In Vivo Cross-Linking Sheds Light on the Salmonella Divisome in Which PBP3 and PBP3SAL Compete for Occupancy.

Bacterial cell division is orchestrated by proteins that assemble in dynamic complexes collectively known as the divisome. Essential monofunctional enzymes with glycosyltransferase or transpeptidase (TPase) activities, FtsW and FtsI respectively, engage in the synthesis of septal peptidoglycan (sPG). Enigmatically, Salmonella has two TPases that can promote cell division independently: FtsI (PBP3) and the pathogen-specific paralogue PBP3SAL. How Salmonella regulates the assembly of the sPG synthase complex with these two TPases, is unknown. Here, we characterized Salmonella division complexes in wild-type cells and isogenic mutants lacking PBP3 or PBP3SAL. The complexes were cross-linked in vivo and pulled down with antibodies recognizing each enzyme. Proteomics of the immunoprecipitates showed that PBP3 and PBP3SAL do not extensively cross-link in wild type cells, supporting the presence of independent complexes. More than 40 proteins cross-link in complexes in which these two TPases are present. Those identified with high scores include FtsA, FtsK, FtsQLB, FtsW, PBP1B, SPOR domain-containing proteins (FtsN, DedD, RlpA, DamX), amidase activators (FtsX, EnvC, NlpD) and Tol-Pal proteins. Other cross-linked proteins are the protease Prc, the elongasome TPase PBP2 and, D,D-endo- and D,D-carboxypeptidases. PBP3 and PBP3SAL localize at midcell and compete for occupying the division complex in response to environmental cues. Thus, a catalytic-dead PBP3SAL-S300A variant impairs cell division in a high osmolarity and acidic condition in which it is produced at levels exceeding those of PBP3. Salmonella may therefore exploit an 'adjustable' divisome to exchange TPases for ensuring cell division in distinct environments and, in this manner, expand its colonization capacities.

The copper (II) complex of salicylate phenanthroline inhibits proliferation and induces apoptosis of hepatocellular carcinoma cells.

In the present study, we investigated the antitumor effect and associated molecular mechanisms of the copper (II) complex of salicylate phenanthroline [Cu(sal)(phen)] against hepatocellular carcinoma (HCC). Cu(sal)(phen) inhibited the proliferation of HCC cells (HepG2 and HCC-LM9) and induced apoptosis of HCC cells in a dose-dependent manner by upregulating mitochondrial reactive oxygen species (ROS) production. The expression of the antiapoptotic proteins survivin and Bcl-2 was decreased, while the expression of the DNA damage marker γ-H2 AX and the apoptotic marker cleaved PARP was upregulated with Cu(sal)(phen) treatment. In vivo, the growth of HepG2 subcutaneous xenograft tumors was greatly attenuated by Cu(sal)(phen) treatment. Immunohistochemistry staining showed that the expression of survivin, Bcl-2, and Ki67 in the tumor was downregulated by Cu(sal)(phen). Toxicity experiments with BALB/c mice revealed that Cu(sal)(phen) is a relatively safe drug. Our results indicate that Cu(sal)(phen) possesses great potential as a therapeutic drug for HCC.

The role of protected areas co-management in enhancing resistance and resilience of deciduous forest ecosystem to extreme climatic events in Bangladesh.

Due to ongoing and projected climate change as well as increasing anthropogenic disturbances, the tropical deciduous forest has been experiencing a decline in its biomass and productivity. To mitigate this adverse effect, many tropical countries have adopted forest co-management engaging local communities. However, the effects of co-management on the resistance and resilience of forest ecosystems to extreme climatic events have rarely been tested. The present study investigates the effects of co-management on resistance and resilience to extreme climatic events in two major tropical deciduous forest protected areas of Bangladesh, namely Madhupur National Park (MNP) and Bhawal National Park (BNP), through remotely sensed satellite data. We used the Google Earth Engine platform to access the Landsat images from 1990 to 2020 for a comprehensive assessment of the forest cover condition under two major management regimes (i.e., traditional and co-management). We find that co-management slows down the rate of forest destruction, where the rate of forest destruction was 108 ha year-1 in MNP and 121 ha year-1 in BNP during the year 1990-2008 under traditional forest management system. Under the co-management regime, forest cover increased by 19 ha year-1 and 41 ha year-1 from 2009 to 2020 respectively in MNP and BNP. Our study finds a highly significant correlation between rainfall (p < 0.001) and forest health, although co-management had poor impacts on forest resistance and resilience in case of extreme climatic events, such as drought and heavy rainfall. We find, no significant impacts of co-management on resistance and resilience to drought in MNP, and on resistance and resilience to heavy rainfall in MNP and BNP. In BNP, the impacts of co-management on resistance (p < 0.05) and resilience (p < 0.01) of forest to drought were highly significant. Forest co-management although have the potentials to reduce the deforestation rate by mitigating anthropogenic disturbances, its capacity to tackle the adverse impact of climate change was limited in our study. An adaptive co-management model, therefore, is crucial for mainstreaming the adverse effect of climate change on the tropical deciduous forest to harness the maximum potential of community participation in forest resources management.

[Correlations between dietary salt intake and 24-h urine sodium excretion in a sample of the urban population of Cáceres]. / Correlaciones entre la ingesta de sal en la dieta y la eliminación de sodio en la orina de 24 h en una muestra de población urbana de Cáceres.

OBJECTIVE: Excessive salt intake is associated with higher levels of AHT and prevalence of cardiovascular diseases. WHO recommends consuming less than 5 g/day of salt (equivalent to 2 g Na+/day). Identifying foods and behaviours with greater contribution to excess intake would facilitate preventive dietary advice. DESIGN: Observational study. SITE: Urban Health Center in Cáceres. MAIN MEASUREMENTS: We estimated salt consumption using two food consumption frequency (FFQ) questionnaires, one generic and one with high Na+ content, and a 24 h follow-up survey. We use the Evalfinut program for nutritional evaluation of the diet. We analyzed correlations between estimated salt intake and 24-h urine sodium elimination (gold standard). RESULTS: 92% of the population had consumption higher than the recommendations with intakes equivalent to 9.5 g/day of salt (3.7 g/d of Na+). When sodium intake is determined by the Na+/K+ ratio, 79.54% have high intakes. Sodium elimination follows a slightly increasing trend with BMI. The perception of salt consumption is low, 56.3% consider it "adequate". 32.4% add salt to food once served. FFQ underestimate Na+ intake and targeted surveys provide higher values. The correlation between FFQ and Na+ urinary elimination is weak. CONCLUSION: We must reduce salt intake by increasing the perception of consumption, improving knowledge about the salt content in food and identifying target patients of the health council.

Transcytosis of IgA Attenuates Salmonella Invasion in Human Enteroids and Intestinal Organoids.

Secretory IgA (SIgA) is the most abundant antibody type in intestinal secretions where it contributes to safeguarding the epithelium from invasive pathogens like the Gram-negative bacterium, Salmonella enterica serovar Typhimurium (STm). For example, we recently reported that passive oral administration of the recombinant monoclonal SIgA antibody, Sal4, to mice promotes STm agglutination in the intestinal lumen and restricts bacterial invasion of Peyer's patch tissues. In this report, we sought to recapitulate Sal4-mediated protection against STm in human Enteroids and human intestinal organoids (HIOs) as models to decipher the molecular mechanisms by which antibodies function in mucosal immunity in the human gastrointestinal tract. We confirm that Enteroids and HIO-derived monolayers are permissive to STm infection, dependent on HilD, the master transcriptional regulator of the SPI-I type three secretion system (T3SS). Stimulation of M-like cells in both Enteroids and HIOs by the addition of RANKL further enhanced STm invasion. The apical addition of Sal4 mouse IgA, as well as recombinant human Sal4 dimeric IgA (dIgA) and SIgA resulted a dose-dependent reduction in bacterial invasion. Moreover, basolateral application of Sal4 dIgA to Enteroid and HIO monolayers gave rise to SIgA in the apical compartment via a pathway dependent on expression of the polymeric immunoglobulin receptor (pIgR). The resulting Sal4 SIgA was sufficient to reduce STm invasion of Enteroid and HIO epithelial cell monolayers by ~20-fold. Recombinant Sal4 IgG was also transported in the Enteroid and HIOs, but to a lesser degree and via a pathway dependent on the neonatal Fc receptor (FCGRT). The models described lay the foundation for future studies into detailed mechanisms of IgA and IgG protection against STm and other pathogens.

A mutation in Arabidopsis SAL1 alters its in vitro activity against IP3 and delays developmental leaf senescence in association with lower ROS levels.

KEY MESSAGE: Our manuscript is the first to find a link between activity of SAL1/OLD101 against IP3 and plant leaf senescence regulation and ROS levels assigning a potential biological role for IP3. Leaf senescence is a genetically programmed process that limits the longevity of a leaf. We identified and analyzed the recessive Arabidopsis stay-green mutation onset of leaf death 101 (old101). Developmental leaf longevity is extended in old101 plants, which coincided with higher peroxidase activity and decreased H2O2 levels in young 10-day-old, but not 25-day-old plants. The old101 phenotype is caused by a point mutation in SAL1, which encodes a bifunctional enzyme with inositol polyphosphate-1-phosphatase and 3' (2'), 5'-bisphosphate nucleotidase activity. SAL1 activity is highly specific for its substrates 3-polyadenosine 5-phosphate (PAP) and inositol 1, 4, 5-trisphosphate (IP3), where it removes the 1-phosphate group from the IP3 second messenger. The in vitro activity of recombinant old101 protein against its substrate IP3 was 2.5-fold lower than that of wild type SAL1 protein. However, the in vitro activity of recombinant old101 mutant protein against PAP remained the same as that of the wild type SAL1 protein. The results open the possibility that the activity of SAL1 against IP3 may affect the redox balance of young seedlings and that this delays the onset of leaf senescence.

A Phase 1 Study To Evaluate Safety and Pharmacokinetics following Administration of Single and Multiple Doses of the Antistaphylococcal Lysin LSVT-1701 in Healthy Adult Subjects.

Thirty-two healthy male subjects (8 per cohort) were randomized 6:2 to active:placebo. LSVT-1701, an antistaphylococcal lysin, was administered intravenously as a 6-mg/kg single dose and as 1.5, 3, and 4.5 mg/kg twice daily for 4 days. LSVT-1701 exposure increased in a greater than dose proportional manner and did not accumulate. Treatment-emergent adverse events (TEAEs) were predominantly of mild intensity. The most common TEAEs were chills, pyrexia, headache, infusion site events, cough, rhinorrhea, and increases in C-reactive protein. (This study has been registered at ClinicalTrials.gov under identifier NCT03446053.).

Integration of water contamination indicators and vulnerability indices on groundwater management in Menzel Habib area, south-eastern Tunisia.

Groundwater salinization is a major problem throughout arid and semi-arid areas due to different natural processes and anthropogenic activities and has caused irreparable environmental and economic effects. The objective of this study is to evaluate groundwater vulnerability of Menzel Habib which has been firstly assessed using multiple methods such as DRASTIC, DRASTIC pesticide, SINTACS and SI models. These indices are based on combination of intrinsic and specific characteristics of the aquifer. Almost the area presents a low to moderate vulnerability with the highest vulnerability on the western region, associated to lower deep of groundwater and evaporation processes, with consequent salinity increase. Total Dissolved solids, chloride, sodium, sulfate, calcium, and magnesium water contents were determined in a total of twenty-five groundwater samples from Menzel Habib aquifer. The accuracy of the best robust model was evaluated by the correlation between the different vulnerability indices and contamination water indicators. The main aim of this study is the development of a modified vulnerability index, DRASTIC_Sal, which includes the contribution of total dissolved solids from Menzel Habib groundwater. DRASTIC_Sal index is a simple approach for aquifer salinization vulnerability assessment, particularly for inland aquifers from arid and semi-arid regions with associated agricultural activities.

Endolysins of bacteriophage vB_Sal-S-S10 can naturally lyse Salmonella enteritidis.

BACKGROUND: The holin-endolysin lysis system plays an essential role in the phage life cycle. Endolysins are promising alternatives to antibiotics, and have been successfully used against Gram-positive bacteria. However, a few endolysins can externally lyse Gram-negative bacteria, due to the inaccessible peptidoglycan layer covered by the envelope. RESULTS: This study investigated the lysis system of a new Siphoviridae bacteriophage vB_Sal-S-S10 (S10), which, that was isolated from broiler farms, was found to be able to infect 51.4% (37/72) of tested S. enteritidis strains. Phage S10 genome had a classic holin-endolysin lysis system, except that one holin and one endolysin gene were functionally annotated. The orf 22 adjacent to the lysis cassette was identified as a new endolysin gene. Antibacterial activity assays showed that holin had an intracellular penetrating activity against S. enteritidis 35; both endolysins acted on the cell envelope of S. enteritidis 35 and showed a natural extracellular antibacterial activity, leading to a ~ 1 log titer decrease in 30 min. Protein characterization of lysin1 and lysin2 revealed that the majority of the N-terminus and the C-terminus were hydrophobic amino acids or positively charged. CONCLUSION: In this study, a new Salmonella phage vB_Sal-S-S10 (S10) was characterized and showed an ideal development prospect. Phage S10 has a classic holin-endolysin lysis system, carrying an overlapping holin-lysin gene and a novel lysin gene. Both endolysins coded by lysin genes could externally lyse S. enteritidis. The natural extracellular antibacterial character of endolysins would provide necessary information for the development of engineering endolysin as the antibiotic alternative against the infection with multidrug-resistant gram-negative bacteria.

Synthesis and characterization of Fe3O4@Sal@Cu as a novel, efficient and heterogeneous catalyst and its application in the synthesis of 2-amino-4H-chromenes.

This research paper presents the synthesis and characterization of the magnetic nanoparticle, Fe3O4@Sal@Cu, [Fe3O4@Si-CH2-CH2-CH2-NH-NH-CO-N=CH-(2-HO-C6H4-)@Cu] as a green and retrievable catalyst. This catalyst was characterized by FTIR, XRD, EDX and TGA analyses. In addition, the catalytic activity of this new catalyst was investigated for the synthesis of 2-amino 4H-chromenes by producing good-to-excellent yields under mild reaction conditions. The other advantages of the developed nanocatalyst are its ecofriendliness, being easy to handle, high reusability and being magnetically separable. The synthesis of some new derivatives of 2-amino-4H-chromenes in the presence of this nanocatalyst is also reported.

Sal synthase induced cytotoxicity of PC12 cells through production of the dopamine metabolites salsolinol and N-methyl-salsolinol.

As a catechol isoquinoline, salsolinol (Sal) is widely distributed in mammalian brains, and is increased in the cerebrospinal fluid (CSF) and urine of Parkinsonian patients. Sal can be metabolized to N-methyl-salsolinol (NM-Sal), an MPP+-like neurotoxin, and impairs the function of dopaminergic neurons, causing the clinical symptoms of Parkinson's disease (PD). Sal synthase, which catalyzes the production of Sal from dopamine and acetaldehyde, may be the important enzyme in the metabolism of catechol isoquinolines (CTIQs). Previously, our work demonstrated the existence of Sal synthase in rat brain and identified its amino acid sequence. However, the biological function of Sal synthase has not been thoroughly explored, especially its role in dopaminergic neuronal degeneration. In this study, we tried to clarify the catalytic role of Sal synthase in the formation of CTIQs which are endogenous neurotoxins in the mammalian brain. Furthermore, the cytotoxicity of Sal synthase was also observed in dopaminergic PC12 cells. The results demonstrated that Sal synthase overexpression can increase the level of Sal and NM-Sal, and ultimately cause mitochondria damage and apoptosis.

Safety and Efficacy of Third-Generation Ultrasound-Assisted Liposuction: A Series of 261 Cases.

BACKGROUND: The introduction of third-generation ultrasound-assisted liposuction (3rd UAL) allows for a less invasive modality of both deep and superficial lipectomy while offering improved skin retraction and reduced rate of complications. This study examined the efficacy and safety profile of this technology over 15 years of clinical experience. METHODS: A consecutive series of patients treated from 2005-2020 by the senior author were reviewed for demographic and anthropometric measurements, intraoperative settings, surgical outcomes, and complications via retrospective chart review. Body-Q survey was used to assess patient satisfaction. RESULTS: A total of 261 patients underwent 3rd UAL in 783 areas. There were 238 female and 23 male patients with an average age of 43.5 years and BMI of 27.4 kg/m2. The most frequently treated areas were the trunk and lower limbs. An average of 2840 mL of wetting solution was used with an average of 2284 mL of lipocrit aspirate. About 65% of the cases were done in conjunction with another procedure. Overall complication rate was 4.6%, contour irregularity (1.9%), seroma (0.8%), cellulitis (0.8%), pigmentation changes (0.4%), and electrolyte imbalance (0.4%), with a minimum follow-up of 6 months. 78% of patient would undergo the procedure again and 86% would recommend it. CONCLUSION: Third-generation ultrasound-assisted liposuction can be used effectively and safely, either alone, or in conjunction with other plastic surgery procedures. VASER liposuction allows surgeons to address superficial fat plane and enhanced skin tightening. Rate of complications are lower than that of traditional liposuction with equivalent or higher patient satisfaction. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Spin-Crossover and Slow Magnetic Relaxation Behavior in Hexachlororhenate(IV) Salts of Mn(III) Complexes [Mn(5-Hal-sal2323)]2[ReCl6] (Hal = Cl, Br).

The cationic complexes of Mn(III) with the 5-Hal-sal2323 (Hal = Cl, Br) ligands and a paramagnetic doubly charged counterion [ReCl6]2- have been synthesized: [Mn(5-Cl-sal2323)]2[ReCl6] (1) and [Mn(5-Br-sal2323)]2[ReCl6] (2). Their crystal structures and magnetic properties have been studied. These isostructural two-component ionic compounds show a thermally induced spin transition at high temperature associated with the cationic subsystem and a field-induced slow magnetic relaxation of magnetization at cryogenic temperature, associated with the anionic subsystem. The compounds are the first examples of the coexistence of spin crossover and field-induced slow magnetic relaxation in the family of known [MnIII(sal2323)] cationic complexes with various counterions.

[Protective effect of salidroside on high fat-induced apoptosis in H9c2 cardiomyocytes through AMPK/mTOR/p70S6K pathway].

The study explored the effect of salidroside(SAL) on high fat-induced apoptosis in H9 c2 cardiomyocytes based on AMPK/mTOR/p70 S6 K pathway.H9 c2 cardiomyocytes were cultured in vitro and the lipotoxicity model of H9 c2 cardiomyocytes was constructed by 0.2 mmol·L~(-1) palmitic acid(PA) treatment for 24 hours.The cells were divided into control group, PA group, and SAL group(20 µmol·L~(-1)).Cell proliferation was detected with cell proliferation kit I(MTT) assay after SAL and PA treatment.Dihydroethidium(DHE) probe, Annexin V-FITC/PI kit, and JC-1 probe were used to estimate reactive oxygen species(ROS) level, cell apoptosis, and mitochondrial membrane potential(MMP) change, respectively.The expression levels of p-AMPK/AMPK, p-mTOR/mTOR, p-p70 S6 K/p70 S6 K and apoptosis-related proteins Bax, Bcl-2, and cleaved caspase-3 were investigated with Western blot.The mRNA levels of AMPK, mTOR and p70 S6 K were determined by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).RESULTS:: showed that compared with control group, PA group had decreased cell proliferation ability, MMP, Bcl-2 protein expression and AMPK protein and mRNA expression, while increased ROS level, Bax and cleaved caspase-3 protein expression, and mTOR and p70 S6 K mRNA and protein expression, and the difference was statistically significant(P<0.05, P<0.01).Compared with PA group, SAL improved cell proliferation ability, MMP level, Bcl-2 protein expression, and AMPK mRNA and protein expression, while down-regulated ROS level, cell apoptosis, Bax and cleaved caspase-3 protein expression, and mTOR and p70 S6 K mRNA and protein expression, and the difference was statistically significant(P<0.05, P<0.01).In conclusion, SAL exerted protective effects on high fat-induced lipotoxicity of H9 c2 cardiomyocytes, alleviated the oxidative stress injury and reduced cell apoptosis via regulating AMPK/mTOR/p70 S6 K signaling pathway.

Carbon storage, flux and mitigation potential of tropical Sal mixed deciduous forest ecosystem in Chhattisgarh, India.

Tropical forests are considered for greater species diversity and ensure climate change mitigation through carbon (C) sink which maintains terrestrial C storage in the world. Tree provides both tangible (timber, fuelwood, etc. For humans) and intangible benefits (as climate security through C sink) that maintains ecosystem processes. Tropical Sal forests are gaining popularity due to its remarkable contribution as C sink, storage, budget and flux. In the present study, an effort has been made to explore vegetational statistics along with C storage, budget and flux in four different site qualities (SQ) of Sal dominating tropical deciduous forest of Chhattisgarh, India. The density (individuals/ha) and basal area (m2/ha) varied from 710 to 1010 and 33.5-46.8 in tree, 2000-2500 and 0.32-0.33 in sapling and 9750-14500 and 17.96-21.43 in seedling, respectively in varying SQ. The total biomass varied from 182.27 to 375.84 t/ha in varying SQ. The total C in trees varied from 79.86 to 163.63 t ha-1. Quantity of C in above ground and below ground portions in trees on different sites were 72.32-143.36 t/ha and 7.54-20.27 t/ha, respectively. Total aboveground tree C sequestration values ranged between 5.12 and 11.68 t C ha-1yr-1 on different SQ. The C storage and net fluxes were represented in compartment models to assess the various SQ. As per models, forest received 14.63, 10.81, 8.19 and 6.83 t/ha/yr of C input through net primary production (NPP) in SQ-I, SQ-II, SQ-III and SQ-IV, respectively which are depleted as 3.55, 3.12, 2.77 and 2.33 t/ha/yr as total C input in the soil. Moreover 1.77 (SQ-I), 1.60 (SQ-II), 1.46 (SQ-III) and 1.30 (SQ-IV) t ha-1 yr-1of C were transferred from foliage to litter compartment, respectively. These dynamics, budgeting and flux of C represents "how C stored and moved within an ecosystem". Similarly, it affects overall terrestrial C pools that is governed by varying SQ.

Assessment of sal (Shorea robusta) forest phenology and its response to climatic variables in India.

Remote sensing-based observation provides an opportunity to study the spatiotemporal variations of plant phenology across the landscapes. This study aims to examine the phenological variations of different types of sal (Shorea robusta) forests in India and also to explore the relationship between phenology metrics and climatic parameters. Sal, one of the main timber-producing species of India, can be categorized into dry, moist, and very moist sal. The phenological metrics of different types of sal forests were extracted from Moderate Resolution Imaging Spectroradiometer (MODIS)-derived Enhanced Vegetation Index (EVI) time series data (2002-2015). During the study period, the average start of season (SOS) was found to be 16 May, 17 July, and 29 June for very moist, moist, and dry sal forests, respectively. The spatial distribution of mean SOS was mapped as well as the impact of climatic variables (temperature and rainfall) on SOS was investigated during the study period. In relation to the rainfall, values of the coefficient of determination (R2) for very moist, moist, and dry sal forests were 0.69, 0.68, and 0.76, respectively. However, with temperature, R2 values were found higher (R2 = 0.97, 0.81, and 0.97 for very moist, moist, and dry sal, respectively). The present study concluded that MODIS EVI is well capable of capturing the phenological metrics of different types of sal forests across different biogeographic provinces of India. SOS and length of season (LOS) were found to be the key phenology metrics to distinguish the different types of sal forests in India and temperature has a greater influence on SOS than rainfall in sal forests of India.

The PAP/SAL1 retrograde signaling pathway is involved in iron homeostasis.

KEY MESSAGE: There is a link between PAP/SAL retrograde pathway, ethylene signaling and Fe metabolism in Arabidopsis. Nuclear gene expression is regulated by a diversity of retrograde signals that travel from organelles to the nucleus in a lineal or classical model. One such signal molecule is 3'-phosphoadenisine-5'-phosphate (PAP) and it's in vivo levels are regulated by SAL1/FRY1, a phosphatase enzyme located in chloroplast and mitochondria. This metabolite inhibits the action of a group of exorribonucleases which participate in post-transcriptional gene expression regulation. Transcriptome analysis of Arabidopsis thaliana mutant plants in PAP-SAL1 pathway revealed that the ferritin genes AtFER1, AtFER3, and AtFER4 are up-regulated. In this work we studied Fe metabolism in three different mutants of the PAP/SAL1 retrograde pathway. Mutant plants showed increased Fe accumulation in roots, shoots and seeds when grown in Fe-sufficient condition, and a constitutive activation of the Strategy I Fe uptake genes. As a consequence, they grew more vigorously than wild type plants in Fe-deficient medium. However, when mutant plants grown in Fe-deficient conditions were sprayed with Fe in their leaves, they were unable to deactivate root Fe uptake. Ethylene synthesis inhibition revert the constitutive Fe uptake phenotype. We propose that there is a link between PAP/SAL pathway, ethylene signaling and Fe metabolism.

Reverse Genetics Assembly of Newcastle Disease Virus Genome Template Using Asis-Sal-Pac BioBrick Strategy.

BACKGROUND: The BioBrick construction as an approach in synthetic biology provides the ability to assemble various gene fragments. To date, different BioBrick strategies have been exploited for assembly and cloning of a variety of gene fragments. We present a new BioBrick strategy, here referred as Asis-Sal-Pac BioBrick, which we used for the assembly of NDV as a candidate for single-stranded non-segmented, negative-sense RNA genome viruses. RESULTS: In the present study, we isolated three NDVs from clinical samples which were classified into the VIId genotype based on their pathogenicity and phylogenetic analyses. Then, SalI, AsisI, and PacI enzymes were used to design and develop a novel BioBrick strategy, which enabled us to assemble the NDV genome, adopting the "rule of six". In this method, in each assembly step, the restriction sites in the newly formed destination plasmid are reproduced, which will be used for the next insertion. In this study using two overlapping PCRs, the cleavage site of the F gene was also modified from 112RRQKRF117to 112GRQGRL117 in order to generate the attenuated recombinant NDV. Finally, in order to construct the recombinant NDV viruses, the plasmids harboring the assembled full-length genome of the NDV and the helper plasmids were co-transfected into T7-BHK cells. The rescue of the recombinant NDVwas confirmed by RT-PCR and HA tests. CONCLUSIONS: These findings suggest that the combination of reverse genetic technology and BioBrick assembly have the potential to be applied for the development of novel vaccine candidates. This promising strategy provides an effective and reliable approach to make genotype-matched vaccines against specific NDV strains or any other virus.

32 Years of Ultrasonic-Assisted Lipoplasty (U.A.L.): From Aesthetic to Obesity.

In this article, we tried to take stock of the state of the art in ultrasonic-assisted lipoplasty focusing on its most recent technologic advances and in the newest clinical applications, such as overweight, obesity, breast surgery and regenerative medicine. Great space was devoted to the analysis and clarification of the most common myths and legends related to this intriguing technique.

Investigating the contribution of climate variables to estimates of net primary productivity in a tropical deciduous forest in India.

Investigating the impact of climate variables on net primary productivity is crucial to evaluate the ecosystem health and the status of forest type response to climate change. The objective of this paper is (1) to estimate spatio-temporal patterns of net primary productivity (NPP) during 2001 to 2010 in a tropical deciduous forest based on the input variable dataset (i.e.meteorological and biophysical) derived from the remote sensing and other sources and (2) to investigate the effects of climate variables on NPP during 2001 to 2010. The study was carried out in Katerniaghat Wildlife Sanctuary that forms a part of a tropical forest and is situated in Uttar Pradesh, India, along the Indo-Nepal border. Mean annual NPP was observed to be highest during 2007 with a value of 878 g C m-2 year-1 and 781.25 g C m-2 year-1 for sal and teak respectively. A decline in mean NPP during 2002-2003, 2005 and 2008-2010 could be attributed to drought, increased temperature and vapour pressure deficit (VPD). The time lag correlation analysis revealed precipitation as the major variables affecting NPP, whereas combination of temperature and VPD showed dominant effect on NPP as revealed by generalized linear modelling. The carbon gain in NPP in sal forest was observed to be marginal higher than that of teak plantation throughout the study period. The decrease in NPP was observed during 2010, pertaining to increased VPD. Contribution of different climatic variables through some link process was revealed in statistical analysis and clearly indicated the co-dominance of all the variables in explaining NPP.