Emotion dysregulation, bullying, and suicide behaviors in adolescents.

Background: Among adolescents, rates for suicide attempts and non-suicidal self-injury (NSSI) remain high. Adolescents also often experience bullying, which has been found to associate with increased risk of suicide attempts and NSSI. Emotion regulation difficulties are associated with both bullying victimization and self-harm behaviors in adolescents. Aims: The current study examined the relationship between emotion dysregulation and suicide attempts and NSSI with bullying as a moderating factor. Method: High school students (n = 804) completed self-report measures on emotion regulation difficulties, suicide attempts, nonsuicidal self-injury, and past-year bullying experiences. Results: Moderation analyses found that the relationships between multiple emotion dysregulation dimensions and suicide attempts were significant regardless of bullying experience, but the relationship was stronger when bullying was present. For NSSI, bullying only moderated the relationships between nonacceptance of emotions, limited emotion regulation strategies and NSSI, with stronger associations when bullying was present. Limitations: Different types of bullying were not assessed, all measures were self-report, and the community sample had relatively low clinical severity. Conclusion: Results indicate that poor emotion regulation and bullying victimization associate with greater frequencies of suicidal and nonsuicidal behaviors. These findings point to the need for self-harm prevention programs to address both bullying and emotion regulation skills.

Fluoride removal in batch and column systems using bonechar produced in a top-lit updraft drum gasifier and furnace.

Hundreds of millions of people are exposed to excessive levels of fluoride in drinking water, predominately in low-resource communities. Activated alumina is recognized as the best available technology for fluoride removal from drinking water by the United States Environmental Protection Agency, but it has substantial economic and environmental costs. Bonechar is a more environmentally friendly and potentially lower cost alternative adsorbent. Here, fluoride adsorption from groundwater (pH 8.1 ± 0.2) by activated alumina was compared with bonechar primarily produced from bovine bones at peak heating temperatures between 400 and 1100 °C in a modular top-lit updraft drum (TLUD) stove (using a bone-wood mixture) and furnace. TLUD and furnace bonechar produced at peak heating temperatures 650-1000 °C and 400-800 °C, respectively, outperformed activated alumina in batch tests (i.e., required smaller doses to achieve 90% fluoride removal). The impact of using bovine versus swine bones to produce bonechar had a negligible impact on fluoride adsorption. A wide range of peak heating temperatures in the TLUD achieved by varying primary air flow rates and fuel selection (e.g., bone-to-wood mass ratios) produced efficient fluoride adsorbents. This finding demonstrates that a TLUD can be a robust, operationally flexible production system. Fluoride removal by TLUD and furnace bonechars showed strong, negative correlations (R2 ≥0.88) with organic matter content. Bonechar pilot column tests indicated that the mass transfer zone was captured (i.e., immediate fluoride breakthrough was not observed) at an empty bed contact time (EBCT) of 5 min, increasing EBCT to 30 min had a minimal impact on adsorption efficiency, and intermittent operation (3-10 d shut-off periods) decreased effluent fluoride concentrations. Furnace bonechars produced at peak heating temperatures 400-700 °C outperformed activated alumina in pilot columns. Differences in adsorption efficiencies in batch and column tests were associated with the linearity of fluoride adsorption. A theoretical model quantifying adsorption linearity with Freundlich 1/n values was able to predict adsorber performance solely based on batch test data.

Regulated disinfection byproduct formation over long residence times.

Design and operation of drinking water treatment plants and associated distribution systems with long residence times are complicated by the formation of regulated disinfection byproducts (DBPs), comprised of total trihalomethanes (TTHM) and five haloacetic acids (HAA5). Treated water dissolved organic carbon (DOC) concentrations, the unit processes required to meet those DOC concentrations, and disinfection strategies (e.g., booster chlorination) are the primary design and operational considerations that can require extensive testing or modeling to determine. In this study, twelve different treated drinking waters were generated at the bench-scale using ferric chloride coagulation and granular activated carbon adsorption from four parent raw waters collected from the San Juan River representing spring runoff, monsoon, and low flow events. Treated drinking waters with DOC concentrations of 0.9, 1.4, and 1.9 mg/L were tested for regulated DBP formation under simulated distribution system (SDS) conditions over residence times as long as 56 days and compared to 7-day formation potential (FP) testing. SDS free chlorine concentrations were maintained between 0.2 and 1.0 mg/L as Cl2 through periodic booster chlorination. Maximum SDS TTHM and HAA5 concentrations were 0.208 and 0.074 mg/L, respectively, with formation consistently varying by approximately ±20% across the four different parent raw waters despite having been treated to the same three DOC concentrations. An average of four existing TTHM models consistently underpredicted TTHM formation by approximatively 20%. Long considered a conservative measure of DBP formation, FP testing also underpredicted SDS DBP formation at 56 days by approximately 40% on average. The DBP testing approach presented in this study allowed for the development of several significant linear relationships for predicting DBP concentrations based on treated water ultraviolet light absorbance at 254 nm, water temperature, and cumulative free chlorine demand.

Pharmacodynamics of the Orotomides against Aspergillus fumigatus: New Opportunities for Treatment of Multidrug-Resistant Fungal Disease.

F901318 is an antifungal agent with a novel mechanism of action and potent activity against Aspergillus spp. An understanding of the pharmacodynamics (PD) of F901318 is required for selection of effective regimens for study in phase II and III clinical trials. Neutropenic murine and rabbit models of invasive pulmonary aspergillosis were used. The primary PD endpoint was serum galactomannan. The relationships between drug exposure and the impacts of dose fractionation on galactomannan, survival, and histopathology were determined. The results were benchmarked against a clinically relevant exposure of posaconazole. In the murine model, administration of a total daily dose of 24 mg/kg of body weight produced consistently better responses with increasingly fractionated regimens. The ratio of the minimum total plasma concentration/MIC (Cmin/MIC) was the PD index that best linked drug exposure with observed effect. An average Cmin (mg/liter) and Cmin/MIC of 0.3 and 9.1, respectively, resulted in antifungal effects equivalent to the effect of posaconazole at the upper boundary of its expected human exposures. This pattern was confirmed in a rabbit model, where Cmin and Cmin/MIC targets of 0.1 and 3.3, respectively, produced effects previously reported for expected human exposures of isavuconazole. These targets were independent of triazole susceptibility. The pattern of maximal effect evident with these drug exposure targets was also apparent when survival and histopathological clearance were used as study endpoints. F901318 exhibits time-dependent antifungal activity. The PD targets can now be used to select regimens for phase II and III clinical trials.IMPORTANCE Invasive fungal infections are common and often lethal. There are relatively few antifungal agents licensed for clinical use. Antifungal drug toxicity and the emergence of drug resistance make the treatment of these infections very challenging. F901318 is the first in a new class of antifungal agents called the orotomides. This class has a novel mechanism of action that involves the inhibition of the fungal enzyme dihydroorotate dehydrogenase. F901318 is being developed for clinical use. A deep understanding of the relationship between dosages, drug concentrations in the body, and the antifungal effect is fundamental to the identification of the regimens to administer to patients with invasive fungal infections. This study provides the necessary information to ensure that the right dose of F901318 is used the first time. Such an approach considerably reduces the risks in drug development programs and ensures that patients with few therapeutic options can receive potentially life-saving antifungal therapy at the earliest opportunity.

Disruption of gel phase lipid packing efficiency by sucralose studied with merocyanine 540.

Sucralose, an artificial sweetener, displays very different behavior towards membranes than its synthetic precursor sucrose. The impact of both sugars on model dipalmitoylphosphatidylcholine model membranes was investigated using absorbance and flourescence spectroscopy and the membrane probe merocyanine 540. This probe molecule is highly sensitive to changes in membrane packing, microviscosity and polarity. This work focuses on the impact of sugars on the outer leaflet of unilamellar dipalmitoyl phosphatidylcholine model membranes. The choice of lipid permits access to the gel phase at room temperature and incorporation of the dye after liposome formation allows us to examine the direct impact of the sugar on the outer leaflet while maximizing the response of the dye to changes in the bilayer. The results demonstrate that sucrose has no impact on the packing efficiency of lipids in unilamellar DPPC vesicles in the gel phase. Conversely sucralose decreases the packing efficiency of lipids in the gel phase and results in decreased microviscosity and increased membrane fluidity, which may be as a result of water disruption at the membrane water interface.

Evaluating Activated Carbon Adsorption of Dissolved Organic Matter and Micropollutants Using Fluorescence Spectroscopy.

Dissolved organic matter (DOM) negatively impacts granular activated carbon (GAC) adsorption of micropollutants and is a disinfection byproduct precursor. DOM from surface waters, wastewater effluent, and 1 kDa size fractions were adsorbed by GAC and characterized using fluorescence spectroscopy, UV-absorption, and size exclusion chromatography (SEC). Fluorescing DOM was preferentially adsorbed relative to UV-absorbing DOM. Humic-like fluorescence (peaks A and C) was selectively adsorbed relative to polyphenol-like fluorescence (peaks T and B) potentially due to size exclusion effects. In the surface waters and size fractions, peak C was preferentially removed relative to peak A, whereas the reverse was found in wastewater effluent, indicating that humic-like fluorescence is associated with different compounds depending on DOM source. Based on specific UV-absorption (SUVA), aromatic DOM was preferentially adsorbed. The fluorescence index (FI), if interpreted as an indicator of aromaticity, indicated the opposite but exhibited a strong relationship with average molecular weight, suggesting that FI might be a better indicator of DOM size than aromaticity. The influence of DOM intermolecular interactions on adsorption were minimal based on SEC analysis. Fluorescence parameters captured the impact of DOM size on the fouling of 2-methylisoborneol and warfarin adsorption and correlated with direct competition and pore blockage indicators.

Distinct membrane properties are differentially influenced by cardiolipin content and acyl chain composition in biomimetic membranes.

Cardiolipin (CL) has a critical role in maintaining mitochondrial inner membrane structure. In several conditions such as heart failure and aging, there is loss of CL content and remodeling of CL acyl chains, which are hypothesized to impair mitochondrial inner membrane biophysical organization. Therefore, this study discriminated how CL content and acyl chain composition influenced select properties of simple and complex mitochondrial mimicking model membranes. We focused on monolayer excess area/molecule (a measure of lipid miscibility), bilayer phase transitions, and microdomain organization. In monolayer compression studies, loss of tetralinoleoyl [(18:2)4] CL content decreased the excess area/molecule. Replacement of (18:2)4CL acyl chains with tetraoleoyl [(18:1)4] CL or tetradocosahexaenoyl [(22:6)4] CL generally had little influence on monolayer excess area/molecule; in contrast, replacement of (18:2)4CL acyl chains with tetramyristoyl [(14:0)4] CL increased monolayer excess area/molecule. In bilayers, calorimetric studies showed that substitution of (18:2)4CL with (18:1)4CL or (22:6)4CL lowered the phase transition temperature of phosphatidylcholine vesicles whereas (14:0)4CL had no effect. Finally, quantitative imaging of giant unilamellar vesicles revealed differential effects of CL content and acyl chain composition on microdomain organization, visualized with the fluorescent probe Texas Red DHPE. Notably, microdomain areas were decreased by differing magnitudes upon lowering of (18:2)4CL content and substitution of (18:2)4CL with (14:0)4CL or (22:6)4CL. Conversely, exchanging (18:2)4CL with (18:1)4CL increased microdomain area. Altogether, these data demonstrate that CL content and fatty acyl composition differentially target membrane physical properties, which has implications for understanding how CL regulates mitochondrial activity and the design of CL-specific therapeutics.

F901318 represents a novel class of antifungal drug that inhibits dihydroorotate dehydrogenase.

There is an important medical need for new antifungal agents with novel mechanisms of action to treat the increasing number of patients with life-threatening systemic fungal disease and to overcome the growing problem of resistance to current therapies. F901318, the leading representative of a novel class of drug, the orotomides, is an antifungal drug in clinical development that demonstrates excellent potency against a broad range of dimorphic and filamentous fungi. In vitro susceptibility testing of F901318 against more than 100 strains from the four main pathogenic Aspergillus spp. revealed minimal inhibitory concentrations of ≤0.06 µg/mL-greater potency than the leading antifungal classes. An investigation into the mechanism of action of F901318 found that it acts via inhibition of the pyrimidine biosynthesis enzyme dihydroorotate dehydrogenase (DHODH) in a fungal-specific manner. Homology modeling of Aspergillus fumigatus DHODH has identified a predicted binding mode of the inhibitor and important interacting amino acid residues. In a murine pulmonary model of aspergillosis, F901318 displays in vivo efficacy against a strain of A. fumigatus sensitive to the azole class of antifungals and a strain displaying an azole-resistant phenotype. F901318 is currently in late Phase 1 clinical trials, offering hope that the antifungal armamentarium can be expanded to include a class of agent with a mechanism of action distinct from currently marketed antifungals.

Effect of DOM Size on Organic Micropollutant Adsorption by GAC.

Granular activated carbon (GAC) adsorption of the micropollutants 2-methylisoborneol (MIB) and warfarin (WFN) at ng/L levels was investigated in five waters with isolated natural dissolved organic matter (DOM) held at a constant dissolved organic carbon concentration. Each water was evaluated for competitive adsorption effects based on the pretreatment of ultrafiltration, coagulation, and additional background micropollutants. Using the breakthrough with unfractionated DOM as a baseline, on average, the water with lower molecular weight (MW) DOM decreased MIB and WFN adsorption capacity by 59%, whereas the water with higher MW DOM increased MIB and WFN adsorption capacity by 64%. All waters showed similar decreasing MIB and WFN adsorption capacity with increasing empty bed contact time (EBCT), with more dramatic effects seen for the more strongly adsorbing WFN. On average, MIB and WFN adsorption kinetics were two times slower in the water with higher MW DOM compared to the water with lower MW DOM, as described by the intraparticle pore diffusion tortuosity. Increased adsorption competition from 27 micropollutants other than MIB and WFN at environmentally relevant concentrations had little to no effect on MIB and WFN breakthrough behavior. Any competitive effect from background micropollutants became indiscernible at longer EBCTs.

Full- and pilot-scale GAC adsorption of organic micropollutants.

Granular activated carbon (GAC) adsorption of 30 environmentally relevant micropollutants (MP) from four surface waters was investigated at the pilot-scale with empty bed contact times (EBCTs) of 7 and 15 min. An increase in background dissolved organic matter resulted in more and earlier MP breakthrough. Compared to an EBCT of 7 min, MP breakthrough at an EBCT of 15 min demonstrated 52% later breakthrough on average for five MPs on a throughput basis. A regression model was developed with data from three waters to predict MP throughput in bed volumes to 10% breakthrough (BV10%) based on the influent dissolved organic carbon concentration and the MP pH-dependent octanol-water partition coefficient, polarizability, and molecular volume. The regression model over predicted full-scale BV10% values when applied to a wastewater-water impacted water source and to GAC with a larger particle diameter, for which a particle size adjustment was able to account for most of the difference.

Discrimination of the hard keratins animal horn and chelonian shell using attenuated total reflection-infrared spectroscopy.

The ability to discriminate between objects manufactured from animal horn and chelonian (turtle, tortoise, or terrapin) shell is important from a cultural and archeological perspective such that it may allow conservators to determine the appropriate treatment and long-term care solution. It would also aid curators in identifying and cataloging items manufactured from these materials. Discrimination and classification is also a valuable tool for those involved in tracking the illegal trade in restricted materials of this nature. Attenuated total reflection infrared (ATR-IR) spectroscopy, using a single reflection diamond internal reflection element (IRE), coupled with discrimination analysis was used to analyze a total of thirty-nine samples (29 calibration samples, 10 validation samples). A discrimination analysis model was constructed using Mahalanobis distances to classify spectra into one of two classes. The model was then subsequently used to successfully classify all validation samples and correctly identify them as animal horn or chelonian shell based on second-derivative spectra of the amide I and II regions. This technique requires minimal to no sample preparation and may be used to nondestructively identify very small samples successfully without performing detailed secondary structural curve-fitting routines. This model should be a valuable resource to museums, conservators, and wildlife management programs for rapidly and reliably discriminating between animal horn and chelonian shell.

Determining the effect of the incorporation of unnatural amino acids into antimicrobial peptides on the interactions with zwitterionic and anionic membrane model systems.

Circular Dichroism (CD), isothermal calorimetry (ITC) and calcein fluorescence leakage experiments were conducted to provide insight into the mechanisms of binding of a series of antimicrobial peptides containing unnatural amino acids (Ac-XF-Tic-Oic-XK-Tic-Oic-XF-Tic-Oic-XK-Tic-KKKK-CONH(2)) to zwitterionic and anionic micelles, SUVs and LUVs; where X (Spacer# 1) is either Gly, ß-Ala, Gaba or 6-aminohexanoic acid. It is the intent of this investigation to correlate these interactions with the observed potency and selectivity against several different strains of bacteria. The CD spectra of these compounds in the presence of zwitterionic DPC micelles and anionic SDS micelles are very different indicating that these compounds adopt different conformations on binding to the surface of anionic and zwitterionic membrane models. These compounds also exhibited very different CD spectra in the presence of zwitterionic POPC and anionic mixed 4:1 POPC/POPG SUVs and LUVs, indicating the formation of different conformations on interaction with the two membrane types. This observation is also supported by ITC and calcein leakage data. ITC data suggested these peptides interact primarily with the surface of zwitterionic LUVs and was further supported by fluorescence experiments where the interactions do not appear to be concentration dependent. In the presence of anionic membranes, the interactions appear more complex and the calorimetric and fluorescence data both imply pore formation is dependent on peptide concentration. Furthermore, evidence suggests that as the length of Spacer# 1 increases the mechanism of pore formation also changes. Based on the observed differences in the mechanisms of interactions with zwitterionic and anionic LUVs these AMPs are potential candidates for further drug development.

Mental health care on the streets: An integrated approach.

BACKGROUND: Mental illness can be both a cause of and a reaction to being homeless. When homelessness co-exists with mental illness, the provision of care for very vulnerable people is significantly complicated. Our initiative built on a model of assertive outreach and embedded mental health staff into the daily operations of Hanover Welfare Services and Sacred Heart Mission welfare services in inner Melbourne. The initiative's aim was to facilitate closer collaboration between mental health and welfare services and develop staff capacity to better identify and support people living homeless with a mental illness. METHOD: The project involved studying the impact of our assertive outreach model on consumer and service outcomes. Demographic, clinical and service usage details for consumers engaged by the initiative were recorded. Changes to the rate of admission of people from both welfare services to The Alfred Inpatient Psychiatry Unit and requests for support from The Alfred Crisis Assessment and Treatment Service were also recorded. RESULTS: People engaged by this initiative had high levels of previous emergency medical or psychiatric service usage, but relatively low levels of current community mental health engagement. There were also high levels (almost 52%) of comorbid substance misuse. The initiative was, however, able to engage more people in ongoing community mental health care, which particularly when provided in collaboration between mental health and welfare staff, achieved improvements in accommodation stability. The initiative also resulted in improved identification and prevention of mental illness crises through supporting a more rapid onsite mental health response. CONCLUSIONS: Embedding mental health staff into the daily operations of two welfare services in inner Melbourne improved inter-service collaboration and the identification and care for people living homeless with a mental illness.

Spectroscopic and thermodynamic evidence for antimicrobial peptide membrane selectivity.

In our laboratory we developed a series of antimicrobial peptides that exhibit selectivity and potency for prokaryotic over eukaryotic cells (Hicks et al., 2007). Circular dichroism (CD), isothermal calorimetry (ITC) and calcein leakage assays were conducted to determine the mechanism of lipid binding of a representative peptide 1 (Ac-GF-Tic-Oic-GK-Tic-Oic-GF-Tic-Oic-GK-Tic-KKKK-CONH(2)) to model membranes. POPC liposomes were used as a simple model for eukaryotic membranes and 4:1 POPC:POPG liposomes were used as a simple model for prokaryotic membranes. CD, ITC and calcein leakage data clearly indicate that compound 1 interacts via very different mechanisms with the two different liposome membranes. Compound 1 exhibits weaker binding and induces less calcein leakage in POPC liposomes than POPC:POPG (4:1 mole ratio) liposomes. The predominant binding mechanism to POPC appears to be limited to surface interactions while the mechanism of binding to 4:1 POPC:POPG most likely involves some type of pore formation.

Pharmacodynamic and efficacy profile of TGN 255, a novel direct thrombin inhibitor, in canine cardiopulmonary bypass and simulated mitral valve repair.

Heparin-induced thrombocytopenia can be a life-threatening sequel to conventional use of unfractionated heparin in cardiopulmonary bypass (CPB). This study evaluated the pharmacokinetic/pharmacodynamic (PK/PD) and efficacy profile of a novel direct thrombin inhibitor, TGN 255, during cardiac surgery in dogs. Point-of-care coagulation monitoring was also compared against the plasma concentrations of TRI 50c, the active metabolite of TGN 255. The study was conducted in three phases using 10 animals: phase 1 was a dose-ranging study in conscious animals (n = 6), phase 2 was a similar but terminal dose-ranging study in dogs undergoing CPB (n = 6), and phase 3 was with animals undergoing simulated mitral valve repair (terminal) using optimal TGN 255 dose regimens derived from phases I and II (n = 4). During the study, PD markers and drug plasma levels were determined. In addition, determinations of hematologic markers and blood loss were undertaken. Phase 1 studies showed that a high-dose regimen of a 5-mg/kg bolus and infusion of 20 mg/kg/h elevated PD markers in conscious animals, at which time there were no measured effects on platelet or red blood cell counts, and the mean plasma concentration of TRI 50C was 20.6 microg/mL. In the phase 2 CPB dose-ranging study, this dosing regimen significantly elevated all the PD markers and produced hemorrhagic and paradoxical thrombogenic effects. In the phase 3 surgical study, a lower TGN 255 dose regimen of a 2.5-mg/kg bolus plus 10 mg/kg/h produced anticoagulation, elevated PD markers, and produced minimal post-operative blood loss in the animals. Plasma levels of TRI 50C trended well with the conventional point-of-care coagulation monitoring. TGN 255 provided effective anticoagulation in a canine CPB procedure, enabling successful completion with minimal blood loss. These findings support further evaluation of TGN 255 as an anticoagulant for CPB.

Physical and thermal properties of blood storage bags: implications for shipping frozen components on dry ice.

BACKGROUND: Frozen blood components are shipped on dry ice. The lower temperature (-70 degrees C in contrast to usual storage at -30 degrees C) and shipping conditions may cause a rent in the storage bag, breaking sterility and rendering the unit useless. The rate of loss can reach 50 to 80 percent. To identify those bags with lower probability of breaking during shipment, the thermal and physical properties of blood storage bags were examined. STUDY DESIGN AND METHODS: Blood storage bags were obtained from several manufacturers and were of the following compositions: PVC with citrate, di-2-ethylhexylphthalate (DEHP), or tri-2-ethylhexyl-tri-mellitate (TEHTM) plasticizer; polyolefin (PO); poly(ethylene-co-vinyl acetate) (EVA); or fluorinated polyethylene propylene (FEP). The glass transition temperature (Tg) of each storage bag was determined. Bag thickness and measures of material strength (tensile modulus [MT] and time to achieve 0.5 percent strain [T0.5%]) were evaluated. M(T) and T0.5% measurements were made at 25 and -70 degrees C. Response to applied force at -70 degrees C was measured using an impact testing device and a drop test. RESULTS: The Tg of the bags fell into two groups: 70 to 105 degrees C (PO, FEP) and -50 to -17 degrees C (PVC with plasticizer, EVA). Bag thickness ranged from 0.14 to 0.41 mm. Compared to other materials, the ratios of M(T) and T0.5% for PVC bags were increased (p < or = 0.001) indicating that structural changes for PVC were more pronounced upon cooling from 25 to -70 degrees C. Bags containing EVA were more shock resistant, resulting in the lowest rate of breakage (10% breakage) when compared with PO (60% breakage, p = 0.0573) or PVC (100% breakage, p = 0.0001). CONCLUSIONS: Blood storage bags made of EVA appear better suited for shipping frozen blood components on dry ice and are cost-effective replacements for PVC bags. For the identification of blood storage bags meeting specific storage requirements, physical and thermal analyses of blood storage bags may be useful and remove empiricism from the process.

Characterization of the main phase transition in 1,2-dipalmitoyl-phosphatidylcholine luvs by 1H NMR.

The main phase transition (Tm) of 100 nm large unilamellar vesicles (LUVs) of 1,2-dipalmitoylphosphatidylcholine (DPPC) was investigated using 1H NMR (proton magnetic resonance) in deuterium oxide, and both DSC (differential scanning calorimetry) and IR (infrared) spectroscopy in water and deuterium oxide. The ability of 1H NMR to determine Tm was demonstrated and the values obtained were in general agreement with those observed with DSC and IR. However, the temperature range of the transition observed by NMR was significantly broader than that observed with either DSC or IR. The effect of deuterium oxide on Tm was studied by comparing results obtained in water and deuterium oxide with DSC and IR. The results showed no significant difference in Tm or temperature range of transition determined in these solvents.