Wall proteins of Vitis vinifera pollen II. Influence of environment and rootstock on the electrophoretic pattern

Soluble wall proteins from Vitis vinifera pollen are genotypically determined and their expression, if not the extent of their expression, is independent from external factors. After we had previously observed that the time of sample collection does not influence their electrophoretic pattern, we demonstrate in the present work that this is true also for environment where the vine is growing and for the rootstock on which the scion is grafted

Wall proteins of Vitis vinifera pollen I. Constancy of the phenotype

Isoelectric focusing of buffer-soluble wall proteins from pollen grains is suggested for the characterization of different Vitis vinifera cultivars. The protein pattern includes several tens of bands, most of which common to all samples. A few clone-specific components are identified, whose expression is stable over time and independent from the influence of environment

some more about dogs proteomics of neglected biological fluids

Abstract We report in this manuscript what is known about the protein makeup of a selection of biological fluids in the domestic dog. The samples we review – amniotic and allantoic fluid, seminal fluid, saliva, bile, synovial fluid, tears – are still very poorly characterized in this species. For some of them we can present results from our own, mainly unpublished experiments. Significance The dog is one of the most widespread companion animals, and also of medical relevance as model species for some human diseases. Still, investigation of body fluids other than serum and urine is not so commonly undertaken, although – like in humans - also these sample types may have potential for diagnostic purposes. We compile published data about proteomes of fetal fluids, seminal plasma, saliva, bile, synovial fluid and tears, enriched by some yet unpublished data of our own (proteins of amniotic and allantoic fluid, tears). Closing gaps in our knowledge on dog proteins will further our understanding of (patho)physiological processes

A chemotaxonomic investigation on Vitis vinifera1. Within-cultivar population analysis

The extent of variability for storage protein subunits as well as the isozymes of AcP, EST and PGM was evaluated by isoelectric focusing on a large number of individual self-pollinated seeds from cvs Chardonnay, Sangiovese and Traminer. Extracts from 35 randomly selected kernels gave reproducible protein patterns and may thus be taken as representative of the average genetic make-up in a given biotype

A chemotaxonomic investigation on Vitis vinifera L. II. Comparison among ssp. sativa traditional cultivars and wild biotypes of ssp. silvestris from various Italian regions

An extensive screening on regional representatives of Vitis vinifera ssp. sativa and silvestris was carried out to look for relationships and differences between different taxa. Total proteins in the pH range 4.0-5.5, and the enzymes AcP, ADH, EST, G-6-PDH, MDH, PGM and POD were recorded. Only patterns of storage protein subunits, AcP, EST and G-6-PDH were taxonomically informative. Dendrograms were computerized on the basis of presence/absence of individual bands; these always distinguish at different levels in homogeneous groups between ssp. sativa and ssp. silvestris. The acidic subunits showed a dichotomy from the first branching of the cladogram. These observations are a demonstration that neither hypotheses of direct or indirect origin of the Italian ssp. sativa from Italian ssp. silvestris via domestication is tenanble. The authors suggest that, the morphological, the ecological and the biochemical differences between the two taxa support the hypothesis that V. sativa and V. silvestris should be regarded as two separate taxa that have had reciprocal interactions such a long period of time that a precise location of origin is no longer possible

Role of α1 Acid Glycoprotein in the In Vivo Resistance of Human BCR-ABL+ Leukemic Cells to the Abl Inhibitor STI571

Background: Chronic myeloid leukemia is caused by a chromosomal translocation that results in an oncogenic fusion protein, Bcr-Abl. Bcr-Abl is a tyrosine kinase whose activity is inhibited by the antineoplastic drug STI571. This drug can cure mice given an injection of human leukemic cells, but treatment ultimately fails in animals that have large tumors when treatment is initiated. We created a mouse model to explore the mechanism of resistance in vivo. Methods: Nude mice were injected with KU812 Bcr-Abl+ human leukemic cells. After 1 day (no evident tumors), 8 days, or 15 days (tumors >1 g), mice were treated with STI571 (160 mg/kg every 8 hours). Cells recovered from relapsing animals were used for in vitro experiments. Statistical tests were two-sided. Results: Tumors regressed initially in all STI571-treated mice, but all mice treated 15 days after injection of tumor cells eventually relapsed. Relapsed animals did not respond to further STI571 treatment, and their Bcr-Abl kinase activity in vivo was not inhibited by STI571, despite high plasma concentrations of the drug. However, tumor cells from resistant animals were sensitive to STI571 in vitro, suggesting that a molecule in the plasma of relapsed animals may inactivate the drug. The plasma protein α1 acid glycoprotein (AGP) bound STI571 at physiologic concentrations in vitro and blocked the ability of STI571 to inhibit Bcr-Abl kinase activity in a dose-dependent manner. Plasma AGP concentrations were strongly associated with tumor load. Erythromycin competed with STI571 for AGP binding. When animals bearing large tumors were treated with STI571 alone or with a combination of STI571 and erythromycin, greater tumor reductions and better long-term tumor-free survival (10 of 12 versus one of 13 at day 180; P<.001) were observed after the combination treatment. Conclusion: AGP in the plasma of relapsed animals binds to STI571, preventing this compound from inhibiting the Bcr/Abl tyrosine kinase. Molecules such as erythromycin that compete with STI571 for binding to AGP may enhance the therapeutic potential of this dru

Infected pancreatic necrosis: outcomes and clinical predictors of mortality. A post hoc analysis of the MANCTRA-1 international study

: The identification of high-risk patients in the early stages of infected pancreatic necrosis (IPN) is critical, because it could help the clinicians to adopt more effective management strategies. We conducted a post hoc analysis of the MANCTRA-1 international study to assess the association between clinical risk factors and mortality among adult patients with IPN. Univariable and multivariable logistic regression models were used to identify prognostic factors of mortality. We identified 247 consecutive patients with IPN hospitalised between January 2019 and December 2020. History of uncontrolled arterial hypertension (p = 0.032; 95% CI 1.135-15.882; aOR 4.245), qSOFA (p = 0.005; 95% CI 1.359-5.879; aOR 2.828), renal failure (p = 0.022; 95% CI 1.138-5.442; aOR 2.489), and haemodynamic failure (p = 0.018; 95% CI 1.184-5.978; aOR 2.661), were identified as independent predictors of mortality in IPN patients. Cholangitis (p = 0.003; 95% CI 1.598-9.930; aOR 3.983), abdominal compartment syndrome (p = 0.032; 95% CI 1.090-6.967; aOR 2.735), and gastrointestinal/intra-abdominal bleeding (p = 0.009; 95% CI 1.286-5.712; aOR 2.710) were independently associated with the risk of mortality. Upfront open surgical necrosectomy was strongly associated with the risk of mortality (p &lt; 0.001; 95% CI 1.912-7.442; aOR 3.772), whereas endoscopic drainage of pancreatic necrosis (p = 0.018; 95% CI 0.138-0.834; aOR 0.339) and enteral nutrition (p = 0.003; 95% CI 0.143-0.716; aOR 0.320) were found as protective factors. Organ failure, acute cholangitis, and upfront open surgical necrosectomy were the most significant predictors of mortality. Our study confirmed that, even in a subgroup of particularly ill patients such as those with IPN, upfront open surgery should be avoided as much as possible. Study protocol registered in ClinicalTrials.Gov (I.D. Number NCT04747990)

Wild-Type Opaque2 and Defective opaque2 Polypeptides Form Complexes in Maize Endosperm Cells and Bind the Opaque2-Zein Target Site1[OA]

The Opaque2 (O2) basic leucine (Leu)-zipper transcriptional activator controls the expression of several genes in maize (Zea mays). We investigated the phosphorylation extent of wild-type O2 and mutant-defective or mutant-truncated o2 polypeptides in endosperm cells, their subcellular localization, participation in complex formation, and involvement in functional activity. Besides wild type, four mutant alleles (o2T, o2-52, o2It, and o2-676) producing o2 polypeptides and a null transcript allele (o2R) were considered. Observing the effects of these mutations, multiphosphorylation events in O2 or o2 proteins were confirmed and further investigated, and the involvement of both the nuclear localization signal (NLS)-B and Leu-zipper domains in proper targeting to the nucleus was ascertained. The absence of these domains in the o2T and o2It-S mutant-truncated forms holds them within the cytoplasm, where they are partially phosphorylated, whereas the presence of NLS-B and a partial Leu-zipper domain in o2-52 distributes this mutant-truncated form in both cytoplasm and nucleus. Although mutated in the NLS-B domain, the o2It-L and o2-676 mutant-defective forms are, respectively, partially or completely distributed into the nucleus. Only wild-type O2 and mutant-defective o2 polypeptides bearing the Leu-zipper are able to form complexes whose components were proven to bind the O2-zein target site by in vitro analyses. The transcription of a subset of H-zein genes as well as H-zein polypeptide accumulation in several o2-mutant-defective genotypes indicate the in vivo involvement of o2-mutant-defective proteins in O2-zein target site recognition. The gathered information broadens our knowledge on O2 functional activity and our view on possible quality protein maize trait manipulation or plant transformation via the utilization of cisgenic elements