Cancer is the result of damage to the genetic system, i.e., dysfunction of the DNA repair system, resulting in dysregulated expression of various molecules, leading to cancer formation, migration, and invasion. In cancer progression, several proteases play a critical role in metastasis; however, their biological mechanism in cancer metastasis is not clearly understood. Among these proteases, cathepsins are a family of lysosomal proteases found in most animal cells. Cathepsins have an important role in protein turnover of mammalian, and are classified into 15 types based on their structure as serine (cathepsin A and G), aspartic (cathepsin D and E), and cysteine cathepsins (cathepsin B, C, F, H, K, L, O, S, V, X, and W). Cysteine cathepsins appear to accelerate the progression of human and rodent cancers, which can be a biomarker of the potency of malignancy or metastasis in mammalian. Overexpression of cyteine cathepsins causes the activation of angiogenesis promoting factor, whereas their downregulation reduces the angiogenesis of cancer progression. Under physiological conditions, cysteine cathepsins are essential in inflammation, infection, and cancer development. Activity of cysteine proteases, i.e., cathepsin B, is required for cancer progression or metastasis. Elevation of cysteine cathepsin is associated with cancer metastasis, angiogenesis, and immunity. Therefore, in this review, we suggest that cysteine cathepsin may be an anticancer target of strong clinical interest, although the exact mechanism of cathepsins in cancer metastasis is under investigation.
Animals ; Cathepsin B ; Cathepsins ; Cysteine ; Cysteine Proteases ; DNA Repair ; Down-Regulation ; Humans ; Inflammation ; Neoplasm Metastasis ; Peptide Hydrolases ; Rodentia ; Serine

Bladder cancer is a common cancer in smoking men and may correlate with mechanosensitive potassium channels because the urinary bladder is a stretch sensing organ. Two-pore K+ channels (K2P), such as TASK3 and TREK1, have recently been shown to play a critical role in both cell apoptosis and tumorigenesis. Of the channels, TREK1 can be activated by many physiological stimuli, including polyunsaturated fatty acids, and intracellular pH, hypoxia, and neurotransmitters. Here we attempted to determine whether TREK1 is functionally expressed in bladder cancer 253J cells. K2P channels, including TREK1, TREK2, TASK1, TASK3, and TWIK1, were quantified in cultured human bladder cancer 253J cells using real time quantitative RT-PCR (qRT-PCR) analysis. Among them, TREK1-like channel was recorded at a single channel level using the patch-clamp technique. The TREKl-like channel, with single-channel conductance of ~90 pS at -80 mV, was recorded in symmetrical 150 mM KCl using an excised inside-out patch configuration. The current-voltage relationships were linear and were insensitive to tetraethylammonium. The channel was activated by membrane stretch, free fatty acids, and intracellular acidosis. These results with electrophysiological properties resemble to those of K2P channel, for instance, TREK1. Therefore, we conclude that TREK1 channel is functionally present in bladder cancer 253J cells.
Acidosis ; Anoxia ; Apoptosis ; Cell Transformation, Neoplastic ; Fatty Acids, Nonesterified ; Fatty Acids, Unsaturated ; Humans* ; Hydrogen-Ion Concentration ; Male ; Membranes ; Neurotransmitter Agents ; Patch-Clamp Techniques ; Potassium Channels* ; Potassium* ; Smoke ; Smoking ; Tetraethylammonium ; Urinary Bladder Neoplasms ; Urinary Bladder*

Uncovering enzyme (UCE), encoded by the human NAGPA, is a trans-Golgi enzyme that adds the mannose-6-phosphate recognition tag on lysosomal enzymes destined for the lysosome. Mutations in NAGPA are known to cause stuttering, a common speech disorder with unknown etiology. The human NAGPA gene is transcribed into two different forms, probably due to alternative splicing. One of them, known as a brain isoform, is lacking exon 8 (102-bp). We performed quantitative real-time PCR for the NAGPA brain and non-brain isoforms in a cDNA panel originating from 16 human tissues and 24 sub-brain regions. According to our findings, the relative quantity of the NAGPA brain isoform in the brain was 4.7 times more than that in the control cDNA, a pooled mixture of equal amounts of cDNAs from the 16 different tissues. Further analysis using the cDNA panel originating from 24 different sub-brain regions revealed that the cerebral cortex contained the largest amount of NAGPA brain isoform. Relative quantity in the cerebral cortex was 8.6 times more than that in the control cDNA (P=0.00004). The lowest quantity of this isoform was detected in cDNA from the pituitary gland. In conclusion, findings of the current study suggest that the cerebral cortex, expressing the highest quantity of the NAGPA brain isoform, might be the region associated with speech function.
Alternative Splicing ; Brain ; Cerebral Cortex ; DNA, Complementary* ; Exons ; Humans* ; Lysosomes ; Mannosephosphates ; Phosphoric Diester Hydrolases ; Pituitary Gland ; Protein Isoforms* ; Real-Time Polymerase Chain Reaction ; Stuttering

Neuronal differentiation is a complex biological process accompanying cytoskeletal reorganization, including neurite outgrowth and growth cone formation. Therefore, neuronal differentiation is critically regulated by actin-related signaling proteins, such as small Rho GTPases, guanine nucleotide exchange factors (GEFs), and myosins. This study will demonstrate the change in activity of three small Rho GTPases, Rac, Cdc42, and Rho A, by treatment with blebbistatin (BBS), a specific inhibitor for myosin, during bFGF-induced neurite outgrowth in PC12 cells. Treatment with BBS induced morphological changes in growth cones and neurites during differentiation. A marked increase in protrusion and filopodia structures in growth cones, the shaft of neuritis, and cell membranes was observed in the cells treated with BBS. Activity of Rho GTPases showed the alterations in response to BBS. Activities of both Rac and Rho A were inhibited by BBS in a time-dependent manner. By contrast, Cdc42 activity was not changed by BBS. These results suggest that inactivation of myosin II by BBS induced morphological changes in neurites and growth cones and distinct regulation of three Rho GTPases during differentiation of PC12 cells.
Animals ; Biological Processes ; Cell Membrane ; Growth Cones ; Guanine Nucleotide Exchange Factors ; Heterocyclic Compounds with 4 or More Rings ; Myosin Type II ; Myosins ; Neurites ; Neuritis ; Neurons ; PC12 Cells* ; Proteins ; Pseudopodia ; rho GTP-Binding Proteins*

Mycoplasma hyopneumoniae (M. hyopneumoniae) is one of the causative bacteria that can induce chronic enzootic pneumonia, resulting in low production in the swine industry. Potentiation of porcine reproductive and respiratory syndrome virus-induced pneumonia by M. hyopneumoniae has also been recognized. Although some available vaccines have been developed for prevention of M. hyopneumoniae infection, protective immunity is still poor. In this study, in order to provide valuable information on vaccine antigen, we investigated the immunogenicity of M. hyopneumoniae on mouse spleen cells. Concanavalin A (ConA) and lipopolysaccharide (LPS) were used for generation of activated T and B lymphocytes. M. hyopneumoniae made clusters of spleen cells and also affected the cellular activity and viability of spleen cells by alone or with mitogens. Of particular interest, it induced a significant increase in production of TNF-alpha in ConA-treated spleen cells, meaning T helper 1 response. In addition, cell size and mitochondrial membrane potential of M. hyopneumoniae-treated spleen cells were measured by flow cytometric analysis. M. hyopneumoniae did not affect the cell size by alone, whereas ConA or LPS profoundly increased the cell size. Taken together, M. hyopneumoniae significantly affect the cellular activity and cytokine production of spleen cells by alone or in a combination of ConA. This study provides valuable information for production of the vaccine against M. hyopneumoniae.
Animals ; B-Lymphocytes ; Bacteria ; Cell Size ; Concanavalin A ; Membrane Potential, Mitochondrial ; Mice* ; Mitogens ; Mycoplasma hyopneumoniae* ; Mycoplasma* ; Pneumonia ; Porcine Reproductive and Respiratory Syndrome ; Spleen* ; Swine ; Tumor Necrosis Factor-alpha ; Vaccines

We would like to report a case of bovine lymphosarcoma. Parous cattle from a livestock farmhouse were examined for mutinodular masses in the abdominal cavity after slaughter. For clinical signs, animals presented mild leukemic signs but did not have viral or bacterial infection. Grossly, whitish to yellowish smooth masses similar to fat tissue were covered with a thin membrane. A multilobulated mass formed around the arteri, and there was a large quantity of reddish fluid on the cut surface. Histopathologically, a monomorphic population of lymphocytes was observed along with small amounts of cytoplasm, round nuclei with coarsely granular chromatin, and numerous mitotic figures in the samples. In the tumor lesion, uniformly round cells had invaded with abundant neovascularization. Especially, the immunohistochemical phenotype of tumor cells was positive for anti-CD3 and negative for anti-CD8 and anti-CD20. Therefore, morphological analysis diagnosed the mass as a multinodular bovine lymphosarcoma of T-cell origin without any sign of infection by a viral agent.
Abdominal Cavity ; Animals ; Bacterial Infections ; Cattle* ; Chromatin ; Cytoplasm ; Enzootic Bovine Leukosis ; Livestock ; Lymphocytes ; Lymphoma, Non-Hodgkin* ; Membranes ; Phenotype ; T-Lymphocytes*

In the livestock feed industry, antibiotics are used to prevent disease, promote growth rate, and improve feed efficiency. However, antibiotic supplementation to animal feed results in increased bacterial resistance to antibiotics as well as antibiotic residues in animal products, which can negatively affect human health. Therefore, alternative sources of antibiotics are needed. Probiotics as an alternative to antibiotics in animal feed have been shown to increase feed efficiency and growth rate by improving microbial balance. Further, Bacillus sp. produces a wide spectrum of antibacterial peptides. The present study was conducted to investigate the effects of dietary supplementation with CS-32 on safety, growth rate, and feed efficiency. Antibacterial substance (5697.9 molecular weights) produced by CS-32 was isolated and purified from culture broth. Moreover, the results of minimal inhibitory concentration (MIC) test confirmed the excellent antibacterial effect of CS-32. In vivo, 0.1% and 1% CS-32 were fed to broiler chickens for 28 days. Feed efficiency was slightly higher in groups of chickens supplemented with 0.1% and 1% CS-32 than those of the control group. CS-32 had no significant effect on necropsy findings, hematology, or serum biochemistry, and there was no mortality. These results suggest that CS-32 among various biologically active substances may be safe and effective as a feed additive to improve growth rate and feed efficiency.
Animal Feed ; Animals ; Anti-Bacterial Agents ; Bacillus ; Biochemistry ; Chickens* ; Dietary Supplements ; Hematology ; Humans ; Livestock ; Mortality ; Peptides ; Probiotics*

The purpose of this study was to explore the morphological characteristics of developing lentiform papilla (LP) in Korean native goats by scanning electron microscopy (SAM). Tongues were removed from fetuses on days 90, 120, neonates, and juveniles on days 30, 60, 90, 120, 150, and 180. In prenatal development, the primordia of LP in 90-day-old fetuses were round and spotted on the inner most part of the torus linguae of the tongue. Primordia of LP in 120-day-old fetuses also had a lens-like shape. In neonates, LP displayed similar features as the adult one. In postnatal juveniles on days 30 and 60, LP continually increased in size without much difference in structure compared to that of neonates. By postnatal day 90, detached pieces of keratinized superficial epithelia were observed. Microridges and microplicae were well developed on the epithelial surface of LP in 60- to 120-day-old goats. The lengths of LP were 476~514 microm in neonates, 687~962 microm in the weaning period of 60-day-old goats, and 1,068~1,567 microm in the maturing period of 180-day-old goats. These findings indicate that goat LP has different sizes and shapes from those of other species during development.
Adult ; Fetus ; Goats* ; Humans ; Infant, Newborn ; Microscopy, Electron, Scanning ; Morphogenesis ; Tongue* ; Virulence Factors, Bordetella ; Weaning

Mycoplasma (M.) hyopneumoniae is the causative agent of swine enzootic pneumonia, a disease that is prevalent in every country where pigs are raised. In this study, we aimed to develop a sensitive and specific PCR assay to detect M. hyopneumoniae in pigs. The suitability of this PCR assay for the detection of mycoplasmal infection was also tested using clinical lung samples from slaughtered pigs. We developed a probe and M. hyopneumoniae-specific primer pairs, MhyoP-F and MhyoP-R, for the new PCR assay based on regions in the Mycoplasma protein P97 gene that are unique to M. hyopneumoniae. The developed PCR assay was very specific and sensitive for the detection of M. hyopneumoniae. The assay was able to detect the equivalent of 10 pg of target template DNA, which indicates that the assay was very sensitive. In addition, the M. hyopneumoniae PCR assay detected only M. hyopneumoniae and no other Mycoplasma spp. or bacterial species of another genera. Further, the newly developed PCR assay effectively detected M. hyopneumoniae infection in pigs. We suggest that this PCR assay using M. hyopneumoniae-specific primer pairs, MhyoP-F and MhyoP-R, will be useful and effective for monitoring M. hyopneumoniae infection in pigs.
Diagnosis ; DNA ; Lung ; Mycoplasma* ; Pneumonia of Swine, Mycoplasmal ; Polymerase Chain Reaction* ; Swine

This study evaluated the efficacy and safety of medical shampoo containing terbinafine hydrochloride and chlorhexidine gluconate in dogs with dermatophytos is complicated with bacterial infection. Although several studies in dogs and cats with fungal dermatitis have found that oral administration of terbinafine is effective for controlling clinical signs, the topical form of terbinafine has rarely been studied in dogs and cats. Therefore, this study evaluated the efficacy of medical shampoo containing terbinafine hydrochloride and chlorhexidine gluconate in dogs with dermatophytos is complicated with bacterial infection. Eight dogs infected with Microsporum spp. complicated with bacterial infection were enrolled in this study. These dogs were further blindly divided into Group 1 (no treatment, fourdogs) and group 2 (treated with medical shampoo with terbinafine/chlorhexidine, four dogs). Clinical improvement was evaluated using bacterial and fungal cultural evaluation combined with clinical evaluation. This study found that the medical shampoo has sufficient efficacy to treat varying degrees of dermatophytosis complicated with bacterial infection in dogs, although the speed of improvement differed according to the degree and type of infection. Our study also found that combined therapy using antifungal and antibacterial agents can improve clinical signs more effectively and rapidly, suggesting that concurrent bacterial infection plays a significant role in the development of dermatitis.
Administration, Oral ; Animals ; Anti-Bacterial Agents ; Bacterial Infections* ; Cats ; Chlorhexidine* ; Dermatitis ; Dogs* ; Microsporum ; Naphthalenes ; Tinea*