Anticancer potential of Ferula assa-foetida and its constituents,a powerful plant for cancer therapy

Cancer is one of the main challenges of the health system around the world.This disease is increasing in developing countries and imposes heavy costs on patients and governments.On the other hand,despite various drugs,the death rate among cancer patients is still high and the current treatments have many harmful effects.In the traditional medicine of different countries,there are many medicinal plants that can be effective in the treatment of cancer.Ferula plants are traditionally used as spices and food or for medicinal purposes.Ferula assa-foetida is one of the famous plants of this genus,which has been used for the treatment of various diseases since ancient times.Among the main compounds of this plant,we can mention monoterpenes,sulfide compounds and polyphenols,which can show different therapeutic effects.This article has been compiled with the aim of collecting evidence and articles related to the anti-cancer effects of extracts,derived compounds,essential oils and nanoparticles containing Ferula assa-foetida.This review article was prepared by searching the terms Ferula assa-foetida and cancer,and relevant information was collected through searching electronic databases such as ISI Web of Knowledge,PubMed,and Google Scholar.Fortunately,the results of this review showed that relatively comprehensive studies have been conducted in this field and shown that Ferula assa-foetida can be very promising in the treatment of cancer.

Comparison of emergency surgical cricothyroidotomy and percutaneous cricothyroidotomy by experienced airway providers in an obese,in vivo porcine hemorrhage airway model

Background: Emergency front-of-neck airway(eFONA) is a life-saving procedure in “cannot intubate, cannot oxygenate”(CICO). The fastest and most reliable method of eFONA has not been determined. We compared two of the most advocated approaches: surgical cricothyroidotomy and percutaneous cricothyroidotomy, in an obese, in vivo porcine hemorrhage model, designed to introduce real-time physiological feedback, relevant and high provider stress. The primary aim was to determine the fastest method to secure airway. Secondary aims were arterial saturation and partial pressure of oxygen, proxy survival and influence of experience.Methods: Twelve pigs [(60.3±4.1) kg] were anesthetized and exposed to 25%–35% total blood volume hemorrhage before extubation and randomization to Seldinger technique “percutaneous cricothyroidotomy”(n=6) or scalpelbougie-tube technique “surgical cricothyroidotomy”(n=6). Specialists in anesthesia and intensive care in a tertiary referral hospital performed the eFONA, simulating an actual CICO-situation.Results: In surgical cricothyroidotomy vs. percutaneous cricothyroidotomy, the median(interquartile range, IQR) times to secure airway were 109(IQR 71–130) s and 298(IQR 128–360) s(P=0.0152), arterial blood saturation(SaO2) were 74.7(IQR 46.6–84.2)% and 7.9(IQR 4.1–15.6)%(P=0.0167), PaO_(2) were 7.0(IQR 4.7–7.7) kPa and 2.0(IQR 1.1–2.9) kPa(P=0.0667), and times of cardiac arrest(proxy survival) were 137–233 s, 190(IQR 143–229) s, from CICO. All six animals survived surgical cricothyroidotomy, and two of six(33%) animals survived percutaneous cricothyroidotomy. Years in anesthesia, 13.5(IQR 7.5–21.3), did not influence time to secure airway.Conclusions: eFONA by surgical cricothyroidotomy was faster and had increased oxygenation and survival, when performed under stress by board certified anesthesiologists, and may be an indication of preferred method in situations with hemorrhage and CICO, in obese patients.

Zataria multiflora and its constituent,carvacrol,counteract sepsis-induced aortic and cardiac toxicity in rat:Involvement of nitric oxide and oxidative stress

Background:Zataria multiflora and carvacrol showed various pharmacological prop-erties including anti-inflammatory and anti-oxidant effects.However,up to now no studies have explored its potential benefits in ameliorating sepsis-induced aortic and cardiac injury.Thus,this study aimed to investigate the effects of Z.multiflora and carvacrol on nitric oxide(NO)and oxidative stress indicators in lipopolysaccharide(LPS)-induced aortic and cardiac injury.Methods:Adult male Wistar rats were assigned to:Control,lipopolysaccharide(LPS)(1 mg/kg,intraperitoneal(i.p.)),and Z.multiflora hydro-ethanolic extract(ZME,50–200 mg/kg,oral)-and carvacrol(25–100 mg/kg,oral)-treated groups.LPS was in-jected daily for 14 days.Treatment with ZME and carvacrol started 3 days before LPS administration and treatment continued during LPS administration.At the end of the study,the levels of malondialdehyde(MDA),NO,thiols,and antioxidant enzymes were evaluated.Results:Our findings showed a significant reduction in the levels of superoxide dis-mutase(SOD),catalase(CAT),and thiols in the LPS group,which were restored by ZME and carvacrol.Furthermore,ZME and carvacrol decreased MDA and NO in car-diac and aortic tissues of LPS-injected rats.Conclusions:The results suggest protective effects of ZME and carvacrol on LPS-induced cardiovascular injury via improved redox hemostasis and attenuated NO pro-duction.However,additional studies are needed to elucidate the effects of ZME and its constituents on inflammatory responses mediated by LPS.

Neuroprotective effects of resveratrol on retinal ganglion cells in glaucoma in rodents:A narrative review

Glaucoma,an irreversible optic neuropathy,primarily affects retinal ganglion cells(RGC)and causes vision loss and blindness.The damage to RGCs in glaucoma occurs by various mechanisms,including elevated intraocular pressure,oxidative stress,inflammation,and other neurodegenerative processes.As the disease progresses,the loss of RGCs leads to vision loss.Therefore,protecting RGCs from damage and promoting their survival are important goals in managing glaucoma.In this regard,resveratrol(RES),a polyphenolic phytoalexin,exerts antioxidant effects and slows down the evolution and progression of glaucoma.The present review shows that RES plays a protective role in RGCs in cases of ischemic injury and hypoxia as well as in ErbB2 protein expression in the retina.Additionally,RES plays protective roles in RGCs by promoting cell growth,reducing apoptosis,and decreasing oxidative stress in H_(2)O_(2)-exposed RGCs.RES was also found to inhibit oxidative stress damage in RGCs and suppress the activation of mitogen-activated protein kinase signaling pathways.RES could alleviate retinal function impairment by suppressing the hypoxia-i nducible factor-1 alpha/vascular endothelial growth factor and p38/p53 axes while stimulating the PI3K/Akt pathway.Therefore,RES might exert potential therapeutic effects for managing glaucoma by protecting RGCs from damage and promoting their survival.

Physiology of bile secretion

The formation of bile depends on the structural and functional integrity of the bile-secretory apparatus and its impairment, in different situations, results in the syndrome of cholestasis. The structural bases that permit bile secretion as well as various aspects related with its composition and flow rate in physiological conditions will first be reviewed. Canalicular bile is produced by polarized hepatocytes that hold transporters in their basolateral (sinusoidal) and apical (canalicular) plasma membrane. This review summarizes recent data on the molecular determinants of this primary bile formation. The major function of the biliary tree is modification of canalicular bile by secretory and reabsorptive processes in bileduct epithelial cells (cholangiocytes) as bile passes through bile ducts. The mechanisms of fluid and solute transport in cholangiocytes will also be discussed. In contrast to hepatocytes where secretion is constant and poorly controlled, cholangiocyte secretion is regulated by hormones and nerves. A short section dedicated to these regulatory mechanisms of bile secretion has been included. The aim of this revision was to set the bases for other reviews in this series that will be devoted to specific issues related with biliary physiology and pathology.

Pacing and predictors of performance during cross-country skiing races:A systematic review

Background: Cross-country skiing(XCS) racing, a popular international winter sport, is complex and challenging from physical, technical, and tactical perspectives. Despite the vast amount of research focusing on this sport, no review has yet addressed the pacing strategies of elite XCS racers or the factors that influence their performance. The aim was to review the scientific literature in an attempt to determine the effects of pacing strategy on the performance of elite XCS racers.Methods: Four electronic databases were searched using relevant subject headings and keywords. Only original research articles published in peerreviewed journals and the English language and addressing performance, biomechanics, physiology, and anthropometry of XCS racers were reviewed.Results: All 27 included articles applied correlative designs to study the effectiveness of different pacing strategies. None of the articles involved the use of an experimental design. Furthermore, potential changes in external conditions(e.g., weather, ski properties) were not taken into consideration. A comparable number of studies focused on the skating or classical technique. In most cases, positive pacing was observed, with certain indications that higher-level athletes and those with more endurance and strength utilized a more even pacing strategy. The ability to achieve and maintain a long cycle length on all types of terrain was an important determinant of performance in all of the included studies, which was not the case for cycle rate. In general, uphill performance was closely related to overall race performance, with uphill performance being most closely correlated to the success of female skiers and performance on flat terrain being more important for male skiers. Moreover, pacing was coupled to the selection and distribution of technique during a race, with faster skiers employing more double poling and kick double poling,less diagonal stride, and more V2(double dance) than V1(single dance) skating across a race.Conclusion: We propose that skiers at all levels can improve their performance with more specific training in techniques(i.e., maintaining long cycles without compromising cycle rate and selecting appropriate techniques) in combination with training for endurance and more strength. Furthermore, we would advise less experienced skiers and/or those with lower levels of performance to apply a more even pacing strategy rather than a positive one(i.e., starting the race too fast).

Immunohistochemical expression of SP-NK-1R-EGFR pathway and VDR in colonic inflammation and neoplasia

AIM:To determine the expression of neurokinin-1receptor(NK-1R),phosphorylated epidermal growth factor receptor(p EGFR),cyclooxygenase-2(Cox-2),and vitamin D receptor(VDR)in normal,inflammatory bowel disease(IBD),and colorectal neoplasia tissues from Puerto Ricans.METHODS:Tissues from patients with IBD,colitisassociated colorectal cancer(CAC),sporadic dysplasia,and sporadic colorectal cancer(CRC),as well as normal controls,were identified at several centers in Puerto Rico.Archival formalin-fixed,paraffin-embedded tissues were de-identified and processed by immunohistochemistry for NK-1R,p EGFR,Cox-2,and VDR.Pictures of representative areas of each tissues diagnosis were taken and scored by three observers using a4-point scale that assessed intensity of staining.Tissues with CAC were further analyzed by photographing representative areas of IBD and the different grades of dysplasia,in addition to the areas of cancer,within each tissue.Differences in the average age between the five patient groups were assessed with one-way analysis of variance and Tukey-Kramer multiple comparisons test.The mean scores for normal tissues and tissues with IBD,dysplasia,CRC,and CAC were calculatedand statistically compared using one-way analysis of variance and Dunnett’s multiple comparisons test.Correlations between protein expression patterns were analyzed with the Pearson’s product-moment correlation coefficient.Data are presented as mean±SE.RESULTS:On average,patients with IBD were younger(34.60±5.81)than normal(63.20±6.13,P<0.01),sporadic dysplasia(68.80±4.42,P<0.01),sporadic cancer(74.80±4.91,P<0.001),and CAC(57.50±5.11,P<0.05)patients.NK-1R in cancer tissue(sporadic CRC,1.73±0.34;CAC,1.57±0.53)and sporadic dysplasia(2.00±0.45)were higher than in normal tissues(0.73±0.19).p EGFR was significantly increased in sporadic CRC(1.53±0.43)and CAC(2.25±0.47)when compared to normal tissue(0.07±0.25,P<0.05,P<0.001,respectively).Cox-2 was significantly increased in sporadic colorectal cancer(2.20±0.23 vs 0.80±0.37 for normal tissues,P<0.05).In comparison to normal(2.80±0.13)and CAC(2.50±0.33)tissues,VDR was significantly decreased in sporadic dysplasia(0.00±0.00,P<0.001 vs normal,P<0.001 vs CAC)and sporadic CRC(0.47±0.23,P<0.001 vs normal,P<0.001 vs CAC).VDR levels negatively correlated with NK-1R(r=-0.48)and p EGFR(r=-0.56)in normal,IBD,sporadic dysplasia and sporadic CRC tissue,but not in CAC.CONCLUSION:Immunohistochemical NK-1R and p EGFR positivity with VDR negativity can be used to identify areas of sporadic colorectal neoplasia.VDR immunoreactivity can distinguish CAC from sporadic cancer.

Optimization of Ultrasonic-Assisted Extraction Process of Polysaccharides from American Ginseng and Evaluation of Its Immunostimulating Activity

Ultrasonic-assisted extraction （UAE） of American ginseng polysaccharides （AGP） was investigated using response surface methodology. Three-factor-three-level Box-Behnken design was employed to optimize the ultrasonic power, extraction time and ratio of water to raw material to obtain a high AGP yield. The analysis of variance and response surface plots indicated that ultrasonic power was the most important factor affecting the extraction yield. The optimal conditions were ultrasonic power 400 W, extraction time 71 min, and ratio of water to raw material 33 mL g-1. Under these conditions, the yield of AGP was 8.09%, which was agreed closely to the predicted value. Gas chromatography （GC） analysis showed that AGP was composed of arabinose, rhamnose, galactose, glucose, and galacturonic acid. Fourier transform infrared spectra revealed the general characteristic absorption peaks of AGP. In addition, AGP exhibited good immunostimulating activities by up-regulating the production of nitric oxide and cytokines. Compared with hot water extraction, UAE required shorter extraction time and gave a higher extraction yield, without changing the structure and immunostimulating activity of AGP. The results indicated that UAE could be an effective and advisable technique for the large scale production of plant polysaccharides.

Leuprolide acetate induces structural and functional recovery of injured spinal cord in rats

Gonadotropin-releasing hormone（Gn RH） and its synthetic analog leuprolide acetate, a Gn RH agonist, have neurotrophic properties. This study was designed to determine whether administration of leuprolide acetate can improve locomotor behavior, gait, micturition reflex, spinal cord morphology and the amount of microglia in the lesion epicenter after spinal cord injury in rats. Rats with spinal cord compression injury were administered leuprolide acetate or saline solution for 5 weeks. At the 5th week, leuprolide acetate-treated rats showed locomotor activity recovery by 38%, had improvement in kinematic gait and exhibited voiding reflex recovery by 60%, as compared with the 1st week. By contrast, saline solution-treated rats showed locomotor activity recovery only by 7%, but voiding reflex did not recover. More importantly, leuprolide acetate treatment reduced microglial immunological reaction and induced a trend towards greater area of white and gray matter in the spinal cord. Therefore, leuprolide acetate has great potential to repair spinal cord injury.

Dystrophic calcification and heterotopic ossification in fibrocartilaginous tissues of the spine in diffuse idiopathic skeletal hyperostosis(DISH)

Diffuse idiopathic skeletal hyperostosis(DISH) is a prevalent noninflammatory spondyloarthropathy characterized by ectopic mineral formation along the anterolateral aspect of the vertebral column, yet little is known about its underlying pathogenesis. Our objective was to evaluate the histopathological features and composition of ectopic mineral within spinal tissues affected by DISH in humans. Thoracic spine segments from six embalmed cadaveric donors(one female and five males;median age 82 years)meeting the radiographic diagnostic criteria for DISH were evaluated using radiological, histological, and physical analyses. Overall,the histological features of ectopic mineralization at individual motion segments were heterogeneous, including regions of heterotopic ossification and dystrophic calcification. Heterotopic ossifications were characterized by woven and lamellar bone,multifocal areas of metaplastic cartilage, and bony bridges along the anterior aspect of the intervertebral disc space. Dystrophic calcifications were characterized by an amorphous appearance, a high content of calcium and phosphorus, an X-ray diffraction pattern matching that of hydroxyapatite, and radiodensities exceeding that of cortical bone. Dystrophic calcifications were found within the anterior longitudinal ligament and annulus fibrosus in motion segments both meeting and not meeting the radiographic criteria for DISH. In summary, our findings indicate that in DISH, ectopic mineral forms along the anterior aspect of the spine by both heterotopic ossification and dystrophic calcification of fibrocartilaginous tissues. Although both types of ectopic mineralization are captured by current radiographic criteria for DISH, dystrophic calcification may reflect a distinct disease process or an early stage in the pathogenesis of DISH.

Phosphatidylserine improves axonal transport by inhibition of HDAC and has potential in treatment of neurodegenerative diseases

Familial dysautonomia （FD） is a rare children neurodegenerative disease caused due to a point mutation in the IKBKAP gene that results in decreased IKK complex-associated protein （IKAP） protein production. The disease affects mostly the dorsal root ganglion （DRG） and the sympathetic ganglion. Recently, we found that the molecular mechanisms underlying neurodegeneration in FD patients are defects in axonal transport of nerve growth factors and microtubule stability in the DRG. Neurons are highly polarized cells with very long axons. In order to survive and maintain proper function, neurons depend on transport of proteins and other cellular components from the neuronal body along the axons. We further demonstrated that IKAP is necessary for axon maintenance and showed that phosphatidylserine acts as an HDAC6 inhib- itor to rescue neuronal function in FD cells. In this review, we will highlight our latest research findings.

Leaky gut, dysbiosis, and enteric glia activation: the trilogy behind the intestinal origin of Parkinson’s disease

Early diagnosis of Parkinson’s disease(PD):how we can improve the therapeutic approach:PD is a neurodegenerative disease characterized by motor dysfunctions(tremor,rigidity,bradykinesia and impaired posture/balance)elicited by selective depletion of dopaminergic(DA)neurons in substantia nigra pars compacta.DA neuron loss is associated with neuronal inclusions of the phosphorylatedα-synuclein protein called Lewy body(Shults,2006).Although the underlying neurodegenerative process is not affected,the management of PD patients has been revolutionized with the introduction of levodopa and DA drugs in the routine therapy,which ensures initial symptomatic relief of motor functions through the DA supply in the nigrostriatal circuit.These drugs are currently the best option for treating PD,although their chronic use is associated with progressive dopamine resistance and loss of effectiveness in the recovery of motor dysfunctions.Alternative therapeutic strategies,including agonists of DA receptors,monoamine oxide B inhibitors,and even deep brain stimulation techniques have been developed to overcome these clinical limitations.Unfortunately,these therapeutic approaches cannot restore PD-compromised functions,as irreversible DA neurodegeneration has occurred in substantia nigra pars compacta when first motor symptoms appear.The nigrostriatal system is traditionally considered as the first region affected by neuronal impairment in Parkinsonisms;however,α-synuclein aggregation appears in a pre-motor stage of the disease in the enteric nervous system(ENS).

A review study on medicinal plants used in the treatment of learning and memory impairments

Alzheimer′s disease(AD) is a progressive brain disorder thai gradual!) impairs the person's memory and ability to learn,reasoning.judgment,communication and daily activities.All is characterized clinically by cognitive impairment and pathologically by the deposition of β amyloid plaques and neurofibrillary tangles,and the degeneration of the cholinergic basal forebrain.During the progression of AD patients may produce changes in personality and behavior,such as anxiety,paranoia,confusion,hallucinations and also to experience delusions and lanlasies.The first neurotransmitter defect discovered in Al) involved acetylcholine as cholinergic function is required for short—term memory.Oxidative stress may underlie the progressive neurodegeneration characteristic of AD.Brain structures supporting memory are uniquely sensitive to oxidative stress due to their elevated demand for oxygen.The neurodegenerative process in AD may involveβ amyloid toxicity.Neurotoxicity of β amyloid appears to involve oxidative stress.Currently,there is no cure for this disease but in new treatments,reveals a new horizon on the biology of this disease.This paper reviews the effects of a number of commonly used types of herbal medicines for the Irealment of AD.The objective of this article was to review evidences from controlled studies in order to determine whether herbs can be useful in the treatment of cognitive disorders in the elderly.

Temperature variable and the efficiency of sperm mediated transfection of HPV16 DNA into cells

Aim:To pretreat sperm at various temperatures before exposure to human papillomavirus (HPV) 16 DNA fragments and to assess the efficiency of HPV carrier sperm to transfect cumulus cells. Methods: Cumulus cells from follicular aspirates were obtained, pooled and divided into culture dishes containing Sybr Gold-stained HPV DNA carrying sperm that were either pretreated at 4℃, 37℃ or 40℃ (n=5). The cells were incubated in 5% CO_2 in air mixture at 37℃ for 24 hours. The efficiency of sperm to take up fluorescent HPV DNA was determined at hour 0. After incubation, cumulus cell viability was assessed using the e0sin method and the percentages of fluorescent cumulus cells determined. Results: Over half of all the cumulus cells became fluorescent with the highest percentage in the 37℃ group. Sperm pretreated at 4℃ had the greatest amount of HPV DNA fragments. Total sperm motility was similar for the 3 pretreatment groups. There were no differences in cumulus viability among the groups. Conclusion: Sperm pretreated at 37℃ transferred the greatest amount of fluorescent HPV DNA fragments to the cumulus cells. The HPV DNA was observed in the nuclear and cytoplasmic compartments. The data suggested the possibility of sperm as a vector for the transmission of HPV DNA to the cumulus cells surrounding ovulated oocytes, which might lead to early implantation failures.

Hydroxycitric acid ameliorates inflammation and oxidative stress in mouse models of multiple sclerosis

Hydroxycitric acid （HCA） is derived primarily from the Garcinia plant and is widely used for its anti-in- flammatory effects. Multiple sclerosis can cause an inflammatory demyelination and axonal damage. In this study, to validate the hypothesis that HCA exhibits therapeutic effects on multiple sclerosis, we established female C57BL/6 mouse models of multiple sclerosis, i.e., experimental autoimmune encephalomyelitis, using Complete Freund＇s Adjuvant （CFA） emulsion containing myelin oligodendrocyte glycoprotein （35-55）. Treatment with HCA at 2 g/kg/d for 3 weeks obviously improved the symptoms of nerve injury of experimental autoimmune encephalomyelitis mice, decreased serum interleulin-6, tumor necrosis factor alpha, nitric oxide, and malondialdehyde levels, and increased superoxide dismutase and glutathione reduc- tase activities. These findings suggest that HCA exhibits neuroprotective effects on multiple sclerosis-caused nerve injury through ameliorating inflammation and oxidative stress.

Sex differences and effects of oestrogen in rat gastric mucosal defence

AIM To evaluate sex differences and the effects of oestrogen administration in rat gastric mucosal defence.METHODS Sex differences in gastric mucus thickness and accumulation rate, absolute gastric mucosal blood flow using microspheres, the integrity of the gastric mucosal epithelium in response to a chemical irritant and the effects of oestrogen administration on relative gastric mucosal blood flow in an acute setting was assessed in an in vivo rat experimental model. Subsequently, sex differences in the distribution of oestrogen receptors and calcitonin gene related peptide in the gastric mucosa of animals exposed to oestrogen in the above experiments was evaluated using immunohistochemistry.RESULTS The absolute blood flow in the GI-tract was generally higher in males, but only significantly different in the corpus part of the stomach (1.12 ± 0.12 m L/min·g in males and 0.51 ± 0.03 m L/min·g in females) (P = 0.002). After removal of the loosely adherent mucus layer the thickness of the firmly adherent mucus layer in males and females was 79 ± 1 μm and 80 ± 3 μm respectively. After 60 min the mucus thickness increased to 113 ± 3 μm in males and 121 ± 3 μm in females with no statistically significant difference seen between the sexes. Following oestrogen administration(0.1 followed by 1 μg/kg·min), mean blood flow in the gastric mucosa decreased by 31% [68 ± 13 perfusion units (PFU)] in males which was significantly different compared to baseline(P = 0.02). In females however, mean blood flow remained largely unchanged with a 4% (5 ± 33 PFU) reduction. The permeability of the gastric mucosa increased to a higher level in females than in males (P = 0.01) after taurocholate challenge. However, the calculated mean clearance increase did not significantly differ between the sexes [0.1 ± 0.04 to 1.1 ± 0.1 m L/min·100 g in males and 0.4 ± 0.3 to 2.1 ± 0.3 m L/min·100 g in females(P = 0.065)]. There were no significant differences between 17β-Estradiol treated males (mean ratio of positive staining ± SEM)(0.06 ± 0.07) and females(0.11 ± 0.11) in the staining of ERα (P = 0.24). Also, there were no significant differences between 17β-Estradiol treated males (0.18 ± 0.21) and females (0.06 ± 0.12) in the staining of ERβ (P = 0.11). Finally, there were no significant differences between 17β-Estradiol treated males (0.04 ± 0.05) and females (0.11 ± 0.10) in the staining of CGRP(P = 0.14).CONCLUSION Gastric mucosal blood flow is higher in male than in female rats and is reduced in male rats by oestrogen administration.

Relaxant effect and possible mechanism of 17-nor-subincanadine E in rabbit corpora cavernosa

Compounds with dual action on cyclic adenosine monophosphate （cAMP） and cyclic guanosine monophosphate （cGMP） may be a treatment option for erectile dysfunction, as they not only promote penile erection but also prevent the upregulation of phosphodiesterase-5. In this study, we examined the possible relaxant effect and mechanism of 17-nor-subincanadine E （SEC, 0.2-200 pmol I^-1）, a plant-derived alkaloid, in rabbit corpus cavernosum （RbCC） strips that had been precontracted by exposure to phenylephrine （10 pmol I^- 1） or a high concentration of K＋ （60 mmol I^- 1） in vitro. In addition to SEC＇s effect on cAMP and cGMP levels, electrical field stimulation （EFS） in phenylephrine-precontracted RbCC and calcium chloride （1-100 mmol I^-1） evoked responses in depolarized RbCC were analysed. SEC relaxed the phenylephrine-precontracted RbCCs in a concentration-dependent manner. Atropine, guanethidine and N-co-nitro-L-arginine methyl ester （L-NAME） did not have any effect on the relaxation of RBCCs. When 1H-1,2,4oxadiazole[4,3-a] quinoxalin-1-one （ODQ） was added, it effectively blocked the relaxant response of SEC. Although SEC enhanced the maximal relaxation produced by sodium nitroprusside （SNP） and forskolin in phenylephrine-precontracted cavernosal smooth muscle, it caused a decrease in the maximal contractile response induced by calcium chloride in depolarized RbCCs. The relaxant effect of SEC was paralleled by an increase in the tissue levels of the Cyclic nucleotides cAMP and cGMP. We conclude that SEC promotes the relaxation of RbCC, possibly favouring cAMP and cGMP accumulation and calcium blockade. This novel mechanism could be useful for patients who do not benefit from phosphodiesterase inhibitors and for those with endothelial and nitrergic dysfunction, such as patients with diabetes, hypertension and dyslipidaemias.

Evaluation of patient visual comfort and repeatability of refractive values in non-presbyopic healthy eyes

AIMTo evaluate the intra-operator repeatability in healthy subjects using the WAM-5500 auto-kerato/refractometer and the iTrace aberrometer, to compare the refractive values and the subjective refraction obtained with both devices and to determine which of these three spherocylindrical corrections allows the subject to achieve the best visual comfort.METHODSForty-two non-presbyopic healthy eyes of 42 subjects were enrolled in this prospective study. Refractive values were compared, evaluating the repeatability, the relationship between the methods and the best visual comfort obtained.RESULTSSphere, cylinder and axis results showed good intraclass correlation coefficients (ICC); the highest ICC was obtained using the spherical refraction with the autorefractometer and the aberrometer, achieving levels of 0.999 and 0.998, respectively. The power vector (PV) was calculated for each refraction method, and the results indicated that there were no statistically significant differences between them (P&#x0003e;0.05). Direct comparison of PV measurements using the three methods showed that aberrometer refraction gave the highest values, followed by the subjective values; the autorefractometer gave the lowest values. The subjective method correction was most frequently chosen as the first selection. Equal values were found for the autorefractometer and the aberrometer as the second selection.CONCLUSIONThe iTrace aberrometer and the WAM-5500 auto-kerato/refractometer showed high levels of repeatability in healthy eyes. Refractive corrections with the aberrometer, the autorefractometer and subjective methods presented similar results, but spherocylindrical subjective correction was the most frequently selected option. These technologies can be used as complements in refractive evaluation, but they should not replace subjective refraction.

How useful are biomarkers for the diagnosis of Alzheimer’s disease and especially for its therapy?

Alzheimer’s disease key facts:Alzheimer’s disease(AD)is a slowly progressive neurodegenerative disease with no available effective treatment.It is possible to distinguish an early-onset AD that affects a limited number of subjects of young age,and a sporadic or late-onset form of the disease that affects the vast majority of subjects who are diagnosed with AD.As life expectancy has increased considerably over the past century,the number of people diagnosed with AD has grown exponentially.So,AD and AD-related pathologies represent a huge social and economic burden.The number of individuals waiting for effective disease-modifying therapy is impressive.It is estimated that 50 million people worldwide live with dementia,the majority of these cases are caused by AD(World Health Organization,2021).In the US about 6 million individuals are living with AD,and more than 9 million people are in EU member states(OECD and European Union,2020).The costs of health care and long-term care are substantial.Given this massive societal impact,enormous efforts have been made to understand the pathogenetic mechanisms of the disease with the hope of identifying new targets and,therefore,developing effective drugs.

Markers for neural degeneration and regeneration:novel highly sensitive methods for the measurement of thrombin and activated protein C in human cerebrospinal fluid

Inflammation and coagulation are tightly interconnected in the pathophysiology of neuronal diseases.Thrombin,a pro-coagulant serine protease is associated with neurodegeneration and its indirect inhibitor,activated protein C(aPC),is considered neuroprotective.While levels of thrombin and aPC activity are readily measured in the blood,similar assays in the cerebrospinal fluid(CSF)have not been described.The aim of this study was to establish a specific and sensitive enzymatic assay to measure both thrombin and aPC activity in the CSF.CSF was collected from 14 patients with suspected normal pressure hydrocephalus served as a control group,while seven patients with central nervous system infections served as an acute neuro-inflammatory study group and one sample of CSF following traumatic lumbar puncture served as a positive control.Thrombin and aPC activities were measured by fluorescence released by specific proteolytic cleavage in the presence of endopeptidase and amino-peptidase inhibitors to ensure specificity.Specificity of the method was verified by thrombin and serine-protease inhibitors N-alpha-((2-naphthylsulfinyl)glycyl)-DL-p-amidinophenylalanylpiperidine and phenylmethanesulfonyl fluoride.Inhibition of thrombin activity by CSF samples and levels of specific thrombin inhibitors were also assessed.Thrombin and aPC activities were reliably measured and were significantly higher in the CSF of patients with central nervous system infections compared to normal pressure hydrocephalus controls,suggesting the involvement of these factors in neuro-inflammation.CSF thrombin activity levels in the presence of known thrombin concentration were high in patients with central nervous system infections,and low in normal pressure hydrocephalus patients.Quantification of endogenous thrombin inhibitors protease nexin 1,amyloid precursor protein and anti-thrombin III in CSF by western blot indicated a significant elevation of amyloid precursor protein in infectious CSF.In conclusion,this study describes a novel and sensitive assay aimed at the detection of thrombin and aPC activity in CSF.This method may be useful for measuring these factors that reflect degenerative and protective influences of coagulation on neurological disorders.The study procedure was approved by the Ethics Committee of the Chaim Sheba Medical Center(approval No.4245-17-SMC)on October 18,2018.