Epitope discovery using bacteriophage display: the minimum epitope of an anti-IRBP antibody.

The purpose of this study was to determine, using random peptide library (RPL) technologies, the minimal epitope requirements of the mouse monoclonal anti-interphotoreceptor-retinoid-binding protein antibody, H3B5. This previously characterized antibody is used as an example to examine whether RPL's offer a relatively easy and rapid route to obtaining detailed epitope mapping data.A pentadecamer random peptide library (RPL) displayed on the major coat protein (gene 8) of filamentous bacteriophage (F88-4-15) was used as a target for selection by the anti-IRBP monoclonal antibody, H3B5. Three rounds of library selection were performed, and 90 of the resultant RPL clones were examined for affinity to H3B5 by enzyme-linked immunosorbent assay (ELISA). DNA sequencing of ELISA positive clones provided sequence of the region encoding the random peptide.After three rounds of selection of the RPL, 76.7% of clones examined interacted with H3B5, 17.7% did not show significant binding and 6.6% bound to control antibody also. The essential elements of the peptide epitope were determined by sequence comparison of 24 clones to be the four amino-acid sequence (Aspartic or glutamic acid)-Proline-Arginine-(Leucine, Isoleucine or Valine). This motif [(D/E) PR (L/I/V)] is in agreement, but at greater resolution, than previous synthetic peptide studies where the motif AASEDPRL was identified. Other motifs were found which bound to H3B5 but did not share primary structure similarities (peptidomimetics). Selection from a RPL has rapidly defined the minimal requirements for the H3B5 epitope in fine detail. Such a process offers great potential for investigating antibody-antigen interactions and core sequences of an epitope, and enables the identification of motifs in other proteins which may be recognized by the antibody, providing information on possible cross-reactivity.     

Hypothemycin inhibits the proliferative response and modulates the production of cytokines during T cell activation

Hypothemycin, a resorcylic acid lactone antibiotic, was identified as active in a screen for inhibitors of T cell activation. It was found to inhibit the proliferation of mouse and human T cells stimulated with anti-CD3 mAb + PMA and of human PBMC stimulated with anti-CD3 mAb alone. This inhibition was partially reversed by exogenous IL-2 indicating that it is not due to non-specific toxicity. Hypothemycin potently suppressed the production of IL-2 (IC50: 9 nM) but affected IL-2-induced proliferation to a lesser extent (IC50: 194 nM). Hypothemycin also inhibited IL-6, IL-10, IFN-gamma and TNF-alpha production. By contrast, it markedly enhanced the production of IL-4, IL-5 and IL-13. These effects were seen both at the mRNA and protein secretion levels. Analysis of the effect of hypothemycin on CD69 induction suggested that it disrupts calcineurin-independent rather than calcineurin-dependent signaling. Furthermore, hypothemycin was able to inhibit the phosphorylation of ERK1/2 induced by PMA treatment of T cells. Therefore, hypothemycin represents an inhibitor of T cell activation with a novel mode of action and unique modulatory activity on cytokine production.     

Calcinosis in juvenile dermatomyositis

A retrospective study of the radiographs and clinical records of 39 children with dermatomyositis, followed up at a referral centre for a mean period of eight years, revealed calcinosis in 29 children (18 giris, 11 boys). Clinical presentation of calcinosis was unpredictable and variable, ranging from four months to 12 years after onset of disease. Subcutaneous calcilication, often associated with subsequent ulceration of overlying skin, was observed more frequently than calcinosis in intermuscular fascial planes (29:7). Subcutaneous calcinosis was demonstrated most commonly about the knees and elbows and in the fingers and toes. Intermuscular calcinosis affected the fascial planes around the quadriceps, deltoid, biceps, and the gastrocnemius/soleus muscles.Calcinosis, predominantly of the subcutaneous lesions, regressed spontaneously in eight of 11 children where this could be assessed over a one to four year period. This high rate of spontaneous resolution must complicate evaluation of the efficacy of proposed anti-calcinotic treatments in juvenile dermatomyositis.While no apparent relationship was established between subcutaneous calcinosis, which was present in all 29 of the affected patients, and disease activity or severity, all seven children with the additional, extensive, and classical intermuscular fascial plane calcification developed this complication early and had severe disease.These findings are discussed in relation to previous studies.     

Increased dopamine receptor sensitivity in the rat following acute administration of sufentanil,U50,488H and d-Ala2-d-Leu5-enkephalin

Summary The effects of acutely administered opioid receptor agonists sufentanil, U50,488H and [d-Ala², d-Leu⁵]-enkephalin (DADL) were observed upon dopamine D₁ and D₂ binding site density in the striatum of the rat. In addition, the functional implications of opioid-induced changes in dopamine receptor sensitivity were studied using the behavioural profile elicited by apomorphine in the rat. The -agonist sufentanil (1 or 20 Erg/kg, i. p.), the -agonist U50,488H (10 mg/kg, i. p.) and DADL (1 g/animal, i. c. v.) all significantly elevated D₂ but not D₁ binding site density in rat striatum. Pretreatment with sufentanil (1 g/kg, i. p.) induced an elevation in apomorphine-induced sterotyped behaviour, but attenuated locomotor activity. Following administration of U50,488H (10 mg/kg, i. p.), both the degree of stereotypy and the intensity of the locomotor activity were enhanced. Contralateral rotation was observed in animals pretreated with DADL (1 g/animal, i. c. v.) following challenge with apomorphine. It is concluded that the opioid agonists studied induce a significant elevation in functional D₂ sites to the exclusion of D₁ sites. However, the precise mechanism by which this effect is elicited remains to be established. Send offprint requests to R. D. E. Sewell at the above address     

Efficacy of Zoladex LA (goserelin) in the treatment of girls with central precocious or early puberty

OBJECTIVE: To assess the efficacy of a longer acting preparation of the gonadotrophin releasing hormone (GnRH) analogue goserelin (Zoladex LA, 10.8 mg) in 12 girls with central precocious or early puberty. METHODS: Two girls started treatment de novo; the remainder had been on suppressive treatment for a median duration of 1.5 (range, 0.2-5.6) years. Assessment comprising auxology, pubertal staging, and pelvic ultrasound examination was carried out at weeks 0, 4, 8, 10, and 12 (first cycle) and weeks 8, 10, and 12 (second cycle) to evaluate the required injection frequency. Thereafter, assessment was performed on the day of injection. Zoladex LA was given every 12 weeks unless pubertal progression occurred. RESULTS: Satisfactory control was achieved in eight patients using this regimen, and three patients required more frequent injections. One girl was removed from the study because of clinical progression and extreme mood swings. No serious adverse effects occurred. Mean height velocity during the study period was 4.5 cm/year (range, 3.1-6.6) compared with 6.5 cm/year (range, 3.8-9.6) before treatment in nine patients for whom data were available. CONCLUSIONS: Zoladex LA was effective in controlling precocious puberty in girls when given at intervals of 9-12 weeks and it is recommended that an initial assessment is made eight weeks after beginning treatment.     

Operando unraveling photothermal-promoted dynamic active-sites generation in NiFe2O4 for markedly enhanced oxygen evolution

The ability to develop highly active and low-cost electrocatalysts represents an important endeavor toward accelerating sluggish water-oxidation kinetics. Herein, we report the implementation and unraveling of the photothermal effect of spinel nanoparticles (NPs) on promoting dynamic active-sites generation to markedly enhance their oxygen evolution reaction (OER) activity via an integrated operando Raman and density functional theory (DFT) study. Specifically, NiFe₂O₄ (NFO) NPs are first synthesized by capitalizing on amphiphilic star-like diblock copolymers as nanoreactors. Upon the near-infrared light irradiation, the photothermal heating of the NFO-based electrode progressively raises the temperature, accompanied by a marked decrease of overpotential. Accordingly, only an overpotential of 309 mV is required to yield a high current density of 100 mA cm⁻², greatly lower than recently reported earth-abundant electrocatalysts. More importantly, the photothermal effect of NFO NPs facilitates surface reconstruction into high-active oxyhydroxides at lower potential (1.36 V) under OER conditions, as revealed by operando Raman spectroelectrochemistry. The DFT calculation corroborates that these reconstructed (Ni,Fe)oxyhydroxides are electrocatalytically active sites as the kinetics barrier is largely reduced over pure NFO without surface reconstruction. Given the diversity of materials (metal oxides, sulfides, phosphides, etc.) possessing the photo-to-thermal conversion, this effect may thus provide a unique and robust platform to boost highly active surface species in nanomaterials for a fundamental understanding of enhanced performance that may underpin future advances in electrocatalysis, photocatalysis, solar-energy conversion, and renewable-energy production.

Motivated by the need for accelerating sluggish reaction kinetics at the anode (1, 2), the focus of water electrolysis has been centered heavily on oxygen evolution reaction (OER) toward sustainable hydrogen fuel production. To date, there has been much effort in developing low-cost yet high-performance earth-abundant transition-metal alternatives to commonly used noble metals for OER. Intriguingly, many Ni-, Co-, Fe-, and Mn-based oxides experience a dynamic surface-reconstruction process to form more active oxyhydroxides, which are recognized as true catalytically active species for OER in alkaline media (3, 4). Among various transition metal-based OER catalysts, bimetal spinel-structured oxides in the form of AB₂O₄ (A and B are different metal ions) have garnered much attention due to their rich compositions, electron configurations, and valence states (5). Interestingly, inverse spinel NiFe₂O₄ (NFO), in principle, exhibits enhanced catalytic activity toward OER because of the presence of multivalent elements (i.e., Ni³⁺/Ni²⁺ and Fe³⁺/Fe²⁺) (6). It is important to note that studies on facilitating the surface reconstruction of NFO to achieve high-performance OER are relatively few, and a fundamental understanding as to what makes the derived OER catalysts perform well remains elusive.Recently, introducing thermal energy to promote electrocatalytic conversion has attracted significant interest (7–10). The thermal energy could accelerate the motion of reactant molecules, promote their effective collisions during the reaction, and thus make it easier to overcome the activation barrier (11, 12). Moreover, the use of thermal energy could also promote the surface reconstruction of catalysts into highly active species and accelerate the electrocatalytic kinetics, thereby leading to improved efficiency (13). Electrocatalysts with photothermal effect (referred to as photothermal electrocatalysts) enable in situ heating due to photo-to-thermal conversion under illumination with visible or near-infrared (NIR) light, thereby dispensing with the need for extra devices required to provide thermal energy. More importantly, in sharp contrast to common approaches where the entire solution is heated (14), the photothermal effect is localized on electrocatalysts themselves (15), thus effectively enabling heat modulation to a defined region (i.e., the working electrode). Despite recent impressive advances in transition metal oxides (e.g., Fe₃O₄ and Co₃O₄) as photothermal agents for cancer therapy (15), their implementations for photothermal-assisted OER, in particular spinel oxides, are comparatively few and limited in scope. Moreover, it has been reported that Ni- and Co-based OER catalysts are prone to surface reconstruction into highly active oxyhydroxides (16, 17). Surprisingly, the photothermal effect on promoting surface reconstruction in spinel oxide catalysts has yet to be explored.Herein, we report an integrated operando Raman and density functional theory (DFT) plus Hubbard U (DFT + U) study to exercise and unveil the photo-to-thermal conversion of inverse spinel oxide nanoparticles (NPs) in promoting the generation of dynamic active sites via surface reconstruction into oxyhydroxides and thus greatly enhancing their OER activity. First, a series of amphiphilic star-like poly(acrylic acid)-block-poly(styrene-co-acrylonitrile) (denoted PAA-b-PSAN) diblock copolymers with a well-defined molecular weight (MW) and low polydispersity index (PDI) are exploited as nanoreactors to synthesize a set of PSAN-ligated NFO NPs with different sizes and PSAN chain lengths. The effects of the NFO NP sizes and the outer PSAN chain lengths on catalytic activity of NFO NPs are then scrutinized. Interestingly, NFO NPs of the largest size (∼12 nm) ligated with the shortest PSAN chains (M_n = 7K) display the best OER reactivity on a glassy carbon (GC) electrode in alkaline media as a result of the high fraction of the exposed electrochemically active surface area and the fastest electrocatalytic kinetics. Subsequently, the photothermal effect of PSAN-ligated NFO NPs is exploited to promote their surface reconstruction and thus boost OER. More importantly, an operando Raman spectroelectrochemistry study is performed to unveil the mechanism of photothermal-assisted enhancement in OER reaction, revealing the emergence of electrocatalytically active γ-NiOOH at a lower potential (1.36 V) during the surface-reconstruction process with a photothermal effect. Finally, the first-principle calculations substantiate that the reconstructed surface, i.e., (Ni,Fe)oxyhydroxides, plays a pivotal role as the active site for electrocatalytic reaction. As such, photothermal electrocatalysts (e.g., metal oxides, sulfides, phosphides, etc.) may render significantly low overpotential and fast OER kinetics, representing an array of important materials that couple the localized heating with electrochemistry for effective renewable-energy production.     

"Early-peak" carbon monoxide production in certain erythropoietic disorders

The "early-labeled" peak (ELP) of 14CO excretion following injection of glycine-2-14C was used to study erythropoiesis in a patient with sideroblastic anemia and in four subjects with myeloproliferative disorders. The ELP was greatly enlarged in all patients, as compared with a normal volunteer. The contour of the peaks from the hematologically abnormal subjects suggested the presence of increased erythroid heme degradation. In the patient with sideroblastic anemia, all hours of the early peak were significantly reduced after transfusion. This was interpreted to mean that even the earliest or "nonerythroid" phase of the peak is influenced by erythropoietic activity, at least under conditions of erythropoietic stress.