Hypopigmentation from UV resin additive

An employee formulating a UV-resin based vehicle developed hypopigmentation limited to the forearms without preceding overt dermatitis. Manual procedures in an automatic process provided opportunity for exposure and contact to various components. A non-resin additive, quinone, alone or in conjunction with a likely contaminant, hydroquinone, was the suspected agent. Patch tests to quinone caused severe irritation. Reactions to acrylic components were not felt to be directly pertinent.     

Effects of enhancing mitochondrial oxidative phosphorylation with reducing equivalents and ubiquinone on 1-methyl-4-phenylpyridinium toxicity and complex I-IV damage in neuroblastoma cells

The effects of increasing mitochondrial oxidative phosphorylation (OXPHOS), by enhancing electron transport chain components, were evaluated on 1-methyl-4-phenylpyridinium (MPP+) toxicity in brain neuroblastoma cells. Although glucose is a direct energy source, ultimately nicotinamide and flavin reducing equivalents fuel ATP produced through OXPHOS. The findings indicate that cell respiration/mitochondrial O(2) consumption (MOC) (in cells not treated with MPP+) is not controlled by the supply of glucose, coenzyme Q(10) (Co-Q(10)), NADH+, NAD or nicotinic acid. In contrast, MOC in whole cells is highly regulated by the supply of flavins: riboflavin, flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN), where cell respiration reached up to 410% of controls. In isolated mitochondria, FAD and FMN drastically increased complex I rate of reaction (1300%) and (450%), respectively, having no effects on complex II or III. MPP+ reduced MOC in whole cells in a dose-dependent manner. In isolated mitochondria, MPP+ exerted mild inhibition at complex I, negligible effects on complexes II-III, and extensive inhibition of complex IV. Kinetic analysis of complex I revealed that MPP+ was competitive with NADH, and partially reversible by FAD and FMN. Co-Q(10) potentiated complex II ( approximately 200%), but not complex I or III. Despite positive influence of flavins and Co-Q(10) on complexes I-II function, neither protected against MPP+ toxicity, indicating inhibition of complex IV as the predominant target. The nicotinamides and glucose prevented MPP+ toxicity by fueling anaerobic glycolysis, evident by accumulation of lactate in the absence of MOC. The data also define a clear anomaly of neuroblastoma, indicating a preference for anaerobic conditions, and an adverse response to aerobic. An increase in CO(2), CO(2)/O(2) ratio, mitochondrial inhibition or O(2) deprivation was not directly toxic, but activated metabolism through glycolysis prompting depletion of glucose and starvation. In conclusion, the results of this study indicate that the mechanism of action for MPP+, involves the inhibition of complex I and and more specifically complex IV, leading to impaired OXPHOS and MOC. Moreover, flavin dervatives control the rate of complex I/cellular respiration and Co-Q10 augments complex II [corrected].     

中药材和饮片的高效液相色谱指纹图谱鉴别

目的：提出中药材和饮片的高效液相色谱指纹图谱(APLU)鉴别的原则和方法。方法：经过HPLC指纹图谱获取、验证和鉴别三个步骤,得出供鉴别用的对照指纹图谱,确定指标成分峰、特征峰的相对保留时间及范围,用于中药材和饮片的鉴别。结论：以丹参药材为例,所提出的原则与方法可以作为药材和饮片高效液相色谱指纹图谱鉴别用途。     

Sun exposure may protect against non-Hodgkin lymphoma: a case-control study

Ultraviolet radiation is a hypothesised risk factor for non-Hodgkin lymphoma (NHL) but no epidemiological study has examined this association using direct measures of sun exposure in individuals. Adults aged 20-74 years living in NSW and ACT, Australia, were the study population. Cases (704 of 829 invited to take part, 85%) were diagnosed January 2000 to August 2001. Controls (694 of 1,136 invited to take part, 61%) were randomly selected from state electoral rolls and frequency-matched to cases by age, sex and state of residence. A self-administered questionnaire and telephone interview measured outdoor hours on working and nonworking days and vacations at 10, 20, 30, 40, 50 and 60 years of age. Logistic regression models of NHL and sun exposure contained the 3 matching variables, ethnicity and sun sensitivity measures as covariates. Contrary to expectations, risk of NHL fell with increasing reported sun exposure hours. Relative to 1.0 for the lowest quarter of total sun exposure hours, the odds ratios (ORs) for successively higher quarters were 0.72 (95% CI 0.53-0.98), 0.66 (0.48-0.91) and 0.65 (0.46-0.91) (p(trend)=0.01). The association of sun exposure on nonworking days with NHL was stronger; OR for highest quarter 0.47 (0.34-0.66) (p(trend)=0.0001). Risk also fell with sun exposure on vacations; OR for highest quarter 0.60 (0.43-0.85) (p(trend)=0.003). These associations appeared strongest in women and in childhood. There was little evident trend in risk with exposure on working day. Our results provide strong statistical evidence for an inverse association between sun exposure and NHL. Increasing evidence that vitamin D may protect against cancer makes UV-mediated synthesis of vitamin D a plausible mechanism whereby sun exposure might protect against NHL.     

Application of triple-probe microdialysis for fast pharmacokinetic/pharmacodynamic evaluation of dopamimetic activity of drug candidates in the rat brain

The technique of microdialysis utilizing three simultaneously implanted probes in the anaesthetized rat enables monitoring of pharmacokinetic (PK) profiles of a tested drug both in blood (1st probe) and brain (2nd probe) compartments and the pharmacodynamic (PD) response of neurotransmitters (3rd probe) released into, or accumulating within the brain extracellular fluid (ECF). In the present study, the PK/PD characteristics of cocaine (psychostimulant, strong abuse potential) and methylphenidate (dopamimetic drug without reinforcing properties) and two novel NeuroSearch (NS) drug candidates, NS-A and NS-B, were examined in blood and brain microdialysates from the anaesthetized rats. The extracellular levels of dopamine (DA) were monitored in the striatum or prefrontal cortex. The NS-A compound entered the brain ECF at a slightly slower rate then methylphenidate; however, both compounds showed about the same effect on the speed of accumulation of extracellular DA concentrations, which gradually increased to about 450% of the basal, predrug levels at the end of the sampling period (180 min). The NS-B compound showed more rapid PK profiles than those observed after methylphenidate and NS-A. The concentrations of NS-B reached the maximal values already 40 min after its administration, while at that time, the corresponding DA values were still unchanged. In fact, the increase in DA concentrations was about two times slower when compared to that of methylphenidate or NS-A-drugs. Faster kinetics of NS-B and its delayed effect on extracellular DA suggests that this compound is metabolized to an active intermediate product, which itself exerts stronger dopamimetic activity in the rat prefrontal cortex that the original NS-B substance. The present study illustrates the feasibility of triple-probe microdialysis to monitor the rate of extracellular accumulation of a drug candidate and DA levels in vivo and compare the resulting PK/PD profiles to those obtained for cocaine and methylphenidate. These measures may serve as initial neurochemical indicators of potential psychomimetic or reinforcing properties of the tested substances.     

Effect of clothing varieties on solar photosynthesis of previtamin D3: an in vitro study

BACKGROUND/PURPOSE: Vitamin D3 plays important roles in the absorption of calcium and phosphorus from the gastrointestinal tract and in the treatment of rickets; in addition, it facilitates the deposition of minerals in bones, thus minimizing the possibility of developing osteomalacia. Sunlight naturally induces vitamin D3 photosynthesis. Such a process is affected by a number of factors such as age, geographical location, skin color, sunscreen application and clothing. It is intended in the present investigation to study in vitro the effect of clothing on the solar photoproduction of vitamin D3. METHODS: Fifteen different fabric samples were tested for their effect on the efficiency of the in vitro solar conversion of 7-dehydrocholesterol (7-DHC) to vitamin D3. 7-DHC was dissolved in methanol to give a concentration of 2.6 x 10(-4) M. Solutions were exposed to sunlight in quartz containers for predetermined periods either uncovered or covered with the fabric sample under test. Changes in the concentrations of 7-DHC and the photoproducts were monitored by HPLC. Fabrics were graded as the number of threads per square inch (in(2)), and their sunlight attenuation was determined. RESULTS: 7-DHC is transformed to previtamin D3 upon exposure to sunlight, and the amount generated exhibited an almost linear relationship. When fabric-covered samples of 7-DHC were irradiated, photoproducts were also detected and their concentrations depended on the degree of sunlight attenuation imposed by the fabric. Generally, the higher the number of threads per in(2) the more the light attenuation produced. CONCLUSION: Clothing plays an important role in attenuating sunlight, thus leading to diminished vitamin D3 production to an extent that would require dietary compensation.     

Application of the HPLC method for benzalkonium chloride determination in aerosol preparations

Benzalkonium chloride (BAC) is a bacteriostatic agent used in the pharmaceutical industry as a preservative. BAC is a mixture of alkylbenzyldimethylammonium chlorides, the three most important of which being those with alkyl substituents C12, C14, C16 at the quaternary ammonium salt.

The purpose of this study was to develop a method for determining benzalkonium chloride identity and content in aerosol preparations in which protein or steroid hormones are the active components. The high performance liquid chromatography (HPLC) method was used for this purpose. In the performed comparison of the influence of selected factors on the process of the separation of BAC homologues, a column with packing modified with cyan groups and mobile phase containing 0.075 M acetate buffer with acetonitrile (45:55), in an isocratic elution, was used for qualitative and quantitative determinations and for method validation. The developed method may be used for the assessment of the identity and content of BAC homologues in various pharmaceutical preparations. It is simple and it does not require particular sample preparation for the tests. It is characterized by good selectivity and high precision of the determinations.     

Analytical and clinical evaluation of the Bio-Rad HPLC kit for measurement of type I collagen cross links

 The measurement of hydroxylysylpyridinoline (PYD) and lysylpyridinoline (DPD), the degradation products of type I collagen, by manual HPLC assay posed practical difficulties. The present study was undertaken to evaluate the first commercially available HPLC kit, which provides a convenient substitute for cumbersome classical HPLC methods. The HPLC procedure is based on an improved sample clean-up chromatography, convenient ready-to-use HPLC reagents, and quicker isocratic elution of PYR and DPD on reverse-phase analytical column. The analytical parameters assessed include sensitivity, within- and between-assay variation, method comparison, recoveries, and interference. Clinical evaluation included discriminatory power of PYD and DPD and response to treatment of osteoporosis patients with Alendronate. DPD and PYD concentrations showed linear (r ² > 0.99) response between 10–400 pmol/ml and 75–4000 pmol/ml, respectively. The average within-assay imprecision, over a range of clinically relevant cross-links concentrations, was CV < 7% (n = 30). The total imprecision (n = 35 days), by ANOVA, for PYD and DPD was CV < 7.5% and CV < 10%, respectively. Average spike recovery was 95.4% ± 6.5%. Comparison with the historical HPLC method exhibited a close correlation (r values between 0.87 and 0.91, P < 0.0001). Creatinine-corrected DPD in postmenopausal (Z score = 2.4, P < 0.05, n = 17) and osteoporotic (Z score = 3.0, P < 0.01, n = 29) women were 44% and 64% higher, respectively, compared to premenopausal samples (n = 15). Similarly, PYD concentration was 26% and 54% higher in postmenopausal and postmenopausal osteoporotic women, respectively. There was a 47% (P < 0.001) decrease in DPD concentration (n = 16), and a 30% decrease in PYD concentration after 90 days of treatment of osteoporotic patients with Alendronate. DPD concentration correlated with N-telopeptide with an r value of 0.69 (n = 67, P < 0.0001). The reported kit method is substantially simpler and precise than the manual method. DPD concentrations determined by the current method reaffirm the clinical value in identifying increased bone resorption in pathological conditions and monitoring response to antiresorptive therapy. Received: June 28, 2002 / Accepted: November 6, 2002 Offprint requests to: D.J. Baylink     

Simultaneous analysis of metabisulfite and sulfate by CE with indirect UV detection. Application to and validation for a pharmaceutical formulation

Metabisulfite is used as an antioxidant agent in a number of pharmaceutical formulations. In order to quantify simultaneously both metabisulfite and its oxidation product (sulfate), a capillary zone electrophoretic (CZE) method with indirect UV detection was developed. Best results were achieved with a background electrolyte (BGE) constituted of 15 mM pyromellitic acid, 15 mM tris-(hydroxymethyl)-aminomethane and 0.2 mM tetradecyltrimethylammonium bromide at pH 8.3 and an applied electrical field of 123 V/cm in a 32.5 cm fused silica capillary. Indirect UV detection was performed at a wavelength of 225 nm. In order to validate this method, an internal standard (IS), namely ammonium formate, was used. Moreover, due to the high chloride concentration in the pharmaceutical formulation, conductivity was adjusted by adding sodium chloride into standard solutions to prevent matrix effect. Linearity and accuracy were successfully tested in a concentration range of 33.3–250 μg/ml for sodium metabisulfite and of 50–375 μg/ml for sodium sulfate. Method precision was determined on six samples each day. Thereby, relative standard deviations (R.S.D.) of 6% and 12–13% were obtained for intra-day and inter-day precision, respectively. Considering the instability of metabisulfite and its use as an antioxidant agent and not as an active principle, the method was accepted and used for routine analyses.     

A size-exclusion HPLC method for the determination of sodium chondroitin sulfate in pharmaceutical preparations

A size-exclusion HPLC method for the determination of sodium chondroitin sulfate (SCS) in pharmaceutical preparations has been developed and validated. The most important feature of this method compared with the previously reported assay methods was improved economical and determinative applications through direct analysis of SCS from pharmaceuticals. The linearity, precision, specificity, and accuracy of the method were established and validated. The intra- and inter-day precision was satisfactory with relative standard deviation lower than 1.0%. The recovery of SCS from multi-components pharmaceutical preparations were from 93.38 to 100.46%. Comparing our HPLC assay results with classical spectrophotometric methods, the developed method was considerably easy, simple and reproducible. As a result, the present method was supposed to be successfully applied to the assay of SCS for the routine quality control in pharmaceutical preparations.