLRP7 (19q13.42) associated with recurrent gestational trophoblastic disease: Data from early embryo development observed during in vitro fertilization / 대한생식의학회지

OBJECTIVE: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. METHODS: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. RESULTS: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. CONCLUSION: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.

Radiation-Induced Giant Cell Granuloma Mimicking Relapsed Hodgkin Lymphoma at FDG-PET/CT / 대한핵의학회잡지

A 22-year-old woman was diagnosed with intermediate risk stage II Hodgkin lymphoma and treated with three cycles of adriamycin, bleomycin, vinblastine, and dacarbazine (ABVD) followed by involved-field radiation therapy. A complete metabolic remission was achieved after two cycles of ABVD, which was maintained until three years after completion of treatment. Follow-up FDG-PET/CT four years after completion of treatment, however, showed a new FDG-avid (Deauville score of 4) lesion in the right scapula, suggesting relapsed disease. Computer tomography (CT)-guided biopsy of this lesion was performed and subsequent histological examination revealed a radiation-induced giant cell granuloma.

Assessment of radiation-induced second cancer risks in proton therapy and IMRT for organs inside the primary radiation field.

There is clinical evidence that second malignancies in radiation therapy occur mainly within the beam path, i.e. in the medium or high-dose region. The purpose of this study was to assess the risk for developing a radiation-induced tumor within the treated volume and to compare this risk for proton therapy and intensity-modulated photon therapy (IMRT). Instead of using data for specific patients we have created a representative scenario. Fully contoured age- and gender-specific whole body phantoms (4 year and 14 year old) were uploaded into a treatment planning system and tumor volumes were contoured based on patients treated for optic glioma and vertebral body Ewing's sarcoma. Treatment plans for IMRT and proton therapy treatments were generated. Lifetime attributable risks (LARs) for developing a second malignancy were calculated using a risk model considering cell kill, mutation, repopulation, as well as inhomogeneous organ doses. For standard fractionation schemes, the LAR for developing a second malignancy from radiation therapy alone was found to be up to 2.7% for a 4 year old optic glioma patient treated with IMRT considering a soft-tissue carcinoma risk model only. Sarcoma risks were found to be below 1% in all cases. For a 14 year old, risks were found to be about a factor of 2 lower. For Ewing's sarcoma cases the risks based on a sarcoma model were typically higher than the carcinoma risks, i.e. LAR up to 1.3% for soft-tissue sarcoma. In all cases, the risk from proton therapy turned out to be lower by at least a factor of 2 and up to a factor of 10. This is mainly due to lower total energy deposited in the patient when using proton beams. However, the comparison of a three-field and four-field proton plan also shows that the distribution of the dose, i.e. the particular treatment plan, plays a role. When using different fractionation schemes, the estimated risks roughly scale with the total dose difference in%. In conclusion, proton therapy can significantly reduce the risk for developing an in-field second malignancy. The risk depends on treatment planning parameters, i.e. an analysis based on our formalism could be applied within treatment planning programs to guide treatment plans for pediatric patients.

In vivo morphological and antigenic characteristics of Photobacterium damselae subsp. piscicida

The present study was conducted to examine the morphology and antigenicity of Photobacterium damselae subsp. piscicida by culturing the bacterium in vivo in the peritoneal cavity of sea bass (Dicentrarchus labrax) within dialysis bags with either a low molecular weight (LMW) cut-off of 25 kDa or a high molecular weight (HMW) cut-off of 300 kDa. Differences were observed in the growth rate between the bacteria cultured in vivo or in vitro. Bacteria cultured in vivo were smaller and produced a capsular layer, which was more prominent in bacteria cultured in the HMW bag. Antigenicity was examined by Western blot analysis using sera from sea bass injected with live Ph. d. subsp. piscicida. The sera recognised bands at 45 and 20 kDa in bacteria cultured in vivo in the LMW bag. Bacteria cultured in vivo in the HMW bag did not express the 45 kDa band when whole cell extracts were examined, although the antigen was present in their extracellular products. In addition, these bacteria had a band at 18 kDa rather than 20 kDa. Differences in glycoprotein were also evident between bacteria cultured in vitro and in vivo. Bacteria cultured in vitro in LMW and HMW bags displayed a single 26 kDa band. Bacteria cultured in the LMW bag in vivo displayed bands at 26 and 27 kDa, while bacteria cultured in vivo in the HMW bag possessed only the 27 kDa band. These bands may represent sialic acid. The significance of the changes observed in the bacterium's structure and antigenicity when cultured in vivo is discussed.

Variation in the molecular weight of Photobacterium damselae subsp. piscicida antigens when cultured under different conditions in vitro

The antigenicity of Photobacterium damselae (Ph. d.)subsp. piscicida, cultured in four different growth media[tryptone soya broth (TSB), glucose-rich medium (GRM),iron-depleted TSB (TSB+IR-), and iron-depleted GRM(GRM+IR-)] was compared by enzyme-linked immuno-sorbent assay (ELISA) and Western blot analysis usingsera obtained from sea bass (Dicentrarchus labrax) raisedagainst live or heat-killed Ph. d. subsp. piscicida. Theantigenic expression of Ph. d. subsp. piscicida was found todiffer depending on the culture medium used. A significantlyhigher antibody response was obtained with iron-depletedbacteria by ELISA compared with non-iron depletedbacteria obtained from the sera of sea bass raised againstlive Ph. d. subsp. piscicida. The sera from sea bass raisedagainst live bacteria showed a band at 22kDa in bacteriacultured in TSB+IR- or GRM+IR- when bacteria thathad been freshly isolated from fish were used for thescreening, while bands at 24 and 47kDa were observedwith bacteria cultured in TSB or GRM. When bacteriawere passaged several times on tryptic soya agar prior toculturing in the four different media, only bands at 24 and47kDa were recognized, regardless of the medium used toculture the bacteria. It would appear that the molecularweight of Ph. d. subsp. piscicida antigens change in thepresence of iron restriction, and sera from sea bassinfected with live bacteria are able to detect epitopes onthe antigens after this shift in molecular weight.

Neuroptera of the Amazon basin. Part 11a. Introduction and chrysopint

33 species of Chrisopini are described and illustrated, of wich 30 are recorded from the Amazon Basin. Ten new species are described in Chrisopodes(duckei, spínella, nebulosa, lineafrons, polygonica, indetata, conisetosa, breviata, mediocris, and tetifera(, and nine in Ceraesochrysa(squalidens, acutipuppis, rafaeli, reddyi, nigripes, teneuícornis, ariasi, falceifera, and michaelmuris). Neosuarus (type species collaris (Schneider), a new subgenus of Chrysopodes, also contains divisa (Walker), escomeli (Navás) porterina (Navás), and flavescens (Blanchard). Orlandsia Navás and Ancylochrysa Navás are synonymized with Chrysopodes limbata (Navás), Chrysoperla asoralis (Banks) and Suarius nesotala (Banks). A neotype is designate for Ceraeochrysa cubana (Hagen). Ceraeochrysa caligata(Banks) and scapularis (Navás) are valid species. C. gloriae Alayo is synonymized with Ceraeochrysa everes (Banks).

As 30 espécies de Chrysopini conhecidas da Bacia Amazônica são descritas, ilustradas, e chaves dadas para suas identificações. Dezenove espécies novas e um subgênero novo de Chrysopodes, Neosuarius, são descritas. Os nomes genéricos Orlandsia Navás e Ancylochrysa Navás são sinonimizados com Chrysopodes.