Listeriosis outbreak caused by contaminated stuffed pork, Andalusia, Spain, July to October 2019.

Between 1 July and 26 October 2019 in Andalusia, Spain, a large outbreak with 207 confirmed cases of listeriosis was identified. Confirmed cases had a median age of 44 years (range: 0-94) and 114 were women (55.1%). Most cases (n = 154) had mild gastroenteritis, 141 (68.1%) required hospitalisation and three died; five of 34 pregnant women had a miscarriage. The median incubation period was 1 day (range: 0-30), and was significantly shorter in cases presenting with gastroenteritis compared to those presenting without gastroenteritis (1 day vs. 3 days, respectively, p value < 0.001). Stuffed pork, a ready-to-eat product consumed unheated, from a single producer contaminated with Listeria monocytogenes ST388 was identified as the source of infection. The outbreak strain was identified in 189 human samples and 87 non-human (82 food and 5 environmental) samples. Notification of new cases declined abruptly after control measures were implemented. These included contaminated food recall, protocols for clinical management of suspected cases and for post-exposure prophylaxis in pregnant women and communication campaigns with concise messages to the population through social media. Given that there were 3,059 probable cases, this was the largest L. monocytogenes outbreak ever reported in Europe.

Circulating microRNAs as potential biomarkers for genetic generalized epilepsies: a three microRNA panel.

BACKGROUND AND PURPOSE: Genetic generalized epilepsies (GGEs) encompass a group of syndromes of mainly genetic causes, characterized by the involvement of both hemispheres. MicroRNAs (miRNAs) are small non-coding RNAs with a critical role in the regulation of neuronal biological processes through gene expression modulation. Dysregulated miRNA expression has been shown in epilepsy. Due to their stability in biological fluids like serum, miRNAs have assumed a prominent role in biomarker research. Our aim was to evaluate circulating levels of three miRNAs in GGE patients and assess their putative diagnostic value. METHODS: MiR-146a, miR-155 and miR-132 were quantified by real-time polymerase chain reaction in the serum of 79 GGE patients (47 women, 32 men, 35.1 ± 12.4 years) and 67 healthy individuals (41 women, 26 men, 42.4 ± 10.1 years). Relative expression values were calculated using the 2-ΔΔCt method. Receiver operating characteristic curve analysis was performed to assess diagnostic value. MiRNA expression was correlated with clinicopathological features. RESULTS: Serum levels of miR-146a and miR-155 were significantly upregulated in GGE patients relative to controls (3.13 and 6.05, respectively). Combined miR-146a, miR-155 and miR-132 serum levels performed well as a diagnostic biomarker, discriminating GGE patients from controls with an area under the curve of 0.85, 80% specificity and 73% sensitivity. CONCLUSIONS: Our results indicate that miR-146a, miR-155 and miR-132 may partake in GGE epileptogenesis. A panel of three circulating miRNAs with potential value as a GGE biomarker is reported for the first time. Novel biomarkers may help to identify new treatment targets and contribute to improved patients' quality of life through earlier diagnosis and a more precise prognosis.

First Measurement of the Total Neutron Cross Section on Argon between 100 and 800 MeV.

We report the first measurement of the neutron cross section on argon in the energy range of 100-800 MeV. The measurement was obtained with a 4.3-h exposure of the Mini-CAPTAIN detector to the WNR/LANSCE beam at LANL. The total cross section is measured from the attenuation coefficient of the neutron flux as it traverses the liquid argon volume. A set of 2631 candidate interactions is divided in bins of the neutron kinetic energy calculated from time-of-flight measurements. These interactions are reconstructed with custom-made algorithms specifically designed for the data in a time projection chamber the size of the Mini-CAPTAIN detector. The energy averaged cross section is 0.91±0.10(stat)±0.09(syst) b. A comparison of the measured cross section is made to the GEANT4 and FLUKA event generator packages, where the energy averaged cross sections in this range are 0.60 and 0.68 b, respectively.

Measurement of Final-State Correlations in Neutrino Muon-Proton Mesonless Production on Hydrocarbon at ⟨E_{ν}⟩=3 GeV.

Final-state kinematic imbalances are measured in mesonless production of ν_{µ}+A→µ^{-}+p+X in the MINERvA tracker. Initial- and final-state nuclear effects are probed using the direction of the µ^{-}-p transverse momentum imbalance and the initial-state momentum of the struck neutron. Differential cross sections are compared to predictions based on current approaches to medium modeling. These models underpredict the cross section at intermediate intranuclear momentum transfers that generally exceed the Fermi momenta. As neutrino interaction models need to correctly incorporate the effect of the nucleus in order to predict neutrino energy resolution in oscillation experiments, this result points to a region of phase space where additional cross section strength is needed in current models, and demonstrates a new technique that would be suitable for use in fine-grained liquid argon detectors where the effect of the nucleus may be even larger.

Eslicarbazepine acetate as adjunctive therapy in clinical practice: ESLADOBA study.

OBJECTIVE: To assess seizure control and tolerability of eslicarbazepine acetate (ESL) as adjunctive therapy to one baseline antiepileptic drug (AED), in adults with partial-onset seizures (POS) with or without secondary generalization. METHODS: Multicenter, non-interventional, prospective cohort study conducted between March 2012 and September 2014 at 12 neurology departments in Portugal. Adults with POS not controlled with one AED who had initiated ESL as adjunctive treatment were enrolled. Retention rate was defined at the final visit (Vfinal) 6-9 months of follow-up. Proportion of responders, seizure-free, changes in seizure frequency were evaluated using patients' diaries. Clinical Global Impression of Change (CGI-C) and Clinical Global Impression of Severity (CGI-S) were assessed by the neurologist. RESULTS: Fifty-two patients (48.1% male) were included with mean age 41.5±13.3 years. Mean epilepsy duration was 18.5±14.8 years; mean seizure frequency in the four previous weeks to baseline was 7.5±12.7. At Vfinal, retention rate was 73.0%; responder rate and seizure-free rates were 71.1% and 39.5%, respectively. The median relative reduction in seizure frequency between baseline and Vfinal was 82.2%. A reduction in epilepsy severity (CGI-S) was observed in 42.1%. According to CGI-C, 73.6% patients had their epilepsy "much improved" or "very much improved". Twelve patients (23.1%) had at least one adverse event (AE), two (3.9%) had one serious AE, and five (9.6%) discontinued due to AE. CONCLUSIONS: Eslicarbazepine acetate showed good retention rates, elicited a significant reduction in seizure frequency, and was well tolerated when used in the clinical practice.

Compact étendue-preserving light-mixing optics.

We present a compact freeform optic, called "Freeform Shell-Mixer", which, when placed on top of a multicolor light source (particularly, a multi-chip LED), turns it into a virtual source in which colors are uniformly mixed. The optic, compatible with injection molding, makes use of étendue-conserving Köhler integration to provide homogeneous mixing of light. Its minimal size (just 2x larger than the source) makes the concept compatible with many luminaries, as ray tracing models show. Simulations indicate that the Freeform Shell-Mixer can reach efficiencies above 95% and both the size of the virtual source and its emission pattern are very similar to the ones of the original source, so the correct performance of the luminaire is secured.

Impact of osteoporosis and bone fracture on health-related quality of life in postmenopausal women.

BACKGROUND: Osteoporosis is responsible for fragility fractures, which are associated with decreased physical and social function. The GINERISK study was a cross-sectional epidemiological study conducted in 4157 Spanish postmenopausal women initially diagnosed with osteoporosis according to WHO criteria within the last 2 years. AIM: The aim of the study was to explore the impact of osteoporosis on health-related quality of life (HRQoL). METHOD: Menopause-specific and generic HRQoL were assessed, respectively, with the specific Cervantes Scale and the generic SF-12v2 Health Survey. The impact of osteoporosis on HRQoL was ultimately evaluated in 3328 (80.1%) women who had measurements for both bone mineral densitometry (BMD) and HRQoL. RESULTS: Menopause-specific or generic HRQoL, respectively, were worse in women with current osteoporosis and prior osteoporotic bone fracture (BF) in comparison with current osteoporosis without BF or whose T-score had increased above -2.5 on the BMD after receiving osteoporosis drug therapy. Impaired HRQoL was found both in Spanish postmenopausal female populations and the Spanish general female population. Women with osteoporosis with BF had physical and mental summary component scores in the 20th and 30th percentiles, respectively, of the Spanish general population. Higher risk for cardiovascular death was also associated with greater HRQoL impairment. The use of selective estrogen receptor modulators in women with a BMD T-score ≤ -2.5 was associated with lower impact of osteoporosis on HRQoL, particularly in the domains of physical health and sexuality. CONCLUSION: The HRQoL analysis results in this study demonstrated a loss of quality of life in postmenopausal women with osteoporosis and confirmed that this loss is greater in women with prior osteoporotic fracture.

Glucose impairment and ghrelin gene variants are associated to cognitive dysfunction.

BACKGROUND AND AIMS: Cognitive state and brain volume have been related to body mass index, abdominal fat, waist-hip ratio, components of metabolic syndrome (MS) and ghrelin. Genetic variations within the ghrelin gene have been recently associated to MS. The aim of our study was to investigate cognitive state by Mini-Mental State Examination (MMSE) in relation to MS components (ATP-III criteria) and ghrelin gene polymorphisms in dwelling individuals aged ≥70. METHODS: 280 subjects (137 men/143 women, age 77.03 ± 5.92) from the Mataró Ageing Study were included. Individuals were phenotypically characterized by anthropometric variables, lipids, glucose, blood pressure and MMSE. SNPs -501AC (rs26802), -994CT (rs26312), -604GA (rs27647), M72L (rs696217) and L90G (rs4684677) of the ghrelin gene were studied. Genotypes were determined by polymerase chain reaction and SNapshot minisequencing. RESULTS: 22.1 % had MMSE <24. MMSE <24 was associated with age (p < 0.001), female gender (p = 0.016), low education (p < 0.001) and glucose impairment or diabetes (p = 0.040). MMSE was influenced by obesity, central obesity, MS and glucose impairment. This latter association remained significant after adjustment by gender, age, alcohol, educational level, GDS and ApoE genotype (p = 0.009). Ghrelin SNPs were associated to MMSE: M72L C/A genotype showed lower score than C/C (p = 0.032, after adjusting for confounders 0.049); L90G A/T genotype showed lower score than A/A (p = 0.054, after adjusting 0.005). MMSE <24 was associated to L90G (39.1 % in A/T genotype vs 19.3 % in A/A, p = 0.026, after adjusting for confounders p = 0.002, OR 6.18 CI 1.93-21.75). CONCLUSIONS: Glucose impairment and L90G Ghrelin gene variant influence cognitive function in old dwelling individuals participating in the Mataró Ageing Study.

A multicenter, randomized phase 2 study to establish combinations of CBP501, cisplatin and nivolumab for ≥3rd-line treatment of patients with advanced pancreatic adenocarcinoma.

BACKGROUND: There is no standard of care for ≥ 3rd-line treatment of metastatic pancreatic adenocarcinoma (PDAC). CBP501 is a novel calmodulin-binding peptide that has been shown to enhance the influx of platinum agents into tumor cells and tumor immunogenicity. This study aimed to (1) confirm efficacy of CBP501/cisplatin/nivolumab for metastatic PDAC observed in a previous phase 1 study, (2) identify combinations that yield 35% 3-month progression-free survival rate (3MPFS) and (3) define the contribution of CBP501 to the effects of combination therapy. METHODS: CBP501 16 or 25 mg/m2 (CBP(16) or CBP(25)) was combined with 60 mg/m2 cisplatin (CDDP) and 240 mg nivolumab (nivo), administered at 3-week intervals. Patients were randomized 1:1:1:1 to (1) CBP(25)/CDDP/nivo, (2) CBP(16)/CDDP/nivo, (3) CBP(25)/CDDP and (4) CDDP/nivo, with randomization stratified by ECOG PS and liver metastases. A Fleming two-stage design was used, yielding a one-sided type I error rate of 2.5% and 80% power when the true 3MPFS is 35%. RESULTS: Among 36 patients, 3MPFS was 44.4% in arms 1 and 2, 11.1% in arm 3% and 33.3% in arm 4. Two patients achieved a partial response in arm 1 (ORR 22.2%; none in other arms). Median PFS and OS were 2.4, 2.1, 1.5 and 1.5 months and 6.3, 5.3, 3.7 and 4.9 months, respectively. Overall, all treatment combinations were well tolerated. Most treatment-related adverse events were grade 1-2. CONCLUSIONS: The combination CBP(25)/(16)/CDDP/nivo demonstrated promising signs of efficacy and a manageable safety profile for the treatment of advanced PDAC. CLINICAL TRIAL REGISTRATION: NCT04953962.

The physiological quality of Vigna unguiculata L. seeds shows tolerance to salinity.

Salinity reduces feijão-caupi production, and the search for tolerant varieties becomes important within the agricultural context, as, in addition to being used in the field, they can be used in genetic improvement. The objective was to for a identify variety that is tolerant to salinity considering the physiological quality of seeds and seedling growth. A 2 × 4 factorial scheme was used, referring to the varieties Pingo-de-ouro and Coruja, and four electrical conductivities of water (0; 3.3; 6.6 and 9.9 dS m-1). The physiological quality of seeds and the growth of seedlings were analyzed, in addition to the cumulative germination. The Pingo-de-ouro variety showed no germination, length of the shoot and root, dry mass of the shoot and root compromised up to electrical conductivity of 6 dS m-1 in relation to 0.0 dS m-1. On the other hand, the Coruja variety showed reduced germination, increased shoot and root length. The creole variety Pingo-de-ouro proved to be tolerant to salinity.

Resistance determinants of emerging pathogens isolated from an intensive care unit as a parameter of population health conditions of the Legal Amazon microregion.

Bacteria responsible for causing infections are common in hospital environments, water, soil, and food products. The infection risk is intensified by the absence of public sanitation, poor quality of life, and food scarcity. These external factors promote the dissemination of pathogens by direct contamination or biofilm formation. In this work, we identified bacterial isolates obtained from intensive care units in the southern region of Tocantins, Brazil. We compared matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) techniques and 16S ribosomal ribonucleic acid (rRNA) molecular analysis; we also performed phenotypic characterization. Fifty-six isolates characterized using morphotinctorial tests were classified as gram-positive (80.4%; n = 45) and gram-negative (19.6%; n = 11) and were resistant to several antibiotic classes; notably, we identified the blaOXA-23 resistance gene in the ILH10 isolate. Microbial identification using MALDI-TOF MS resulted in the identification of Sphingomonas paucimobilis and Bacillus circulans. 16S rRNA sequencing revealed four isolates belonging to the genera Bacillus and Acinetobacter. The similarity was superior to 99% for Acinetobacter schindleri in the Basic Local Alignment Search Tool (BLAST), grouped in the clade superior to 90%. Several strains isolated from intensive care units (ICU) were resistant to various antibiotic classes. These techniques allowed for the identification of several microorganisms of importance in public health, enabling improvements in human infection control and proving the quality of inputs, food, and water.

Origin and population history of a recent colonizer, the yellow warbler in Galápagos and Cocos Islands.

The faunas associated with oceanic islands provide exceptional examples with which to examine the dispersal abilities of different taxa and test the relative contribution of selective and neutral processes in evolution. We examine the patterns of recent differentiation and the relative roles of gene flow and selection in genetic and morphological variation in the yellow warbler (Dendroica petechia aureola) from the Galápagos and Cocos Islands. Our analyses suggest aureola diverged from Central American lineages colonizing the Galápagos and Cocos Islands recently, likely less than 300 000 years ago. Within the Galápagos, patterns of genetic variation in microsatellite and mitochondrial markers suggest early stages of diversification. No intra-island patterns of morphological variation were found, even across steep ecological gradients, suggesting that either (i) high levels of gene flow may be homogenizing the effects of selection, (ii) populations may not have had enough time to accumulate the differences in morphological traits, or (iii) yellow warblers show lower levels of 'evolvability' than some other Galápagos species. By examining genetic data and morphological variation, our results provide new insight into the microevolutionary processes driving the patterns of variation.

Keratinolytic activity of Bacillus subtilis AMR using human hair.

AIMS: To determine the ability of a novel Bacillus subtilis AMR isolated from poultry waste to hydrolyse human hair producing peptidases including keratinases and hair keratin peptides. METHODS AND RESULTS: The Bacillus subtilis AMR was identified using biochemical tests and by analysis of 16S rDNA sequence. The isolate was grown in medium containing human hair as the sole source of carbon and nitrogen. The supplementation of hair medium (HM) with 0.01% yeast extract increased the keratinolytic activity 4.2-fold. B. subtilis AMR presented high keratinase production on the 8th day of fermentation in hair medium (HM) supplemented with 0.01% yeast extract (HMY) at pH 8.0. Keratinase yield was not correlated with increase in biomass. Zymography showed keratin-degrading peptidases migrating at c. 54, 80 and 100 kDa and gelatin-degrading bands at c. 80, 70 63, 54 32 and 15 kDa. Keratinases were optimally active at 50 degrees C and pH 9.0 and was fully inhibited by the serine proteinase inhibitor (PMSF). Scanning electron microscopy showed complete degradation of the hair cuticle after exposure to B. subtilis AMR grown in HMY. MALDI-TOF analysis of culture supernatant containing peptides produced during enzymatic hydrolysis of hair by B. subtilis AMR revealed fragments in a range of 800-2600 Da. CONCLUSIONS: This study showed that B. subtilis AMR was able to hydrolyse human hair producing serine peptidases with keratinase and gelatinase activity as well as hair keratin peptides. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report describing the production and partial characterization of keratinases by a B. subtilis strain grown in a medium containing human hair. These data suggest that peptides obtained from enzymatic hair hydrolysis may be useful for future applications on pharmaceutical and cosmetic formulations.

RIVER: Portuguese registry to monitor unnecessary right ventricular pacing.

The aim of this prospective registry is to evaluate a new algorithm designed to reduce the percentage of unnecessary ventricular pacing (%VP) in patients implanted with a dual-chamber pacemaker, through a dedicated pacing mode (called AAISafeR2) operating in AAI mode with back-up ventricular pacing in DDD mode, and to describe the incidence and distribution of atrioventricular (AV) block in this population. Investigators were free to assign patients to AAISafeR2 mode or to standard DDD (if AAISafeR was contraindicated, mainly due to permanent high-degree AV block). Patients underwent routine follow-up visits at 3, 6, 12, 18 and 24 months after implantation. At each follow-up visit, data were retrieved from pacemaker memories and analyzed to extract %VP and incidence of AV block. Up to December 2006, 158 patients (94 men, mean age 69 +/- 14 years) from nine Portuguese centers had been consecutively included. We also determined the distribution of AV block (according to the criteria used by the pacemaker to classify AV block and switch to DDD mode). AAISafeR was shown to be effective in reducing unnecessary VP in our patient population. The analysis also reveals a high incidence of paroxysmal AV block, often unknown at the time of implantation. There were no complications associated with AAISafeR programming.

Truncated human betaB1-crystallin shows altered structural properties and interaction with human betaA3-crystallin.

The purpose of the study was to determine the effects of truncation of various regions of betaB1-crystallin on its structural properties and stability of heterooligomers formed by wild-type (WT) betaB1 or its deletion mutants with WT betaA3-crystallin. For these analyses, seven deletion mutants of betaB1-crystallin were generated with the following sequential deletions of either N-terminal arm [betaB1(59-252)], N-terminal arm + motif I [betaB1(99-252)], N-terminal arm + motif I + motif II [betaB1(144-252)], N-terminal arm + motif I + motif II + connecting peptide [betaB1(149-252)], C-terminal extension [betaB1(1-234)], C-terminal extension plus motif IV [betaB1(1-190)], or C-terminal extension + motif III + motif IV [betaB1(1-148)]. The betaB1-crystallin became water insoluble on the deletion of C-terminal extension and subsequent deletions of the C-terminal domain (C-terminal extension plus motifs III and IV) while it remained partially soluble on the deletion of the N-terminal domain (N-terminal arm plus motifs I and II). However, circular dichroism spectral analysis showed that the deletion of the N-terminal domain but not the C-terminal domain exhibited relatively greater structural changes in the crystallin. The deletion of the C-terminal domain resulted in a greater exposure and disturbance in the microenvironment of Trp-100, Trp-123, and Trp-126 (localized in the motif II), suggesting a relatively greater role of the C-terminal domain than the N-terminal domain in the structural stability of the crystallin. The deletion of the N-terminal extension in betaB1 resulted in maximum exposure of hydrophobic patches and compact structure and in a maximum loss of subunit exchange with WT betaA3-crystallin compared to deletion of either the C-terminal extension, the N-terminal domain, or the C-terminal domain. The thermal stability results of the heterooligomer of betaB1- plus betaA3-crystallins suggested that oligomers lose their stability on deletion of the C-terminal domain. Together, the results suggested that the N-terminal arm of betaB1-crystallin plays a major role in interaction with betaA3-crystallin during heterooligomer formation, and the solubility of betaB1-crystallin per se and that of the heterooligomer with betaA3-crystallin are dependent on the intact C-terminal domain of betaB1-crystallin.

Profiling of patients with non-valvular atrial fibrillation and moderate-to-high risk of stroke not receiving oral anticoagulation in Spain.

OBJECTIVE: In non-valvular atrial fibrillation (NVAF) with embolic risk, the guidelines recommend oral anticoagulation (OAC), although not all patients receive it. In this study, an attempt is made to identify these patients, and to study factors related to non-anticoagulation. MATERIAL AND METHODS: Non-interventional, cross-sectional, multicentre study was performed on a population of patients ≥18 years with a NVAF diagnosis, moderate-high embolic risk (CHADS2 score≥2), not treated with OAC. Atrial fibrillation (AF) prevalence was also collected. RESULTS: AF prevalence was 4.5%, and 80.7% of the patients had NVAF (20.0% did not receive OAC). A total of 1310 non-OAC-treated patients were included (51.8% male, mean age: 76.0 years). The mean time since AF diagnosis was 58.4 months. The main therapeutic decision for stroke prevention was prescription of antiplatelet agents (82.4%, n=1078), and the main reasons were: patient refusal to monitoring (37.3%), high bleeding risk (31.1%), uncontrolled hypertension (27.9%), and frequent falls (27.6%). The mean CHA2DS2-VASc score was 4.6, and the HAS-BLED was 2.7 (55.9% of patients scoring HAS-BLED≥3). The most common thromboembolic risk factors were: hypertension (89.1%), age≥75 years (61.5%); the haemorrhagic factors: use of drugs increasing the bleeding risk (41.2%), uncontrolled blood pressure (33.7%). CONCLUSIONS: About 20% of Spanish NVAF patients do not receive OAC in the clinical practice and are treated with antiplatelet agents, which do not reduce haemorrhagic risk. Most patients do not clearly show a contraindication to OACs, particularly considering that there are other available options (direct oral anticoagulant drugs [DOACs]).

Deadbeat-fuzzy controller for the power control of a Doubly Fed Induction Generator based wind power system.

This paper proposes a Fuzzy Logic Controller for improvement of the steady-state response of a Doubly Fed Induction Generator used in a wind energy system, and governed by means of a Deadbeat Power Controller. The generator mathematical model is consistent with the Stator Flux oriented strategy in the synchronous reference frame. Different simulation scenarios were developed in Matlab/Simulink to evaluate the dynamic and the steady-state responses. In order to obtain experimental results, the simulated scenarios were repeated by means of a test bench and a Digital Signal Processor board. These results demonstrate that the response still follows the power references imposed, despite the fact that the generator parameters ( Rr, Ls and Lm) were varied in a 30%. A lower steady-state error is also achieved when compared with a Deadbeat and a classical PI controller. All the aforementioned evidence the proper application of this Fuzzy Controller in a wind power system based on a Doubly Fed Induction Generator.

Magnetoliposomes as model for signal transmission.

Liposomes containing magnetic nanoparticles (magnetoliposomes) have been extensively explored for targeted drug delivery. However, the magnetic effect of nanoparticles movement is also an attractive choice for the conduction of signals in communication systems at the nanoscale level because of the simple manipulation and efficient control. Here, we propose a model for the transmission of electrical and luminous signals taking advantage of magnetophoresis. The study involved three steps. Firstly, magnetite was synthesized and incorporated into fusogenic large unilamellar vesicles (LUVs) previously associated with a fluorescent label. Secondly, the fluorescent magnetite-containing LUVs delivered their contents to the giant unilamellar vesicles (GUVs), which were corroborated by magnetophoresis and fluorescence microscopy. In the third step, magnetophoresis of magnetic vesicles was used for the conduction of the luminous signal from a capillary to an optical fibre connected to a fluorescence detector. Also, the magnetophoresis effects on subsequent transmission of the electrochemical signal were demonstrated using magnetite associated with CTAB micelles modified with ferrocene. We glimpse that these magnetic supramolecular systems can be applied in micro- and nanoscale communication systems.

Isolation and characterization of betaA3-crystallin associated proteinase from alpha-crystallin fraction of human lenses.

PURPOSE: The purpose was to characterize the properties of a proteinase activity associated with betaA3-crystallin, which was isolated from the alpha-crystallin fraction of human lenses. METHODS: An inactive, Arg-bond hydrolyzing proteinase in the alpha-crystallin fraction, which was isolated from the water soluble (WS) protein fraction of 60- to 70-year-old human lenses, was activated by sodium deoxycholate treatment. The activated enzyme was purified by a three-step procedure that included a size-exclusion Agarose A1.5 m chromatography, non-denaturing preparative gel-electrophoresis, and size-exclusion HPLC. The purified proteinase was characterized for the proteinase type, proteolysis of bovine recombinant gammaB-, gammaC-, and gammaD-crystallins, and its presence in three different protein fractions of human lenses (i.e., alpha-crystallin, beta(H)-crystallin, and membrane fractions). RESULTS: An inactive, Arg-bond hydrolyzing proteinase present in the alpha-crystallin fraction showed activity on treatment with detergents such as sodium deoxycholate, Triton X-100, octyl beta-D-glucopyranoside, and CHAPS (3-[(3-cholamido propyl) dimethylammonio]-1-propanesulfonate). The sodium deoxycholate-activated enzyme was released from the alpha-crystallin fraction since it eluted at a lower molecular weight species than alpha-crystallin during size-exclusion Agarose A1.5 m chromatography. Following a three-step purification procedure, the enzyme showed three species between 22 kDa and 25 kDa during sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The three protein bands were identified as betaA3-, betaB1-, and betaB2-crystallin by the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and tandem mass spectrometric (ES-MS/MS) methods. Inhibitor studies revealed that the enzyme was a serine-type proteinase. Among the recombinant betaA3-, betaB1-, or betaB2-crystallins, only the betaA3-crystallin exhibited the proteinase activity following detergent treatment and size-exclusion chromatography. The proteinase also exhibited proteolysis of gammaC- and gammaD- crystallins, and the cleavage of gammaD-crystallin at M(1)-G(2), Q(54)-Y(55), M(70)-G(71), and Q(103)-M(104) bonds. Further, the enzyme was also present in three fractions of human lenses (alpha-crystallin, beta(H)-crystallin, and membrane fractions). CONCLUSIONS: A serine-type betaA3-crystallin proteinase existed in an inactive state in the alpha-crystallin fraction and was activated by detergents. The enzyme proteolyzed alphaA-, alphaB-, gammaC-, and gammaD-crystallins and was present in three fractions (alpha-crystallin, beta(H)-crystallin, and membrane-fractions) of 60 to 70-year-old human lenses.