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Flow cytometry was used to study the genome sizes and ploidy levels for four thrips species: Franklinothrips orizabensis Johansen (Thysanoptera: Aeolothripidae), Frankliniella occidentalis Pergande, Frankliniella fusca Hinds, and Thrips tabaci Lindeman (Thysanoptera: Thripidae). F. orizabensis males and females had 1C genome sizes of 426 Mb and 422 Mb, respectively. Male and female F. fusca had 1C genome sizes of 392 Mb and 409 Mb, whereas F. occidentalis males and females had smaller 1C genomes that were 345 Mb and 337 Mb, respectively. Male F. orizabensis, F. occidentalis and F. fusca were haploid and females diploid. Five isofemale lines of T. tabaci, initiated from parthenogenetic, thelytokous females and collected from different locations in North Carolina, were included in this study; no males were available. One isofemale line was diploid with a genome size of 1C = 310 Mb, and the other four had a mean genome size of 1C = 482 Mb, which is consistent with evidence from microsatellite data of diploidy and polyploidy, respectively, in these same five thelytokous lines. This is the first study to produce genome size estimates for thysanopteran species, and report polyploidy in T. tabaci populations.

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As red imported fire ant, Solenopsis invicta Buren, continues to expand its range into the southwestern United States, it can be easily confused with the native southern fire ant, Solenopsis xyloni McCook. Variability in the morphological characteristics commonly used to differentiate these ant species was quantified by examining the length of the clypeal tooth, striations of the mesopleuron, length of antennal scape, area of the petiolar process, number and size of mandibular teeth, and color by using both scanning electron and light microscopy. Given enough samples, the average values of each of these characteristics is different between the two species; however, significant morphological variability occurs in both S. xyloni and S. invicta populations, creating an area of overlap where either of the two species could exhibit similar characteristics. Better differentiation of these two species is achieved using a combination of characteristics, but morphological techniques are not dependable unless numerous ants from each population are analyzed by a taxonomist familiar with Solenopsis. For situations requiring a more accurate identification, such as before quarantining a county or a portion of a county, a molecular technique using mitochondrial DNA and polymerase chain reaction techniques was developed.

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The thermal denaturation of wild-type beta-galactosidase and two beta-galactosidases with substitutions at the active site was studied by kinetics, differential scanning calorimetry, electrophoresis, molecular exclusion chromatography, and circular dichroism. From the results, a model is developed for thermal denaturation of beta-galactosidase which includes the reversible dissociation of ligands, reversible formation of an inactive tetramer, irreversible dissociation of the inactive tetramer to inactive monomers, and subsequent aggregation of inactive monomers to dimers and larger aggregates. Under some conditions, partial reversibility of the activity loss could be demonstrated, and several intermediates in the thermal denaturation process were trapped by quenching and observed by electrophoresis and molecular exclusion chromatography. The ligands Mg2+ and phenylethyl thio-beta-D-galactoside increase the stability of beta-galactosidase to heat denaturation by shifting the ligand binding equilibrium according to Le Chatelier's principle, thus decreasing the concentration of the ligand-free tetramer which can proceed to subsequent steps. Circular dichroism results indicated that beta-galactosidase is dominated by beta-sheet with lower amounts of alpha-helix. Large changes in secondary structure begin to occur only after activity has been lost. Single amino acid changes at the active site can have significant effects on thermal stability of beta-galactosidases. Some of the effects result from increased thermal stability of the ligand-free enzyme itself. Other effects result from changes in ligand binding, but the magnitude of the resulting changes in stability is not related to the strength of ligand binding in a simple fashion.

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Cranial neuralgias are relatively uncommon but the excruciating nature of the pain they cause make them the focus of much clinical and research interest. The underlying mechanism of the disease remains to be uncovered. The complexities of the trigeminal system make the quest for comprehensive understanding a difficult one. Fortunately, pharmacotherapy is quite effective for many patients. For those patients who cannot be managed medically, there are a variety of surgical options. Criteria have been proposed for evaluating surgical efficacy in trigeminal neuralgia. These parameters should be used when considering the surgical options for each patient. Above all, the Latin motto, Primum non nocerum (first, do not harm) should prevail. With this in mind, the physician and surgeon will make the best choice for the care of each patient.

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Patients with AIDS or ARC may present with parotid gland enlargement either initially or during the course of their disease. Of nine such patients, eight had unilateral gland enlargement, and one had bilateral disease. CT and MR scanning revealed that all patients except one had bilateral multiple intraparotid cysts. In addition, four of the nine patients clinically had cervical adenopathy, while all patients had adenopathy on scanning. We believe that the association of parotid cysts and cervical adenopathy is another manifestation of AIDS or ARC and, when found on scanning, should alert the clinician to the probable underlying condition.

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Thermal transitions were measured by differential scanning calorimetry for rabbit cardiac sarcolemma in 3-(N-morpholino)propanesulfonic acid buffer at pH 7.5, in glycerol-buffer and dimethyl sulfoxide - buffer mixtures, after heat denaturation, and after enzymatic degradation of the proteins. Specific solvent effects on the protein transitions were observed. Glycerol stabilized some of the four protein transitions, while dimethyl sulfoxide destabilized all protein transitions. The thermal transitions in the lower temperature range were studied for both the membranes and the lipid extracted from the membranes. A very small endotherm was observed for both the lipid extracted from the sarcolemma and the intact membrane (0.1-0.2 cal/g; 1 cal = 4.1868 J). A larger endotherm was observed in both the glycerol-buffer and dimethyl sulfoxide - buffer mixtures. Major perturbation of the protein by enzymatic degradation (papain or trypsin digestion), by heat denaturation, or by reaction with excess N-ethylmaleimide all produced larger endotherms near 20 degrees C. The very small magnitude of the endotherm near 20 degrees C suggests that it is not a typical gel - liquid crystalline transition of the bilayer. However, the occurrence of an endotherm in the extracted lipid suggests that some reorientation of lipid is involved.

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The paranasal sinuses are a common source of maxillofacial pain. When the sphenoid sinus is involved, 13 different structures can also be affected. The various neuralgias are usually easily recognized by their classic symptoms. Temporal arteritis must be diagnosed early in order to prevent blindness. The temporomandibular joint syndrome remains the most misdiagnosed and misunderstood orofacial pain.

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Ionic calcium, calcium binding sites, and other urinary variables were measured in 58 patients with idiopathic calcium nephrolithiasis and 36 normal subjects. The patients showed higher urinary concentrations of calcium. The mean calcium excretion (mmole/24 hr) was 4.45 +/- 0.56 (+/- 1 SEM) in patients and 2.19 +/- 0.22 (+/- 1 SEM) in normal subjects. This difference was highly significant (P less than 0.001). The mean ionic calcium excretion (mmole/24 hr) was 1.90 +/- 0.21 (+/- 1 SEM) for patients and 0.97 +/- 0.12 (+/- 1 SEM) for control subjects. The normal subjects showed significantly higher (P less than 0.01) concentrations and total excretions of magnesium and citrate. Excretory patterns for sodium, potassium, phosphate, and oxalate were not significantly different. The normal subjects had higher mean urinary concentrations of binding sites for calcium ions (23.2 +/- 4.8 mM) than the patients (18.5 +/- 2.9 mM). However, as the patients had higher urinary volumes the difference in the 24-hr excretion of calcium binding sites was not significant statistically. Out of 58 patients 43 (74%) were hypercalciuric. Twenty patients (46%) were categorized as an absorptive group and one patient as a resorptive type, and for the rest of the patients (51%) the mechanism of hypercalciuria remained unidentified. Only two of the control subjects (5%) were found to be hypercalciuric under calcium restricted diet conditions. Though these "control" subjects excreted a high amount of calcium there was no associated increase in the fraction of the calcium in the ionic form (0.37). Patients, however, still had relatively high fractions of ionic calcium (0.48 +/- 0.03).

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Differential scanning calorimetry has been used to study the thermal denaturation of ribonuclease in aqueous dimethyl sulfoxide and acqueous p-dioxane. A two-state reversible denaturation occurs in acqueous dimethyl sulfoxide. The denaturation is irreversible in aqueous p-dioxane. The transition temperatuire decreases in both organic solvents, with p-dioxane producing a considerably greater effect. The enthalpy of the denaturation initially increases with increasing dimethyl sulfoxide concentration and then decreases at high concentrations. Similar behavior is observed in the entropy in the dimethyl sulfoxide solutions. The enthalpy of the denaturation decreases markedly with low p-dioxane concentrations. Changes in pH due to presence of organic solvent cannot account for the changes in enthalpy of denaturation. Addition of organic solvent tends to increase the pH of the ribonuclease solution. Higher pH in aqueous solutions gives a high transition temperature, whereas addition of organic solvent results in the opposite bahavior. The relative changes in the two solvents systems suggest that specific solvent effects occur and that destablization of the native state relative to the denatured state is greater with p-dioxane than with dimethyl sulfoxide.

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Total bindings sites, ionic calcium, and other crystalloids were measured in 41 normocalciuric patients with idopathic calcium urolithiasis and in 34 normocalciuric healthy subjects. Twenty-six stoneformers were treated with thiazide to determine the effect of the drug. The average total calcium excretion was higher in patients than in controls and patients receiving therapy. The fraction of total calcium ionized was higher in patients than in normal subjects; thiazide therapy did not seem to cause a decrease in this fraction in stoneformers. The 24-hr excretion of ionic calcium was equal in normal subjects and in patients receiving thiazide therapy, but was higher in patients without therapy. Normal subjects have a significantly higher concentration and total excretion of magnesium and citrate. Apparent binding constants for calcium were significantly different for all three groups. The concentration of total binding sites was lower in patients with and without therapy than in normal subjects; this was partly attributed to the higher urinary concentration of citrate in normal subjects. However, citrate accounted for less than 20 per cent of the total bindings sites in urine. There was no significant differential 24-hr excretion of binding sites. Measurement of the specific excreting pattern of binding sites on proteins, peptides, nucleotides, and amino acid may emerge with a characteristic pattern in patients with urolithiasis.

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The relationship between calcium binding and ATPase activity has been investigated for guinea pig cardiac sarcolemma. The concentrations of calcium ions and of ATP were the main factors in determining the amount of calcium bound. With a saturating concentration of ATP, at low calcium concentrations (0.1 mM) more than 50% of the total calcium bound was ATP dependent while at high concentrations of calcium (10 mM) only 20% of the calcium bound was ATP dependent. The ATP-dependent calcium binding process involves one class of calcium binding sites while the non-ATP-dependent calcium binding process involves two classes of binding sites. Inhibitor studies of Ca2+-stimulated MgATPase, MgATPase, and CaATPase activities suggest Ca2+ and Mg2+ are either interacting with different sites on the same enzyme or with different enzymes.

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Sarcolemma isolated from guinea pig heart binds calcium in an ATP-dependent manner. Sodium ions decrease the total amount of calcium bound by the membranes. ATP-dependent calcium binding is more sensitive to sodium than the non-ATP-dependent calcium binding. The ATPase active during calcium binding is affected by sodium ions to the same extent as the ATP-dependent calcium binding process. The inhibition of the calcium binding process and of ATPase activity by sodium was more pronounced when the membranes were preincubated with sodium. The effect of sodium on calcium binding is dependent on both the time of contact between sodium and the membranes and the concentration of sodium. It is suggested that the effect of sodium on the calcium binding system in the sarcolemma may be a link between the inhibition of Na+K+-ATPase (EC 3.6.1.3) by cardiac glycosides and the subsequent increase in intracellular calcium.

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1. Differential scanning calorimetry has been used to study the thermal denaturation of lactate dehydrogenase. At pH 7.0 in 0.1 M potassium phosphate buffer, only one transition was observed. Both the enthalpy of denaturation and the melting temperature are linear function of heating rate. The enthalpy is 430 kcal/mol and the melting temperature 61 degrees C at 0 degrees C/min heating rate. The ratio of the calorimetric heat to the effective enthalpy indicated that the denaturation is highly cooperative. Subunit association does not appear to significantly contribute to the enthalpy of denaturation. 2. Both cofactor and sucrose addition stabilized the protein against thermal denaturation. Pyruvate addition produced no changes. Only a small time-dependent destabilization was observed at low concentrations of urea. Large effects were observed in concentrated NaCl solutions and with sulfhydryl-modified lactate dehydrogenase.

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Infrared absorption spectroscopy has been used to study the effect of organic solvents on the conformation of myoglobin, apomyoglobin, hemoglobin, lysozyme and ribonuclease. Beta structure can easily be induced by specific solvent effects. Films prepared from a 50% (v/v) mixture of alcohol, acetone, pyridine, tetrahydrofuran or dimethylsulfoxide/water mixtures show a high proportion of beta structure. The degree of induction of beta structure depends on the hydrocarbon content of the alcohol in the order methanol greater than ethanol greater than butanol. No beta structure was observed in films prepared from aqueous octanol solutions. Lyophilization tends to decrease secondary structure. The conformation of the proteins depends on the particular solvent system and the solvent composition. Solution studies of myoglobin in pure dimethylsulfoxide show that the conformation is a mixture of random and beta forms while in dimethylsulfoxide/2H2O mixtures the conformation is a mixture of alpha-helical and beta forms.

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