Effect of cochlear implantation on cognitive decline and quality of life in younger and older adults with severe-to-profound hearing loss.

PURPOSE: (a) To measure the change in cognition, the improvement of speech perception, and the subjective benefit in people under and over 60 years following cochlear implantation. (b) To assess the relationship between cognition, demographic, audiometric, and subjective outcomes in both age groups. METHODS: 28 cochlear implant (CI) users were assigned to the < 60y group and 35 to the ≥ 60y group. Cognition was measured using the Repeatable Battery for the Assessment of Neuropsychological Status for Hearing impaired individuals (RBANS-H); subjective benefit was measured using the Nijmegen Cochlear Implant Questionnaire (NCIQ); the Glasgow Benefit Inventory (GBI); the Hearing Implant Sound Quality Index (HISQUI19); Speech, Spatial and Qualities of Hearing Scale (SSQ12); and the Hospital Anxiety and Depression Scale (HADS). RESULTS: Prior to surgery: the RBANS-H total score positively correlated with the domains "Advanced sound", "Self-esteem", and "Social functioning" of NCIQ, and negatively with HADS scores. 12 months post-implantation: the RBANS-H total score increased in the < 60y (p = 0.038) and in the ≥ 60y group (p < 0.001); speech perception and subjective outcomes also improved; RBANS-H total score positively correlated with "Self-esteem" domain in NCIQ. Age and the RBANS-H total score correlated negatively in the ≥ 60y group (p = 0.026). CONCLUSIONS: After implantation, both age groups demonstrated improved cognition, speech perception and quality of life. Their depression scores decreased. Age was inversely associated with cognition.

A state-of-the-art implementation of a binaural cochlear-implant sound coding strategy inspired by the medial olivocochlear reflex.

Cochlear implant (CI) users find it hard and effortful to understand speech in noise with current devices. Binaural CI sound processing inspired by the contralateral medial olivocochlear (MOC) reflex (an approach termed the 'MOC strategy') can improve speech-in-noise recognition for CI users. All reported evaluations of this strategy, however, disregarded automatic gain control (AGC) and fine-structure (FS) processing, two standard features in some current CI devices. To better assess the potential of implementing the MOC strategy in contemporary CIs, here, we compare intelligibility with and without MOC processing in combination with linked AGC and FS processing. Speech reception thresholds (SRTs) were compared for an FS and a MOC-FS strategy for sentences in steady and fluctuating noises, for various speech levels, in bilateral and unilateral listening modes, and for multiple spatial configurations of the speech and noise sources. Word recall scores and verbal response times in a word recognition test (two proxies for listening effort) were also compared for the two strategies in quiet and in steady noise at 5 dB signal-to-noise ratio (SNR) and the individual SRT. In steady noise, mean SRTs were always equal or better with the MOC-FS than with the standard FS strategy, both in bilateral (the mean and largest improvement across spatial configurations and speech levels were 0.8 and 2.2 dB, respectively) and unilateral listening (mean and largest improvement of 1.7 and 2.1 dB, respectively). In fluctuating noise and in bilateral listening, SRTs were equal for the two strategies. Word recall scores and verbal response times were not significantly affected by the test SNR or the processing strategy. Results show that MOC processing can be combined with linked AGC and FS processing. Compared to using FS processing alone, combined MOC-FS processing can improve speech intelligibility in noise without affecting word recall scores or verbal response times.

Initial findings in chest X-rays as predictors of worsening lung infection in patients with COVID-19: correlation in 265 patients.

BACKGROUND AND AIMS: We aimed to analyze the relationship between the initial chest X-ray findings in patients with severe acute respiratory syndrome due to infection with SARS-CoV-2 and eventual clinical worsening and to compare three systems of quantifying these findings. MATERIAL AND METHODS: This retrospective study reviewed the clinical and radiological evolution of 265 adult patients with COVID-19 attended at our center between March 2020 and April 2020. We recorded data related to patients' comorbidities, hospital stay, and clinical worsening (admission to the ICU, intubation, and death). We used three scoring systems taking into consideration 6 or 8 lung fields (designated 6A, 6B, and 8) to quantify lung involvement in each patient's initial pathological chest X-ray and to classify its severity as mild, moderate, or severe, and we compared these three systems. We also recorded the presence of alveolar opacities and linear opacities (fundamentally linear atelectasis) in the first chest X-ray with pathologic findings. RESULTS: In the χ2 analysis, moderate or severe involvement in the three classification systems correlated with hospital admission (P = .009 in 6A, P = .001 in 6B, and P = .001 in 8) and with death (P = .02 in 6A, P = .01 in 6B, and P = .006 in 8). In the regression analysis, the most significant associations were 6B with alveolar involvement (OR 2.3; 95%CI 1.1.-4.7; P = .025;) and 8 with alveolar involvement (OR 2.07; 95% CI 1.01.-4.25; P = .046). No differences were observed in the ability of the three systems to predict clinical worsening by classifications of involvement in chest X-rays as moderate or severe. CONCLUSION: Moderate/severe extension in the three chest X-ray scoring systems evaluating the extent of involvement over 6 or 8 lung fields and the finding of alveolar opacities in the first pathologic X-ray correlated with mortality and the rate of hospitalization in the patients studied. No significant difference was found in the predictive ability of the three classification systems proposed.

[Initial findings in chest X-rays as predictors of worsening lung infection in patients with COVID-19: correlation in 265 patients]. / Hallazgos iniciales en la radiografía de tórax como predictores de empeoramiento en la infección pulmonar por SARS-CoV-2. Correlación en 265 pacientes.

Background and aims: We aimed to analyze the relationship between the initial chest X-ray findings in patients with severe acute respiratory syndrome due to infection with SARS-CoV-2 and eventual clinical worsening and to compare three systems of quantifying these findings. Material and methods: This retrospective study reviewed the clinical and radiological evolution of 265 adult patients with COVID-19 attended at our center between March 2020 and April 2020. We recorded data related to patients' comorbidities, hospital stay, and clinical worsening (admission to the ICU, intubation, and death). We used three scoring systems taking into consideration 6 or 8 lung fields (designated 6 A, 6 B, and 8) to quantify lung involvement in each patient's initial abnormal chest X-ray and to classify its severity as mild, moderate, or severe, and we compared these three systems. We also recorded the presence of alveolar opacities and linear opacities (fundamentally linear atelectasis) in the first chest X-ray with pathologic findings. Results: In the χ2 analysis, moderate or severe involvement in the three classification systems correlated with hospital admission (p = 0.009 in 6 A, p = 0.001 in 6 B, and p = 0.001 in 8) and with death (p = 0.02 in 6 A, p = 0.01 in 6 B, and p = 0.006 in 8). In the regression analysis, the most significant associations were 6 B with alveolar involvement (OR 2.3; 95%CI 1.1.-4.7; p = 0.025;) and 8 with alveolar involvement (OR 2.07; 95% CI 1.01.-4.25; p = 0.046). No differences were observed in the ability of the three systems to predict clinical worsening by classifications of involvement in chest X-rays as moderate or severe. Conclusion: Moderate/severe extension in the three chest X-ray scoring systems evaluating the extent of involvement over 6 or 8 lung fields and the finding of alveolar opacities in the first abnormal X-ray correlated with mortality and the rate of hospitalization in the patients studied. No significant difference was found in the predictive ability of the three classification systems proposed.

Speech-in-Noise Recognition With More Realistic Implementations of a Binaural Cochlear-Implant Sound Coding Strategy Inspired by the Medial Olivocochlear Reflex.

OBJECTIVES: Cochlear implant (CI) users continue to struggle understanding speech in noisy environments with current clinical devices. We have previously shown that this outcome can be improved by using binaural sound processors inspired by the medial olivocochlear (MOC) reflex, which involve dynamic (contralaterally controlled) rather than fixed compressive acoustic-to-electric maps. The present study aimed at investigating the potential additional benefits of using more realistic implementations of MOC processing. DESIGN: Eight users of bilateral CIs and two users of unilateral CIs participated in the study. Speech reception thresholds (SRTs) for sentences in competition with steady state noise were measured in unilateral and bilateral listening modes. Stimuli were processed through two independently functioning sound processors (one per ear) with fixed compression, the current clinical standard (STD); the originally proposed MOC strategy with fast contralateral control of compression (MOC1); a MOC strategy with slower control of compression (MOC2); and a slower MOC strategy with comparatively greater contralateral inhibition in the lower-frequency than in the higher-frequency channels (MOC3). Performance with the four strategies was compared for multiple simulated spatial configurations of the speech and noise sources. Based on a previously published technical evaluation of these strategies, we hypothesized that SRTs would be overall better (lower) with the MOC3 strategy than with any of the other tested strategies. In addition, we hypothesized that the MOC3 strategy would be advantageous over the STD strategy in listening conditions and spatial configurations where the MOC1 strategy was not. RESULTS: In unilateral listening and when the implant ear had the worse acoustic signal-to-noise ratio, the mean SRT was 4 dB worse for the MOC1 than for the STD strategy (as expected), but it became equal or better for the MOC2 or MOC3 strategies than for the STD strategy. In bilateral listening, mean SRTs were 1.6 dB better for the MOC3 strategy than for the STD strategy across all spatial configurations tested, including a condition with speech and noise sources colocated at front where the MOC1 strategy was slightly disadvantageous relative to the STD strategy. All strategies produced significantly better SRTs for spatially separated than for colocated speech and noise sources. A statistically significant binaural advantage (i.e., better mean SRTs across spatial configurations and participants in bilateral than in unilateral listening) was found for the MOC2 and MOC3 strategies but not for the STD or MOC1 strategies. CONCLUSIONS: Overall, performance was best with the MOC3 strategy, which maintained the benefits of the originally proposed MOC1 strategy over the STD strategy for spatially separated speech and noise sources and extended those benefits to additional spatial configurations. In addition, the MOC3 strategy provided a significant binaural advantage, which did not occur with the STD or the original MOC1 strategies.

Towards the quantitative characterisation of piglets' robustness to weaning: a modelling approach.

Weaning is a critical transition phase in swine production in which piglets must cope with different stressors that may affect their health. During this period, the prophylactic use of antibiotics is still frequent to limit piglet morbidity, which raises both economic and public health concerns such as the appearance of antimicrobial-resistant microbes. With the interest of developing tools for assisting health and management decisions around weaning, it is key to provide robustness indexes that inform on the animals' capacity to endure the challenges associated with weaning. This work aimed at developing a modelling approach for facilitating the quantification of piglet resilience to weaning. A total of 325 Large White pigs weaned at 28 days of age were monitored and further housed and fed conventionally during the post-weaning period without antibiotic administration. Body weight and diarrhoea scores were recorded before and after weaning, and blood was sampled at weaning and 1 week later for collecting haematological data. A dynamic model was constructed based on the Gompertz-Makeham law to describe live weight trajectories during the first 75 days after weaning, following the rationale that the animal response is partitioned in two time windows (a perturbation and a recovery window). Model calibration was performed for each animal. Our results show that the transition time between the two time windows, as well as the weight trajectories are characteristic for each individual. The model captured the weight dynamics of animals at different degrees of perturbation, with an average coefficient of determination of 0.99, and a concordance correlation coefficient of 0.99. The utility of the model is that it provides biologically meaningful parameters that inform on the amplitude and length of perturbation, and the rate of animal recovery. Our rationale is that the dynamics of weight inform on the capability of the animal to cope with the weaning disturbance. Indeed, there were significant correlations between model parameters and individual diarrhoea scores and haematological traits. Overall, the parameters of our model can be useful for constructing weaning robustness indexes by using exclusively the growth curves. We foresee that this modelling approach will provide a step forward in the quantitative characterisation of robustness.

Association of genetic variants and expression levels of porcine FABP4 and FABP5 genes.

The FABP4 and FABP5 genes, coding for fatty acid transport proteins, have long been studied as positional candidate genes for SSC4 QTL affecting fat deposition and composition traits in pigs. Polymorphisms in these genes, FABP4:g.2634_2635insC and FABP5:g.3000T>G, have previously been associated with fatness traits in an Iberian by Landrace cross (IBMAP). The aim of the present work was to evaluate the functional implication of these genetic variants. For this purpose, FABP4 and FABP5 mRNA expression levels in 114 BC1_LD animals (25% Iberian × 75% Landrace) were analyzed using real-time quantitative PCR in backfat and muscle. FABP4 gene expression in backfat, but not in muscle, was associated with FABP4:g.2634_2635insC. In contrast, FABP5:g.3000T>G was not associated with gene expression levels. An expression-based genome-wide association study highlighted the FABP4:g.2634_2635insC polymorphism as the polymorphism most associated with FABP4 gene expression in backfat. Furthermore, other genomic regions associated in trans with the mRNA expression of FABP4 in backfat and FABP5 in muscle were also identified. Finally, two putative transcription binding sites for PPARG and NR4A2 may be affected by the FABP4:g.2634_2635insC polymorphism, modifying FABP4 gene expression. Our results reinforce FABP4 as a candidate gene for fatness traits on SSC4.

Systemic Besnoitiosis in a Juvenile Roe Deer (Capreolus capreolus).

Herein, we report the first incidence of systemic besnoitiosis in a male juvenile roe deer Capreolus capreolus. The animal was found dead in an area where bovine besnoitiosis is endemic and showed cachexia and multiple skin erosions in the metacarpal and metatarsal areas. Moreover, round and elevated white structures suggestive of Besnoitia spp. tissue cysts were also present. Twenty-eight tissue samples from different anatomical locations were collected for microscopic lesion and parasite detection through histopathology and PCR. Immunohistochemistry was performed to confirm Besnoitia-positive reaction in the tissue cysts. In addition, the identity of Besnoitia spp. in PCR-positive tissue samples was also investigated using microsatellite (MS) markers, and the comparison of protein disulphide isomerase gene sequences (BbPDI) of B. besnoiti and B. tarandi isolated from cattle and reindeer, respectively. Besnoitia cysts were detected in the skin (several parts), respiratory and upper digestive tracts, eyes, kidney, liver, testicle, cardiac muscle and lymphoid tissue. Remarkably, the presence of tissue cysts in the brain confirmed the capacity of Besnoitia spp. to form tissue cysts in the central nervous system (CNS). Finally, the Besnoitia species detected showed the same MS genotype as B. besnoiti, and BbPDI sequences from roe deer and two B. besnoiti isolates were genetically identical throughout multiple sequence alignment. Thus, for the first time, there is evidence that roe deer might act as an intermediate host of B. besnoiti. Further molecular analyses and parasite isolations are needed to corroborate these findings.

Analysis of the porcine APOA2 gene expression in liver, polymorphism identification and association with fatty acid composition traits.

APOA2 is a protein implicated in triglyceride, fatty acid and glucose metabolism. In pigs, the APOA2 gene is located on pig chromosome 4 (SSC4) in a QTL region affecting fatty acid composition, fatness and growth traits. In this study, we evaluated APOA2 as a candidate gene for meat quality traits in an Iberian × Landrace backcross population. The APOA2:c.131T>A polymorphism, located in exon 3 of APOA2 and determining a missense mutation, was associated with the percentage of hexadecenoic acid [C16:1(n-9)], linoleic acid [C18:2(n-6)], α-linolenic acid [C18:3(n-3)], dihomo-gamma-linolenic acid [C20:3(n-6)] and polyunsaturated fatty acids (PUFAs) in backfat. Furthermore, this SNP was associated with the global mRNA expression levels of APOA2 in liver and was used as a marker to determine allelic expression imbalance by pyrosequencing. We determined an overexpression of the T allele in heterozygous samples with a mean ratio of 2.8 (T/A), observing a high variability in the allelic expression among individuals. This result suggests that complex regulatory mechanisms, beyond a single polymorphism (e.g. epigenetic effects or multiple cis-acting polymorphisms), may be regulating APOA2 gene expression.

New insight of functional molecular imaging into the atheroma biology: 18F-NaF and 18F-FDG in symptomatic and asymptomatic carotid plaques after recent CVA. Preliminary results.

AIM: The identification of the vulnerable atheroma plaque could allow a more effective treatment of cerebrovascular accident (CVA). Active calcification and inflammation of the carotid atheroma were assessed and compared in symptomatic and asymptomatic plaques by 18F-NaF and 18F-FDG PET/CT. METHODS: Nine patients investigated for recent CVA and no preventive treatment with statins were enrolled. In each patient, at least one atheroma plaque was detected by CT angiogram. In total, 18 plaques were available: 9 symptomatic and 9 asymptomatic. 18F-NaF uptake and 18F-FDG uptake by each plaque were assessed visually and semiquantitatively by calculating target/background ratios (TBRs) and TBR indexes (TBR symptomatic/TBR asymptomatic and 18F-NaF TBR/18F-FDG TBR within each of the 2 clinical groups of plaques). RESULTS: All plaques showed 18F-NaF and 18F-FDG uptake, and semiquantitation showed higher 18F-NaF uptake by 11 of the 18 plaques, 6 symptomatic and 5 asymptomatic. In the symptomatic group, the mean 18F-NaF TBR was 2·12 ± 0·44, and in the asymptomatic group, it was 1·85 ± 0·46. The 18F-NaF/18F-FDG showed that, overall, 18F-NaF uptake is higher than 18F-FDG. In the symptomatic plaques, the 18F-NaF was higher for the low calcium content and the lowest for the high. CONCLUSION: Active calcification and inflammation are simultaneous processes in the symptomatic and asymptomatic carotid atheroma. However, active calcification seems predominant over inflammation in both groups. In the symptomatic plaques, the highest 18F-NaF uptake does not correspond with the largest calcium content. These patterns open new insights on the role of 18F-NaF in the study of calcification and in the identification of the vulnerable carotid atheroma.

Domain-averaged exchange-correlation energies as a physical underpinning for chemical graphs.

A novel solution to the problem of assigning a molecular graph to a collection of nuclei (i.e. how to draw a molecular structure) is presented. Molecules are universally understood as a set of nuclei linked by bonds, but establishing which nuclei are bonded and which are not is still an empirical matter. Our approach borrows techniques from quantum chemical topology, which showed for the first time the construction of chemical graphs from wave functions, shifting the focus on energetics. This new focus resolves issues surrounding previous topological analyses, in which domain-averaged exchange-correlation energies (V(xc)), quantities defined in real space between each possible atom pair, hold the key. Exponential decay of V(xc) in non-metallic systems as the intercenter distance increases guarantees a well-defined hierarchy for all possible V(xc) values in a molecule. Herein, we show that extracting the set of atom pairs that display the largest V(xc) values in the hierarchy is equivalent to retrieving the molecular graph itself. Notably, domain-averaged exchange-correlation energies are transferable, and they can be used to calculate bond strengths. Fine-grained details resulted to be related to simple stereoelectronic effects. These ideas are demonstrated in a set of simple pilot molecules.

Contribution of 18F-sodium fluoride PET/CT to the study of the carotid atheroma calcification.

AIM: To assess the calcification process of the carotid plaque by (18)F-sodium fluoride PET/CT imaging. MATERIAL AND METHODS: A prospectively designed study including 15 patients in whom an atheroma plaque was detected by contrast enhanced CT scan during a neurological work-up was performed. A total of 29 plaques, 19 asymptomatic and 10 symptomatic, were studied. An (18)F-sodium fluoride PET/CT scan was acquired 180min after the i.v. injection of 370 MBq of (18)F-sodium fluoride in all the patients. The images obtained were analyzed visually according to the intensity of the uptake. RESULTS: All the plaques showed (18)F-sodium fluoride uptake, regardless of the intensity. However, the plaques of the symptomatic group showed a level of 2 or greater intensity while the intensity in 6 of the 19 in the asymptomatic group was lower than 2. CONCLUSIONS: Although the study is limited by the small number of cases, the results show the feasibility of the technique to study the calcification of the atheroma using (18)F-sodium fluoride and suggest an association between symptomatology and higher uptake of (18)F-sodium fluoride. Thus, these results encourage us to continue this study, with the inclusion of a larger number of patients.

Chemical Interactions and Spin Structure in (O2)4: Implications for the ε-O2 Phase.

The chemical interactions and spin structure of (O2)4 in its ground singlet state are analyzed by means of Quantum Chemical Topology descriptors. The energetic contributions of the Interacting Quantum Atoms approach are used to obtain information about the class of interactions displayed along the dissociation path of (O2)4. The exchange-correlation contribution to the binding energy is non-negligible for the O2-O2 interactions at intermolecular distances close to those found for the pressure induced ε phase of solid (O2) and this strengthening of the intermolecular bonding is built up from a simultaneous weakening of the intramolecular bond. This result is of interest in connection with the observed softening of the IR vibron frequency in the lower pressure range of the ε phase. The spin structure in the real space along the dissociation process is interpreted with the help of the so-called electron number distribution functions. At large distances, the four triplet O2 molecules are arranged in a way consistent with an antiferromagnetic structure, whereas at short distances, a significant spin redistribution is driven by the exchange process and it involves a propensity toward a null magnetic moment per molecule. Such probability behavior can be related with the magnetic evolution of solid oxygen across the δ → ε phase transition. Additional calculations of (O2)4 excited states support the conclusion that the relative stabilization and magnetic features of the ground singlet state are due to the onset of the new intermolecular bonds, and not to an exclusive modification of the electronic character within the O2 molecules.

Water quality assessment using satellite-derived chlorophyll-a within the European directives, in the southeastern Bay of Biscay.

The implementation of water quality European Directives requires an intensification of water quality monitoring, within the limits of the Exclusive Economic Zone. Remote sensing technologies can provide a valuable tool for frequent, synoptic, water-quality observations, over large areas. The aim of this study is to assess the ecological status of Basque coastal water bodies using satellite imagery from MODIS sensor, together with optical and chlorophyll-ain situ measurements. Thus, sea surface satellite-derived chl-a algorithms, the OC3 M, OC5 and a Local empirical algorithm, were compared against in situ measurements using satellite in situ match-ups, 90th Percentile (P90) monthly values for the 2005-2010 period. The OC5 algorithm corresponded most accurately with in situ measurements performed in the area, hence, it was selected. A P90 chlorophyll-a map was created with this algorithm to apply the classification scheme required by the directives. The classification of water bodies, based upon satellite-derived chlorophyll-a, could improve considerably the assessment of water quality.

Disección arterial carotídea: cuándo se hace necesario replantearse el tratamiento / Dissection of the carotid artery: when it becomes necessary to reconsider the treatment

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Selective labeling of lipid droplets in aldehyde fixed cell monolayers by lipophilic fluorochromes.

We evaluated a number of lipophilic dyes and fluorochromes, including oxazone and thiazone derivatives of oxazine and thiazine dyes, scintillator agents, a carotenoid and a metal-porphyrin complex for visualization of lipid droplets within aldehyde fixed cultured HeLa and BGC-1 cells. Observation under ultraviolet, blue or green exciting light revealed selective fluorescence of lipid droplets, particularly after treatment with aqueous solutions of Nile blue and brilliant cresyl blue oxazones, toluidine blue thiazone, or propylene glycol solutions of canthaxanthin, ethyl-BAO, and ZnTPyP. Mounting in water was required to maintain the fluorescence of lipids; the use of glycerol, Mowiol or Vectashield was not adequate. The effect of dye structure on staining intensity was assessed with the aid of numerical structure parameters modeling lipophilicity (HI and log P), overall size (MW) and the size of the conjugated system (conjugated bond number; CBN). The best stains for lipid droplets were relatively lipophilic (HI > 4.0, log P > 5.0), of small size overall (MW < 370), with small conjugated systems (CBN < 24), and not significantly amphiphilic. The two hydrophobic-hydrophilic parameters (the classic log P and the hydrophobic index, HI; values calculated by molecular modeling software) were highly correlated; however, HI was a more suitable hydrophobicity index for the dyes studied here.

Occurrence of plant growth regulators in Psilotum nudum.

The endogenous content of the auxin indole-3-acetic acid (IAA), the cytokinins trans-zeatin (tZ), trans-zeatin riboside (tZR), dihydrozeatin (DHZ), dihydrozeatin riboside (DHZR), isopentenyladenine (iP) and isopentenyladenosine (iPR), the gibberellins GA(1), GA(3), GA(4), GA(7), GA(9) and GA(20) in the rhizome and aerial shoots during and after sporogenesis were measured by high-performance liquid chromatography-tandem mass spectrometry in the fern Psilotum nudum. The present study shows, for the first time, the presence of the auxin IAA, the cytokinins tZR, DHZR and iP, and the gibberellins GA(4), GA(9) and GA(20) in the rhizome and aerial shoots of this species and suggests a possible role of gibberellins in the evolution of ferns.

Epigenetic changes detected in micropropagated hop plants.

Micropropagation is a widely used technique in hops (Humulus lupulus L.). However, to the best of our knowledge, the genetic and epigenetic stability of the microplants has never been tested before. In the present study, two hop accessions were established in vitro and micropropagated for 2 years. The genetic and epigenetic stability of the in vitro plants was analyzed with several molecular techniques: random amplified DNA polymorphism (RAPD), retrotransposon microsatellite amplified polymorphism (REMAP), and methylation-sensitive amplification polymorphism (MSAP). No genetic variation among control and treated plants was found, even after 12 cycles of micropropagation. Epigenetic variation was detected, first, when field and in vitro samples were compared. Nearly a 30% of the detected fragments presented the same pattern of alterations in all the vitroplants. Second, lower levels of epigenetic variation were detected among plants from the different subcultures. Part of this detected variation seemed to be accumulated along the 12 sequential subcultures tested.

Genetic and epigenetic stability of cryopreserved and cold-stored hops (Humulus lupulus L.).

Conventional cold storage and cryopreservation methods for hops (Humulus lupulus L.) are available but, to our knowledge, the genetic and epigenetic stability of the recovered plants have not been tested. This study analyzed 51 accessions of hop using the molecular techniques, Random Amplified DNA Polymorphism (RAPD) and Amplified Fragment Length Polymorphism (AFLP), revealing no genetic variation among greenhouse-grown controls and cold stored or cryopreserved plants. Epigenetic stability was evaluated using Methylation Sensitive Amplified Polymorphism (MSAP). Over 36% of the loci were polymorphic when the cold and cryo-treated plants were compared to greenhouse plants. The main changes were demethylation events and they were common to the cryopreserved and cold stored plants indicating the possible effect of the in vitro establishment process, an essential step in both protocols. Protocol-specific methylation patterns were also detected indicating that both methods produced epigenetic changes in plants following cold storage and cryopreservation.