Elevated expression of endocan in the development of cervical squamous neoplasia of the uterus.

Accumulated evidence has shown that endocan, which was originally called endothelial cell-specific molecule-1, is an attractive prognostic factor in a variety of cancers. However, the relevance of endocan expression in human malignancies remains to be clarified. In the present study, the expression of endocan in cervical squamous neoplasia of the uterus, including low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), as well as in invasive squamous cell carcinoma was examined by immunohistochemistry. Endocan was not sufficiently expressed in the normal cervical epithelium. Endocan expression was present in LSIL cases but was limited to basal and parabasal areas of the cells. HSIL cases exhibited strong expression of endocan with widely distributed expression toward the epithelial surface. In contrast, further strong expression of endocan was not observed in patients with invasive carcinoma. This study is the first study showing increased expression of endocan in precancerous dysplastic lesions and malignancy of the cervix. The data suggest that a high expression level of endocan potentially contributes to the development of cervical squamous neoplasia of the uterus.

TRIM22 negatively regulates MHC-II expression.

The development of cancer treatment has recently achieved a remarkable breakthrough, and checkpoint blockade immunotherapy has received much attention. To enhance the therapeutic efficacy of checkpoint blockade immunotherapy, recent studies have revealed the importance of activation of CD4+ T cells via an increase in major histocompatibility complex (MHC) class II molecules in cancer cells. Here, we demonstrate that tripartite motif-containing (TRIM) 22, negatively regulates MHC-II expression. Gene knockout of TRIM22 using Cas9-sgRNAs led to an increase of MHC-II proteins, while TRIM22 overexpression remarkably decreased MHC-II proteins. mRNA levels of MHC-II and class II transactivator (CIITA), which plays an essential role in the regulation of MHC-II transcription, were not affected by TRIM22. Furthermore, TRIM22 knockout did not suppress the degradation of MHC-II protein but rather promoted it. These results suggest that TRIM22 decreases MHC-II protein levels through a combination of multiple mechanisms other than transcription or degradation. We showed that inhibition of TRIM22 can increase the amount of MHC-II expression in cancer cells, suggesting a possibility of providing the biological basis for a possible therapeutic target to potentiate checkpoint blockade immunotherapy.

R-spondin1 plays an essential role in ovarian development through positively regulating Wnt-4 signaling.

In mammals, female development has traditionally been considered a default process in the absence of the testis-determining gene, Sry. Recently, it has been documented that the gene for R-spondin1 (RSPO1), a novel class of soluble activator for Wnt/beta-catenin signaling, is mutated in two Italian families with female-to-male (XX) sex reversal. To elucidate the role of Rspo1 as a candidate female-determining gene in a mouse model, we generated Rspo1-null (Rspo1(-/-)) mice and found that Rspo1(-/-) XX mice displayed masculinized features including pseudohermaphroditism in genital ducts, depletion of fetal oocytes, male-specific coelomic vessel formation and ectopic testosterone production in the ovaries. Thus, although Rspo1 is required to fully suppress the male differentiation program and to maintain germ cell survival during the development of XX gonads, the loss of its activity has proved to be insufficient to cause complete XX sex reversal in mice. Interestingly, these partial sex-reversed phenotypes of Rspo1(-/-) XX mice recapitulated those of previously described Wnt-4(-/-) XX mice. In accordance with this finding, the expression of Wnt-4 and its downstream genes was deregulated in early Rspo1(-/-) XX gonads, suggesting that Rspo1 may participate in suppressing the male pathway in the absence of Sry and maintaining oocyte survival through positively regulating Wnt-4 signaling.

Influence of Artificial Inoculation with Pseudomonas fluorescens on Enzymatic Browning Reactions of Fresh-cut Potatoes.

We initially correlated fluorescent pseudomonads and severity of enzymatic browning on fresh-cut potatoes. Subsequently, we determined the influence of inoculation with Pseudomonas fluorescens following its isolation from the brown tissues on the browning response on fresh-cut potatoes. Bacterial counts on potato slices were higher on browning tissues than on non-browning tissues. P. fluorescens that has been isolated only from the severely browning tissues developed brown discoloration on surface tissues when inoculated onto potato slices. When potato slices were initially inoculated with 103 colony-forming unit (CFU) per mL of P. fluorescens and then stored at 5ºC, bacterial counts, polyphenol oxidase (PPO) activity, phenolic content, and browning severity increased after 3 days of storage. We observed plant PPO derived from potatoes and bacterial PPO released by P. fluorescens and dictated that the plant PPO contributed to browning reactions because only the plant PPO was activated at pH 6-7 that lies in potato tissues. The PPO1 gene that contributed to browning on potatoes was expressed prominently in potato tissues following inoculation with P. fluorescens. These results indicated that P. fluorescens enhanced browning of fresh-cut potatoes by inducing the plant PPO gene, plant PPO activity, and accumulation of phenolics as a biocontrol agent.

Viability of sublethally injured coliform bacteria on fresh-cut cabbage stored in high CO2 atmospheres following rinsing with electrolyzed water.

The extent of sublethally injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, was evaluated during storage in air and high CO2 controlled atmospheres (5%, 10%, and 15%) at 5°C and 10°C using the thin agar layer (TAL) method. Sublethally injured coliform bacteria on nonrinsed shredded cabbage were either absent or they were injured at a 64-65% level when present. Rinsing of shredded cabbage with electrolyzed water containing 25ppm available chlorine reduced the coliform counts by 0.4 to 1.1 log and caused sublethal injury ranging from 42 to 77%. Pantoea ananatis was one of the species injured by chlorine stress. When shredded cabbage, nonrinsed or rinsed with electrolyzed water, was stored in air and high CO2 atmospheres at 5°C for 7days and 10°C for 5days, coliform counts on TAL plates increased from 3.3-4.5 to 6.5-9.0 log CFU/g during storage, with the increase being greater at 10°C than at 5°C. High CO2 of 10% and 15% reduced the bacterial growth on shredded cabbage during storage at 5°C. Although injured coliform bacteria were not found on nonrinsed shredded cabbage on the initial day, injured coliforms at a range of 49-84% were detected on samples stored in air and high CO2 atmospheres at 5°C and 10°C. Injured cells were detected more frequently during storage at both temperatures irrespective of the CO2 atmosphere when shredded cabbage was rinsed with electrolyzed water. These results indicated that injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, exhibited different degrees of injury during storage regardless of the CO2 atmosphere and temperature tested.

Viability of Chlorine-injured Escherichia Coli O157:H7 on Fresh-cut Cabbage during Cold Storage in High CO2 Atmospheres.

Viability of chlorine-injured E. coli O157:H7 inoculated onto shredded cabbage was evaluated during storage in air or high CO2 controlled atmospheres (CA) of 5%, 10%, and 15% at 10℃ and in a modified atmosphere packaging (MAP) at 5℃ and 10℃. When shredded cabbage was inoculated with chlorine-injured E. coli O157:H7 (% injury = 65%) and then stored in air or CA at 10℃, counts of E. coli O157:H7 increased during storage and injured E. coli O157:H7 (% injury = 34-66%) were detected on samples throughout the storage regardless of the CO2 atmosphere. When shredded cabbage inoculated with chlorine injured E. coli O157:H7 (% injury = 45-59%) were stored in a MAP using either a high or low oxygen transmission permeability (OTR) package film, the counts of E. coli O157:H7 increased during storage at 10℃ and they remained constant during storage at 5℃. Injured E. coli O157:H7 were detected on shredded cabbage at a 54-56% level in a low OTR film at 10℃ and a 73-74% level in a high OTR film at 5℃. These results indicated that chlorine-injured E. coli O157:H7 inoculated on fresh-cut cabbage exhibited different degrees of injury during storage in a high CO2 CA and MAP at 5℃ or 10℃.

Enumeration and Identification of Coliform Bacteria Injured by Chlorine or Fungicide Mixed with Agricultural Water.

Chemical sanitizers may induce no injury (bacteria survive), sublethal injury (bacteria are injured), or lethal injury (bacteria die). The proportion of coliform bacteria that were injured sublethally by chlorine and fungicide mixed with agricultural water (pond water), which was used to dilute the pesticide solution, was evaluated using the thin agar layer (TAL) method. In pure cultures of Enterobacter cloacae , Escherichia coli , and E. coli O157:H7 (representing a human pathogen), the percentage of chlorine-injured cells was 69 to 77% for dilute electrolyzed water containing an available chlorine level of 2 ppm. When agricultural water was mixed with electrolyzed water, the percentage of injured coliforms in agricultural water was 75%. The isolation and identification of bacteria on TAL and selective media suggested that the chlorine stress caused injury to Enterobacter kobei . Of the four fungicide products tested, diluted to their recommended concentrations, Topsin-M, Sumilex, and Oxirane caused injury to coliform bacteria in pure cultures and in agricultural water following their mixture with each pesticide, whereas Streptomycin did not induce any injury to the bacteria. The percentage of injury was 45 to 97% for Topsin-M, 80 to 87% for Sumilex, and 50 to 97% for Oxirane. A comparison of the coliforms isolated from the pesticide solutions and then grown on either TAL or selective media indicated the possibility of fungicide-injured Rahnella aquatilis , Yersinia mollaretii , and E. coli . These results suggest the importance of selecting a suitable sanitizer and the necessity of adjusting the sanitizer concentration to a level that will kill the coliforms rather than cause sanitizer-induced cell injury that can result in the recovery of the coliforms.