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A response by Paul G. Abel and Alan W. Heath to a letter to the editor about their letter on astronomical observations in the June 2010 issue is presented.

A letter to the editor is presented in response to the article regarding the current apparition of Saturn in the previous issue.

The article presents an obituary for Edward L. Ellis, a reliable observer at the British Astronomical Association.

A response by Alan W. Heath to a letter to the editor about the article on the emphasis of the Webcam technology in astronomical observation in the August 2006 issue of the journal is presented.

Abstract: Two candidate preparations of human sequence recombinant Interleukin-2 (IL-2) were formulated and lyophilized at NIBSC prior to evaluation in a collaborative study for their suitability to serve as a replacement international standard. The preparations were tested by eight laboratories using in vitro bioassays and immunoassays. The candidate preparation 86/500 was judged suitable to serve as a replacement international standard based on the data obtained for activity and stability. On the basis of the results reported here, the preparation coded 86/500 was established by the WHO Expert Committee on Biological Standardisation (ECBS) in 2012 as the WHO 2nd IS for human IL-2 with an assigned value for IL-2 activity of 210IU/ampoule. Calibration of the 2nd IS is primarily based on the bioassay in use in various laboratories and relies exclusively on the estimates calculated relative to the WHO 1st IS for continuity of the IU. [Copyright &y& Elsevier]

Abstract: Current HPV vaccines and vaccine candidates are based on recombinant virus capsid proteins, so called virus-like particles (VLPs). Standardisation of assays for HPV capsid antibody will assist with epidemiology studies and future vaccine development. A World Health Organization international collaborative study was undertaken to assess the suitability of a freeze-dried serum, obtained from women naturally infected with HPV 16 and reactive against HPV 16 only, to serve as the International Standard for antibodies to HPV 16 in immunoassays and pseudovirion neutralisation assays. Eleven laboratories from nine countries participated in the collaborative study in which the candidate (NIBSC code 05/134) was assayed alongside samples from both vaccinees and naturally infected individuals. 05/134 had titres which were comparable to those obtained with serum from a naturally infected individual. Overall the variation between laboratories is similar to that observed in the previous study for samples from naturally infected individuals although slightly wider for sera from vaccinees. 05/134 has been established by the WHO Expert Committee on Biological Standardization as the 1st International Standard for antibodies to HPV 16, human serum, with an assigned potency of 5IUper ampoule. [Copyright &y& Elsevier]

Abstract: An international standard for the quantitation of HIV-2 RNA in plasma samples was developed. A collaborative study involving 29 laboratories from 15 countries was carried out in order to evaluate HIV-2 RNA candidate materials for use with nucleic acid-based tests (NATs). Candidate reference standards consisted of duplicate copies of two HIV-2 genotype A viruses, HIV-2 CAM2 and HIV-2 ROD and were coded S1–S4. Each laboratory assayed all four candidates on at least three separate occasions and data were collated and analysed at NIBSC. Of the data sets returned the majority were from qualitative assays. All assays detected both candidate standards with the exception of one commercial assay, the Nuclisens Easy Q, which was designed primarily for HIV-1 detection which did not detect HIV-2 CAM2 but showed good detection of HIV-2 ROD. This highlighted possible cross reactivity with HIV-2 ROD with some NAT primer/probe combinations; as a result the HIV-2 CAM2 material was established as the 1st international standard for HIV-2 RNA with an assigned unitage of 1000 International Units (IU) per ampoule and is available upon request from the National Institute for Biological Standardisation and Control (NIBSC) (www.nibsc.ac.uk). [Copyright &y& Elsevier]

Abstract: Twenty-eight laboratories from 16 countries participated in a collaborative study to evaluate an HIV-1 RNA Genotype Reference Panel for use with nucleic acid-based tests (NAT). The Reference Panel consisted of 11 coded samples representing different HIV-1 genotypes (subtypes A–D, AE, F, G, AA-GH, groups N and O) as well as a negative diluent control. Each laboratory assayed the eleven panel members concurrently with the 1st International Standard for HIV-1 RNA (NIBSC Code 97/656) on at least three separate occasions and the data collated and analysed at NIBSC. Twenty-nine sets of data from NAT were received, 19 from quantitative and 10 from qualitative assays, with six different commercial assays and five “in-house” assays represented. The results showed that viruses from subtypes A–D and recombinant virus AE [CRF01_AE] were detected consistently, but that some assays had difficulty with the detection and quantification of viruses from subtypes F and G, a mixed recombinant virus AA-GH and a representative of group N. Furthermore, most assays failed to detect the group O representative. The study illustrated the limitations of some molecular assays particularly in detection of certain non-B genotypes which are important viruses in the global AIDS pandemic and illustrated the value of a well-characterised genotype panel. The panel has been established by the World Health Organisation’s Expert Committee on Biological Standardisation as the 1st International Reference Panel HIV-1 RNA Genotypes (code 01/466). [Copyright &y& Elsevier]

This Report discusses ten successive morning and evening elongations. Data concerning phase anomaly, bright and dark atmospheric markings, cusp extensions and the Ashen Light are discussed. Systematic visual observations over days and weeks again provided definitive evidence for the 'four-day' retrograde 'weather' period, and measurements over a longer, eight-year epoch yielded a reliable average period of 3.99524 ± 0.00027 days, closely comparable with the long-term average derived by C. Boyer and others. The phase anomaly was never very large, but 1994 E yielded a significantly higher anomaly than the other elongations. Occasional records were made of the blunting of the S cusp near dichotomy; the less commonly blunted N cusp was well observed at the 1995 W elongation. High resolution data for two elongations suggest that cusp-blunting may simply be due to the presence of high latitude dark bands at such times, or strong polar turbulence. Of the other discrete bright features (recorded mostly at the limb), there was a definite preponderance of the southern over the northern hemisphere. During 1991 to 1998 there were somewhat more records of the true Ashen Light (i.e., when it appeared brighter than the surrounding sky) compared with the equivalent period from 1999 to 2006, with the 1991, 1993 and 1996 evening elongations yielding a significant number of independently confirmed sightings. [ABSTRACT FROM AUTHOR]

The year 2004 was exceptional in producing the first solar transit of Venus since the late Victorian era. The bright aureole and atmospheric ring were re-observed, and the entire phenomenon was witnessed for the first time ever in hydrogen alpha light. Although routine observations throughout 2004 were unexceptional, patterns of visibility of bright and dark markings, cusp extensions and cusp caps were recorded. No correlation was found between the latitude of the sub-Earth point and the visibility of either cusp cap, with the S. cap predominating for most of the year. It was possible to accurately follow individual ultraviolet dark markings over many consecutive rotations, extending from the E. to W. elongations, and thereby to make a current measurement of the synodic atmospheric rotation period for the near-equatorial features: 3.996 ± 0.001 days. The true Ashen Light was reported visually on only a few occasions, but these correspond closely to times when infrared emission from the surface of the dark side was recorded in 1-micron waveband images. Some of the stable dark side albedo features were also visible on the 1-micron images, and have been tentatively identified with known surface features. Infrared imaging at the same waveband showed little detail on the sunlit disk, but a few bright spots were sufficiently well observed to suggest a synodic rotation period close to 5.0 days, not atypical for the lower cloud decks. [ABSTRACT FROM AUTHOR]

Abstract: Regulatory requirements to control the level of anti-D in intravenous immunoglobulin (IVIG) products with European and United States (US) licences are to be introduced. A reference preparation of IVIG containing anti-D at 0.0475 IU/ml and having a nominal titre of 8 using the proposed direct haemagglutination reference method was deemed suitable to define the anti-D limit. This preparation, code 02/228, and a negative control IVIG preparation, code 02/226, were established by the World Health Organization as International Reference Reagents (IRRs). As stocks of the IRRs are limited, new larger fill stocks of positive and negative reference preparations, codes 04/132 and 04/140, respectively, were produced. The results from an international collaborative study involving 16 laboratories showed that preparations 04/132 and 04/140 are indistinguishable from the corresponding IRRs 02/228 and 02/226, respectively, using the proposed direct haemagglutination reference method. Stocks of 04/132 and 04/140 have been shared with the European Directorate for the Quality of Medicines (re-coded as 23613 and 23614, respectively) and with the Center for Biologics Evaluation and Research of the United States Food and Drug Administration (re-coded as CBER Lots 1B and 1N-b, respectively) for use as European and US Biological Reference Preparations, respectively. [Copyright &y& Elsevier]

A nucleic acid test (NAT) assay reference reagent for West Nile virus (WNV) RNA, consisting of heat-inactivated WNV grown in tissue culture and diluted in pooled, negative human plasma, was evaluated and quantitated in a collaborative study in which 14 laboratories participated.Participants were requested to assay serial half-log and log dilutions of the reagent to determine the RNA endpoint. A single endpoint for each such dilution series was calculated with the maximum likelihood method, which assumes that the probability of a positive result at a given dilution follows a Poisson distribution. The calculated endpoint was used to give an estimated“NAT-detectable units per mL” (not necessarily equivalent to genome equivalents/mL or copies/mL). The assays used by participants included qualitative and quantitative NAT assays and both the commercial WNV assays (Chiron and Roche).The estimated number of detectable units per mL for the 14 laboratories varied from log 2.0 to 3.0 with the exception of two outliers. The overall mean titer for all the assays was log 2.52 detectable units per mL (330 detectable units/mL). Multiple testing of individual vials by two laboratories indicated that there was no evidence of vial-to-vial variation in WNV content of the reference reagent.A reference reagent for WNV NAT assays has been established. The mean titer of the reagent, with the results from 14 laboratories, was 330 detectable units per mL. [ABSTRACT FROM AUTHOR]

Folate measurements, particularly for whole blood, show wide inter-laboratory and inter-methodology variability. This variability appears to be due in part to the lack of internationally accepted reference materials. A whole blood haemolysate, lyophilised in ampoules and designated 95/528, was therefore evaluated by 15 laboratories in five countries for its suitability as an International Standard (IS) for whole blood folate. The preparation was assayed using a variety of microbiological and protein-binding methodologies against local standards and calibrators. A consensus folate content was assigned to 95/528. The inclusion of three whole blood samples in the study with widely differing folate levels demonstrated a considerable reduction in inter-laboratory variability when the folate content of the samples was determined relative to the proposed IS 95/528 rather than to laboratories' local standards and calibrators. Accelerated degradation studies indicated that the folate content of 95/528 is stable when stored at -20°C. On the basis of the results presented here, the World Health Organization Expert Committee on Biological Standardization established 95/528 as an IS for whole blood folate. [ABSTRACT FROM AUTHOR]

• We reconstruct a molecular phylogeny for all 68 taxa of Chrysoritis butterflies. • The genus is 32 Mya, but the most diverse clade (thysbe – 28 species) is ~2 Mya. • Two main lineages in the genus diversified in western and eastern South Africa. • Fynbos is the origin of the thysbe radiation. • Reticulation within thysbe suggest a complex of incipient species and/or taxonomic oversplitting. Although best known for its extraordinary radiations of endemic plant species, the South African fynbos is home to a great diversity of phytophagous insects, including butterflies in the genus Chrysoritis (Lepidoptera: Lycaenidae). These butterflies are remarkably uniform morphologically; nevertheless, they comprise 43 currently accepted species and 68 currently valid taxonomic names. While many species have highly restricted, dot-like distributions, others are widespread. Here, we investigate the phylogenetic and biogeographic history underlying their diversification by analyzing molecular markers from 406 representatives of all described species throughout their respective ranges. We recover monophyletic clades for both C. chrysaor and C. thysbe species-groups, and identify a set of lineages that fall between them. The estimated age of divergence for the genus is 32 Mya, and we document significantly rapid diversification of the thysbe species-group in the Pleistocene (~2 Mya). Using ancestral geographic range reconstruction, we show that West Fynbos is the most likely region of origin for the radiation of the thysbe species-group. The colonization of this region occurred 9 Mya and appears to have been followed by a long period of relative stasis before a recent increase in diversification. Thus, the thysbe radiation does not appear to have resulted from the colonization of new biogeographic areas. Rather, the impact of species interactions (with ants and plants), the appearance of key innovations, and/or the opening of new ecological niche space in the region might explain the sudden burst of speciation that occurred in this group 2 Mya. The biogeographic model suggests two different diversification processes with few historical cross-colonisations, one in eastern South Africa for the C. chrysaor group and the other in western South Africa for the remaining taxa. Distributional range assessments and ecological niche models for each species show important niche overlap, and in a few cases, complete overlap. However, these shared traits are not explained by phylogenetic history. Chrysoritis taxa frequently fly in sympatry and gene tree reticulation appears to be widespread at the species level, suggesting that several episodes of range shifts might have led to secondary sympatries, allowing limited gene flow that challenges species delimitation efforts. In addition, the unusually high diversification rate for the thysbe clade of 1.35 [0.91–1.81] lineages per million years also suggests the possibility of taxonomic oversplitting. The phylogeny presented here provides a framework for a taxonomic revision of the genus. We highlight cases of potential synonymy both in allopatry and sympatry, and stress the importance of dedicated studies to assess potential pre- and post-zygotic barriers giving rise to species delimitations of the thysbe group. [ABSTRACT FROM AUTHOR]

It is imperative to ensure biological products are free of contaminating pyrogenic material prior to administration to patients. Historically the rabbit pyrogen test (RPT) was used to screen for such contamination in medicines for intravenous delivery. This test was adapted for use to screen vaccines. However, some, including meningococcal vaccines containing outer membrane vesicles, are intrinsically pyrogenic. Indeed, this is the case for Bexsero which contains relatively high levels of endotoxin and other potential pyrogens such as lipoproteins and porins. The RPT proved a difficult method for measuring the pyrogenic content of Bexsero and differences between laboratories in different countries made repeat testing at the control laboratories problematic resulting in batches being wrongly identified as unsafe. At NIBSC a monocyte activation test (MAT) was adapted and validated as an alternative. This required setting of a specification in-house and deciding on a decisional procedure using multiple donors, allowing batches equally pyrogenic or less, than those batches shown to be safe in a clinical trial, to be certified as safe. The resulting format was a reference comparison method with an upper limit of 1.8 relative pyrogen units (RPU). The batch passed if an initial four donors had a response equal to or less than 1.8 RPU, if one donor is above this limit the batch was tested in a further four donors and seven of the eight must be equal to or below 1.8 RPU. If two donors have a response greater than 1.8 the batch failed. [ABSTRACT FROM AUTHOR]

Background The WHO International Standard (IS) for hepatitis B surface antigen (HBsAg) is used to standardize HBsAg assays. Stocks of the 2nd IS for HBsAg are depleted. The proposal to establish its replacement was endorsed by WHO in 2012. Objective Preparation of a freeze-dried candidate 3rd IS (NIBSC 12/226); evaluation of its suitability in a WHO international collaborative study; calibration of its potency in International Units (IU). Study design The 3rd IS is based on plasma-derived, purified, inactivated HBsAg from Vietnam. Qualitative and quantitative HBsAg assays were used to evaluate 12/226 alongside the 2nd IS and 1st IS. Blinded study samples included a duplicate of 12/226, a negative control and two diluted plasma samples representing hepatitis B virus (HBV) genotypes A and B. Results Twelve laboratories from 9 countries returned 22 data sets from 15 methods. The overall geometric mean potency of 12/226 is 47.3 IU/mL (±13% CV) when compared to the 2nd IS with HBV subgenotype A2. The 3rd IS has HBV subgenotype B4 with a heterogeneous HBsAg subtype population of ayw1 and adw2 . Some genotype-dependent effects on the inter-laboratory variability were observed but overall mean potencies were virtually identical irrespective of the IS used for calibration. Stability studies indicate that the candidate is stable for long-term use. Conclusions 12/226 was established in October 2014 by the WHO Expert Committee on Biological Standardization as the 3rd IS for HBsAg with a potency of 47.3 IU per ampoule maintaining the continuity in the standardization of HBsAg assays. [ABSTRACT FROM AUTHOR]

Background. Interassay harmonization of cytomegalovirus (CMV) DNA measurement is important for infection management. Uncertainty exists regarding the result harmonization achievable in patient plasma samples using quantitative polymerase chain reaction (qPCR) assays with calibrators now traceable to the First World Health Organization International Standard (IS) for CMV DNA. Method. Serial dilutions of the IS and a blinded panel of 40 genotyped CMV DNA-positive pooled plasma samples and 10 negative plasma samples were tested by 6 laboratories using 10 qPCR assays calibrated to the IS. Each clinical sample was constructed using plasma from a single unique transplant recipient. Results. The variance for individual CMV DNA-positive samples was greater for clinical samples (median, 1.50 [range, 1.22-2.82] log10 IU/mL) than for IS dilutions (median, 0.94 [range, 0.69-1.35] log10 IU/mL) (P< .001); 58.9% of all clinical sample results and 93.6% of IS dilution results fell within ±0.5 log10 IU/mL of the mean viral load of each sample. Result variability was not impacted by either genotype or quantitative levels of CMV DNA. Testing procedure differences can significantly influence results, even when analyte-specific reagents are identical. For clinical samples, all assays demonstrated result bias (P< .008). Assays with amplicon sizes ≤86 bp had significantly higher results compared to assays with larger amplicon sizes (≥105 bp) (P< .001). Conclusions. The variability in CMV DNA results reported on individual samples has been reduced by the IS, but ongoing clinically relevant variability persists, preventing meaningful interassay result comparison. [ABSTRACT FROM AUTHOR]

Abstract: One candidate preparation of human sequence recombinant transforming growth factor-β3 (TGF-β3) was formulated and lyophilized at NIBSC prior to evaluation in a collaborative study for its suitability to serve as an international standard. The preparation was tested by 8 laboratories using in vitro bioassays and immunoassays. The candidate preparation 09/234 was judged suitable to serve as an international standard based on the data obtained for biological activity and stability. On the basis of the results reported here, the preparation coded 09/234 was established by the WHO Expert Committee on Biological Standardisation (ECBS) as the WHO 1st IS for human TGF-β3 with an assigned value for TGF-β3 activity of 19, 000IU/ampoule. [Copyright &y& Elsevier]

Abstract: Five candidate preparations of human sequence recombinant granulocyte-colony stimulating factor (G-CSF) were formulated and lyophilized at NIBSC prior to evaluation in a collaborative study for their suitability to serve as a replacement International Standard (IS). The preparations were tested by 13 laboratories using in vitro bioassays. The candidate preparation 09/136 was judged suitable to serve as a replacement international standard based on the data obtained for biological activity and stability. On the basis of the results reported here, the preparation coded 09/136 was established by the WHO Expert Committee on Biological Standardization (ECBS) as the 2nd IS for human G-CSF with an assigned value for G-CSF activity of 95,000IU/ampoule. Calibration of the 2nd IS is primarily based on the bioassay in use in various laboratories and relies entirely on the estimates calculated relative to the WHO 1st IS for continuity of the IU. [Copyright &y& Elsevier]

Background: Ferritin standardisation is problematical due to the heterogeneity of ferritin isoforms and the antibodies used in its immunoassay, and the lack of a reference measurement procedure. We investigated the performance of the 1st (liver), 2nd (spleen) and 3rd (recombinant) International Standards (ISs) for ferritin in major assays. Methods: The ferritin in a serum pool ‘spiked’ with either the 2nd or 3rd IS for ferritin was measured by 52 laboratories using five automated methods and the recovery of the target values calculated. A smaller serum pool was ‘spiked’ with the 1st IS for a limited recovery exercise. The ferritin values of five serum samples were also measured and recalculated relative to the ISs. Results: Recoveries of each of the 2nd and 3rd ISs were 90%–110% for four of five methods; recoveries of the 1st IS were 104% and 111% for two of three methods claiming traceability to this IS. One method significantly over-recovered each of the IS (124%–155%). Recalculating the ferritin values of the serum samples relative to the IS reduced the overall inter-method agreement, largely because of the anomalous over-recovery of the IS by one method. Conclusions: The use of the 3rd IS to standardise assays will minimise assay drift due to manufacturers adopting a ‘harmonisation’ approach in which the calibration is adjusted to conform to overall mean values. Standardisation against the current IS also ensures compliance with the European Union In-Vitro Diagnostic Directive which requires traceability of assay calibrators to reference materials of a higher order. Assay drift may result in poor sensitivity and specificity in the diagnosis of iron status, and would require laboratories to continually re-evaluate reference intervals. Clin Chem Lab Med 2008;46:1450–7. [ABSTRACT FROM AUTHOR]

Abstract: Japanese encephalitis (JE) viruses are grouped into four genotypes. Although currently available vaccines are derived only from viruses in genotype III, vaccines are known to protect against naturally occurring strains. Studies were undertaken to assess the suitability of a freeze-dried pool of human anti-JE plasma, collected from recipients of Biken (Nakayama-NIH) killed vaccine, to serve as an International Standard for antibodies to JE virus. Five participants in five countries submitted data from 11 assays on the candidate International Standard and seven coded samples including sera from recipients of vaccines containing a range of virus strains. The results of the study indicated that the 50% plaque reduction neutralization test (PRNT50titres) obtained for serum from recipients of killed vaccines, including the candidate standard, vary depending on the virus strain used in the neutralization tests, namely higher PRNT50titres were obtained when the challenge virus was homologous to the vaccine strain compared to use of a heterologous virus. Potencies expressed relative to the candidate standard are therefore affected by the strain of virus used in assays and the use of a standard would therefore not facilitate direct comparison of data from laboratories that have used different challenge strains. [Copyright &y& Elsevier]

Abstract: The in vivo biological activity of erythropoietin (Epo) is dependent on its being adequately sialylated. Current in vitro bioassays for Epo do not correlate with the in vivo bioassays as the former do not take into account the role the liver plays in clearing desialylated glycoproteins from the circulation. Here we describe a sialylation-sensitive cell-based Epo bioassay. In the first instance, Epo activity in vitro was measured using proliferation of AS-E2 cells, and in vivo using the polycythaemic mouse bioassay. Activity in vivo was progressively abolished by controlled desialylation, whereas activity in vitro was essentially unaffected. Incorporation of an incubation step with a solid-phase galactose-binding lectin (Erythrina crista-galli), effectively mimicking passage through the liver in vivo, renders the in vitro bioassay sensitive to desialylation, such that Epo desialylated almost to completion had &#60;10% of the activity of untreated Epo. These studies offer proof of principle, that rational manipulation of in vitro bioassays can allow prediction of activity in vivo without the use of live animals. [Copyright &y& Elsevier]

We have examined the quantity and composition of melanin in both photoprotected (volar upper arm) and chronically photoexposed (dorsal forearm) skin from a range of different ethnic skin types including African, Indian, Mexican, Chinese and European. The most lightly pigmented (European, Chinese and Mexican) skin types have approximately half as much epidermal melanin as the most darkly pigmented (African and Indian) skin types. However, the composition of melanin in these lighter skin types is comparatively more enriched with lightly coloured, alkali-soluble melanin components (up to three-fold). Regardless of ethnicity, epidermal melanin content is significantly greater in chronically photoexposed skin than it is in corresponding photoprotected skin (up to two-fold). However, by comparison there is only a modest enrichment of lightly coloured, alkali soluble melanin components in photoprotected skin (up to 1.3-fold). Analysis of melanosomes extracted from the epidermis in these subjects indicates that the proportion of spheroidal melanosomes is low in all skin types examined (<10%). This suggests that in human skin, pheomelanin is a very minor component of epidermal melanin, even in the lightest (European) skin types. Analysis of melanosome size revealed a significant and progressive variation in size with ethnicity: African skin having the largest melanosomes followed in turn by Indian, Mexican, Chinese and European. On the basis of these findings, we propose that variation in skin pigmentation is strongly influenced by both the amount and the composition (or colour) of the melanin in the epidermis. Variation in melanosome size may also play a significant role. However, the data also suggest that in human skin there are subtle differences in the mechanisms associated with the maintenance of constitutive pigmentation and facultative hyperpigmentation, respectively. [ABSTRACT FROM AUTHOR]

Objective measurements of human skin colour were made with a tristimulus (L *a *b *) chromameter in a range of different ethnic skin types. These were compared with biochemical measurements of melanin content, melanin composition and melanosome size in skin biopsies obtained from the same sites. L *, a * and b * values were found to vary significantly with ethnicity. In general, constitutively dark skin types have lower L * values, higher a * values and higher b * values than constitutively light skin types. Total epidermal melanin content appears to be the primary determinant of L * values in human skin (r =-0.88; P < 0.00001), whilst melanosome size also has a significant but more subtle influence on L * values (r =-0.73; P < 0.00001). There is also a strong positive contribution to a * values from epidermal melanin (r =0.66, P < 0.00001), which accounts for the ethnic variation in a * values observed in this study. Melanin is also a major contributor to b * values in lighter skin types (r =0.71, P < 0.00001). However, this relationship breaks down in darker skin types where b * values actually reach a maximum and then decrease as the concentration of melanin in the skin increases. This appears to be because of optical masking of yellow light by high concentrations of melanin in the epidermis. Analysis of the relationships between L *, a * and b * values in human skin indicate that they are very closely interrelated, and suggest that the optical properties of melanin in the epidermis are very similar to those of a dye on a fabric substrate. [ABSTRACT FROM AUTHOR]

Aims: Pancreatic beta-cell lipo-dysfunction decreases insulin secretion and predisposes to the development of type 2 diabetes. Through targeted Pex11β knockdown and peroxisome depletion, our aim was to investigate the specific contribution of peroxisomes to palmitate mediated pancreatic beta-cell dysfunction.Methods: MIN6 cells were transfected with probes targeted against Pex11β, a regulator of peroxisome abundance, or with scrambled control probes. Peroxisome abundance was measured by PMP-70 protein expression. 48 h post transfection, cells were incubated with 250 μM palmitate or BSA control for a further 48 h before measurement of glucose stimulated insulin secretion and of reactive oxygen species.Results: Pex11β knockdown decreased target gene expression by >80% compared with the scrambled control (P<0.001). This led to decreased PMP-70 expression (p<0.01) and a 22% decrease in peroxisome number (p<0.05). At 25 mM glucose, palmitate treatment decreased insulin secretion by 64% in the scrambled control cells (2.54±0.25 vs 7.07±0.83 [mean±SEM] ng/h/μg protein; Palmitate vs BSA P<0.001), but by just 37% in the Pex11β knockdown cells. Comparing responses in the presence of palmitate, insulin secretion at 25 mM glucose was significantly greater in the Pex11β knockdown cells compared with the scrambled controls (4.04±0.46 vs 2.54±0.25 ng/h/μg protein; p<0.05). Reactive oxygen species generation with palmitate was lower in the Pex11β knockdown cells compared with the scrambled controls (P<0.001).Conclusion: Pex11β knockdown decreased peroxisome abundance, decreased palmitate mediated reactive oxygen species generation, and reversed the inhibitory effect of palmitate on insulin secretion. These findings reveal a distinct role of peroxisomes in palmitate mediated beta-cell dysfunction. [ABSTRACT FROM AUTHOR]