Exploratory Multisite MR Spectroscopic Imaging Shows White Matter Neuroaxonal Loss Associated with Complications of Type 1 Diabetes in Children.

BACKGROUND AND PURPOSE: Type 1 diabetes affects over 200,000 children in the United States and is associated with an increased risk of cognitive dysfunction. Prior single-site, single-voxel MRS case reports and studies have identified associations between reduced NAA/Cr, a marker of neuroaxonal loss, and type 1 diabetes. However, NAA/Cr differences among children with various disease complications or across different brain tissues remain unclear. To better understand this phenomenon and the role of MRS in characterizing it, we conducted a multisite pilot study. MATERIALS AND METHODS: In 25 children, 6-14 years of age, with type 1 diabetes across 3 sites, we acquired T1WI and axial 2D MRSI along with phantom studies to calibrate scanner effects. We quantified tissue-weighted NAA/Cr in WM and deep GM and modeled them against study covariates. RESULTS: We found that MRSI differentiated WM and deep GM by NAA/Cr on the individual level. On the population level, we found significant negative associations of WM NAA/Cr with chronic hyperglycemia quantified by hemoglobin A1c (P < .005) and a history of diabetic ketoacidosis at disease onset (P < .05). We found a statistical interaction (P < .05) between A1c and ketoacidosis, suggesting that neuroaxonal loss from ketoacidosis may outweigh that from poor glucose control. These associations were not present in deep GM. CONCLUSIONS: Our pilot study suggests that MRSI differentiates GM and WM by NAA/Cr in this population, disease complications may lead to neuroaxonal loss in WM in children, and deeper investigation is warranted to further untangle how diabetic ketoacidosis and chronic hyperglycemia affect brain health and cognition in type 1 diabetes.

Pharmacognostic studies on Neuracanthus sphaerostachyus Dalz. (Acanthaceae) leaves.

BACKGROUND: Neuracanthus sphaerostachyus Dalz. species shows the spherically arranged flowering head. Mostly flowers are observed in purple colour. It is an annual herb with 30-60 cm high. Flowers arranged in rounded clusters without a stalk. Leaves are 5-10 cm in length, and are almost without a stalk. N.sphaerostachyus has been traditionally used to treat skin diseases, cough and asthma. OBJECTIVE: The present study was undertaken to study the Pharmacognostic parameters for the rapid identification and authentication of the plant. MATERIALS AND METHODS: The macroscopic and microscopic characteristics with intensive quantitative microscopy of N.sphaerostachyus Dalz leaves were done using different chemicals and reagents. RESULTS: The plant leaves show single layered, wavy walled cells in upper epidermis. Powder study of leaves shows light greenish colour with multicellular covering trichomes and sessile glandular trichomes with fibres passing through parenchymatous cells. CONCLUSION: The macroscopic and microscopic characteristics of N.sphaerostachyus Dalz leaves serves as a tool for low cost, rapid identification and authentication of this plant.

Tooth wear risk assessment and care-planning in general dental practice.

Objective To assess charting, risk assessment and treatment-planning of tooth wear between recently qualified and experienced dentists in general dental practice.Design Service evaluation.Setting Multi-setting evaluation of three mixed NHS/Private general dental practices in North-East London.Methods The clinical notes of new patient examinations on dentate adults presenting from the 1 October 2016 to 31 December 2016 were audited collecting data on tooth wear charting, risk assessment and treatment planning. Data were analysed using descriptives, chi square and logistic regressions in SPSS. Significance was inferred at p <0.05.Results Foundation dentists and experienced dentists performed 85 and 200 new patient examinations, respectively, during the evaluation period. Tooth wear was charted for 48% of those attending foundation dentists and 5% of those attending experienced dentists. Diet was assessed in 50.6% of patients examined by foundation dentists and 1.0% of patients examined by experienced dentists. Foundation dentists were more likely to chart tooth wear, risk assess and preventively manage tooth wear compared to experienced dentists (p <0.001).Conclusion This service evaluation highlights that improvements are required in recording, risk assessing and preventive treatment planning of erosive tooth wear. Experienced dentists were less likely to risk assess tooth wear and less likely to provide preventive treatment. Experienced GDPs may benefit from re-training in this area.

Development of a selective androgen receptor modulator for transdermal use in hypogonadal patients.

We have identified a non-steroidal selective androgen receptor modulator (SARM), termed LY305, that is bioavailable through a transdermal route of administration while highly cleared via hepatic metabolism to limit parent compound exposure in the liver. Selection of this compound and its transdermal formulation was based on the optimization of skin absorption properties using both in vitro and in vivo skin models that supported PBPK modeling for human PK predictions. This molecule is an agonist in perineal muscle while being a weak partial agonist in the androgenic tissues such as prostate. When LY305 was tested in animal models of skeletal atrophy it restored the skeletal muscle mass through accelerated repair. In a bone fracture model, LY305 remained osteoprotective in the regenerating tissue and void of deleterious effects. Finally, in a small cohort of healthy volunteers, we assessed the safety and tolerability of LY305 when administered transdermally. LY305 showed a dose-dependent increase in serum exposure and was well tolerated with minimal adverse effects. Notably, there were no statistically significant changes to hematocrit or HDL after 4-week treatment period. Collectively, LY305 represents a first of its kind de novo development of a non-steroidal transdermal SARM with unique properties which could find clinical utility in hypogonadal men.

Brain uptake of S-(1,2-dichlorovinyl)glutathione and S-(1,2-dichlorovinyl)-L-cysteine, the glutathione and cysteine S-conjugates of the neurotoxin dichloroacetylene.

Dichloroacetylene causes trigeminal neuropathy in humans and animals. Glutathione conjugation of dichloroacetylene affords S-(1,2-dichlorovinyl)glutathione (DCVG), which is hydrolyzed to S-(1,2-dichlorovinyl)-L-cysteine (DCVC). This study was undertaken to test the hypothesis that the neurotoxicity of dichloroacetylene may be associated with glutathione S-conjugate formation and brain uptake and bioactivation of the dichloroacetylene-derived S-conjugates. With the Oldendorf technique, the Brain Uptake Index for [35S]DCVC and [35S]DCVG was determined and compared with the uptake of [35S]methionine and [14C]sucrose. Brain uptake of DCVC exceeded uptake of methionine and DCVG uptake was comparable to methionine uptake. Both [35S]DCVC and [35S]DCVG were recovered intact in brain tissue. The uptake of the 35S-labeled S-conjugates was inhibited by unlabeled DCVC and DCVG in a concentration-dependent manner. The data indicated that DCVC, but not DCVG, was transported by the sodium-independent system-L transporter for neutral amino acids. In vitro studies revealed that DCVG can be hydrolyzed to DCVC by brain tissue in a concentration-dependent manner.

Rubidium (86Rb+) influx into dorsal root ganglia and sciatic nerve endoneurium of control and streptozotocin-diabetic rats: comparison with enzymatic Na,K-ATPase activity.

Streptozotocin (STZ)-induced diabetes in the rat causes a significant reduction in ouabain-sensitive Na,K-ATPase pumping activity measured by 86Rb+ influx, in sciatic endoneurium (by 54%) and dorsal root ganglia (by 22%). For endoneurium, the change is similar to that of ouabain-sensitive enzymatic Na,K-ATPase activity (42%), but in dorsal root ganglia, the decrease in enzymatic Na,K-ATPase activity was much greater. 86Rb+ efflux from dorsal root ganglia showed no difference between diabetic and control animals, confirming that the abnormal 86Rb+ influx reflects Na,K-ATPase function and not abnormal membrane permeability. The significance of these findings to pathogenetic mechanisms in diabetic neuropathy is discussed.

Protein synthesis in cells isolated from the developing rat cerebellum.

The rate of protein synthesis was estimated in structurally preserved perikaryal preparations from 8-day-old rat cerebellum under conditions which overcome the problems of intracellular compartmentation. The rates were lower than the in vivo estimates at comparable ages, but they were of similar magnitude, and very much higher than previous estimates on isolated cells. Protein synthesis rate depended on the cell type. When expressed per cell the rank order in the preparations enriched in the indicated classes of cells was: Purkinje cells > astrocytes > granule cells in the S, G2 and M phase of the cell cycle > granule cells in G1 and G0. However, after normalizing the results for size differences between cell types, by expressing the rates in terms of unit protein or as a percentage replacement of the protein bound amino acid, astrocytes and replicating granule cells displayed greater rates than the Purkinje cells. The resolution of labelled proteins using SDS-PAGE indicated marked differences in the rate of synthesis of particular proteins. The results were consistent with the view that certain polypeptides are uniquely expressed in particular cell classes.

Polypeptide and glycoprotein abnormalities in dorsal root ganglia of streptozotocin-diabetic rats.

Observations were made on the polypeptide and glycoprotein composition of dorsal root ganglia from streptozotocin-induced diabetic rats by sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE). Silver staining of one-dimensional gels failed to demonstrate any differences between diabetic and control animals. In two-dimensional studies, good resolution of polypeptides with a mass greater than 70 kDa was not obtained, but a number of important abnormalities in the polypeptide composition of diabetic ganglia were detected. Some polypeptides recognized in the gels from control ganglia were present in high concentrations in diabetic ganglia; other polypeptides, particularly a number of basic polypeptides of low molecular mass, were only identified in the diabetic rats. Three major polypeptides showed a small shift in their isoelectric point in the diabetic animals. The glycoprotein content of the ganglia was examined by lectin binding to both one- and two-dimensional gel separations. An increase in total glycoprotein content was evident in the diabetic ganglia. A number of polypeptides with a molecular mass between 70 and 110 kDa showed heavy glycosylation. Altered glycosylation of some specific polypeptides of lower molecular mass was also seen, 7 of these showing increased and 3 reduced glycosylation. The significance of these findings is discussed.

Glucose and leucine uptake by rat dorsal root ganglia is not insulin sensitive.

Dorsal root ganglia from rats were incubated with 3-O-methyl-[14C]glucose, and [3H]leucine in the presence or absence of insulin in order to determine whether insulin influences the uptake of glucose and amino acids by the cells of the ganglion. No effect was detected. A significant proportion (38%) of the uptake of [3H]leucine was shown to be inhibited by ouabain and therefore energy dependent, utilizing Na+K(+)-ATPase. The activity of this enzyme is known to be impaired in dorsal root ganglia in diabetic rats, as is the uptake of amino acids; these phenomena are therefore unlikely to be due to a direct effect of reduced circulating insulin levels. The relevance of these findings to theories as to the causation of diabetic neuropathy is discussed.

Induced moderate hypotensive anesthesia for spinal fusion and Harrington-rod instrumentation.

The effect of moderate hypotensive anesthesia on blood loss, need for transfusion, and length of surgery of forty-nine patients who underwent posterior spinal fusion and Harrington-rod instrumentation was compared retrospectively. Twenty-seven patients were given enflurane as the main anesthetic agent, with fentanyl supplementation, and their blood pressure was maintained at twenty to thirty millimeters of mercury less than the preoperative systolic blood pressure. These patients were compared with twenty-two patients who had been anesthetized with nitrous oxide, oxygen, and narcotic technique and were normotensive throughout the duration of the anesthesia. The results were analyzed by the unpaired Student t test. Moderate hypotensive anesthesia was found to significantly decrease the average blood loss by nearly 40 per cent, reduce the need for transfusion by nearly 45 per cent, and shorten the average operating time by nearly 10 per cent. No complications attributable to the anesthetic technique occurred. The findings of this study suggest that moderate hypotensive anesthesia with enflurane and fentanyl supplementation may be of benefit in scoliosis surgery by reducing blood loss, the need for blood replacement, and operating time.

Paired-ion high-performance liquid chromatographic assay for sulfinpyrazone in plasma.

A specific and sensitive liquid chromatographic method is reported for the assay of sulfinpyrazone in plasma utilizing ion pairing between the tetrabutylammonium cation and the sulfinpyrazone anion. The method is rapid in that conventional extraction procedures are avoided in favor of using disposable cartridges packed with an octade-cylsilane bonded phase as a means of separating the drug from plasma. The samples were chromatographed on a C18 reversed-phase column using a mobile phase consisting of 0.005 M tetrabutylammonium phosphate in methanol-water (56:44). The coefficient of variation obtained was 4.5% and the response was linear over a range of 0.2-80 micrograms/ml.

Arthropathy and HIV infection. A muddle of mimicry.

Several rheumatic diseases are associated with human immunodeficiency virus (HIV) infection. The most common are reactive and psoriatic arthritis. Classic septic arthritis caused by Staphylococcus aureus and other common organisms is very rare: Instead, infectious arthritis caused by unusual organisms is the rule. Some of the HIV-related rheumatic syndromes behave like classic rheumatic diseases, while others may actually be new forms of disease. Often, one of the rheumatic syndromes is the presenting manifestation of underlying HIV infection. HIV-infected patients and patients with rheumatic disease often have similar laboratory abnormalities. Systemic lupus erythematosus, in particular, may be mistaken for HIV infection, in part because of cross-reactivity of antibodies. However, coexistence of systemic lupus erythematosus and rheumatoid arthritis with HIV infection is a rare occurrence. Traditional therapy for rheumatic diseases may not be indicated in HIV-infected patients and in fact may even be contraindicated.

A clofarabine-based bridging regimen in patients with relapsed ALL and persistent minimal residual disease (MRD).

In patients with relapsed ALL, minimal residual disease (MRD) identified prior to allogeneic hematopoietic cell transplantation (HCT) is a strong predictor of relapse. We report our experience using a combination of reduced-dosing clofarabine, CY and etoposide as a 'bridge' to HCT in eight patients with high risk or relapsed ALL and pre-HCT MRD. All patients had detectable MRD (>0.01%, flow cytometry) at the start of therapy with all eight achieving MRD reduction following one cycle. The regimen was well tolerated with seven grade 3/4 toxicities occurring among four of the eight patients. Five patients (62.5%) are alive, one died from relapse (12.5%) and two from transplant-related mortality (25%). The combination of reduced-dose clofarabine, CY and etoposide as bridging therapy appears to be well tolerated in patients with relapsed ALL and is effective in reducing pre-HCT MRD.

RP-HPLC Method for the Estimation of Dutasteride in Tablet Dosage Form.

A simple, sensitive and precise RP-HPLC method was developed for the determination of dutasteride in tablet dosage form. The RP-HPLC separation was achieved on phenomenex C(18) column (250 mm, id 4.6 mm, 5 mum) using mobile phase methanol:water (90:10 v/v) at a flow rate of 1 ml/min at an ambient temperature. Quantification was achieved with photodiode array detection at 235 nm over the concentration range 1-12 mug/ml. The method was validated statistically and was applied successfully for the determination of dutasteride in tablets.

Simultaneous RP-HPLC and HPTLC Estimation of Fluoxetine Hydrochloride and Olanzapine in Tablet Dosage Forms.

A binary mixture of fluoxetine HCl and olanzapine was determined by two different methods. The first method involved determination of fluoxetine HCl and olanzapine using reversed-phase liquid chromatography using acetonitrile:methanol:0.032 M ammonium acetate buffer (45:05:50, v/v/v) as the mobile phase at a flow rate of 1.5 ml/min. Quantitation was achieved with ultraviolet detection at 235 nm over concentration ranges of 0.2-4 and 0.1-2 mug/ml; mean accuracies were 101.16+/-0.59 and 99.79+/-0.56% for fluoxetine HCL and olanzapine, respectively. The second method was based on the high performance thin layer chromatography separation of the two drugs followed by densitometric measurements of their spots at 235 nm. The separation was carried out on Merck TLC aluminium sheets of silica gel 60 F254 using acetone:methanol:triethyleamine (5:3:0.5, v/v/v), as mobile phase. The linearity was found to be in the range of 300-1000 and 150-500 ng/spot; mean accuracies were 100.95+/-0.52 and 99.31+/-0.51% for fluoxetine HCl and olanzapine, respectively. The method was successively applied to tablets because no chromatographic interferences from the tablet excipients were found. The methods retained their accuracy and precision when the standard addition technique was applied. The results obtained by applying the proposed methods were statistically analyzed.

Spectrophotometric and chromatographic simultaneous estimation of amitriptyline hydrochloride and chlordiazepoxide in tablet dosage forms.

A binary mixture of amitriptyline HCl and chlordiazepoxide was determined by three different methods. The first method involved determination of amitriptyline HCl and chlordiazepoxide using the first derivative spectrophotometric technique at 219 and 230 nm over the concentration ranges of 1-20 and 2-24 mug/ml with mean accuracies 100.9+/-0.87 and 99.2+/-1.0%, respectively. The second method was reversed-phase high performance liquid chromatography using methanol: acetonitrile: 0.065 M ammonium acetate buffer (50:20:30, v/v/v), final pH adjust to 5.5 +/- 0.02 with ortho phosphoric acid as the mobile phase and was pumped at a flow rate of 1.0 ml/min. Quantification was achieved with ultraviolet detection at 240 nm over concentration ranges of 0.25-4 and 0.1-1.6 mug/ml; mean accuracies were 100.55+/-0.62 and 100.71+/-0.81%, respectively. The third method utilized high performance thin layer chromatography method in tablet dosage form. The method was based on separation of the two drugs followed by densitometric measurements of their spots at 240 nm. The separation was carried out on Merck thin layer chromatographic aluminium sheets of silica gel 60 F254 using carbon tetrachloride: acetone: triethylamine (6:3:0.2, v/v/v) as mobile phase. The linearity was found to be in the range of 50-600 and 20-240 ng/spot for amitriptyline hydrochloride and chlordiazepoxide, respectively. The methods were successively applied to pharmaceutical formulation because no chromatographic interferences from the tablet excipients were found. The suitability of these methods for the quantitative determination of the compounds was proved by validation.

Simultaneous RP-HPLC Estimation of Trifluoperazine Hydrochloride and Chlordiazepoxide in Tablet Dosage Forms.

A binary mixture of trifluoperazine HCl and chlordiazepoxide was determined using reversed-phase liquid chromatography method using methanol:water (97:03, v/v) pumped at a flow rate of 1.0 ml/min. Quantification was achieved with ultraviolet detection at 262 nm over concentration ranges of 0.1-1 and 0.5-5 mug/ml; mean accuracies were 101.05+/-0.47 and 98.97+/-0.33 %, respectively. The method was successively applied to tablet dosage forms as no chromatographic interferences from the tablet excipients were observed. The method retained its accuracy and precision when the standard addition technique was applied.

Simultaneous spectrophotometric estimation of imipramine hydrochloride and chlordiazepoxide in tablets.

A binary mixture of imipramine HCl and chlordiazepoxide was determined by three different spectrophotometric methods. The first method involved determination of imipramine HCl and chlordiazepoxide using the simultaneous equations and the second method involved absorbance ratio method. Imipramine has absorbance maxima at 251 nm, chlordiazepoxide has absorbance maxima at 264.5 nm and isoabsorptive point is at 220 nm in methanol. Linearity was obtained in the concentration ranges of 1-25 and 1-10 mug/ml for Imipramine HCL and Chlordiazepoxide, respectively. The third method involved determination of these two drugs using the first-derivative spectrophotometric technique at 219 and 231.5 nm over the concentration ranges of 1-20 and 2-24 mug/ml with mean accuracies 99.46+/-0.78 and 101.43+/-1.20%, respectively. These methods were successively applied to pharmaceutical formulations because no interferences from the tablet excipients were found. The suitability of these methods for the quantitative determination of the compounds was proved by validation.