Insights into cellular crosstalk regulating cytoplasmic male sterility and fertility restoration.

Cytoplasmic male sterility has been a popular genetic tool in development of hybrids. The molecular mechanism behind maternal sterility varies from crop to crop. An understanding of underlying mechanism can help in development of new functional CMS gene in crops which lack effective and stable CMS systems. In crops where seed or fruit is the commercial product, fertility must be recovered in F1 hybrids so that higher yield gains can be realized. This necessitates the presence of fertility restorer gene (Rf) in nucleus of male parent to overcome the effect of sterile cytoplasm. Fertility restoring genes have been identified in crops like wheat, maize, sunflower, rice, pepper, sugar beet, pigeon pea etc. But in crops like eggplant, bell pepper, barley etc. unstable fertility restorers hamper the use of Cytoplasmic genic male sterility (CGMS) system. Stability of CGMS system is influenced by environment, genetic background or interaction of these factors. This review thus aims to understand the genetic mechanisms controlling mitochondrial-nuclear interactions required to design strong and stable restorers without any pleiotropic effects in F1 hybrids.

Genome-wide identification of the restorer-of-fertility-like (RFL) gene family in Brassica napus and expression analysis in Shaan2A cytoplasmic male sterility.

BACKGROUND: Cytoplasmic male sterility (CMS) is very important in hybrid breeding. The restorer-of-fertility (Rf) nuclear genes rescue the sterile phenotype. Most of the Rf genes encode pentatricopeptide repeat (PPR) proteins. RESULTS: We investigated the restorer-of-fertility-like (RFL) gene family in Brassica napus. A total of 53 BnRFL genes were identified. While most of the BnRFL genes were distributed on 10 of the 19 chromosomes, gene clusters were identified on chromosomes A9 and C8. The number of PPR motifs in the BnRFL proteins varied from 2 to 19, and the majority of BnRFL proteins harbored more than 10 PPR motifs. An interaction network analysis was performed to predict the interacting partners of RFL proteins. Tissue-specific expression and RNA-seq analyses between the restorer line KC01 and the sterile line Shaan2A indicated that BnRFL1, BnRFL5, BnRFL6, BnRFL8, BnRFL11, BnRFL13 and BnRFL42 located in gene clusters on chromosomes A9 and C8 were highly expressed in KC01. CONCLUSIONS: In the present study, identification and gene expression analysis of RFL gene family in the CMS system were conducted, and seven BnRFL genes were identified as candidates for the restorer genes in Shaan2A CMS. Taken together, this method might provide new insight into the study of Rf genes in other CMS systems.

Enhancing the Assessment of Resiliency to Suicide Ideation among Older Adults: The Development and Initial Validation of the Reasons for Living-Suicide Resiliency Scale (RFL-SR).

Objectives: To derive a brief late-life suicide resiliency scale from the 69-item Reasons for Living Scale-Older Adult version (RFL-OA).Methods: We conducted a series of secondary analyses of RFL-OA data (N = 204) from a dataset combining: 1. A follow-up assessment of nursing home residents in the Geriatric Suicide Ideation Scale (GSIS) development study; 2. A trial of Interpersonal Psychotherapy (IPT) with suicidal older adults; 3. A longitudinal study of risk and resiliency to late-life suicide ideation. We specifically assessed the distributions of RFL-OA items and their associations with suicide ideation and behavior to create an RFL-Suicide Resiliency subscale (RFL-SR); we then tested the psychometric properties of this measure's items drawn from the larger RFL-OA.Results: Nine RFL-OA items were significantly associated with suicide ideation and history of suicide behavior and were not highly correlated with social desirability. Psychometric analyses supported the internal consistency, test-retest reliability, and construct validity of this scale.Conclusions: The items of the RFL-SR demonstrated strong psychometric properties with older adults in clinical and community settings.Clinical Implications: The RFL-SR may make a useful addition to suicide risk assessment in gerontological research and clinical practice.

The far-upstream regulatory region of RFL is required for its precise spatial-temporal expression for floral development in rice.

KEY MESSAGE: A rice mutant aberrant floral organ 1 (afo1) was identified, showing increased floral organ number, aberrant floral organ identity and loss of floral meristem determinacy. A disruption of sequence integrity at 6292-bp upstream of RFL by a T-DNA insertion led to varied RFL expression patterns in floral meristem and floret in afo1 and caused the mutant phenotype. The LEAFY (LFY) transcription factor and its homologs affect many aspects of plant development, especially floral development. RICE FLORICAULA/LEAFY (RFL), the rice ortholog of LFY, has complicated expression patterns and different functions in floral development. However, the mechanisms regulating the spatial-temporal expression of RFL remain largely unknown. Here, we describe a rice aberrant floral organ 1 (afo1) mutant that was produced by a T-DNA insertion at 6292-bp upstream of the start codon of RFL. This insertion altered the expression of RFL in floral meristem (FM) and floret. The in situ hybridization result showed that, when florets appear, RFL was expressed almost exclusively at the palea/lemma adaxial base adjacent to lodicules in the wild-type panicle. However, in afo1 florets, RFL mRNA signals were detected in the region between lodicule and stamen, and strong signals persisted in FM. The altered pattern of RFL expression in afo1 resulted in enlarged FMs, more floral organs, aberrant floral organ identity, and loss of FM determinacy. Transformation of rice with an RFL construct driven by the 6292-bp upstream genomic sequence re-built the mutant phenotype similar to afo1. The results suggest that the far-upstream region of RFL may contain potential cis element(s) that are critical to define the precise spatial-temporal expression pattern of RFL for its function in floral development.

Comparative genomic analysis of the compound Brassica napus Rf locus.

BACKGROUND: The plant trait of cytoplasmically-inherited male sterility (CMS) and its suppression by nuclear restorer-of-fertility (Rf) genes can be viewed as a genetic arms race between the mitochondrial and nuclear genomes. Most nuclear Rf genes have been shown to encode P-type pentatricopeptide repeat proteins (PPRs). Phylogenetic analysis of P-class PPRs from sequenced plants genomes has shown that Rf-proteins cluster in a distinct clade of P-class PPRs, RFL-PPRs, that display hallmarks of positive evolutionary selection. Genes encoding RFL-PPRs (RFLs) within a given plant genome tend to be closely related both in sequence and position, but a detailed understanding of how such species-specific expansion occurs is lacking. In the canola, (oilseed rape) species Brassica napus, previous work has indicated the nuclear restorer genes for the two native forms of CMS, Rfn (for nap CMS) and Rfp (pol CMS), represent alternate haplotypes, or alleles, of a single nuclear locus. RESULTS: Fine genetic mapping indicates that Rfn does indeed localize to the same genomic region as Rfp. We find this region is enriched in RFL genes, three of which, based on their position and expression, represent potential candidates for Rfn; one of these genes, designated PPR4, is a preferred candidate in that it is not expressed in the nap CMS line. Comparison of the corresponding regions of the genomes of B. rapa, B. oleracea, Arabidopsis thaliana and A. lyrata provides insight into the expansion of this group of RFL genes in different lines of evolutionary descent. CONCLUSIONS: Unlike other nuclear restorer loci containing multiple RFL genes, the RFL genes in the Rf region of B. napus are not present in tandem arrays but rather are dispersed in genomic location. The genes do not share similar flanking non-coding regions and do not contain introns, indicating that they have duplicated primarily through a retrotransposition-mediated process. In contrast, segmental duplication has been responsible for the distribution of the 10 sequences we annotated as RFL genes in the corresponding region of the A. lyrata genome. Our observations define the Brassica Rf locus and indicate that different mechanisms may be responsible for the proliferation of RFL genes even among closely related genomes.

Reasons for living, meaning in life, and suicide ideation: investigating the roles of key positive psychological factors in reducing suicide risk in community-residing older adults.

OBJECTIVES: To investigate the roles of reasons for living (RFL) and meaning in life (MIL) in potentially promoting mental health and well-being and protecting against suicide ideation among community-residing older adults and to investigate the psychometric properties of the Reasons for Living Scale-Older Adult version (RFL-OA). METHOD: Of 173 older adults initially recruited into a longitudinal study on late-life suicide ideation, 109 completed the RFL-OA and measures of cognitive and physical functioning and positive and negative psychological factors at a two-year follow-up assessment. We tested a model in which RFL and MIL protect against suicide ideation, controlling for demographic and clinical factors. We also assessed the psychometric properties of the RFL-OA in community-residing older adults, investigating its internal consistency and its convergent (MIL, perceived social support, and life satisfaction), divergent (loneliness, depressive symptom severity, and suicide ideation), and discriminant validity (cognitive and physical functioning). RESULTS: RFL-OA scores explained significant variance in suicide ideation, controlling for age, sex, depressive symptom severity, and loneliness. MIL explained significant unique variance in suicide ideation, controlling for these factors and RFL, and MIL significantly mediated the association between RFL and suicide ideation. Psychometric analyses indicated strong internal consistency (α = .94), convergent, divergent, and discriminant validity for the RFL-OA relative to positive and negative psychological factors and cognitive and physical functioning. CONCLUSION: These findings add to a growing body of literature suggesting merit in investigating positive psychological factors together with negative factors when assessing suicide risk and planning psychological services for older adults.

Functions for rice RFL in vegetative axillary meristem specification and outgrowth.

Axillary meristems (AMs) are secondary shoot meristems whose outgrowth determines plant architecture. In rice, AMs form tillers, and tillering mutants reveal an interplay between transcription factors and the phytohormones auxin and strigolactone as some factors that underpin this developmental process. Previous studies showed that knockdown of the transcription factor gene RFL reduced tillering and caused a very large decrease in panicle branching. Here, the relationship between RFL, AM initiation, and outgrowth was examined. We show that RFL promotes AM specification through its effects on LAX1 and CUC genes, as their expression was modulated on RFL knockdown, on induction of RFL:GR fusion protein, and by a repressive RFL-EAR fusion protein. Further, we report reduced expression of auxin transporter genes OsPIN1 and OsPIN3 in the culm of RFL knockdown transgenic plants. Additionally, subtle change in the spatial pattern of IR4 DR5:GFP auxin reporter was observed, which hints at compromised auxin transport on RFL knockdown. The relationship between RFL, strigolactone signalling, and bud outgrowth was studied by transcript analyses and by the tillering phenotype of transgenic plants knocked down for both RFL and D3. These data suggest indirect RFL-strigolactone links that may affect tillering. Further, we show expression modulation of the auxin transporter gene OsPIN3 upon RFL:GR protein induction and by the repressive RFL-EAR protein. These modified forms of RFL had only indirect effects on OsPIN1. Together, we have found that RFL regulates the LAX1 and CUC genes during AM specification, and positively influences the outgrowth of AMs though its effects on auxin transport.

'Give Me a Reason to Live!' Examining Reasons for Living Across Levels of Suicidality.

Scholarly research focusing on social psychological factors (e.g. mental health) and social environmental factors (e.g. childhood trauma) has found these measures to be correlated with suicidality. However, such literature has tended to overlook what may impact one's reasons for living. Using a sample of over 1,200 students from a Canadian university, the goal of the current study is to empirically test, by employing multivariate nested regression models (by levels of suicidal behaviour), known and relative unknown correlates with reasons for living, with a particular focus on strength of religious faith, which is a well-known predictor for suicidality, but less studied as a reason for living. Results show that, among students with serious suicidal ideation and/or a previous suicide attempt, the strongest predictor for student's reasons for living was strength of religious faith. Strength of religious faith has seldom been acknowledged or identified as an important measure in assessing one's reasons to live. These findings have implications for the role of religiosity among suicidality research, especially studies that focus on reasons for living.

The role of neutrophils and neutrophil extracellular traps (NETs) in stages, outcomes and pregnancy complications.

Neutrophils are the main components of innate immunity to eliminate infectious pathogens. Neutrophils play a role in several stages of the reproductive cycle, and their presence in the female reproductive system is highly regulated, so their function may change during pregnancy. Emerging evidence suggests that neutrophils are important at all stages of pregnancy, from implantation, placentation, and connective tissue regeneration to birth, as well as birth itself. Neutrophil extracellular traps (NETs) are defined as extracellular strands of unfolded DNA together with histone complexes and neutrophil granule proteins. NET formation is a new mechanism of these cells for their defense function. These strands containing DNA and antimicrobial peptides were initially recognized as one of the defense mechanisms of neutrophils, but later it was explained that they are involved in a variety of non-infectious diseases. Since the source of inflammation and tissue damage is the irregular activity of neutrophils, it is not surprising that NETosis are associated with a number of inflammatory conditions and diseases. The overexpression of NET components or non-principled NET clearance is associated with the risk of production and activation of autoantibodies, which results in participation in autoinflammatory and autoimmune disorders (SLE, RA), fibrosis, sepsis and other disorders such as vascular diseases, for example, thrombosis and atherosclerosis. Recent published articles have shown the role of neutrophils and extracellular traps (NETs) in pregnancy, childbirth and pregnancy-related diseases. The aim of this study was to identify and investigate the role of neutrophils and neutrophil extracellular traps (NETs) in the stages of pregnancy, as well as the complications caused by these cells.

Induction of Male Sterility by Targeted Mutation of a Restorer-of-Fertility Gene with CRISPR/Cas9-Mediated Genome Editing in Brassica napus L.

Brassica napus L. (canola, oil seed rape) is one of the world's most important oil seed crops. In the last four decades, the discovery of cytoplasmic male-sterility (CMS) systems and the restoration of fertility (Rf) genes in B. napus has improved the crop traits by heterosis. The homologs of Rf genes, known as the restoration of fertility-like (RFL) genes, have also gained importance because of their similarities with Rf genes. Such as a high non-synonymous/synonymous codon replacement ratio (dN/dS), autonomous gene duplications, and a possible engrossment in fertility restoration. B. napus contains 53 RFL genes on chromosomes A9 and C8. Our research aims to study the function of BnaRFL11 in fertility restoration using the CRISPR/Cas9 genome editing technique. A total of 88/108 (81.48%) T0 lines, and for T1, 110/145 (75%) lines carried T-DNA insertions. Stable mutations were detected in the T0 and T1 generations, with an average allelic mutation transmission rate of 81%. We used CRISPR-P software to detect off-target 50 plants sequenced from the T0 generation that showed no off-target mutation, signifying that if the designed sgRNA is specific for the target, the off-target effects are negligible. We also concluded that the mutagenic competence of the designed sgRNAs mediated by U6-26 and U6-29 ranged widely from 31% to 96%. The phenotypic analysis of bnarfl11 revealed defects in the floral structure, leaf size, branch number, and seed production. We discovered a significant difference between the sterile line and fertile line flower development after using a stereomicroscope and scanning electron microscope. The pollen visibility test showed that the pollen grain had utterly degenerated. The cytological observations of homozygous mutant plants showed an anther abortion stage similar to nap-CMS, with a Orf222, Orf139, Ap3, and nad5c gene upregulation. The bnarfl11 shows vegetative defects, including fewer branches and a reduced leaf size, suggesting that PPR-encoding genes are essential for the plants' vegetative and reproductive growth. Our results demonstrated that BnaRFL11 has a possible role in fertility restoration. The current study's findings suggest that CRISPR/Cas9 mutations may divulge the functions of genes in polyploid species and provide agronomically desirable traits through a targeted mutation.

Longitudinal Development of Reasons for Living and Dying With Suicide Attempters: A 2-Year Follow-Up Study.

Background: Clinical interventions for patients after a suicide attempt might include a focus on Reasons for Living (RFL) and/or Reasons for Dying (RFD). The present study examined the longitudinal development of RFL and RFD in patients with and without a suicide-specific intervention - the Attempted Suicide Short Intervention Program (ASSIP). Methods: In this secondary analysis of a 2-year follow-up randomized controlled study, participants completed the Suicide Status Form II to assess RFL and RFD, at baseline, as well as at 6-, 12-, 18-, and 24-months follow-up. Growth models and latent class analysis were used to investigate longitudinal developments in RFL and RFD. Regression models were used to test the association between RFL, RFD and suicidal reattempts and ideation. Results: Cross-sectionally and longitudinally, RFD, but not RFL, were associated with suicide reattempts and suicidal ideation. The number of RFD decreased significantly across the 24 month period (from 1.90 at t1 to 1.04 at t5 in the control group and from 2.32 at t1 to 0.51 at t5 in the intervention group), and this decrease was stronger (b = -0.02; p = 0.004) in the ASSIP group than in the control group. There was no overall change in RFL. Three latent trajectories of RFD were identified: a decreasing (n = 77), a steady high (n = 17) and a trajectory with first increasing and then decreasing RFD (n = 26). The proportion of patients in the ASSIP intervention was highest in the decreasing trajectory and lowest in the steady high trajectory. Patients in the steady high trajectory were characterized by worse mental health and fewer social obligations (partner, children) at baseline. Conclusion: The results confirm the importance of RFD within the suicidal process and show that the number of RFD can be further reduced over the period of 24 months with short interventions such as ASSIP. The relevance of number of RFL in the suicidal process, as protective factor, was not confirmed. In the subgroup of patients whose RFD did not decrease over a long period of time, there is a particularly high risk of suicidal ideation/behavior. Clinical interventions should focus more closely on RFD, their etiology and maintenance.

Triticeae genome sequences reveal huge expansions of gene families implicated in fertility restoration.

Breakthroughs in assembly of whole-genome sequencing and targeted sequence capture data have accelerated comparative genomics analyses in cereals with big and complex genomes such as wheat. This newly acquired information has revealed unexpected expansions in two large gene families linked to restoration of fertility in species that exhibit cytoplasmic male sterility. Extreme levels of copy-number and structural variation detected within and between species illustrate the genetic diversity among the family members and reveal the evolutionary mechanisms at work. This new knowledge will greatly facilitate the development of hybrid production strategies in wheat and related species.

Analysis of the association between polymorphisms in Vitamin D receptor gene and dental caries.

BACKGROUND: The influence of environmental factors on dental caries is a well-established fact; however, little is known about the impact of genetics on the caries process. AIMS: This study was aimed to analyse the association between the vitamin D receptor (VDR) gene polymorphism and dental caries. It also compared the risk of dental caries between individuals with homozygous (tt and TT) genotype and heterozygous (Tt) genotype. Further, this study also aimed to compare VDR gene polymorphism by gender. METHODOLOGY: A total of 196 subjects were selected based on their decayed, missing, filled teeth (DMFT) index according to the WHO guidelines (1997), and were divided into two groups based on caries experience. The case group was further classified into low and high caries groups. The saliva samples were collected, DNA was extracted, and VDR Taq1 (T/C) gene polymorphism was analysed by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) and visualized under 2% agarose gel. RESULTS: There was no significant difference in the VDR Taq1 polymorphism and dental caries between the case and control group. However, further stratification showed a statistically significant difference based on caries experience in the study group. The heterozygous mutant genotype (Tt) was associated with a higher caries experience (62.1%) (P = 0.008) with the frequency of 't' allele being 77% as compared to 'T' allele (63.6%) in the high caries group. CONCLUSION: VDR Taq1 gene polymorphism does not seem to have a role in the incidence of dental caries in the studied population; however, it may have a role in the severity of the caries process.

IGF1R Deficiency Modulates Brain Signaling Pathways and Disturbs Mitochondria and Redox Homeostasis.

Insulin-like growth factor 1 receptor (IGF1R)-mediated signaling pathways modulate important neurophysiological aspects in the central nervous system, including neurogenesis, synaptic plasticity and complex cognitive functions. In the present study, we intended to characterize the impact of IGF1R deficiency in the brain, focusing on PI3K/Akt and MAPK/ERK1/2 signaling pathways and mitochondria-related parameters. For this purpose, we used 13-week-old UBC-CreERT2; Igf1rfl/fl male mice in which Igf1r was conditionally deleted. IGF1R deficiency caused a decrease in brain weight as well as the activation of the IR/PI3K/Akt and inhibition of the MAPK/ERK1/2/CREB signaling pathways. Despite no alterations in the activity of caspases 3 and 9, a significant alteration in phosphorylated GSK3ß and an increase in phosphorylated Tau protein levels were observed. In addition, significant disturbances in mitochondrial dynamics and content and altered activity of the mitochondrial respiratory chain complexes were noticed. An increase in oxidative stress, characterized by decreased nuclear factor E2-related factor 2 (NRF2) protein levels and aconitase activity and increased H2O2 levels were also found in the brain of IGF1R-deficient mice. Overall, our observations confirm the complexity of IGF1R in mediating brain signaling responses and suggest that its deficiency negatively impacts brain cells homeostasis and survival by affecting mitochondria and redox homeostasis.

A Novel Approach of Promoting Adhesion of Reinforcing Cord to Elastomers by Plasma Polymerization.

Adhesion of cords to elastomers is crucial for many elastomeric products, such as tires and V-belts. The best adhesion system so far is based on a combination of resorcinol, formaldehyde, and a latex (RFL). However, this cord treatment has serious disadvantages in terms of processing and toxicity. A promising alternative is a plasma treatment of the cords prior to be embedded in the elastomer. For rayon cords, a plasma polymerization of sulfur-containing precursors results in adhesion levels close to RFL treatment. However, for polyethylene terephthalate (PET) cords, this treatment is not satisfactory. For this type of cords, a water-plasma activation followed by a silane dip is more promising, as 72% of the adhesion level of RFL treatment could be achieved. For rayon, an even higher adhesion level was realized.

In Silico Identification of Candidate Genes for Fertility Restoration in Cytoplasmic Male Sterile Perennial Ryegrass (Lolium perenne L.).

Perennial ryegrass (Lolium perenne L.) is widely used for forage production in both permanent and temporary grassland systems. To increase yields in perennial ryegrass, recent breeding efforts have been focused on strategies to more efficiently exploit heterosis by hybrid breeding. Cytoplasmic male sterility (CMS) is a widely applied mechanism to control pollination for commercial hybrid seed production and although CMS systems have been identified in perennial ryegrass, they are yet to be fully characterized. Here, we present a bioinformatics pipeline for efficient identification of candidate restorer of fertility (Rf) genes for CMS. From a high-quality draft of the perennial ryegrass genome, 373 pentatricopeptide repeat (PPR) genes were identified and classified, further identifying 25 restorer of fertility-like PPR (RFL) genes through a combination of DNA sequence clustering and comparison to known Rf genes. This extensive gene family was targeted as the majority of Rf genes in higher plants are RFL genes. These RFL genes were further investigated by phylogenetic analyses, identifying three groups of perennial ryegrass RFLs. These three groups likely represent genomic regions of active RFL generation and identify the probable location of perennial ryegrass PPR-Rf genes. This pipeline allows for the identification of candidate PPR-Rf genes from genomic sequence data and can be used in any plant species. Functional markers for PPR-Rf genes will facilitate map-based cloning of Rf genes and enable the use of CMS as an efficient tool to control pollination for hybrid crop production.

Use of cost effective and rapid molecular tools for identification of Candida species, opportunistic pathogens.

BACKGROUND AND PURPOSE: Candidiasis is a widespread fungal infection caused by different Candida species. Rapid identification of Candida species in clinical laboratory is becoming increasingly important since the identification and discrimination of ethological agents for early treatment. We aimed at molecular identification of commonly Candida species isolated from clinical samples by using both PCR-RFLP assay and amplification of hwp1 gene. MATERIALS AND METHODS: Clinical samples comprising of vaginal specimens ,cutaneous, sputum, bronchoalveolar lavage(BAL,( and blood cultures were recovered from suspected patients. Candida isolates were initially identified phenotypically and confirmed by molecular approaches based on restriction fragment length polymorphism (PCR-RFLP (with MspI restriction enzyme. Amplification of hwp1 gene was performed for discrimination of C. albicans from C. dubliniensis and C.africana. RESULTS: The most abundant species were C. albicans (n=67; 44.6 %), C. glabrata (n=10; 20 %), C.tropicalis (n=20; 13.3 %), C. krusei (n=12; 8 %), C.parapsilosis (n=11; 7.3 %). Out of 67 C.albicans species, 6 species identified as C. dubliniensis and 4 species identified as C. africana. CONCLUSION: High frequency of non-albicansCandida species and differences in levels of susceptibility to the antifungal agents are important issues in medicine .Therefore, to manage the Candida-related infections properly, molecular diagnostic methods would be fast, reliable and even cost-effective approaches for identification of Candida species.

Long-term efficacy and safety of internal neurolysis for trigeminal neuralgia without neurovascular compression.

OBJECT Trigeminal neuralgia (TN) occurs and recurs in the absence of neurovascular compression (NVC). While microvascular decompression (MVD) is the most effective treatment for TN, it is not possible when NVC is not present. Therefore, the authors sought to evaluate the safety, efficacy, and durability of internal neurolysis (IN), or "nerve combing," as a treatment for TN without NVC. METHODS This was a retrospective review of all cases of Type 1 TN involving all patients 18 years of age or older who underwent evaluation (and surgery when appropriate) at Oregon Health & Science University between July 2006 and February 2013. Chart reviews and telephone interviews were conducted to assess patient outcomes. Pain intensity was evaluated with the Barrow Neurological Institute (BNI) Pain Intensity scale, and the Brief Pain Inventory-Facial (BPI-Facial) was used to assess general and face-specific activity. Pain-free survival and durability of successful pain relief (BNI pain scores of 1 or 2) were statistically evaluated with Kaplan-Meier analysis. Prognostic factors were identified and analyzed using Cox proportional hazards regression. RESULTS A total of 177 patients with Type 1 TN were identified. A subgroup of 27 was found to have no NVC on high-resolution MRI/MR angiography or at surgery. These patients were significantly younger than patients with classic Type 1 TN. Long-term follow-up was available for 26 of 27 patients, and 23 responded to the telephone survey. The median follow-up duration was 43.4 months. Immediate postoperative results were comparable to MVD, with 85% of patients pain free and 96% of patients with successful pain relief. At 1 year and 5 years, the rate of pain-free survival was 58% and 47%, respectively. Successful pain relief at those intervals was maintained in 77% and 72% of patients. Almost all patients experienced some degree of numbness or hypesthesia (96%), but in patients with successful pain relief, this numbness did not significantly impact their quality of life. There was 1 patient with a CSF leak and 1 patient with anesthesia dolorosa. Previous treatment for TN was identified as a poor prognostic factor for successful outcome. CONCLUSIONS This is the first report of IN with meaningful outcomes data. This study demonstrated that IN is a safe, effective, and durable treatment for TN in the absence of NVC. Pain-free outcomes with IN appeared to be more durable than radiofrequency gangliolysis, and IN appears to be more effective than stereotactic radiosurgery, 2 alternatives to posterior fossa exploration in cases of TN without NVC. Given the younger age distribution of patients in this group, consideration should be given to performing IN as an initial treatment. Accrual of further outcomes data is warranted.

Nitrosopersulfide (SSNO(-)) accounts for sustained NO bioactivity of S-nitrosothiols following reaction with sulfide.

Sulfide salts are known to promote the release of nitric oxide (NO) from S-nitrosothiols and potentiate their vasorelaxant activity, but much of the cross-talk between hydrogen sulfide and NO is believed to occur via functional interactions of cell regulatory elements such as phosphodiesterases. Using RFL-6 cells as an NO reporter system we sought to investigate whether sulfide can also modulate nitrosothiol-mediated soluble guanylyl cyclase (sGC) activation following direct chemical interaction. We find a U-shaped dose response relationship where low sulfide concentrations attenuate sGC stimulation by S-nitrosopenicillamine (SNAP) and cyclic GMP levels are restored at equimolar ratios. Similar results are observed when intracellular sulfide levels are raised by pre-incubation with the sulfide donor, GYY4137. The outcome of direct sulfide/nitrosothiol interactions also critically depends on molar reactant ratios and is accompanied by oxygen consumption. With sulfide in excess, a 'yellow compound' accumulates that is indistinguishable from the product of solid-phase transnitrosation of either hydrosulfide or hydrodisulfide and assigned to be nitrosopersulfide (perthionitrite, SSNO(-); λ max 412 nm in aqueous buffers, pH 7.4; 448 nm in DMF). Time-resolved chemiluminescence and UV-visible spectroscopy analyses suggest that its generation is preceded by formation of the short-lived NO-donor, thionitrite (SNO(-)). In contrast to the latter, SSNO(-) is rather stable at physiological pH and generates both NO and polysulfides on decomposition, resulting in sustained potentiation of SNAP-induced sGC stimulation. Thus, sulfide reacts with nitrosothiols to form multiple bioactive products; SSNO(-) rather than SNO(-) may account for some of the longer-lived effects of nitrosothiols and contribute to sulfide and NO signaling.

Reflux finding score: análisis de la concordancia inter-observador / Reflux finding score: analysis of the interobserver concordance / Reflux finding score: análise de concordância interobservador

Introducción: el reflujo laringofaríngeo (RLF) es una manifestación extradigestiva del Reflujo Gastroesofágico (RGE). Actualmente, el algoritmo diagnóstico usado en la mayoría de los Servicios de Otorrinolaringologìa (ORL) consiste en identificar los síntomas sugestivos de reflujo laringofaríngeo y realizar una fibronasolaringoscopia flexible para determinar si existen cambios estructurales identificables en la laringe o en los tejidos vecinos. Se ha desarrollado una escala subjetiva para cuantificar la severidad de los cambios inflamatorios de la laringe, identificando los hallazgos laringoscópicos asociados con reflujo laringofaríngeo. Los mismos son clasificados por el índice de hallazgos de reflujo “Reflux Finding Score” (RFS), desarrollado por Belafky y cols. El puntaje máximo es de 26 y un puntaje de 7 o mayor a 7 es considerado altamente sospechoso de RLF. Objetivos: Determinar el índice Kappa de Resultados Interobservador usando el RFS en pacientes con RLF en el Servicio de Otorrinolaringología del Sanatorio Allende. Determinar las características de los pacientes con diagnóstico de RLF según: Sexo, edad, síntomas presentes y tratamiento indicado en pacientes que consultaron ambulatoriamente en el Servicio de ORL del Sanatorio Allende sucursal Nueva Córdoba. Material y método: Se determinó el índice Kappa de Resultados entre observadores usando el RFS en paciente con RLF entre 2 médicos del staff de Servicio de ORL del Sanatorio Allende, se analizó frecuencia de sexo, edad, síntomas, conducta médica. Conclusión: El índice Kappa del RFS para cada uno de los 8 ìtems del RFS es leve, medido como < 0,2.

Introduction: the Laryngopharyngeal Reflux (LPR) is recognized as an extra-digestive manifestation of gastroesophageal reflux (GERD). At present, the diagnostic algorithm used in most of the otorhinolaryngology (ORL) services involves identifying suggestive symptoms of laryngopharyngeal reflux and carrying out a flexible fibronasolaryngoscopy to determine if there are identifiable structural changes in the larynx or neighboring tissues. A subjective scale has been developed to quantify the severity of inflammatory changes in the larynx by identifying endoscopic findings related to laryngopharyngeal reflux. These subjective findings for patients with LPR are classified using the “Reflux Finding Score” (RFS) developed by Belafsky et al.The maximum score is 26 and a score of 7 or greater is considered to indicate that LPR is highly likely. Objetive: To determine the Kappa Coefficient of the interobserver agreement using the RFS in patients with LPR in the Otorhinolaryngology Service of the Allende Hospital. To determine the characteristics of patients with LPR diagnosis according to: Gender, age, manifested symptoms and treatment indicated in patients that were treated as an outpatient in the ORL Service of the Allende Hospital, Nueva Córdoba’s branch. Material and method: The Kappa Coefficient was determined by the interobserver agreement using the RFS in patients with LPR between two Staff doctors of the ORL Service of the Allende Hospital; it was investigated the sex frequency, age, symptoms and medical behavior. Conclusions: The Kappa Coefficient of LPR for each of the eight items of LPR is poor, valued as < 0.2.

Introdução: o refluxo laringofaríngeo (RFL) é uma manifestação extra-digestiva do refluxo gastroesofágico (RGE). Atualmente o algoritmo diagnóstico usado pela maioria dos Serviços de Otorrinolaringología (ORL) consistem em identificar os sintomas sugestivos do RFL e realizar uma fibronasolaringoscopia flexível para determinar se existem mudanças estruturais identificáveis na laringe ou nos tecidos vizinhos. Existe uma escala subjetiva para quantificar a severidade das alterações inflamatórias na laringe, identificando os achados laringoscópicos que têm relação com o RFL. Eles são classificados pelo índice de achados de refluxo “Reflux Finding Score” (RFS), desenvolvido por Belafky e cols. A pontuação maxima é de 26 e uma pontuação de 7 ou maior é considerado quase sugestivo de RFL. Objetivos: Determinar o índice Kappa dos resultados interobservador usando o RFS em pacientes com RFL no Serviço de Otorrinolaringologia do Sanatorio Allende. Determinar as características dos pacientes com diagnóstico de RFL Segundo: Sexo, idade, sintomas presentes e tratamento indicado em pacientes que fizeram consultas externas no Serviço de ORL do Sanatorio Allende em Nueva Córdoba. Material e método: Determinou-se o índice Kappa dos resultados entre observadores usando o RFS em pacientes com RFL entre 2 médicos do staff do Serviço de ORL do Sanatorio Allende. Analizou-se a frequência entre o sexo, a idade, os sintomas, as doenças associadas e a terapeútica médica. Conclusões: O índice Kappa do RFS para cada item do RFS é insuficiente, medida como < 0,2.