In recent years, there has been a rapid decrease in amphibian populations worldwide, and infectious diseases have been associated with this decline. Diseased frogs inhabiting Korea were collected from fields, and the diseases were identified by morphological and molecular analyses. Two fungal diseases-saprolegniasis and chromomycosis-were detected in the frogs. Saprolegniasis caused by Saprolegnia spp. was found in Rana plancyi chosenica from Gangwon-do and Rana huanrenensis from Chungbuk. Chromomycosis, which is caused by infection with Cladosporium cladosporioides, was detected in Rana catesbeiana from Busan.
Amphibians ; Chromoblastomycosis ; Cladosporium ; Communicable Diseases ; Korea ; Rana catesbeiana ; Ranidae ; Saprolegnia

Previously, we have isolated authentic bombesin and another bombesin like peptide named bombesin like immunoreactivity (BLI)-K2 from the skin of Korean fire-bellied toad, Bombina orientalis. In the present study, we have newly purified three heterogeneous forms of BLI named BLI-K3, BLI-K4, and BLI-K5 from side fractions obtained in previous isolation of bombesin like peptide. The BLIs were separated into five peaks on a column of C18 preparative HPLC. Among them, three minor peaks containing BLI-K3, K4, and K5 were purified by means of sequential chromatography on the columns of SP cation exchange HPLC and C18 reverse phase HPLC. The purified BLI-K3 and K4 showed high binding affinity to an anti-bombesin serum (LBE 2G-2) with binding potency of 72 and 95%, respectively, relative to that of bombesin. However, they did not possess any distinctive biological activity of bombesin like peptide. On the contrary, the biological activity of BLI-K5 was similar to that of bombesin but its binding affinity to an anti-bombesin serum was low. The results indicate that three heterogeneous forms of BLI were coexpressed with bombesin and BLI-K2 in the skin of B. orientalis. All forms of the purified BLI in the present study were immunologically active but only BLI-K5 possessed the distinctive biological activity of bombesin like peptide.
Anura* ; Bombesin* ; Chromatography ; Chromatography, High Pressure Liquid ; Population Characteristics ; Skin*

The drug-resistance of malaria parasites is the main problem in the disease control. The huge Brazilian biodiversity promotes the search for new compounds, where the animal kingdom is proving to be a promising source of bioactive compounds. The main objective of this study was to evaluate the antiplasmodial and cytotoxic activity of the compounds obtained from the toad venoms of Brazilian Amazon. Toad venoms were collected from the secretion of Rhinella marina and Rhaebo guttatus in Mato Grosso State, Brazil. The powder was extracted at room temperature, yielding 2 extracts (RG and RM) and a substance ('1') identified as a bufadienolide, named telocinobufagin. Growth inhibition, intraerythrocytic development, and parasite morphology were evaluated in culture by microscopic observations of Giemsa-stained thin blood films. Cytotoxicity was determined against HepG2 and BGM cells by MTT and neutral red assays. The 2 extracts and the pure substance ('1') tested were active against chloroquine-resistant Plasmodium falciparum strain, demonstrating lower IC₅₀ values. In cytotoxic tests, the 2 extracts and substance '1' showed pronounced lethal effects on chloroquine-resistant P. faciparum strain and low cytotoxic effect, highlighting toad parotoid gland secretions as a promising source of novel lead antiplasmodial compounds.
Amphibian Venoms* ; Animals ; Biodiversity ; Brazil* ; Bufo marinus ; Malaria ; Neutral Red ; Parasites ; Plasmodium falciparum

OBJECTIVETo determine the origin of Rana temporaria for quality Oviduetus Ranae in the light of historical documents and modern researches on the classification of Rana temporaria chensinensis.

METHODWorks of Chinese meteria medica of all ages, related historical documents and reports from home and abroad on researches of R. temporaria chensinensis were consulted, sorted out, analyzed and summarized.

RESULTThe original Shange recorded in the works of Chinese meteria medica is R. temporaria chensinensis, which is the independent species, not one of species of European forest frogs. R. temporaria chensinensis is divided into 4 subspecies: R. temporaria chensinensis, Lanzhou, Kangding, and Changbaishan. The origin of R. temporaria is Changbaishan subspecies of R. temporaria chensinensis.

CONCLUSIONChangbaishan subspecies of R. temporaria chensinensis is determined as the origin for quality Oviduetus Ranae.

Animals ; Female ; History, 18th Century ; History, 19th Century ; History, 20th Century ; History, Ancient ; Materia Medica ; history ; isolation & purification ; Oviducts ; chemistry ; Rana temporaria ; anatomy & histology ; Ranidae ; anatomy & histology ; classification ; Species Specificity ; Terminology as Topic

The furcocercus cercariae of Neodiplostomum seoulense (Digenea: Neodiplostomidae) penetrate the skins of tadpoles and shed their tails. The speculated mechanism of this tail loss was physical efforts required to produce a vigorous zigzag motion during skin penetration; no other mechanism has been proposed. We examined the relationship between the host serum and cercarial tail loss. Cercariae of N. seoulense were collected from experimentally infected Segmentina hemisphaerula, and lots of 300 cercariae were cultured in medium 199 contained several types of sera. Cercarial tail degradation was induced in all media, but all the cercariae cultured except those cultured in media containing fetal bovine serum (FBS) died within 48 hr. After 72 hr cultivation in media containing FBS, cercarial tail degradation was induced in 67.0%; in continuous cultivation 13.3% of larvae survived for 7 days. Tail degradation did not occur in the absence of serum and when serum was heat inactivated at 56 degrees C for 30 min. The addition of 20 mM ethylenediaminetetraacetic acid (EDTA) blocked cercarial tail degradation completely. Moreover, the addition of 20 mM MgCl2 restored tail degradation blocked by EDTA. These results suggest that the alternative complement pathway is related with the N. seoulense cercarial tail degradation induced by serum.
Trematoda/*physiology ; Tail/*physiology ; Larva/parasitology ; Complement System Proteins/immunology/*physiology ; Anura/parasitology ; Animals

Electron microscopy on the skin of young frogs, Rana temporaria, has been carried out with particular reference to cellular attachment sites. For the first time now several technical developments allow a more detailed visualization of the fine structure within the cellular attachment sites as well as making it possible to show the ultrastructural morphology of the junctional complexes, and to demonstrate that the desmosomes are regularly distributed around each skin cell, especially in the S. granulosum. The relations of these findings to those of previous investigations concerning the functional organization of the junctional complexes and to the findings in skin cancer from a cellular adhesion view point have been briefly discussed.
Animal ; Anura ; Cell Membrane ; Epithelial Cells* ; Microscopy, Electron ; Skin/cytology* ; Skin Neoplasms/etiology

This study was carried out to observe the ultrastructure of tongue mast cells in frog, Bombina orientalis. In the light microscopy, mast cells were round, oval, spindle-like and irregular shape and occasionally had one or more tail-like long cytoplasmic processes. Mast cells had many red granules stained with toluidine blue in the cytoplasm. In the electron microscopy, mast cells had membrane-bounded granules and poorly developed organelles. The granules were composed of two parts, low and dense electron density. In high magnification the dense part in the middle region of granule had honey comb-like crystalline structure.
Anura* ; Crystallins ; Cytoplasm ; Honey ; Mast Cells* ; Microscopy ; Microscopy, Electron ; Organelles ; Tolonium Chloride ; Tongue*

OBJECTIVE: To quantitatively investigate the effects of Ringer's solution with different concentrations of alcohol (1%～80%) on biphasic compound action potentials (AP) from frog sciatic nerve trunk, and their recoveries from alcohol effects. METHODS: Individual segments of frog sciatic nerve trunk with a length of 6 to 8 cm were prepared. Ringer's solution with different concentrations of alcohol (0%, 1%, 2%, 4%, 8%, 16%, 32%, 48%, 64% and 80%) was applied onto the segment of the trunk between the stimulus and ground electrodes via an agent reservoir which was newly armed in a nerve trunk shielded chamber for 5 minutes. The nerve trunk was respectively electro-stimulated to generate the biphasic compound AP which was recorded using the experimental system of BL-420F. This was followed by 5 times washout plus 5 min administration with Ringer's solution before recovery recording of AP. RESULTS: Compared to normal Ringer's solution, Ringer's solution with alcohol at ≤4% did not have dramatic impacts on the AP amplitude and conduction velocity, while Ringer's solution with alcohol at ≥8% there was significant decrease in these two parameters. Ringer's solution with alcohol at the conentrations of 16%, 32% and ≥48% could prevent a small proportion (30%), a large proportion (90%) and all (100%) of sciatic nerve trunks, respectively, from generating AP. Washout with normal Ringer's solution after alcohol application at the concentration of ≤32%, AP could totally recover to normal status. While alcohol at the concentration of 48%, 64% and 80%, the probabilities to regenerate APs were 90%, 40% and 0%, and the AP amplitudes were decreased to 60%, 36% and 0%, respectively. After washout, AP conduction velocity showed no difference with alcohol at the concentration of ≤8% when compared with that before washout, while it could not be recovered to normal under alcohol at ≥16%. CONCLUSION Ringer's solution with different concentrations of alcohol exerts different effects on biphasic compound AP amplitude and conduction velocity. Hopefully, our findings could be helpful for the alcoholic usage and its recovery from alcoholic damage.
Action Potentials ; Animals ; Anura ; Ethanol ; pharmacology ; Ringer's Solution ; pharmacology ; Sciatic Nerve ; drug effects

The purpose of the present study was to examine the kinetic process of hemoglobin (Hb) carrying and releasing oxygen. Under the standard conditions (pH 7.4, Po(2) 20 mmHg, 20 °C) the blood samples of chicken, rabbit, frog and carp were equilibrated in oxygen content analyzer with calibrated gas mixture A (0.5% CO(2) and 99.5% N(2)). Then the blood samples were exposed to gas mixture B (21% O(2), 0.5% CO(2) and 78.5% N(2)). After equilibration, the blood samples were exposed to gas mixture A again. During the whole process, Po(2) of blood samples was detected in real-time. The time spent in blood Po(2) changing from 0 to 21 kPa was recorded carefully. The results indicated that the kinetic curve of Hb carrying oxygen presented a shape of "S". It was similar to the Hb oxygen dissociation curve (Hb ODC). Based on the curve, T(50), a new kinetic parameter, was established. T(50) is the time of 50% O(2) saturation of Hb. It can reflect the efficiency of Hb carrying oxygen. Through comparing of T(50), the efficiency of Hb carrying oxygen among 4 species of animals was: frog < carp < rabbit < chicken. In the phase I of Hb carrying and releasing oxygen kinetic curve, the slope in carp was much larger than that in rabbit; the time [(1 411±6) s] of Hb releasing oxygen in chicken was longer than that in other 3 animals. These differences reflected the variety of efficiency of Hb carrying and releasing oxygen. In addition, the kinetic features of Hb carrying oxygen were likely to become an important index to evaluate the function of Hb carrying oxygen, especially in evaluating the ability of artificial blood substitute. On the basis of the analysis of the kinetic curve of Hb carrying oxygen and Hb ODC, another new important efficacy parameter E(50) was proposed. E(50) reflects the relationship between the time of 50% O(2) saturation of Hb and environmental Po(2). E(50) can be used as a synthetic index to assess the efficiency of Hb carrying oxygen.
Animals ; Anura ; Carps ; Chickens ; Hemoglobins ; metabolism ; Hydrogen-Ion Concentration ; Kinetics ; Oxygen ; metabolism ; Rabbits

Full evidence and obvious reasons made it possible to arrive at the conclusion that the nature of transmission upon cerebrospinal neurons is overwhelmingly mechanical, not only in the periphery- between various receptors and afferent nerve terminals, and between surrounding tissues and free nerve endings- but also in the cerebral cortex. When viewed from the standpoint of the everchanging patterns of natural mechanical stimuli, the neurons in the conscious cerebral cortex and the pain endings in an acute inflammatory locus have the same situation very much in common. It is quite likely that natural mechanical stimuli dominate over cerebrospinal nervous phenomena and physiologists have been watching the missing mechanism at work in every experiment upon afferent nerve terminals and cerebral cortex that they have done. The terms "psychic tension" and "central excitatory state" comparable to muscular tonus are of interest because they involve the use of mathematical techniques in psychology and neurophysiology. They are capable of becoming weak or strong, and they serve as an inner stimulus to give impetus to behavior. Unfortunately, however, it is an elusive inner stimulus, and it defies a lucid definition. But natural mechanical stimuli embody the psychic tension and the central excitatory state ultimately. It seems now that we just found a place where constant complaints against neurophysiology and physiological psychology are ventilated. We may conclude that natural mechanical stimuli are the leading direct stimuli to cerebrospinal neurons in the human body, and the plastic and developmental nervous phenomena and mental phenomena can be explained objectively by a familliar datum of mechanical energy and that we can reasonably expect the day of regarding material world and spiritual world in the monistic conception of matter-energy system.
Animals ; Anura ; Cerebral Cortex/*physiology ; Human ; In Vitro ; Motor Neurons/physiology ; Nerve Endings/physiology ; Receptors, Sensory/*physiology ; Spinal Cord/*physiology