Cryopreservation of seeds of Neonotonia wightii Wight and Arn: a strategy for conservation, dormancy breaking and preservation of nutritional status.

BACKGROUND: N. wightii (Leguminosae) is valued as a cover crop and as a potential source of protein in food insecure countries. However, plantlet establishment is limited by physical dormancy. Our previous work has shown that exposure of N. wightii seeds to cryogenic temperatures is able to overcome physical dormancy. OBJECTIVE: The current study is an extension of that work where the field performance and nutritional composition of plants regenerated from N. wightii seeds was investigated. RESULTS: It was evident that plants regenerated from cryopreserved seeds displayed faster growth rates than those from control seeds. In addition, cryopreservation did not alter the nutritional profile of plants produced from cryo-stored seeds. CONCLUSION: Collectively, the results indicate that cryopreservation serves as a suitable strategy for the preservation of seeds of N. wightii with the added benefit of also serving as a dormancy breaking mechanism upon retrieval from cryogenic temperatures. Doi.org/10.54680/fr23510110712.

Exposure of maize seeds to liquid nitrogen modifies the morphology and hormonal response of young plants.

BACKGROUND: Cryopreservation currently represents the most suitable strategy for the long-term conservation of plant germplasm. While much effort has focused on the development of protocols to enable successful cryostorage, there are few, if any reports, that consider the effect of cryogenic temperatures on the phytohormone status of the seed and developing seedlings. OBJECTIVE: To investigate the effect of cryopreservation on external seed coat features as well as levels of indole-3-acetic acid (IAA), abscisic acid (ABA) and 1-aminocyclopropane-1-carboxylic acid (ACC) in maize. MATERIALS AND METHODS: Two groups of seeds at 6% moisture content were compared: one was maintained at 4 degree C (control) while the other was exposed to LN within cryo-vials. RESULTS: Seeds exposed to cryogenic temperatures were characterized by the presence of large cracks in the seed coat compared with control seeds. Cryogenic exposure also resulted in a reduction in biomass and plant height. Results from the phytohormone analysis showed an initial reduction in the levels of IAA, ABA and ACC after 7 days of growth followed by sharp increase in levels relative to the control by 14 days. Whilst the roles of ABA and ethylene (and by extension, its precursor ACC) are well studied as stress response molecules, much less is known about the potentially vital role of auxins in regulating plant growth under conditions of low temperature stress. CONCLUSION: It is postulated that the interaction of all three hormones modulate crosstalk between various stress responses and recovery pathways to ameliorate the damage caused by freezing stress and enable plant survival. Given the dearth of information on phytohormones in cryobiology, more studies are needed to fully elucidate these relationships in the context of freezing stress caused by liquid nitrogen. Doi.org/10.54680/fr23610110612.

Exposure of Calophyllum antillanum seeds to liquid nitrogen delays seedling emergence and decreases leaf anthraquinones.

BACKGROUND: Trees within the Calophyllum genus are multi-use trees that produce valuable wood, phytochemicals with a range of biological activities, and seed oil as a source of biodiesel. As a consequence of climate change, there is a need to develop strategies to preserve valuable plant genetic resources. Cryopreservation represents the most suitable option for the long-term storage of germplasm with minimal space and maintenance requirements. OBJECTIVE: To determine appropriate methods to cryopreserve seeds of Calophyllum antillanum and maintain secondary compound production. MATERIALS AND METHODS: Seeds at a moisture content of 6% were used to evaluate two treatments: seeds immersed in liquid nitrogen and control seeds. Biosynthetic pathway efficiency was assessed post-cryo by determining anthraquinone contents in roots, stems and leaves following 30 and 75 d of seedling growth. RESULTS: The results indicated that exposure to liquid nitrogen delayed germination and seedling emergence for a period of up to 45 d after seed sowing. By 60 d of cultivation, no significant differences in plant growth were observed for cryostored and control seeds. The levels of anthraquinones, which were also measured in seeds and seedlings, were lower in plants regenerated from cryostored seeds following 30 d of growth, but there were no differences in roots and stems by 75 d of growth. Furthermore, the difference in leaf anthraquinone levels for cryopreserved and control seeds at 75 d was much smaller than at 30 d. CONCLUSION: The low initial anthraquinone levels in emerging seedlings correlated with the initial slow growth of cryopreserved seeds.

Cryopreservation of seeds of the highly valued tropical timber species Swietenia mahagoni.

BACKGROUND: Swietenia mahagoni wood is one of the most valuable in world trade and, as a result, natural populations have been decimated due to unsustainable harvesting. The decline in natural population levels is being exacerbated by climate change. In order to ensure the preservation of valuable genotypes, there is an urgent need to develop strategies to conserve the genetic diversity present within this species. At present, cryopreservation is the most viable option for the long-term storage of plant germplasm, particularly for long-lived species which are challenging to maintain in the field. OBJECTIVE: To cryopreserve intact seeds of S. mahagoni, with the dual goal of retaining the biosynthetic capacity of plants, which is critical since this species is highly valued for medicinal purposes. MATERIALS AND METHODS: Seeds at a moisture content of 6% were immersed in liquid nitrogen (LN) before warming and recovery. Plantlet establishment and growth were assessed over a period of 70 days and anthraquinone synthesis was determined in roots, stems and leaves. RESULTS: The results showed an initial lag in the germination rate of cryopreserved seeds compared with control seeds; however, this difference disappeared over time. The lag in seedling emergence observed in cryostored seeds was also evident in the plant characteristics measured following 30 days of culture when all plant parameters measured were significantly higher in plants produced from control than cryostored seeds. However, after 70 days of growth, these differences were no longer apparent. Anthraquinone levels were also initially lower (at 30 days) in plants regenerated from cryopreserved seeds than those from control seeds, however, this difference was substantially reduced by 70 days thereby indicating the ability of these plants to accumulate secondary metabolites, albeit at a reduced rate, during the early stages of development. CONCLUSION: In S. mahagoni, the delay in anthraquinone production in plants regenerated from cryostored seeds during the early stages of development may have occurred as a consequence of the preferential allocation of resources towards the initiation of recovery processes in response to the stresses imposed by cryopreservation. Once the stresses were overcome and plant growth resumed, resources could be directed to secondary processes such as anthraquinone synthesis. doi.org/10.54680/fr22610110412.

Exposure of Teramnus labialis (L.F.) Spreng Seeds to Liquid Nitrogen Does Not Affect Nutritional Status of Field Grown Adult Plants.

BACKGROUND: Teramnus labialis (L.f.) Spreng is a potentially important legume species, and can be used as an animal feed and to enhance soil physicochemical characteristics. Despite the biological and agricultural importance, the low availability of seeds, their small size and the low percentage germination limit their large-scale use by farmers. We previously reported a method to cryopreserve seeds of T. labialis which also allowed for the breaking of seed dormancy. OBJECTIVE: The current study reports on the nutritional status of 5 month old field grown plants regenerated from cryostored and control seeds. MATERIALS AND METHODS: Biomass (fresh and dry mass of leaves and stems) and contents of ash, neutral detergent fiber, acid detergent fiber, lignin, cellulose, crude protein, P, Ca, Mg and K were measured. RESULTS: Seeds germinated and emerged faster following immersion in liquid nitrogen (LN) which was supported by quantitative evaluations of fresh and dry weights per m2. However, the ratio of leaf:stem mass were not altered by seed exposure to LN. CONCLUSION: The results showed that exposure of seeds to cryogenic temperatures did not alter the nutritional composition of regenerated plants.

Seed Cryostorage Enhances Subsequent Plant Productivity in the Forage Species Teramnus Labialis (L.F.) Spreng.

BACKGROUND: Teramnus labialis is an herbaceous legume that serves as a source of carbohydrates and proteins for animals and humans, and is valued for its nitrogen contribution to soil. The benefits of this species are, however, limited by low seed availability, small seed size and low in situ seed germination levels, due to physical dormancy. Cryostorage has been shown to be beneficial for both seed storage and breaking physical dormancy in seeds of various species. However, its potential effects on subsequent seedling emergence, plant growth and seed production need to be studied before large-scale implementation for T. labialis. OBJECTIVE: To record agricultural traits of T. labialis after seed exposure to liquid nitrogen. MATERIALS AND METHODS: Seeds were maintained at 5 degree C (control) or stored in LN before sowing. Seedling emergence percentage and traits related to plant growth and seed production were evaluated for 6 months. RESULTS: Except for seed weight, all traits differed significantly between seedlings generated from cryostored and control seeds. Except for pod number, seedling emergence and plant growth traits were enhanced by cryostorage to a greater extent than seed production traits. Cryostorage resulted in cracks and breaks in the seed coat which were absent in control seeds (scanning electron microscopy), and in breaking physical dormancy may have facilitated more rapid seedling emergence than for control seeds. CONCLUSION: Seed cryostorage enhances subsequent plant productivity in terms of growth and to a lesser extent seed production in Teramnus labialis, validating its use for commercial growth of this species.

Phenotypic and molecular characterization of plants regenerated from non-cryopreserved and cryopreserved wild Solanum lycopersicum mill. Seeds.

BACKGROUND: Before cryopreservation is routinely used, its effect on the trueness-to-type of the regenerated plant material needs to be evaluated. OBJECTIVE: In this work, we studied the effect of seed cryopreservation on the phenotypic and molecular characteristics of wild Solanum lycopersicum Mill. plants. METHODS: Thirty-five morphological traits of plants regenerated from cryopreserved seeds were compared to those measured on plants regenerated from non-cryopreserved seeds. RESULT: No statistically significant differences were observed between cryopreserved and non-cryopreserved samples, either in the first or in the second generation post-liquid nitrogen exposure. However, at the molecular level, the genetic analyses performed on the second generation plants germinated from control and cryopreserved seeds using 14 nuclear Simple Sequences Repeats (SSR) markers uncovered some changes in microsatellite length between control and cryopreserved samples. These results confirm at the botanical phenotype level the effectiveness of seed cryostorage for conservation and regeneration of true-to-type S. lycopersicum plants. CONCLUSION: Further experiments are required to clarify potential phenotypic effects of the changes observed in the DNA.

Biochemical characterization of ecuadorian wild Solanum lycopersicum Mill. plants produced from non-cryopreserved and cryopreserved seeds.

This paper presents some of the effects of cryopreservation of wild Solanum lycopersicum Mill. seeds on the early stages of germination post liquid nitrogen exposure. Percentage of germination, conversion into plantlets and plant fresh mass were evaluated after cryostorage. Levels of chlorophyll pigments (a, b, total), malondialdehyde, other aldehydes, phenolics (cell wall-linked, free, and total) and proteins were determined. Peroxidase and superoxide dismutase activities were recorded. Liquid nitrogen exposure increased the percentage of seed germination at 5 days but at 7 days, the conversion into plantlets and the plant fresh mass were not statistically different between non-cryopreserved and cryopreserved samples. Several significant effects of cryopreservation were recorded at the biochemical level at 7 days of germination under controlled conditions. Highly significant effects due to liquid nitrogen exposure were observed in leaves: increased levels of peroxidase enzymatic and specific activities and cell wall-linked phenolics. Very remarkable effects were also recorded in roots: decreased contents of chlorophylls and cell wall-linked phenolics.

[Updated report on STOPP-START criteria in Primary Care]. / Evaluación de la actualización de los criterios STOPP-START en atención primaria.

INTRODUCTION: The aim of this study is to determine the rate of potentially inappropriate prescriptions (PIP) in people older than 64 years of age in Avilés, Asturias, Spain. MATERIALS AND METHODS: A descriptive cross-sectional study was conducted in six Health Care Centres. A sample of 400 people was selected, obtained by a random sampling proportional to the population registered in each Health Centre. A review was made of the computerised clinical records, and the STOPP-START (version 2014 with 114 items) criteria were applied to evaluate the PIP. RESULTS: The study contained 378 (95.5%) patients with a mean age of 75.4 (SD: 7.4) and of which 57.7% were women. Almost all (94.2%: 95% CI; 91.7-96.7) met some PIP criteria. Taking only the STOPP criteria into consideration, 52.4% (95%CI: 47.2-57.6) met at least one breach, and in the START criteria a 90.5% (95%CI; 87.4-93.6), which was reduced to 40.5% (95%CI; 36.4-45.6) if criteria on vaccination were removed. In the STOPP criteria, the most frequent PIP was taking benzodiazepines followed by the use of medication without indications based on the evidence; in the START, the criteria was the anti-pneumococcus vaccination, and the lack of taking vitamin D supplements and calcium in osteoporosis. CONCLUSIONS: There were high levels of PIP, very superior to the previous version, especially for the START criteria. There is a high level of PIP related to the use of benzodiazepines and the use of medication without any clinic evidence. The STOPP-START criteria are useful in Primary Care to assess the PIP.

[Rates of surgical wound infection in hip arthroplasty (2007-2015)]. / Tasas de infección de herida quirúrgica en artroplastia de cadera (2007-2015).

OBJECTIVE: To estimate the incidence of surgical site infection in the hip replacement procedure during nine years of follow-up in a local public hospital. MATERIAL AND METHODS: A prospective study conducted between 2007 and 2015, using the Clinical Indicators of Continuous Quality Improvement (INCLIMECC) tool. All primary hip prostheses and revisions were studied. The definitions, criteria and categories for surgical site infection were those estimated for the risk index of the National Nosocomial Infections Surveillance of Centres for Disease Control and Prevention. Crude and adjusted rates were calculated by risk index. RESULTS: A total of 999 interventions were included. The overall infection rate was 1.5% (95%CI: 0.75-2.25). Surgical prophylaxis was prescribed in 98.3% of the cases, and was adequate in 70.7%, with the main cause of inadequacy being its prolonged duration in 71.3%. The infection rate was 53%, being better than the rates published for Spanish hospitals INCLIMECC in the period 1997-2012. But they were 19% higher than those published for the USA according to the National Healthcare Safety Network, and 50% higher than expected according to data published for Europe by the European Centre for Disease Prevention and Control. DISCUSSION: The infection rate in the present study is better than those described in the Spanish literature, but it is higher than those published for Europe and North America. Strategies for surveillance and control of infections associated with health care allow the evaluation of trends and the impact of prevention measures.

[Potentially inappropriate prescription according to the "STOPP" Criteria in heavily polymedicated elderly patients]. / Prescripción potencialmente inadecuada en pacientes mayores grandes polimedicados según criterios "STOPP".

INTRODUCTION: Appropriate prescribing is especially relevant in elderly people. The objective of this study is to analyse the potentially inappropriate prescription (PIP) in heavily polymedicated elderly patients according to the criteria Screening Tool of Older Person's Prescriptions. PATIENTS AND METHODS: A descriptive cross-sectional study was conducted in Primary Care on patients assigned to the Siero-Sariego (Asturias) Health Centre, who were over 64 years old and heavily polymedicated (consumption >10 drugs for six months). The 65 Screening Tool of Older Person's Prescriptions criteria were reviewed in the electronic Primary Care patient records, collecting sociodemographic variables, prescribed medications, and chronic diseases. Frequency distributions were made for each criterion, and causes related to PIP were explored using contingency tables, the Spearman correlation coefficient, and logistic regression. RESULTS: A total of 349 polymedicated elderly patients were analysed with a prevalence of 6.4 (95% CI:5.76-7.08), a mean age of 79.2 years (SD:3.7), 62.2% were female, 14% institutionalised, a Charlson index of 2.9. The mean of number of drugs was 11.5 (SD:1.7), and the most frequent pathologies were high blood pressure (64%), diabetes (46%), and osteoarticular diseases (41%). There was at least one PIP in 72.9% of heavily polymedicated elderly patients [(Mean: 1.32 (SD:1.2)]. The Screening Tool of Older Person's Prescriptions criteria least complied with were: therapeutic duplication (25.2%), use of long-acting benzodiazepines (15.8%), and inappropriate use of aspirin (10.9%). An association was found between having any inappropriate prescription and the number of medications prescribed (OR=1.22 [95% CI:1.04-1.43]) and inversely to the Charlson index (OR=0.76 [95% CI:0.65-0.89]). CONCLUSIONS: PIP is common in heavily polymedicated elderly patients. Special attention must be paid to the use of psychotropic drugs, which are implicated in a high volume of PIP.

Paclitaxel resistance in non-small-cell lung cancer associated with beta-tubulin gene mutations.

PURPOSE: The mechanisms that cause chemoresistance in non-small-cell lung cancer (NSCLC) patients have yet to be clearly elucidated. Paclitaxel is a tubulin-disrupting agent that binds preferentially to beta-tubulin. Tubulins are guanosine triphosphate (GTP)-binding proteins. Beta-tubulin is a GTPase, whereas alpha-tubulin has no enzyme activity. We reasoned that polymerase chain reaction (PCR) and DNA sequencing of the beta-tubulin gene could reveal more information regarding the connection between beta-tubulin mutations and primary paclitaxel resistance. PATIENTS AND METHODS: Constitutional genomic DNA and paired tumor DNA were isolated from 49 biopsies from 43 Spanish and six American stage IIIB and IV NSCLC patients who had been treated with a 3-hour, 210 mg/m(2) paclitaxel infusion and a 24-hour, 200 mg/m(2) infusion, respectively. Oligonucleotides specific to beta-tubulin were designed for PCR amplification and sequencing of GTP- and paclitaxel-binding beta-tubulin domains. RESULTS: Of 49 patients with NSCLC, 16 (33%; 95% confidence interval [CI], 20.7% to 45.3%) had beta-tubulin mutations in exons 1 (one patient) or 4 (15 patients). None of the patients with beta-tubulin mutations had an objective response, whereas 13 of 33 (39.4%; 95% CI, 22.8% to 56%; P = 0.01) patients without beta-tubulin mutations had complete or partial responses. Median survival was 3 months for the 16 patients with beta-tubulin mutations and 10 months for the 33 patients without beta-tubulin mutations (P =.0001). CONCLUSION: We have identified beta-tubulin gene mutations as a strong predictor of response to the antitubulin drug paclitaxel; these mutations may represent a novel mechanism of resistance and should be examined prospectively in future trials of taxane-based therapy in NSCLC.

Microsatellite alterations at 5q21, 11p13, and 11p15.5 do not predict survival in non-small cell lung cancer.

We investigated the clinical implications of allelic deletions at three common sites of loss of heterozygosity (LOH) in regions 5q21, 11p15.5, and 11p13 in 86 patients with non-small cell lung cancer (NSCLC). We performed a PCR-based microsatellite polymorphism assay for detection of LOH. The microsatellite markers used were D5S82 (proximal to the APC gene), MCC (within the MCC gene), D11S904 (11p13), HRAS (within the H-ras gene), and D11S860 (11p15.5). Of the 68 informative cases at 5q21 loci, LOH was found in 14 cases (20%), whereas LOH frequency in 11p15.5 and 11p13 was 31% (19 of 61 informative cases) and 19% (12 of 63 informative cases), respectively. There was a significant correlation between 5q21 LOH and mediastinal lymph node involvement (P = 0.03). However, no differences were observed in median survival times (26 months in patients with 5q21 LOH versus 37 months in the remainder; P = 0.33) nor in patients with 11p LOH (38 months versus 32 months, respectively; P = 0.72). Cox's proportional hazards model predicted that stage was the only independent poor prognostic marker in the entire cohort of NSCLC patients. Thus, the present study revealed two important abnormalities, LOH at chromosome 5q21 and LOH at chromosome 11p, both implied in NSCLC development.

Giant-cell reparative granuloma of short tubular bones of the hands and feet.

Eight cases of giant-cell reparative granuloma were encountered in the short tubular bones of the hands and feet. This entity was originally described by Jaffe as a jaw lesion. Subsequently the lesion was found in the skull and facial bones. The name "giant cell reaction" was applied to similar lesions found in the short tubular bones of hands and feet. Thirteen prior cases have been reported and none recurred. Of the eight new cases described herein, four have recurred one or more times, requiring further treatment. No association with hyperparathyroidism was found. We prefer to designate these lesions as giant cell reparative granulomas of extragnathic sites. Because of overlapping ultrastructural and light-microscopic features, as well as similar biologic behavior, we feel that aneurysmal bone cyst and these giant-cell reparative lesions represent related responses to intraosseous hemorrhage.

Pseudotumoral articular amyloidosis.

We present a 71-year-old man with primary amyloidosis characterized by several outstanding clinical features, namely hip involvement of a pseudotumoral appearance, periarticular osteolytic lesions and a pathological fracture. Kappa light chains in the synovial fluid were observed. The patient survived for 90 months without developing plasma cell dyscrasia, and with preservation of vital functions despite parenchymal involvement of multiple organs.

Field performance of sugarcane (Saccharum sp.) plants derived from cryopreserved calluses.

This study compared the field performance of sugarcane plants originating from three different sources: control, non-cryopreserved embryogenic calluses, cryopreserved embryogenic calluses and macropropagated material of the same commercial hybrid. Several agronomic traits were evaluated on 100 plants per treatment over a 27-month period covering the growth of the stool and of the first ratoon. Significant differences between treatments were observed only during the first six months of field growth of sugarcane stools. Stems produced from in vitro cultured material, irrespective of their cryopreservation status, had a smaller diameter and a shorter height than those produced from macropropagated material. These differences disappeared by 12 months of stool field growth.

Effect of cryopreservation on the structural and functional integrity of cell membranes of sugarcane (Saccharum sp.) embryogenic calluses.

In this paper, we investigated if the differences consistently noted in survival and plantlet production between cryopreserved and non-cryopreserved, control sugarcane embryogenic calluses were related to modifications induced during cryopreservation in the structural and functional integrity of cell membranes. For this, the evolution of electrolyte leakage, lipid peroxidation products and cell membrane protein contents was measured during 5 d after cryopreservation. Differences between control and frozen calluses were observed only during the first 2 (electrolyte leakage) or 3 d (lipid peroxidation products and membrane protein content) after freezing. It was not possible to link these differences with the differences noted in survival and plant production between control and cryopreserved calluses. Additional studies are thus needed to elucidate which biochemical factors, linked to survival and plantlet regeneration, are affected by cryopreservation.

[Pubertal idiopathic gynecomastia]. / Ginecomastia idiopática puberal.

14 boys with pubertal gynecomastia, aged 9 to 16 years, were presented. Physical assessment, including pubertal maturation, height, weight, bone age, pubic hair stage, testicular and penis calibre and degree of gynecomastia was determined. Sexual steroids, LH-FSH, prolactin, were studied. All patients had psychological stress. 11 patients had been treated surgically by mastectomy with success.

[Pharmacological cost impact of the osteoarthritis of the knee in a primary care health centre]. / Impacto del gasto farmacológico de la artrosis de rodilla en un centro de salud de atención primaria.

OBJECTIVE: To establish the pharmacological cost incurred by an urban Primary Health Care centre for treating osteoarthritis (OA) of the knee, including the main drugs used in its treatment and its contribution to the total cost of each group studied. PATIENTS AND METHODS: A cross-sectional, descriptive study of 188 patients diagnosed with OA of the knee. After counting the total number of pre-packaged pharmaceutical products assigned to each patient over one year, the average cost of each product was calculated. The ratio between the total cost and the total number of patients was then calculated, which gave the cost per patient per year. RESULTS: NSAIDs (non-steroids anti-inflammatory drugs) were the most frequently prescribed. SYSADOAs (symptomatic slow-acting drugs for osteoarthritis) rated second in both frequency and cost. Acetaminophen rated third. Topical treatments, "other analgesics" and proton pump inhibitors (PPIs), cost much less. The total pharmacological cost per patient per year thus amounted to 151.6 € (SD:±101,9). CONCLUSIONS: The pharmacological cost for treatment of osteoarthritis of the knee incurs an increase in cost of health resources in Primary Care. The total pharmacological cost per patient per year was 151.60 €. In our centre, NSAIDs account for most of the pharmacological costs in the treatment of osteoarthritis of the knee.