Transcriptomic and physiological approaches to decipher cold stress mitigation exerted by brown-seaweed extract application in tomato.

Chilling temperatures represent a challenge for crop species originating from warm geographical areas. In this situation, biostimulants serve as an eco-friendly resource to mitigate cold stress in crops. Tomato (Solanum lycopersicum L.) is an economically important vegetable crop, but quite sensitive to cold stress, which it encounters in both open field and greenhouse settings. In this study, the biostimulant effect of a brown-seaweed extract (BSE) has been evaluated in tomato exposed to low temperature. To assess the product effects, physiological and molecular characterizations were conducted. Under cold stress conditions, stomatal conductance, net photosynthesis, and yield were significantly (p ≤ 0.05) higher in BSE-treated plants compared to the untreated ones. A global transcriptomic survey after BSE application revealed the impact of the BSE treatment on genes leading to key responses to cold stress. This was highlighted by the significantly enriched GO categories relative to proline (GO:0006560), flavonoids (GO:0009812, GO:0009813), and chlorophyll (GO:0015994). Molecular data were integrated by biochemical analysis showing that the BSE treatment causes greater proline, polyphenols, flavonoids, tannins, and carotenoids contents.The study highlighted the role of antioxidant molecules to enhance tomato tolerance to low temperature mediated by BSE-based biostimulant.

Brown Seaweed Extract (BSE) Application Influences Auxin- and ABA-Related Gene Expression, Root Development, and Sugar Yield in Beta vulgaris L.

The molecular and phenotypic effects of a brown seaweed extract (BSE) were assessed in sugar beet (Beta vulgaris L.). Transcript levels of BSE-treated and untreated plants were studied by RNA-seq and validated by quantitative real-time PCR analysis (RT-qPCR). Root morphology, sugar yield, and processing quality traits were also analyzed to better elucidate the treatment effects. RNA-seq revealed 1019 differentially expressed genes (DEGs) between the BSE-treated and untreated plants. An adjusted p-value < 0.1 and an absolute value of log2 (fold change) greater than one was used as criteria to select the DEGs. Gene ontology (GO) identified hormone pathways as an enriched biological process. Six DEGs involved in auxin and ABA pathways were validated using RT-qPCR. The phenotypic characterization indicated that BSE treatment led to a significant increase (p < 0.05) in total root length and the length of fine roots of plants grown under hydroponics conditions. The sugar yield of plants grown under field conditions was higher (p < 0.05) in the treated field plots compared with the control treatment, without impacting the processing quality. Our study unveiled the relevant effects of BSE application in regulating auxin- and ABA-related gene expression and critical traits related to sugar beet development and yield.

Novel Effects of Leonardite-Based Applications on Sugar Beet.

The present study aimed to explore the effects of foliar application of a leonardite-based product on sugar beet (Beta vulgaris L.) plants grown in the field. The approach concerned the evaluation of the community compositional structure of plant endophytic bacteria through a metabarcoding approach, the expression level of a gene panel related to hormonal metabolism and signaling, and the main sugar beet productivity traits. Results indicated that plants treated with leonardite (dosage of 2,000 ml ha-1, dilution 1:125, 4 mg C l-1) compared with untreated ones had a significant increase (p < 0.05) in (i) the abundance of Oxalicibacterium spp., recognized to be an endophyte bacterial genus with plant growth-promoting activity; (ii) the expression level of LAX2 gene, coding for auxin transport proteins; and (iii) sugar yield. This study represents a step forward to advance our understanding of the changes induced by leonardite-based biostimulant in sugar beet.

Managing a Tertiary Orthopedic Hospital during the COVID-19 Epidemic, Main Challenges and Solutions Adopted.

The present paper is a review of the main challenges faced by the management of a tertiary specialty hospital during the COVID-19 pandemic in the northern Italian region of Lombardy, an area of extremely high epidemic impact. The article focuses on the management of patient flows, access to the hospital, maintaining and reallocating staffing levels, and managing urgent referrals, information, and communications from the point of view of the hospital managers over a seven-week period. The objective of the article is to provide beneficial insights and solutions to other hospital managers and medical directors who should find themselves in the same or a similar situation. In such an epidemic emergency, in the authors' opinion, the most important factors influencing the capability of the hospital to maintain operations are (1) sustaining the strict triage of patients, (2) the differentiation of flows and pathways to create what could be regarded as "a hospital inside a hospital", (3) tracing and sharing all available information to face the rapidly changing environment, (4) being able to maintain staffing levels in critical areas by flexibly allocating the workforce, and (5) from a regional perspective, being organized along a hub-and-spoke system for critical and time-sensitive networks was key for focusing the hospital's resources on the most needed services.

Expression Profiling of Candidate Genes in Sugar Beet Leaves Treated with Leonardite-Based Biostimulant.

Leonardite-based biostimulants are a large class of compounds, including humic acid substances. Foliar application of biostimulants at field level improves plant growth, yield and quality through metabolic changes and stimulation of plant proton pumps. The present study aimed at identifying optimum dosage of BLACKJAK, a humic acid-based substance, which is able to modify genes involved in sugar beet growth. Thirty-three genes belonging to various biochemical pathway categories were tested in leaves of treated sugar beet (Beta vulgaris L.) samples to assess gene expression profiling in response to BLACKJAK. Seedlings of a diploid and multigerm variety were grown in plastic pots and sprayed with two dilutions of BLACKJAK (dilution 1:500-1.0 mg C L-1 and dilution 1:1000-0.5 mg C L-1). Leaf samples were collected after 24, 48, and 72 h treatment with BLACKJAK for each dilution. RNA was extracted and the quantification of gene expression was performed while using an OpenArray platform. Results of analysis of variance demonstrated that, 15 genes out of a total of 33 genes tested with OpenArray qPCR were significantly affected by treatment and exposure time. Analysis for annotation of gene products and pathways revealed that genes belonging to the mitochondrial respiratory pathways, nitrogen and hormone metabolisms, and nutrient uptake were up-regulated in the BLACKJAK treated samples. Among the up-regulated genes, Bv_PHT2;1 and Bv_GLN1 expression exerted a 2-fold change in 1:1000 and 1:500 BLACKJAK concentrations. Overall, the gene expression data in the BLACKJAK treated leaves demonstrated the induction of plant growth-related genes that were contributed almost to amino acid and nitrogen metabolism, plant defense system, and plant growth.

Molecular and Morphological Changes Induced by Leonardite-based Biostimulant in Beta vulgaris L.

Humic substances extracted from leonardite are widely considered to be bioactive compounds, influencing the whole-plant physiology and the crop yield. The aim of this work was to evaluate the effect of a new formulate based on leonardite in the early stage of growth of sugar beet (Beta vulgaris L.). A commercial preparation of leonardite (BLACKJAK) was characterized by ionomic analysis, solid-state 13C MAS NMR spectroscopy. Seedlings of sugar beet were grown in Hoagland's solution under controlled conditions. After five days of growth, an aliquot of the concentrated BLACKJAK was added to the solution to obtain a final dilution of 1:1000 (0.5 mg C L-1). The sugar beet response in the early stage of growth was determined by evaluating root morphological traits as well as the changes in the expression of 53 genes related to key morphophysiological processes. Root morphological traits, such as total root length, fine root length (average diameter < 0.5 mm), and number of root tips, were significantly (p < 0.001) increased in plants treated with BLACKJAK, compared to the untreated plants at all sampling times. At the molecular level, BLACKJAK treatment upregulated many of the evaluated genes. Moreover, both Real Time PCR and digital PCR showed that genes involved in hormonal response, such as PIN, ARF3, LOGL 10, GID1, and BRI1, were significantly (p < 0.05) upregulated by treatment with BLACKJAK. Our study provides essential information to understand the effect of a leonardite-based formulate on plant growth hormone metabolism, although the molecular and physiological basis for these complicated regulatory mechanisms deserve further investigations.

[Did household parental smoking attitude change over the last 15 years? A survey among primary school children in the city of Agrigento, Italy]. / È cambiato qualcosa nelle abitudini dei genitori fumatori in ambito domestico negli ultimi 15 anni? Indagine epidemiologica tra gli alunni della città di Agrigento.

BACKGROUND: Exposure of children to passive tobacco smoking in the household setting has been demonstrated to cause respiratory diseases. Early atherosclerosis has also been demonstrated in young people previously exposed to passive tobacco smoking. Functional damage can initiate at the fetal age due to maternal smoking, with a tendency towards over-time progress. To date only scant data are available about indoor parental smoking attitudes and their changes after campaigns against smoke and risk factors in exposed youths. Questionnaires are useful tools in order to search for information on cigarette smoking and parental household lifestyle. In this study, we asked pupils of primary schools for providing information on their relatives' every 5 years throughout the period 1994-2009. METHODS: A multiple-choice answer questionnaire about sharing household parental smoking was administered to all primary school children (mean age 10.5 ± 0.5 years) of the city of Agrigento (Sicily, Italy). RESULTS: A total of 2221 questionnaires were collected from 637 children in 1994, 687 in 1999, 516 in 2004, and 381 in 2009. Important findings show a significant reduction in both smoking parents and exposure to passive tobacco smoking (from 64% in 1994 to 45% in 2009, p<0.00001). In 2009, 22% of mothers and 39% of fathers resulted to be smokers in the household setting yet. CONCLUSIONS: In agreement with the 2011 Italian DOXA survey on the general population, the present study demonstrates a negative trend in the prevalence of household smoker parents (predominantly mothers) and exposure of children to passive tobacco smoking. These results likely reflect such a greater awareness about smoke-related risks in children, and the effectiveness of medical campaigns against cardiovascular risk factors as well.

Natura abhorret a vacuo--use of fibrin glue as a filler and sealant in neurosurgical "dead spaces". Technical note.

OBJECTIVE: The objective of this study is to report our experience and illustrate our technique in the use of fibrin glue in the treatment of post-operatory cerebrospinal fluid (CSF) leaks and collections following different neurosurgical procedures. METHODS: In a 3-year period, 40 subjects underwent endoscopic endonasal approach for different sellar and skull base lesions (three tuberculum sellae meningiomas, six craniopharyngiomas, three Rathke's cleft cysts and 28 pituitary macroadenomas), in which an intraoperative CSF leakage was evident. In such subjects, the fibrin glue was used as a first step of the final phase of the procedure-i.e. the reconstruction of the skull base defect-followed by the other materials employed. Furthermore, ten other patients, who had undergone transsphenoidal (four cases), spinal (two cases), posterior fossa (three cases) and transcortical intraventricular tumour removal (one case) neurosurgical procedures and developed CSF leaks or collections, were conservatively treated by single or repeated in situ injections of "modified" fibrin glue under local anaesthesia according to different described techniques. In total, 50 patients constitute the clinical material of the present study. TECHNIQUE: In the cases where the fibrin glue was used during the reconstruction phase of the procedure (40 cases), the glue was injected inside the tumour cavity to fill the dead space left by the removal of the lesion. In case of post-operative CSF leak or CSF fluid collection (ten cases), after discarding 50-80% of the thrombin solution to obtain prevalence of the product's adhesive properties, fibrin glue was injected directly in the path of the CSF leak or into the collection cavity after aspiration of the collection's content. This was performed with the provided application system or through lumbar or Tuohy needles. Applications were repeated every 48 h until the disappearance of the leak. In all the treated cases, the disappearance of CSF leaks or collections was obtained with a number of applications ranging from one to five. Successful results are stable with a follow-up ranging from 6 months to 3 years. CONCLUSIONS: In our experience, the injection of fibrin glue has proved to be effective in filling or sealing post-operative "dead spaces" and treating minor or initial CSF leaks resulting from procedures of transsphenoidal, cranial and spinal surgery, adding another possibility in the management of many of these dreadful complications.

Combined endoscopic and microsurgical removal of a giant cystic craniopharyngioma in a six-year-old boy.

Giant cystic craniopharyngiomas are rare lesions whose clinical and surgical management is extremely challenging, often requiring more than one craniotomy before obtaining a satisfying removal. We report one case of a giant cystic craniopharyngioma completely excised with a two-step combined use of a minimally invasive endoscopic approach followed by a single microsurgical transcranial procedure. A six-year-old boy presented with symptoms of increased intracranial pressure and posterior fossa involvement. Preliminary imaging revealed a large para- and suprasellar cystic tumor bulging superiorly into the third ventricle, and extending posteriorly from the retroclival region into the posterior fossa to the level of the foramen magnum. The suprasellar cystic quota was initially approached endoscopically through a right precoronal-transventricular approach and the cyst drained, while the remaining tumor was macroscopically totally removed one week later by a right pterional approach. A combined technique--endoscopic drainage followed by microsurgical removal--can be a valid alternative for the removal of giant cystic craniopharyngiomas in cases in which the cystic compartment bulges within the ventricular spaces, and may avoid multiple craniotomies.

Anatomical study of the pterygopalatine fossa using an endoscopic endonasal approach: spatial relations and distances between surgical landmarks.

OBJECT: The pterygopalatine fossa is an area that lies deep within the skull base. The recent extensive use of the endoscopic endonasal approach has provided neurosurgeons with a method to reach various areas of the skull base through a less invasive approach than traditional transcranial or transfacial approaches. This study aims to provide neurosurgeons with new data concerning direct endoscopic measurements and precise anatomical topography features of the pterygopalatine fossa. METHODS: An anatomical dissection of six fixed cadaver heads (12 pterygopalatine fossae) was performed to analyze spatial relationships and distances between the most important neurovascular structures in this region, and to estimate the size of the endoscopic surgical field for operations in this area. The endoscopic endonasal approach offers direct access to the pterygopalatine fossa through its anteromedial walls. CONCLUSIONS: Using an endoscopic endonasal approach makes it possible to identify all of the anatomical landmarks of the pterygopalatine fossa and almost all of the contiguous skull base areas.

The endoscopic endonasal approach to the lateral recess of the sphenoid sinus via the pterygopalatine fossa: comparison of endoscopic and radiological landmarks.

OBJECTIVE: The endoscopic endonasal approach offers the opportunity to reach the pterygopalatine fossa, the lateral recess of the sphenoid sinus, and other areas of the cranial base through a minimally invasive approach. This study compares the anatomy of these areas when observed through an endoscopic endonasal view with the anatomy of the same regions as they appear in computed tomographic scans. The aim was to identify and correlate the corresponding anatomic structures, providing the surgeons with anatomic landmarks to guide them when operating in these areas through an endoscopic endonasal approach. METHODS: An anatomic dissection of six fixed cadaver heads was performed by an endoscopic endonasal approach. A step-by-step comparison of endoscopic and radiological images was made to identify the landmarks of the surgical field. RESULTS: The step-by-step comparison of endoscopic and radiological images acquired during the endoscopic endonasal approach to the lateral recess of the sphenoid sinus via the pterygopalatine fossa allowed the identification of all the relevant anatomic landmarks of the procedure. CONCLUSION: The endoscopic endonasal approach via the pterygopalatine fossa offers direct, minimally invasive access to the lateral recess of the sphenoid sinus, which can be monitored in each phase through consistent radiological imagery.