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Color-pattern modifications and speciation in butterflies of the genus Vanessa and its related genera Cynthia and Bassaris.

We have previously shown that the systemic injection of sodium tungstate, a protein-tyrosine phosphatase (PTPase) inhibitor, to pupae immediately after pupation efficiently produces characteristic color-pattern modifications on the wings of many species of butterflies including Vanessa indica and Cynthia cardui. In these species, the black spots reduced in size in response to the treatment. Similar modifications are occasionally seen in the field-caught aberrant individuals. Exceptionally, however, a C. cardui individual with enlarged black spots ("reversed" modification pattern) has been reported. Here we show that these modified patterns of V. indica and C. cardui are quite similar to the normal color-patterns of other Vanessa species. V. indica with tungstate-induced modifications resembled V. tameamea, V. samani, and Bassaris itea, whereas V. dilecta, V. atalanta, and V. dejeanii are similar to the "reversed" individual. Most features seen in the experimentally-modified V. indica were observed throughout the fore- and hindwings of V. samani. In contrast, the experimentally-induced color-patterns of C. cardui did not parallel variation of Cynthia butterflies. Since it has been proposed that a hypothetical transduction pathway with a PTPase for the scale-cell differentiation globally coordinates the wing-wide color-patterns, our findings suggest that spontaneous mutations in genes in this hypothetical pathway might have played a major role in creating new color-patterns and species in the Vanessa genus but not in the Cynthia genus. This evolutionary mechanism may probably be shared more widely in Lepidoptera, although this would not be a sole determinant for the color-pattern development and evolution.

Animals↗

Tungstate-induced color-pattern modifications of butterfly wings are independent of stress response and ecdysteroid effect.

Systemic injections of sodium tungstate, a protein-tyrosine phosphatase (PTPase) inhibitor, to pupae immediately after pupation have been shown to efficiently produce characteristic color-pattern modifications on the wings of many species of butterflies. Here we demonstrated that the tungstate-induced modification pattern was entirely different from other chemically-induced ones in a species of nymphalid butterfly Junonia (Precis) orithya. In this species, the systemic injections of tungstate produced characteristic expansion of black area and shrinkage of white area together with the move of parafocal elements toward the wing base. Overall, pattern boundaries became obscure. In contrast, an entirely different modification pattern, overall darkening of wings, was observed by the injections of stress-inducing chemicals, thapsigargin, ionomycin, or geldanamycin, to pupae under the rearing conditions for the adult summer form. On the ventral wings, this darkening was due to an increase of the proportion of peppered dark scales, which was reminiscent of the natural fall form of this species. Under the same rearing conditions, the injections of ecdysteroid, which is a well-known hormone being responsible for the seasonal polyphenism of nymphalid butterflies, yielded overall expansion of orange area especially around eyespots. Taken together, we conclude that the tungstate-induced modifications are clearly distinguishable from those of stress response and ecdysteroid effect. This conclusion then suggests that the putative PTPase signaling pathway that is sensitive to tungstate uniquely contributes to the wing-wide color-pattern development in butterflies.

Animals↗

Interaction between isopoly-tungstic acid and berberine hydrochloride by resonance Rayleigh scattering spectrum.

In a weak acid medium, when an isopoly-tungstic acid reacts with berberine to form an ion-association complex, strong resonance Rayleigh scattering (RRS) appears and obvious frequency-doubling scattering (FDS) as well as second-order scattering (SOS) come into being. The scattering peaks are located at 288 and 369 nm for RRS, at 290, 370 and 390 nm for FDS, and at 480, 540 and 740 nm for SOS. Under optimum conditions, all three scattering intensities are directly proportional to the concentration of berberine and can be applied to the determination of berberine. The sensitivity of the RRS method is the highest; the detection limit (3sigma) for berberine hydrochloride is 7.6 ng/ml. The method also has good selectivity. It has been applied to the determination of berberine in pharmaceuticals and goldthread extracts with satisfactory results. In the paper, studies of the existing species of isopoly-acid and the component of the ion-association complex are reported and the reasons of enhancement of RRS are discussed.

Berberine↗

Simple and rapid determination of hydrogen peroxide using phosphine-based fluorescent reagents with sodium tungstate dihydrate.

A simple batch method for the fluorometric determination of hydrogen peroxide using phosphine-based fluorescent reagents has been developed. A rapid, mild and selective derivatization reaction was achieved by adding sodium tungstate dihydrate to the reaction mixture of hydrogen peroxide and a phosphine-based fluorescent reagent. When 4-diphenylphosphino-7-methylthio-2,1,3-benzoxadiazole was used as a reagent, the derivatization reaction was completed after 2 min at room temperature. The calibration curve was linear between 12.5 and 500 ng hydrogen peroxide in a 10 microL sample solution. This method is accurate and has potential for on-line applications.

Calibration↗

Tungstate treatment improves Leydig cell function in streptozotocin-diabetic rats.

Oral administration of sodium tungstate to adult male streptozotocin-diabetic rats for 3 months normalized serum levels of glucose, insulin, luteinizing hormone, and follicle-stimulating hormone. These effects were accompanied by an increase in reproductive performance, which was related to a strong improvement in Leydig cell function markers, such as the recovery of the number of Leydig cells and serum testosterone levels. Moreover, this in vivo recovery was related to a concomitant increase in the cell expression of insulin receptors. Tungstate treatment did not modify Leydig cell function in healthy rats. Furthermore, the addition of tungstate or insulin to the mTLC-1 cell line from Leydig cell origin increased the phosphorylation states of MAP-kinase and glycogen synthase kinase-3. Our results indicate that tungstate treatment in diabetic rats leads to a recovery of reproductive performance by increasing the number of Leydig cells. This increase contributes to the recovery of their functionality, thereby improving the overall function of these cells. We propose that this improvement is caused by the combined effect of the tungstate-induced normalization of insulin glucose and luteinizing hormone serum levels and a direct action of the effector on Leydig cells through modulation of at least MAP-kinase and glycogen synthase kinase-3 activities.

Animals↗

Antiviral therapy in AIDS. Clinical pharmacological properties and therapeutic experience to date.

The rapid spread of human immunodeficiency virus (HIV) infections and the grim outcome of these infections have focused interest on the possibilities for medical intervention. The end-stage of these infections, acquired immune deficiency syndrome (AIDS), was first recognised in 1981, and the causative agent isolated in 1983. Already several antiviral drugs have been investigated. One initially promising drug, suramin, was found to have a net harmful effect but another, zidovudine (azidothymidine) has been shown to prolong life in AIDS patients. The properties of these and several other antiviral drugs such as antimoniotungstate (HPA-23), foscarnet (phosphonoformate) ribavirin, dideoxynucleotides, and interferons, are reviewed. The role of immunomodulating modalities such as plasmapheresis, bone marrow transplantation, thymosin, interleukin-2, inosine pranobex (isoprinosine), and cyclosporin are also discussed. None of the currently available drugs hold promise as monotherapy. Through analysis of the experience with these drugs and the increasing knowledge of HIV pathogenesis, new drugs can be designed. It seems increasingly clear that drugs will eventually have to be used in combination in order to reduce toxicity, exploit therapeutic synergy, and reduce the risk of HIV resistance. The theoretical and experimental background for such combinations are currently being elucidated.

Acquired Immunodeficiency Syndrome↗

Polyoxotungstates reduce the beta-lactam resistance of methicillin-resistant Staphylococcus aureus.

Bacterial strains isolated from clinical specimens have become more and more resistant to many anti-microbials. This is because we have consumed large amounts of strong antimicrobials over long periods of time and thus bacterial cells are able to survive by altering the target(s) of antimicrobial agents. A good example of this phenomenon is methicillin-resistant Staphylococcus aureus (MRSA). One of the cell wall-synthesizing enzymes (known as PBP2') of this pathogen has low affinity to beta-lactams, and therefore the bacterial cells continue to grow even under high concentrations of the agents. However, this drug resistance does not seem to be total. They seem to have some weak spots and several substances are known to sensitize strains of MRSA to beta-lactams. This review discusses the ability of polyoxotungstates (POTs) to sensitize MRSA to beta-lactams by reducing the expression of PBP2'. It is also possible that the sensitization is a type of stress response of MRSA to POTs. This idea may provide a hint for the development of a new antimicrobial agent.

Anti-Bacterial Agents↗

Effects of tungstate, a new potential oral antidiabetic agent, in Zucker diabetic fatty rats.

Tungstate was orally administered to 7.5-week-old male Zucker diabetic fatty (ZDF) rats that already showed moderate hyperglycemia (180 +/- 16 mg/dl). The animals became normoglycemic for approximately 10 days. Then, glycemia started to rise again, although it did not reach the initial values until day 24, when levels stabilized at approximately 200 mg/dl for the duration of the experiment. Untreated ZDF rats showed steadily increased blood glucose levels between 7.5 and 10 weeks of age, when they reached a maximum value of 450 +/- 19 mg/dl, which was maintained throughout the experiment. In addition, tolerance to intraperitoneal glucose load improved in treated diabetic rats. Serum levels of triglycerides were elevated in untreated diabetic rats compared with their lean counterparts (ZLC). In the liver of diabetic animals, glucokinase (GK), glycogen phosphorylase a (GPa), liver-pyruvate kinase (L-PK), and fatty acid synthase (FAS) activities decreased by 81, 30, 54, and 35%, respectively, whereas phosphoenolpyruvate carboxykinase (PEPCK) levels increased by 240%. Intracellular glucose-6-phosphate (G6P) decreased by 40%, whereas glycogen levels remained unaffected. Tungstate treatment of these rats induced a 42% decrease in serum levels of triglycerides and normalized hepatic G6P concentrations, GPa activity, and PEPCK levels. GK activity in treated diabetic rats increased to 50% of the values of untreated ZLC rats. L-PK and FAS activity increased to higher values than those in untreated lean rats (1.7-fold L-PK and 2.4-fold FAS). Hepatic glycogen levels were 55% higher than those in untreated diabetic and healthy rats. Tungstate treatment did not significantly change the phosphotyrosine protein profile of primary cultured hepatocytes from diabetic animals. These data suggest that tungstate administration to ZDF rats causes a considerable reduction of glycemia, mainly through a partial restoration of hepatic glucose metabolism and a decrease in lipotoxicity.

Administration, Oral↗

Efficiency of the polymer bur SmartPrep compared with conventional tungsten carbide bud bur in dentin caries excavation.

SmartPrep is a rotating instrument for dentin caries excavation made from a special polymer. The manufacturer's product information stated that SmartPrep removes carious dentin selectively. This in vitro study compared the efficiency of SmartPrep with conventional tungsten carbide bud burs. Fifty extracted teeth were split in the center of a carious lesion. The 100 specimens were randomly divided into five groups. Five dentists were asked to excavate 10 teeth each: one half with SmartPrep and the corresponding half with conventional bud burs. The time needed for the caries excavation was measured. Subsequently, histological specimens were produced from all cavities and analyzed by light-microscope after Mallory-Azan-staining. The thickness of the remaining caries was measured (< 1 mm or > 1 mm). The time expended was analyzed using the paired t-test. The results were analyzed for the remaining caries and thickness of the carious layer for every tooth, using the non-parametric Wilcoxon test for combined random samples. A binary logistical regression was performed to determine the influence of the three variables (tooth, sections or bur) on the criteria "caries" or "carious layer thickness (> 1 mm)." The average time to excavate a cavity with SmartPrep was 208.1 seconds, and it was 228.32 seconds with conventional bud burs. The difference between the recorded times was not statistically significant (p > 0.05). In 37 of 50 teeth, the number of carious sections was higher in the SmartPrep group than in the bud bur group. In nine teeth, the quantity of carious sections was higher in the bud bur group than in the SmartPrep group. Four teeth showed no difference in the number of carious sections. The results were statistically significantly different (p < 0.001). In 30 teeth, the number of carious sections with a carious layer thicker than 1 mm was higher in the SmartPrep group compared with the bud bur group. In nine teeth, the number of carious sections was higher in the bud bur group than in the SmartPrep group. Eleven teeth showed no difference in thickness of the carious dentin layer. These results were statistically significantly different (p = 0.003). Binary logistical regression showed that only the variable "bur" (bud bur or SmartPrep) influenced the results concerning the criterion "caries" (p < 0.001).

Dental Caries↗

Review of the effects of pH on the synthesis of heteropoly tungstate anions.

The reaction of the WO4(2-) ion in the pH range 2 to 8 was carried out with the following metal ions individually in combination: Mn2+, Mn3+, Fe2+, Fe3+, Co2+, Co3+, Ni2+, Cu1+, Cu2+, Zn2+, Ga3+, Mg2+, In3+, RAsO3(2-), RSbO3(2-). Some 36 heteropolyanions have been isolated in the crystalline form characterized fully. The antiviral activity of these anions has been determined against HIV, herpes respiratory viruses. The redox potentials for these anions have been measured by cyclic voltammetry documented. At pH 8, the anions adopt a Weakley structure at pH 7, the Keggin structure is formed. At pH 6 to 7, the distorted Keggin structure, 1:1:11, is formed. Below pH 6, the distorted Dawson structure is formed. Some anions showed good antiviral activity with positive redox potentials and others with negative redox potentials. When the metal ion is attached to an organic group, a ring-type structure is formed: a 2:5, 2:6, and 1:7 heteropolytungstate.

Anti-HIV Agents↗

Cryofixation combined with physical dehydration for quantitative immunoelectron cytochemistry.

Common methods for preparing samples for immunoelectron microscopy involve glutaraldehyde fixation (GA) followed by chemical dehydration (CD) or cryofixation (CF) succeeded by physical dehydration, i.e., freeze drying (FD) or freeze substitution (FS). The effects of these techniques have been evaluated with regard to the sizes of epoxy resin embedded rat somatotrophic secretory granules as well as the immunolabeling densities over these granules. The measurements were performed by computerized image analysis using electron microscopy in transmission (TEM) and scanning transmission (STEM) modes, which allowed us to define the immunolabeling in detail. The embedded secretory granules showed the same diameters after GA (2 hr) with CD and GA (15 min) with CF and FS, but were smaller after CF-FS, and smallest after GA (15 min) with CF and FD. The highest labeling density appeared after GA (15 min) and physical dehydration, in particular after freeze substitution. Based on our STEM pictures a new factor for evaluating and interpreting immunolabeling of granules is introduced; the "accessible immunogold labeling surface." It defines the fraction of the epoxy resin surface that is labeled and varies with the preparation methods. By using this factor, an order of labeling densities/micron 2 over the accessible areas could be established for the different techniques: GA-CF-FS > CF-FS > GA-CF-FD > GA-CD. The high labeling after GA-CF-FS may be due to the combination of a large accessible area and accurate preservation of the antigenicity of the hormones in the granules.

Animals↗

Durability of tungsten carbide burs for the fabrication of titanium crowns using dental CAD/CAM.

The purpose of this study was to evaluate the durability of tungsten carbide burs for the fabrication of titanium crowns using two dental CAD/CAM systems (DECSY, Digital Process, Japan and Cadim, Advance, Japan). A tungsten carbide bur in each system was examined and used without fracture to fabricate 51 titanium crowns. For both systems tiny chips were found on the bur blade at the 11th machining. These chips gradually enlarged as the number of machining times increased. At the first machining no significant difference in the average surface roughness was found on the crown between the two systems (1.6 microm for DECSY and 1.2 microm for Cadim). The cutting grooves became dull and the average surface roughness increased as the number of machining times increased. It is concluded that the tungsten carbide burs for both systems can be used to fabricate up to 51 titanium crowns.

Computer-Aided Design↗

Immunologic reconstitution in the acquired immunodeficiency syndrome.

Although effective therapies are available for many of the infections and tumors that occur in patients with the acquired immunodeficiency syndrome (AIDS), no therapy exists for the underlying immunodeficiency. Three approaches used to treat this immunodeficiency have included wholescale immune replacement through lymphocyte transfers, bone marrow transplantation, and thymic implantation; immunologic enhancement with biologic-response modifiers, such as gamma interferon, interleukin-2, and some drugs; and antiretroviral therapy. Because of the persistence of the etiologic virus, both immune replacement and enhancement will probably be ineffective unless effective strategies are developed against the virus. Agents that have shown antiviral activity in vitro which are currently in clinical trials include suramin, heteropolyanion-23 (HPA-23), ribavirin, and alpha interferon. Foscarnet and monoclonal antibodies have yet to enter clinical trials in the United States. It is still too early to tell if any of the antiviral agents will prove of value in the management of patients with AIDS.

Acquired Immunodeficiency Syndrome↗

Prospects of therapy for infections with human T-lymphotropic virus type III.

Human T-lymphotropic virus type III is susceptible to attack at various sites during its replicative cycle. Inhibitors of reverse transcriptase activity, including suramin, antimoniotungstate (HPA-23), and trisodium phosphonoformate, have shown in-vitro activity against the virus in early clinical trials. Other significant antiviral agents are recombinant interferon alpha-A, ribavirin, and ansamycin. Double-blind, placebo-controlled clinical trials of interferon alpha, which inhibits viral replication at easily achievable serum levels, are underway. The development of optimal therapeutic regimens will require carefully controlled, multicenter collaborative trials with standardized criteria for evaluating responses. Prolonged treatment with combinations of antiviral and immunomodulating agents may be necessary for control of HTLV-III replication and for effective treatment of the acquired immunodeficiency syndrome.

Acquired Immunodeficiency Syndrome↗