Downregulation of thromboxane A2 and angiotensin II type 1 receptors associated with aging-related decrease in internal anal sphincter tone.

Aging-associated decrease in internal anal sphincter (IAS) tone (AADI) is a major contributor in the rectoanal incontinence (RI). To determine the pathogenesis of AADI, we investigated the effect of aging on GPCR activation and related downstream signaling. We particularly investigated two GPCRs that characterize IAS smooth muscle cells (SMCs): thromboxane A 2 and angiotensin II type 1. Two groups of Fischer 344 rats (6-month-old [young group] and 26-month-old [old group]) were employed to determine the GPCR function by isometric contraction, the expressions of GPCRs, and their downstream regulatory signaling proteins (regulator of G-protein signaling 2, RGS2; GPCR Kinase 5, GRK5; and β-arrestin, Arrb2) using RT-PCR, qPCR, and western blot analyses. We used reversible biotinylation to monitor the GPCR trafficking using SMCs. Aging selectively attenuated thromboxane A 2 and Ang II-induced IAS contraction. RT-PCR, qPCR, and WB data revealed a significant decrease in the expressions of the GPCRs and increase in the expression of RGS2, GRK5, and Arrb2. The increased GPCR internalization and decreased recycling under aging were validated by reversible biotinylation. We conclude that downregulation of GPCR, accompanied by upregulation of regulatory proteins, plays an important role in receptor desensitization and may be important underlying mechanisms of RI in certain aging patients

Observations on some motility disturbances of the human distal bowel and pelvic floor

The following paragraphs summarise the principal findings and conclusions drawn from results of the studies described in the previous Sections. | 1. The use of a microtransducer in anorectal manometry avoids many of the drawbacks of conventional fluid-filled systems. The data obtained is highly reproducible and probably more accurate. | 2. The proctometrogram provides a reproducible and useful method of evaluating the pressure-volume distension characteristics of the rectum in health and disease. The expected correlation between compliance and volume is confirmed. There do not appear to be significant variations in rectal volume or compliance with age in healthy subjects. | 3. The electrophysiological latency of the pudendo-anal reflex in health is 38.5±5.8 ras ( Mean±SD, n=38 ). It is completely reproducible and does not appear to vary with age or sex. | 4. In symptomatic chronic radiation anorectal injury: | 4.1 The manometric function of the internal anal sphincter is compromised with diminution in sphincter length, basal pressure and abnormalities of the rectosphincteric reflex. | 4.2 The external anal sphincter remains manoraetrically normal. Its mean motor unit potential duration is increased although the latency of the pudendo-anal reflex is unaffected. These findings suggest possible minor damage to its terminal innervation. | 4.3 The volume and compliance of the rectum are severely diminished, there being a significant positive correlation between these parameters. The manometric changes correlate well with symptomatic and sigmoidoscopic findings. | 5. Following colo-anal sleeve anastomosis for severe chronic radiation injury to the rectum the organic complications are resolved but some of the functional abnormalities persist. These are in part due to: | 5.1 Persisting internal anal sphincter dysfunction. | 5.2 External anal sphincter dysfunction on rectal distension and defaecation straining. | 5.3 Persisting severe reduction in rectal volume and compliance. | 5.4 Reduction in rectal sensory threshold volume, although rectal 'proprioception' remains intact. | 6. Some of the sphincteric and rectal manoraetric and symptomatic disturbances in chronic radiation injury and those following colo-anal sleeve anastomosis may be secondary to damage to the myenteric plexus which is histologically demonstrable. | 7. Following complete transection of the spinal cord above the sacral segments, severe constipation may result. The differential effects on colorectal and pelvic floor motility of sacral anterior root stimulators in five such patients may be summarised as follows: | 7.1 There is an increasing influence on striated pelvic floor muscle activity from S2 to S4. | 7.2 S2 stimulation results in occasional low pressure phasic colorectal contractions. | 7.3 S3 stimulation results in high-pressure phasic colorectal contractions reminiscent of peristaltic activity. This response is frequency dependent. | 7.4 S4 stimulation results in tonic pressure increases in the colon and rectum. These may simply be the cosequence of increase in intra-abdominal pressure due to pelvic floor contraction. | 8. Two groups of elderly patients who suffer from chronic constipation are identifiable as defined by the proctometrogram. One group has a significantly increased rectal capacity and compliance - a 'megarectum' syndrome. The second group shows significant reduction in rectal volume and compliance demonstrating hypertonicity of the distal bowel, both groups have blunting of rectal sensation to distension. Increased gastrointestinal transit times seem mainly due to rectal stasis. Abnormalities of the pudendo-anal reflex suggest that neurogenic deficits of the sacral spinal cord may contribute to these disturbances of colorectal function. | 9. Electrophysiological studies in young women with chronic idiopathic constipation suggest the possibility of a neurogenic deficit in the conus medullaris in the presence of normal sphincteric innervation. | 10. In neurogenic faecal incontinence the latency of the pudendo-anal reflex is significantly prolonged and its amplitude diminished. It may be absent in some patients. A significant correlation exists between the latency of the reflex and the corresponding mean motor unit potential duration of the external anal sphincter. These findings confirm the its usefulness as an index of neuropathy in neurogenic faecal incontinence. | 11. In genuine female stress urinary incontinence the latency of the pudendo-anal reflex is significantly prolonged, as are the latencies of two other polysynaptic sacral reflexes ( dorsal nerve to urethral sphincter, urethral mucosa to external anal sphincter ). The electrosensitivity of both the dorsal nerve and urethral mucasa is significantly blunted. These observations point to a strong neurogenic element in this disorder. The clinical and manometric response to pelvic floor physiotherapy is disappointing

RhoA/ROCK Pathway is the Major Molecular Determinant of Basal Tone in Intact Human Internal Anal Sphincter

Poster presented at: Digestive Disease Week (DDW) International meeting in San Diego, California. Background and Aims Knowledge of molecular control mechanisms underlying the basal tone in the intact human IAS is critical for the pathophysiology and rational therapy for debilitating rectoanal motility disorders

Biochar-based organic amendments on soil health, nutrient status and quality of potato (Solanum tuberosum)

Cultivation of quality potatoes ensures a good earning compared to low quality, especially in terms of tuber weight and starch content. Therefore, an experiment was laid out to find out the impact of an organic amendment based on the combinations of biochar, vermicompost, poultry manure, and bone meal for the enrichment of soil health and quality of potato crops at the research farm of Lovely Professional University, Punjab. The parameters of pH, EC, organic carbon, soil microbial biomass, nitrogen, phosphorus, potassium, labile carbon, and particulate organic carbon (POC) were used to understand the soil health improvement, while starch content and grading systems ranging from A to C were used for the quality of the potato. Among the treatments, most of the parameters were recorded as statistically significant at p>0.05. The highest improvement in terms of pH, EC, organic carbon, soil microbial biomass, nitrogen content, labile carbon, and particulate organic carbon (POC) was recorded in T3 (7.58, 0.39 dSm-1, 0.53 %, 333.3 µg g-1, 198.3 kg ha-1, 3.71 and 7.0 g kg-1 of soil) compared to T0 (7.38, 0.32 dSm-1, 0.44 %, 325.33 µg g-1, 171.5 kg ha-1, 2.33 and 3.0 g kg-1 g kg-1), while the phosphorus and potassium contents were estimated highest in T2 (17.4 and 255 kg ha-1). The quality parameters like starch content and grading quality of potato tubers were also influenced positively and estimated significantly highest in T3 (53.60 % and 153.7 q ha-1 of A grade potato). This study has shown the potential to improve the quality of potato tuber by providing a desirable soil environment to coordinate with potato plants

La formation des enseignants

En Inde, dans un contexte de vaste expansion éducative, les systèmes de formation des enseignants sont insuffisants, malgré les nombreuses recommandations exprimées après l’indépendance. Certains domaines prioritaires ont été définis dans la perspective d’une évolution : professionnalisation de l’éducation, développement des approches scientifiques, contribution à la cohésion sociale, sensibilisation à l’environnement, prise en compte des spécificités culturelles. L’enseignant doit devenir un agent de la modernisation, du développement et du progrès tout en préservant les liens avec la culture locale.Despite the repeated recommendations made since independence, India’s efforts at expanding its national education system have been consistently hampered by inadequate teacher training. To redress this imbalance, a number of priority areas have been defined, among them fostering greater professionalism in education, the development of scientific approaches, promoting social cohesion, raising awareness for the environment and making provisions for cultural differences. Teachers must help consolidate modernisation, economic development and progress while preserving links with their native cultures.En India, en un entorno de amplia expansión educativa, los sistemas de formación de los docentes son insuficientes, a pesar de las numerosas recomendaciones expresadas tras la independencia. Algunos sectores prioritarios han sido determinados en la perspectiva de una evolución : profesionalización de la educación, desarrollo de los enfoques científicos, contribución a la cohesión social, sensibilización al medio ambiente e integración de las especifidades culturales. El docente debe convertirse en un agente de la modernización, del desarrollo y del progreso y debe conservar los vínculos con la cultura local

Tied with a single thread : children, community and teachers

We have often heard this said, ‘पेट में नहीं होगा अन्न तो पढ़ने में कैसे लगेगा मन’, loosely translated, it means, ‘when the stomach is empty, how will the mind focus on learning?’ What else do such aphorisms tell us if not that, surely, survival comes before any formal schooling? Children from the families whose primary concern is to quell the hunger in their stomachs or to look for a livelihood to silence this hunger are often called ‘first generation learners’ in academic circles. Who knows why they are called first-generation learners? Perhaps because schools are considered to be the sole dispensers of learning. Perhaps because their parents are not able to deduce that, if their children learn to read and write now, they will be able to reap the benefits of their learning in 15-20 years by working in better and less physically taxing conditions. Who knows if it is the lack of an ability to think or an effect of it that prevents one from considering this option? Be that as it may, the consequence remains the same: education is always a secondary priority

Aging-associated changes in microRNA expression profile of internal anal sphincter smooth muscle: Role of microRNA-133a.

A comprehensive genomic and proteomic, computational, and physiological approach was employed to examine the (previously unexplored) role of microRNAs (miRNAs) as regulators of internal anal sphincter (IAS) smooth muscle contractile phenotype and basal tone. miRNA profiling, genome-wide expression, validation, and network analyses were employed to assess changes in mRNA and miRNA expression in IAS smooth muscles from young vs. aging rats. Multiple miRNAs, including rno-miR-1, rno-miR-340-5p, rno-miR-185, rno-miR-199a-3p, rno-miR-200c, rno-miR-200b, rno-miR-31, rno-miR-133a, and rno-miR-206, were found to be upregulated in aging IAS. qPCR confirmed the upregulated expression of these miRNAs and downregulation of multiple, predicted targets (Eln, Col3a1, Col1a1, Zeb2, Myocd, Srf, Smad1, Smad2, Rhoa/Rock2, Fn1, Tagln v2, Klf4, and Acta2) involved in regulation of smooth muscle contractility. Subsequent studies demonstrated an aging-associated increase in the expression of miR-133a, corresponding decreases in RhoA, ROCK2, MYOCD, SRF, and SM22α protein expression, RhoA-signaling, and a decrease in basal and agonist [U-46619 (thromboxane A2 analog)]-induced increase in the IAS tone. Moreover, in vitro transfection of miR-133a caused a dose-dependent increase of IAS tone in strips, which was reversed by anti-miR-133a. Last, in vivo perianal injection of anti-miR-133a reversed the loss of IAS tone associated with age. This work establishes the important regulatory effect of miRNA-133a on basal and agonist-stimulated IAS tone. Moreover, reversal of age-associated loss of tone via anti-miR delivery strongly implicates miR dysregulation as a causal factor in the aging-associated decrease in IAS tone and suggests that miR-133a is a feasible therapeutic target in aging-associated rectoanal incontinence

ShoreTension, Dynamic Mooring System

A lo largo de esta última década, el mundo marítimo se ha visto afectado cada vez más por las nuevas tecnologías. Este desarrollo exige que los profesionales encargados del mantenimiento de la instalación marítima, deban estar cada vez más preparados. En los buques la maquinaria es uno de los temas más importantes en el atraque y el desatraque, en el atraque es necesario un sistema de amarre y el buque queda a manos de las condiciones atmosféricas de viento y corriente del lugar que le obligan a ser amarrado al muelle. Hoy en día, la mayoría de los buques son amarrados con estachas como lo eran tradicionalmente durante miles de años. Pero debido a la dinámica y las cargas inherentes que afectan a los cuerpos grandes, como los barcos, cualquier cambio que se quiera aplicar a esta práctica requerirá una innovación significativa para que los nuevos sistemas sean más rentables y más seguros que sus predecesores. Por lo tanto, la empresa llamada KRVE ha desarrollado un sistema muy sencillo junto con la Autoridad Portuaria de Róterdam, que ofrece una tensión permanente sin necesidad de energía externa constante. El sistema se llama ShoreTension. Este, lo que hace es reducir el movimiento del buque provocado por el viento, la corriente y los buques que pasan durante la estancia en puerto

Bridge-overlap-Extension PCR method for constructing chimeric genes

52-56We understand that this bridge-overlap-extension PCR approach can have several applications, such as insertion of epitope tags, nuclear localization signals, secretory or proteolytic signals, insertion of DNA-binding regions or other functional elements into proteins of interest.The sequence element to be inserted can be of small size or long both being within the limits of efficient oligonucleotide synthesis. A distinct feature of this method is that by using overlapping oligonucleotides and bringing about a bridging step, we carry out the synthesis of a longer primer that contains an overlap with the hybrid PCR product, in the absence of any wild-type template. This helps in avoiding the use of template DNA and, therefore, the problem of re-annealing by oligonucleotides to wild-type template during the PCR steps altogether. The success of the technique is indicated by the 100% efficiency of obtaining the correct clones. Moreover, a high level of expression of precursor (fusion of P-factor and SK) and mature SK was observed when the hybrid gene construct, was cloned into an expression vector and transformed in suitable strains of S-pombe- Therefore, this new approach could serve as an alternative method for construction of chimeric genes