Response to comment by Puppels et al. on “A modification for the calculation of water depth profiles in oil‐treated skin by in vivo Raman microscopy”

The presence of penetrated oils in the stratum corneum (SC), oil‐induced occlusion of the SC and formation of occluding homogeneous film on the skin surface are discussed in relation to their influence on results of water profile calculations using conventional and newly proposed extended methods. It is shown that the conventional method does not determine the water profiles in treated skin correctly due to the superposition of Raman bands of SC's proteins and penetrated and remnant oils.     

Purification and characterization of human prostatic acid phosphatase.

Human prostatic acid phosphatase (orthophosphoric monoester phosphohydrase, EC 3.1.3.2) is purified to homogeneity by standard procedures which include CM-Sephadex, Con A affinity chromatography and gel filtration. The purified enzyme is antigenically specific and has a M.W. of 100,000 with subunit M.W. of 48,000. However, the enzyme exhibited charge heterogeneity. Two major electrophoretic or chromatographic isozymic forms of PAP were separated by DEAE-Sephadex chromatography and their immunochemical identity was studied by immunodiffusion before and after the neuraminidase digestion. Quantitative precipitin and inhibition experiments showed immunological identity of the two chromatographic isozymes. Immunologic specificity of this enzyme resides on the protein moiety rather than the carbohydrate residue, although the latter group is mostly responsible for the charge group heterogeneity of the enzyme.     

Wolbachia infection in the Loxoblemmus complex (Orthoptera: Gryllidae) in Korea

The Wolbachia bacterium is one of the most prevalent intracellular symbionts of invertebrates, particularly insects. This bacterium induces four distinct reproductive anomalies such as cytoplasmic incompatibility, feminization, male killing, and parthenogenesis of its hosts. Here we report that three closely related cricket species, Loxoblemmus doenitzi, L. campestris, and L. equestris can become infected with Wolbachia. Based on the 16s rRNA sequences, all three species were single infections. However, Wolbachia infecting L. campestris showed diverse Wolbachia surface protein gene sequences resembling multiple infections. In addition, all Wolbachia strains in the three host species harbored the Wolbachia specific bacteriophage.     

First Clinical Cases of Spirometrosis in Two Cats in Korea

This study reports the first two clinical cases of spirometrosis caused by Spirometra sp. in cats in Korea. In these two cases, the cats vomited, and long proglottids of tapeworm were recovered. The sick cats presented with anorexia and lethargy. However, they unexpectedly showed no diarrhea, which is the main symptom of spirometrosis. Based on a fecal floatation test as well as morphological and molecular analyses, the parasite was diagnosed as Spirometra sp. The 2 cases were treated with praziquantel. This study suggests regular monitoring of health and deworming in companion animals, even when animals are well cared for, with regular preventive medication. Additionally, spirometrosis should be considered in the differential diagnosis in cases of gastrointestinal symptoms in Spirometra endemic areas.     

Analytical characterization of host-cell-protein-rich aggregates in monoclonal antibody solutions

Host-cell proteins (HCPs) and high molecular weight (HMW) species have historically been treated as independent classes of impurities in the downstream processing of monoclonal antibodies (mAbs), but recent indications suggest that they may be partially linked. We have explored this connection with a shotgun proteomic analysis of HMW impurities that were isolated from harvest cell culture fluid (HCCF) and protein A eluate using size-exclusion chromatography (SEC). As part of the proteomic analysis, a cross-digest study was performed in which samples were analyzed using both the standard and native digest techniques to enable a fair comparison between bioprocess pools. This comparison reveals that the HCP profiles of HCCF and protein A eluate overlap substantially more than previous work has suggested, because hundreds of HCPs are conserved in aggregates that may be up to ~50 nm in hydrodynamic radius and that persist through the protein A capture step. Quantitative SWATH proteomics suggests that the majority of the protein A eluate's HCP mass is found in such aggregates, and this is corroborated by ELISA measurements on SEC fractions. The SWATH data also show that intra-aggregate concentrations of individual HCPs are positively correlated between aggregates that were isolated from HCCF and protein A eluate, and species that have generally been considered difficult to remove tend to be more concentrated than their counterparts. These observations support prior hypotheses regarding aggregate-mediated HCP persistence through protein A chromatography and highlight the importance of this persistence mechanism.     

DNA-free genome editing with preassembled CRISPR/Cas9 ribonucleoproteins in plants

Transgenic Research - Processes of traditional trait development in plants depend on genetic variations derived from spontaneous mutation or artificial random mutagenesis. Limited availability of...     

Region-specific changes in the immunoreactivity of TRPV4 expression in the central nervous system of SOD1G93A transgenic mice as an in vivo model of amyotrophic lateral sclerosis

Transient receptor potential vanilloid 4 (TRPV4) is a broadly expressed Ca²⁺-permeable cation channel in the vanilloid subfamily of transient receptor potential channels. It is activated by warm temperature, lipids downstream of arachidonic acid metabolism, hypoosmolarity, or mechanical stimulation. In the present study, we used SOD1^G93A mutant transgenic mice as the animal model of amyotrophic lateral sclerosis (ALS) and investigated the changes of TRPV4 immunoreactivity in the central nervous system of these mice by immunohistochemical studies. An increased expression of TRPV4 was pronounced in the cerebral cortex, hippocampal formation, thalamus, cerebellum and spinal cord of symptomatic SOD1^G93A transgenic mice. In the cerebral cortex, TRPV4 immunoreactivity was significantly increased in pyramidal cells of SOD1^G93A transgenic mice. In the hippocampal formation, pyramidal cells of the CA1-3 areas and in the granule cells of the dentate gyrus demonstrated increased TRPV4 immunoreactivity. In addition, TRPV4 immunoreactivity was increased in the spinal cord, thalamus and cerebellum of the symptomatic SOD1^G93A transgenic mice. This study, which showed increased TRPV4 in different brain and spinal cord regions of SOD1^G93A transgenic mice, may provide clues to the understanding of many basic neuronal functions in ALS. These findings suggest a role for TRPV4 in the neuronal functions in ALS but the mechanisms and functional implications of increased TRPV4 require elucidation.     

Effects of bisphenol A and light conditions on the circadian rhythm of the goldfish Carassius auratus

We investigated how exposure to bisphenol A (BPA) under different photoperiodic conditions affected the expression of clock genes in the brain and liver of the goldfish, Carassius auratus. Three photoperiodic conditions were used: control, LD; continuous light, LL; and continuous dark, DD; the fish were exposed to three concentrations of BPA, namely 0, 10, or 100 μg/L. We measured changes in the expression of cryptochrome 1 (Cry1), period 2 (Per2), and melatonin receptor 1 (MT-R1). The levels of Cry1, Per2, and MT-R1 mRNAs decreased with increasing BPA concentration and with increasing exposure time. Expression of Cry1 and Per2 increased more in the LL group than in the LD and DD groups. However, for MT-R1, the DD group showed increased expression compared to the LL and LD groups. Our analysis shows that circadian rhythms in goldfish can be disrupted by exposure to BPA and that the response can be modified by regulating the photoperiod.     

Esophageal striated muscle hypertrophy and muscle fiber type transformation in MSTN knockout pigs

Transgenic Research - Myostatin (MSTN) is a member of the transforming growth factor-β superfamily that inhibits skeletal muscle growth and development. The esophagus is composed of skeletal...