Recent Developments in Synthesis, Properties, Applications and Recycling of Bio-Based Elastomers.

In 2021, global plastics production was 390.7 Mt; in 2022, it was 400.3 Mt, showing an increase of 2.4%, and this rising tendency will increase yearly. Of this data, less than 2% correspond to bio-based plastics. Currently, polymers, including elastomers, are non-recyclable and come from non-renewable sources. Additionally, most elastomers are thermosets, making them complex to recycle and reuse. It takes hundreds to thousands of years to decompose or biodegrade, contributing to plastic waste accumulation, nano and microplastic formation, and environmental pollution. Due to this, the synthesis of elastomers from natural and renewable resources has attracted the attention of researchers and industries. In this review paper, new methods and strategies are proposed for the preparation of bio-based elastomers. The main goals are the advances and improvements in the synthesis, properties, and applications of bio-based elastomers from natural and industrial rubbers, polyurethanes, polyesters, and polyethers, and an approach to their circular economy and sustainability. Olefin metathesis is proposed as a novel and sustainable method for the synthesis of bio-based elastomers, which allows for the depolymerization or degradation of rubbers with the use of essential oils, terpenes, fatty acids, and fatty alcohols from natural resources such as chain transfer agents (CTA) or donors of the terminal groups in the main chain, which allow for control of the molecular weights and functional groups, obtaining new compounds, oligomers, and bio-based elastomers with an added value for the application of new polymers and materials. This tendency contributes to the development of bio-based elastomers that can reduce carbon emissions, avoid cross-contamination from fossil fuels, and obtain a greener material with biodegradable and/or compostable behavior.

Effect of low dietary concentrations of Acacia mearnsii tannin extract on chewing, ruminal fermentation, digestibility, nitrogen partition, and performance of dairy cows.

The supplementation of dairy cows with tannins can reduce the ruminal degradation of dietary protein and urine N excretion, but high concentration in the diet can impair ruminal function, diet digestibility, feed intake, and milk yield. This study evaluated the effect of low concentrations (0, 0.14, 0.29, or 0.43% of diet in DM basis) of a tannin extract from the bark of Acacia mearnsii (TA) on milking performance, dry matter intake (DMI), digestibility, chewing behavior, ruminal fermentation, and N partition of dairy cows. Twenty Holstein cows (34.7 ± 4.8 kg/d, 590 ± 89 kg, and 78 ± 33 d in lactation) were individually fed a sequence of 4 treatments in 5, 4 × 4 Latin squares (with 21-d treatment periods, each with a 14-d adaptation period). The TA replaced citrus pulp in the total mixed ration and other feed ingredients were kept constant. Diets had 17.1% crude protein, mostly from soybean meal and alfalfa haylage. The TA had no detected effect on DMI (22.1 kg/d), milk yield (33.5 kg/d), and milk components. The proportions in milk fat of mixed origin fatty acids (16C and 17C) and the daily secretion of unsaturated fatty acids were linearly reduced and the proportion of de novo fatty acids was increased by TA. Cows fed TA had linear increase in the molar proportion of butyrate and linear reduction in propionate in ruminal fluid, whereas acetate did not differ. There was a tendency for the ratio of acetate to propionate to be linearly increased by TA. Cows fed TA had a linear reduction in the relative ruminal microbial yield, estimated by the concentrations of allantoin and creatinine in urine and body weight. The total-tract apparent digestibility of neutral detergent fiber, starch, and crude protein also did not differ. The TA induced a linear increase in meal size and duration of the first daily meal and reduced meal frequency. Rumination behavior did not differ with treatment. Cows fed 0.43% TA selected against feed particles >19 mm in the morning. There were tendencies for linear decreases in milk urea N (16.1-17.3 mg/dL), urine N (153-168 g/d and 25.5-28.7% of N intake), and plasma urea N at 6, 18, and 21 h postmorning feeding, and plasma urea N 12 h postfeeding was reduced by TA. The proportion of N intake in milk (27.1%) and feces (21.4%) did not differ with treatment. Reductions in urine N excretion and milk and plasma urea N suggest that TA reduced ruminal AA deamination, whereas lactation performance did not differ. Overall, TA up to 0.43% of DM did not affect DMI and lactation performance, while there was a tendency to reduce urine N excretion.

Deletion of the phosphatase INPP5E in the murine retina impairs photoreceptor axoneme formation and prevents disc morphogenesis.

INPP5E, also known as pharbin, is a ubiquitously expressed phosphatidylinositol polyphosphate 5-phosphatase that is typically located in the primary cilia and modulates the phosphoinositide composition of membranes. Mutations to or loss of INPP5E is associated with ciliary dysfunction. INPP5E missense mutations of the phosphatase catalytic domain cause Joubert syndrome in humans-a syndromic ciliopathy affecting multiple tissues including the brain, liver, kidney, and retina. In contrast to other primary cilia, photoreceptor INPP5E is prominently expressed in the inner segment and connecting cilium and absent in the outer segment, which is a modified primary cilium dedicated to phototransduction. To investigate how loss of INPP5e causes retina degeneration, we generated mice with a retina-specific KO (Inpp5eF/F;Six3Cre, abbreviated as retInpp5e-/-). These mice exhibit a rapidly progressing rod-cone degeneration resembling Leber congenital amaurosis that is nearly completed by postnatal day 21 (P21) in the central retina. Mutant cone outer segments contain vesicles instead of discs as early as P8. Although P10 mutant outer segments contain structural and phototransduction proteins, axonemal structure and disc membranes fail to form. Connecting cilia of retInpp5e-/- rods display accumulation of intraflagellar transport particles A and B at their distal ends, suggesting disrupted intraflagellar transport. Although INPP5E ablation may not prevent delivery of outer segment-specific proteins by means of the photoreceptor secretory pathway, its absence prevents the assembly of axonemal and disc components. Herein, we suggest a model for INPP5E-Leber congenital amaurosis, proposing how deletion of INPP5E may interrupt axoneme extension and disc membrane elaboration.

In Vivo Photocontrol of Microtubule Dynamics and Integrity, Migration and Mitosis, by the Potent GFP-Imaging-Compatible Photoswitchable Reagents SBTubA4P and SBTub2M.

Photoswitchable reagents are powerful tools for high-precision studies in cell biology. When these reagents are globally administered yet locally photoactivated in two-dimensional (2D) cell cultures, they can exert micron- and millisecond-scale biological control. This gives them great potential for use in biologically more relevant three-dimensional (3D) models and in vivo, particularly for studying systems with inherent spatiotemporal complexity, such as the cytoskeleton. However, due to a combination of photoswitch isomerization under typical imaging conditions, metabolic liabilities, and insufficient water solubility at effective concentrations, the in vivo potential of photoswitchable reagents addressing cytosolic protein targets remains largely unrealized. Here, we optimized the potency and solubility of metabolically stable, druglike colchicinoid microtubule inhibitors based on the styrylbenzothiazole (SBT) scaffold that are nonresponsive to typical fluorescent protein imaging wavelengths and so enable multichannel imaging studies. We applied these reagents both to 3D organoids and tissue explants and to classic model organisms (zebrafish, clawed frog) in one- and two-protein imaging experiments, in which spatiotemporally localized illuminations allowed them to photocontrol microtubule dynamics, network architecture, and microtubule-dependent processes in vivo with cellular precision and second-level resolution. These nanomolar, in vivo capable photoswitchable reagents should open up new dimensions for high-precision cytoskeleton research in cargo transport, cell motility, cell division, and development. More broadly, their design can also inspire similarly capable optical reagents for a range of cytosolic protein targets, thus bringing in vivo photopharmacology one step closer to general realization.

Chlorophyll-a Pigment Measurement of Spirulina in Algal Growth Monitoring Using Portable Pulsed LED Fluorescence Lidar System.

Chlorophyll-a measurement is important in algal growth and water quality monitoring in natural waters. A portable pulsed LED fluorescence lidar system based on the preliminary algal organic matter and pigments excitation-emission matrix (EEM) of commercialized AZTEC Spirulina powder at varying concentrations was developed. Fluorescence peaks from EEMs showed increasing intensity as the Spirulina concentration increases. Using this information, an LED fluorescence lidar with a wavelength of 385 nm, pulse width of 10 ns, and repetition frequency of 500 kHz was constructed for chlorophyll detection at 680 nm. Turbidity measurements were also conducted at 700 nm emission wavelength at the same excitation wavelength. Range-resolved fluorescence lidar signals from the portable pulsed LED fluorescence lidar system are highly correlated with the standard methods such as optical density at 680 nm (R2 = 0.87), EEM fluorescence chlorophyll-a pigment at 680 nm (R2 = 0.89), and corrected chlorophyll-a concentration (R2 =0.92). The F680/F700 lidar ratio was measured to provide a linear relationship of chlorophyll-a and turbidity in waters. The F680/F700 measurement showed strong correlations with Spirulina concentration (R2 = 0.94), absorbance at 680 nm (R2 = 0.84), EEM chlorophyll-a pigment at 680 nm (R2 = 0.83), and corrected chlorophyll-a concentration (R2 = 0.86). Results revealed that this new technique of chlorophyll-a measurement can be used as an alternative to other standard methods in algal growth monitoring.

Species loss due to nutrient addition increases with spatial scale in global grasslands.

The effects of altered nutrient supplies and herbivore density on species diversity vary with spatial scale, because coexistence mechanisms are scale dependent. This scale dependence may alter the shape of the species-area relationship (SAR), which can be described by changes in species richness (S) as a power function of the sample area (A): S = cAz , where c and z are constants. We analysed the effects of experimental manipulations of nutrient supply and herbivore density on species richness across a range of scales (0.01-75 m2 ) at 30 grasslands in 10 countries. We found that nutrient addition reduced the number of species that could co-occur locally, indicated by the SAR intercepts (log c), but did not affect the SAR slopes (z). As a result, proportional species loss due to nutrient enrichment was largely unchanged across sampling scales, whereas total species loss increased over threefold across our range of sampling scales.

The small GTPase RAB28 is required for phagocytosis of cone outer segments by the murine retinal pigmented epithelium.

RAB28, a member of the RAS oncogene family, is a ubiquitous, farnesylated, small GTPase of unknown function present in photoreceptors and the retinal pigmented epithelium (RPE). Nonsense mutations of the human RAB28 gene cause recessive cone-rod dystrophy 18 (CRD18), characterized by macular hyperpigmentation, progressive loss of visual acuity, RPE atrophy, and severely attenuated cone and rod electroretinography (ERG) responses. In an attempt to elucidate the disease-causing mechanism, we generated Rab28-/- mice by deleting exon 3 and truncating RAB28 after exon 2. We found that Rab28-/- mice recapitulate features of the human dystrophy (i.e. they exhibited reduced cone and rod ERG responses and progressive retina degeneration). Cones of Rab28-/- mice extended their outer segments (OSs) to the RPE apical processes and formed enlarged, balloon-like distal tips before undergoing degeneration. The visual pigment content of WT and Rab28-/- cones was comparable before the onset of degeneration. Cone phagosomes were almost absent in Rab28-/- mice, whereas rod phagosomes displayed normal levels. A protein-protein interaction screen identified several RAB28-interacting proteins, including the prenyl-binding protein phosphodiesterase 6 δ-subunit (PDE6D) and voltage-gated potassium channel subfamily J member 13 (KCNJ13) present in the RPE apical processes. Of note, the loss of PDE6D prevented delivery of RAB28 to OSs. Taken together, these findings reveal that RAB28 is required for shedding and phagocytosis of cone OS discs.

Neural Network Direct Control with Online Learning for Shape Memory Alloy Manipulators.

New actuators and materials are constantly incorporated into industrial processes, and additional challenges are posed by their complex behavior. Nonlinear hysteresis is commonly found in shape memory alloys, and the inclusion of a suitable hysteresis model in the control system allows the controller to achieve a better performance, although a major drawback is that each system responds in a unique way. In this work, a neural network direct control, with online learning, is developed for position control of shape memory alloy manipulators. Neural network weight coefficients are updated online by using the actuator position data while the controller is applied to the system, without previous training of the neural network weights, nor the inclusion of a hysteresis model. A real-time, low computational cost control system was implemented; experimental evaluation was performed on a 1-DOF manipulator system actuated by a shape memory alloy wire. Test results verified the effectiveness of the proposed control scheme to control the system angular position, compensating for the hysteretic behavior of the shape memory alloy actuator. Using a learning algorithm with a sine wave as reference signal, a maximum static error of 0.83° was achieved when validated against several set-points within the possible range.

Tyro3 Modulates Mertk-Associated Retinal Degeneration.

Inherited photoreceptor degenerations (IPDs) are the most genetically heterogeneous of Mendelian diseases. Many IPDs exhibit substantial phenotypic variability, but the basis is usually unknown. Mutations in MERTK cause recessive IPD phenotypes associated with the RP38 locus. We have identified a murine genetic modifier of Mertk-associated photoreceptor degeneration, the C57BL/6 (B6) allele of which acts as a suppressor. Photoreceptors degenerate rapidly in Mertk-deficient animals homozygous for the 129P2/Ola (129) modifier allele, whereas animals heterozygous for B6 and 129 modifier alleles exhibit an unusual intermixing of degenerating and preserved retinal regions, with females more severely affected than males. Mertk-deficient mice homozygous for the B6 modifier allele display degeneration only in the far periphery, even at 8 months of age, and have improved retinal function compared to animals homozygous for the 129 allele. We genetically mapped the modifier to an approximately 2-megabase critical interval that includes Tyro3, a paralog of Mertk. Tyro3 expression in the outer retina varies with modifier genotype in a manner characteristic of a cis-acting expression quantitative trait locus (eQTL), with the B6 allele conferring an approximately three-fold higher expression level. Loss of Tyro3 function accelerates the pace of photoreceptor degeneration in Mertk knockout mice, and TYRO3 protein is more abundant in the retinal pigment epithelium (RPE) adjacent to preserved central retinal regions of Mertk knockout mice homozygous for the B6 modifier allele. Endogenous human TYRO3 protein co-localizes with nascent photoreceptor outer segment (POS) phagosomes in a primary RPE cell culture assay, and expression of murine Tyro3 in cultured cells stimulates phagocytic ingestion of POS. Our findings demonstrate that Tyro3 gene dosage modulates Mertk-associated retinal degeneration, provide strong evidence for a direct role for TYRO3 in RPE phagocytosis, and suggest that an eQTL can modify a recessive IPD.

Novel Automated Approach to Predict the Outcome of Laser Peripheral Iridotomy for Primary Angle Closure Suspect Eyes Using Anterior Segment Optical Coherence Tomography.

Develop an algorithm to predict the success of laser peripheral iridotomy (LPI) in primary angle closure suspect (PACS), using pre-treatment anterior segment optical coherence tomography (ASOCT) scans. A total of 116 eyes with PACS underwent LPI and time-domain ASOCT scans (temporal and nasal cuts) were performed before and 1 month after LPI. All the post-treatment scans were classified to one of the following categories: (a) both angles open, (b) one of two angles open and (c) both angles closed. After LPI, success is defined as one or more angles changed from close to open. In this proposed method, the pre and post-LPI ASOCT scans were registered at the corresponding angles based on similarities between the respective local descriptor features and random sample consensus technique was used to identify the largest consensus set of correspondences between the pre and post-LPI ASOCT scans. Subsequently, features such as correlation co-efficient (CC) and structural similarity index (SSIM) were extracted and correlated with the success of LPI. We included 116 eyes and 91 (78.44%) eyes fulfilled the criteria for success after LPI. Using the CC and SSIM index scores from this training set of ASOCT images, our algorithm showed that the success of LPI in eyes with narrow angles can be predicted with 89.7% accuracy, specificity of 95.2% and sensitivity of 36.4% based on pre-LPI ASOCT scans only. Using pre-LPI ASOCT scans, our proposed algorithm showed good accuracy in predicting the success of LPI for PACS eyes. This fully-automated algorithm could aid decision making in offering LPI as a prophylactic treatment for PACS.

Arf-like Protein 3 (ARL3) Regulates Protein Trafficking and Ciliogenesis in Mouse Photoreceptors.

Arf-like protein 3 (ARL3) is a ubiquitous small GTPase expressed in ciliated cells of plants and animals. Germline deletion ofArl3in mice causes multiorgan ciliopathy reminiscent of Bardet-Biedl or Joubert syndromes. As photoreceptors are elegantly compartmentalized and have cilia, we probed the function of ARL3 (ADP-ribosylation factor (Arf)-like 3 protein) by generating rod photoreceptor-specific (prefix(rod)) and retina-specific (prefix(ret))Arl3deletions. In predegenerate(rod)Arl3(-/-)mice, lipidated phototransduction proteins showed trafficking deficiencies, consistent with the role of ARL3 as a cargo displacement factor for lipid-binding proteins. By contrast,(ret)Arl3(-/-)rods and cones expressing Cre recombinase during embryonic development formed neither connecting cilia nor outer segments and degenerated rapidly. Absence of cilia infers participation of ARL3 in ciliogenesis and axoneme formation. Ciliogenesis was rescued, and degeneration was reversed in part by subretinal injection of adeno-associated virus particles expressing ARL3-EGFP. The conditional knock-out phenotypes permitted identification of two ARL3 functions, both in the GTP-bound form as follows: one as a regulator of intraflagellar transport participating in photoreceptor ciliogenesis and the other as a cargo displacement factor transporting lipidated protein to the outer segment. Surprisingly, a farnesylated inositol polyphosphate phosphatase only trafficked from the endoplasmic reticulum to the Golgi, thereby excluding it from a role in photoreceptor cilia physiology.

Validation of the AFP model as a predictor of HCC recurrence in patients with viral hepatitis-related cirrhosis who had received a liver transplant for HCC.

BACKGROUND & AIMS: The AFP model was shown to be superior to the Milan criteria for predicting hepatocellular carcinoma (HCC) recurrence after liver transplantation in a French population. Our aim was to test the AFP model in a non-French, post-hepatitic cirrhosis-based population of HCC candidates. METHODS: 574 patients transplanted for HCC in four Italian centers were studied. AFP score was assessed at the last evaluation before liver transplantation (LT). Probabilities of recurrence and survival were estimated by the log-rank test or competing risk analysis and compared according to the AFP model. RESULTS: 24.7% patients were beyond Milan criteria. HCC complicated hepatitis C virus (HCV) and hepatitis B virus (HBV) cirrhosis in 58.7% and 24% of the cases, respectively. Five-year probabilities of recurrence differed according to AFP score ⩽2 vs. >2 in the whole population (13.2±1.8% vs. 49.8±8.7%, p<0.001, HR=4.98), in patients within Milan criteria (12.8±2.0% vs. 32.4±12.1%, p=0.009, HR=3.51), beyond Milan criteria (14.9±4.2% vs. 58.9±11.5%, p<0.001, HR=4.26), HCV patients (14.9±2.5% vs. 67.6±14.7%, p<0.001, HR=6.56) and HBV patients (11.6±3.4% vs. 34.3±12.5%, p=0.012, HR=3.49). By net reclassification improvement analysis AFP score significantly improved prediction of non-recurrence compared to Milan criteria. Overall five-year survival rates according to AFP score ⩽2 or >2 were 71.7±2.2% vs. 42.2±8.3% (p<0.001, HR=2.14). CONCLUSIONS: The AFP model identifies HCC candidates at low risk of recurrence, otherwise excluded by Milan criteria in a population with a predominance of post-hepatitic-related HCC. The AFP score can be proposed for selection of HCC candidates in programs with a high proportion of viral/HCV-related cirrhosis. LAY SUMMARY: Selection criteria for liver transplantation of patients affected with hepatocellular carcinoma (HCC) are based on the Milan criteria, which have been shown to be too restrictive, precluding access to liver transplantation for some patients who might be cured by this operation. Recently, a French group of researchers developed a new selection model called the AFP model, or AFP score, allowing some patients with HCC not meeting Milan criteria to be transplanted with excellent results. In the present work, the AFP score was tested in a population of non-French patients transplanted for HCC occurring mainly on post-hepatitic (HCV or HBV) cirrhosis. The results confirm that in this specific population, as in the original French population of patients, the AFP model better selects patients with HCC eligible for transplantation, compared to Milan criteria. We conclude that the AFP score, which has been officially adopted by the French organization for Organ Sharing for HCC patients, can also be implemented in countries with an important burden of HCC occurring on post-hepatitic cirrhosis.

Seroprevalence of dengue in a rural and an urbanized village: A pilot study from rural western India.

BACKGROUND & OBJECTIVES: Dengue is highly prevalent in tropical and subtropical regions. The prevalence of dengue is influenced by number of factors, i.e. host, vector, virus and environmental conditions including urbanization and population density. A cross sectional study was undertaken to determine the seroprevalence of dengue in two selected villages that differed in the level of their urbanization and population density. METHODS: Two villages with demographically well-defined populations close to Pune, a metropolitan city of western India, were selected for the study. Age stratified serosurvey was carried out during February to May 2011 in the two villages-a rural village A, located 6 km from the national highway with a population density of 159/km2 ; and an urbanized village B, located along the highway with a population density of 779/km2 . Assuming a low seroposi- tivity of 10%, 702 serum samples were collected from village A. Sample size for village B was calculated on the basis of seropositivity obtained in village A, and 153 samples were collected. Serum samples were tested for the presence of dengue virus (DENV)-specific IgG. Simple proportional analyses were used to calculate and compare the seroprevalence. RESULTS: Of the 702 samples collected from village A, 42.8% were found positive for anti-DENV IgG. A significantly higher seropositivity for DENV (58.8%) was found in village B. In village A, there was an age dependent increase in seroprevalence; whereas, in village B, there was a steep increase from 17% positivity in 0-10 yr age group to 72% in the 11-20 yr age group. The seroprevalence was almost similar in the older age groups. INTERPRETATION & CONCLUSION: The observations suggested that prevalence of dengue is probably associated with urbanization and host population density. Areas that are in the process of urbanization needs to be monitored for prevalence of dengue and its vector, and appropriate vector control measures may be implemented.

Knockdown of unc119c results in visual impairment and early-onset retinal dystrophy in zebrafish.

PURPOSE: UNC119 proteins are involved in G protein trafficking in mouse retinal photoreceptors and Caenorhabditis elegans olfactory neurons. An Unc119 null allele is associated with cone-rod dystrophy in mouse, but the mechanism leading to disease is not understood. We studied the role of Unc119 paralogs and Arl3l2 in zebrafish vision and retinal organization resulting from unc119c and arl3l2 knockdown. METHODS: Zebrafish unc119c was amplified by PCR from retina and pineal gland cDNA. Its expression pattern in the eye and pineal gland was determined by whole-mount in-situ hybridization. unc119c and arl3l2 were knocked down using morpholino-modified oligonucleotides (MO). Their visual function was assessed with a quantitative optomotor assay on 6 days post-fertilization larvae. Retinal morphology was analyzed using immunohistochemistry with anti-cone arrestin (zpr-1) and anti-cone transducin-α (GNAT2) antibodies. RESULTS: The zebrafish genome contains four genes encoding unc119 paralogs located on different chromosomes. The exon/intron arrangements of these genes are identical. Three Unc119 paralogs are expressed in the zebrafish retina, termed Unc119a-c. Based on sequence similarity, Unc119a and Unc119b are orthologs of mammalian UNC119a and UNC119b, respectively. A third, Unc119c, is unique and not present in mammals. Whole mount in-situ hybridization revealed that unc119a and unc119b RNA are ubiquitously expressed in the CNS, and unc119c is specifically expressed in photoreceptive tissues (pineal gland and retina). A Unc119 interactant, Arl3l2 also localizes to the pineal gland and the retina. As measured by the optomotor response, unc119c and arl3l2 knockdown resulted in significantly lower vision compared to wild-type zebrafish larvae and control morpholino (MO). Immunohistological analysis with anti-cone transducin and anti-cone arrestin (zpr-1) indicates that knockdown of unc119c leads to photoreceptor degeneration mostly affecting cones. CONCLUSIONS: Our results suggest that Unc119c is the only Unc119 paralog that is highly specific to the retina in zebrafish. Unc119c and Arl3l2 proteins are important for the function of cones.

Mistrafficking of prenylated proteins causes retinitis pigmentosa 2.

The retinitis pigmentosa 2 polypeptide (RP2) functions as a GTPase-activating protein (GAP) for ARL3 (Arf-like protein 3), a small GTPase. ARL3 is an effector of phosphodiesterase 6 Δ (PDE6D), a prenyl-binding protein and chaperone of prenylated protein in photoreceptors. Mutations in the human RP2 gene cause X-linked retinitis pigmentosa (XLRP) and cone-rod dystrophy (XL-CORD). To study mechanisms causing XLRP, we generated an RP2 knockout mouse. The Rp2h(-/-) mice exhibited a slowly progressing rod-cone dystrophy simulating the human disease. Rp2h(-/-) scotopic a-wave and photopic b-wave amplitudes declined at 1 mo of age and continued to decline over the next 6 mo. Prenylated PDE6 subunits and G-protein coupled receptor kinase 1 (GRK1) were unable to traffic effectively to the Rp2h(-/-) outer segments. Mechanistically, absence of RP2 GAP activity increases ARL3-GTP levels, forcing PDE6D to assume a predominantly "closed" conformation that impedes binding of lipids. Lack of interaction disrupts trafficking of PDE6 and GRK1 to their destination, the photoreceptor outer segments. We propose that hyperactivity of ARL3-GTP in RP2 knockout mice and human patients with RP2 null alleles leads to XLRP resembling recessive rod-cone dystrophy.

Altered profile of regulatory T cells and associated cytokines in mild and moderate dengue.

The pathogenesis of dengue is immune-mediated. Regulatory T cells suppress immune response and may contribute to better prognosis. The present study evaluates Tregs and cytokines in dengue patients in the context of disease severity, time of sampling and immune status. The cohort included 90 patients (51 mild, 39 moderate) and 27 healthy controls. Frequencies of Tregs, CD4(+)CD25(-)Foxp3(+) T cells and CD3(+), CD3(+)CD4(+) and CD3(+)CD8(+) T cells were enumerated by flow cytometry. Circulating levels of 15 cytokines/chemokines were measured using Luminex technology and mRNA levels of Foxp3, IL-10 and TGF-ß were assessed by real-time polymerase chain reaction (PCR). Significantly higher frequencies of Tregs were observed in mild cases, especially during post-defervescence. The difference between mild and moderate cases was more evident in secondary infections. Frequencies of T cells were higher in mild cases but during pre-defervescence. On the other hand, the levels of IL-6, IL-7, IL-8, TNF-α and IL-10 were significantly higher in moderate cases. IL-6 and IL-8 levels correlated negatively with Treg frequencies during post-defervescence and in secondary infections. Higher levels of IL-10 and TGF-ß in moderate cases were not reflected by their corresponding mRNA levels. Platelet counts correlated positively with Treg frequencies and TGF-ß levels, and negatively with IL-10 levels. Higher Treg frequencies may favour a beneficial outcome in dengue. Higher cytokine levels may indirectly contribute to disease severity by exerting an inhibitory influence on Tregs. The dichotomy between mRNA and proteins levels for IL-10 and TGF-ß is suggestive of increased translational efficiency.

Centrin 2 is required for mouse olfactory ciliary trafficking and development of ependymal cilia planar polarity.

Centrins are ancient calmodulin-related Ca(2+)-binding proteins associated with basal bodies. In lower eukaryotes, Centrin2 (CETN2) is required for basal body replication and positioning, although its function in mammals is undefined. We generated a germline CETN2 knock-out (KO) mouse presenting with syndromic ciliopathy including dysosmia and hydrocephalus. Absence of CETN2 leads to olfactory cilia loss, impaired ciliary trafficking of olfactory signaling proteins, adenylate cyclase III (ACIII), and cyclic nucleotide-gated (CNG) channel, as well as disrupted basal body apical migration in postnatal olfactory sensory neurons (OSNs). In mutant OSNs, cilia base-anchoring of intraflagellar transport components IFT88, the kinesin-II subunit KIF3A, and cytoplasmic dynein 2 appeared compromised. Although the densities of mutant ependymal and respiratory cilia were largely normal, the planar polarity of mutant ependymal cilia was disrupted, resulting in uncoordinated flow of CSF. Transgenic expression of GFP-CETN2 rescued the Cetn2-deficiency phenotype. These results indicate that mammalian basal body replication and ciliogenesis occur independently of CETN2; however, mouse CETN2 regulates protein trafficking of olfactory cilia and participates in specifying planar polarity of ependymal cilia.

Interaction of host cell microRNAs with the HCV RNA genome during infection of liver cells.

It has remained an enigma how hepatitis C viral (HCV) RNA can persist in the liver of infected patients for many decades. With the recent discovery of roles for microRNAs in gene expression, it was reported that the HCV RNA genome subverts liver-specific microRNA miR-122 to protect its 5' end from degradation by host cell exoribonucleases. Sequestration of miR-122 in cultured liver cells and in the liver of chimpanzees by small, modified antisense RNAs resulted in dramatic loss of HCV RNA and viral yield. This finding led to the first successful human trial in which subcutaneous administration of antisense molecules against miR-122 lowered viral yield in HCV patients, without the emergence of resistant virus. In this review, the authors summarize the molecular mechanism by which miR-122 protects the HCV RNA genome from degradation by exoribonucleases Xrn1 and Xrn2 and discuss the application of miR-122 antisense molecules in the clinic.

Acquisition and loss of desiccation tolerance in seeds: from experimental model to biological relevance.

MAIN CONCLUSION: Besides being an important model to study desiccation tolerance, the induction of desiccation tolerance in germinated seeds may also play an ecological role in seedling establishment. Desiccation tolerance (DT) is the ability of certain organisms to survive extreme water losses without accumulation of lethal damage. This was a key feature in the conquering of dry land and is currently found in all taxa including bacteria, fungi, roundworms and plants. Not surprisingly, studies in various fields have been performed to unravel this intriguing phenomenon. In flowering plants, DT is rare in whole plants (vegetative tissues), yet is common in seeds. In this review, we present our current understanding of the evolution of DT in plants. We focus on the acquisition of DT in seeds and the subsequent loss during and after germination by highlighting and comparing research in two model plants Medicago truncatula and Arabidopsis thaliana. Finally, we discuss the ability of seeds to re-establish DT during post-germination, the possible ecological meaning of this phenomenon, and the hypothesis that DT, in combination with dormancy, optimizes seedling establishment.